42 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 42-47 research article survey on wireless indoor positioning systems wisam hasan ali*, amnah abas kareem, manal jasim department of electrical engineering, university of technology, baghdad, iraq abstract indoor positioning has finally testified a rise in interest, thanks to the big selection of services it is provided, and ubiquitous connectivity. there are currently many systems that can locate a person, be it wireless or by mobile phone and the most common systems in outdoor environments is the gps, the most common in indoor environments is wi-fi positioning technique positioning. the improvement of positioning systems in indoor environments is desirable in many areas as it provides important facilities and services, such as airports, universities, factories, hospitals, and shopping malls. this paper provides an overview of the existing methods based on wireless indoor positioning technique. we focus in this survey on the strengths of these systems mentioned in the literature discordant with the present surveys; we also assess to additionally measure various systems from the scene of energy efficiency, price, and following accuracy instead of comparing the technologies, we also to additionally discuss residual challenges to correct indoor positioning. keywords: indoor localization, indoor positioning, received signal strength indicator, wi-fi introduction the widespread proliferation of smartphones and different wireless device within past years has resulted in an exceeding range selection of services as well as indoor positioning. indoor positioning technique is the method of finding a device or user position in an interior setting. indoor device positioning technique has been achieving a widespread the previous few decades; positioning technique has largescale applications in the health sector, disaster, industry, management,[1] surveillance, building management, and a number of several other sectors. so as to currently the studies and also the business spotlight is on rising technologies, like wi-fi, ultra-wideband (uwb), zigbee, bluetooth, etc. due to, wi-fi fingerprinting is based on the (received signal strength indicator [rssi]) that oblivious from each of the wireless access points that are available and on comparisons to a reference database. therefore, the position of a device is commonly determined by calculating the distance or the likeness between a fingerprint collected by the device and the fingerprints contained in the reference database.[2] wi-fi fingerprinting is a complex subject and suffers from received signal strength (rss) difference problem causes by environmental changes that are ingrained in both the training and positioning phases.[3] due to, they proposed a smartphone-based, indoor pedestrian-tracking system and they analyze various sides of the rss difference when using smartphones.[4] they propose a new algorithm based on an improved double-peak gaussian distribution for wi-fi fingerprinting, and the results show the algorithm can improve the positioning accuracy at about 20%.[5] however, they present magicol, an indoor positioning and tracking system, and they did an in-depth study on the incorporation of magnetic and wi-fi signals. the incorporation with wi-fi leads to 90 percentile accuracy.[6] they provided a survey of the existing methods based on smartphone that includes wi-fi, bluetooth, and magnetic field fingerprinting and described the technologies and position estimation methods of these technology components.[7] they used deep neural networks and employ the dotnetnuke system for classification building/floor while still attained satisfactory results.[8] this leads to obtain a more accurate position they combine the method of fingerprint and k-nearest neighbor (knn) method, where the classification of positions is made from some rss data that obtained from some ap that subsist around the user’s position.[9] therefore, they attempted to establish a bidirectional link between internet of things and indoor positioning also they discussed challenges to accurate indoor positioning.[10] therefore, they presented wideep combines a stacked reduce the noise of the autoencoder deep learning the model and a probabilistic framework to controlled the noise in the received wi-fi signal. this survey focuses on wireless technology indoor positioning system. corresponding author: wisam hasan ali, department of electrical engineering, university of technology, baghdad, iraq. e-mail: 30088@uotechnology.edu.iq received: apr 20, 2019 accepted: apr 24, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp42-47 copyright © 2019 wisam hasan ali, amnah abas kareem, manal jasim. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) ali, et al.: survey on wireless indoor positioning systems 43 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 42-47 this paper aims to make an available general overview of past work at the side of a beginning platform for future adds this promising however difficult space. in section 2, we discuss different techniques for indoor positioning. section 3, we offer different technologies specializing in wireless technologies for indoor positioning. sections 4, we offer many of the criteria which will be used to evaluating the performance of an indoor positioning system. finally, section 5, we provide the conclusion of the survey. table 1 presented a list of abbreviations used in this paper. indoor positioning metrics during this section, we discuss different criteria measurements for signal in the positioning system. rss it is one of the most commonly used standards in indoor environments positioning.[11,12] is the actual signal strength received in the receiver and measured in milliwatt or ordbmw? as for rssi is that the rss indicator, a relative measurement of the rss that has generally defined by each chip vendor, for example, the cisco uses a range of wi-fi chipset between 0 and 100, while atheros uses rssi values between 0 and 60. it is potential to understand the distance by employing a variety of various signal propagation models given the ability at the reference or transmission power that is famed by utilize, the rssi and an easy path-loss propagation model,[13] the spaced between transmitter (tx) and receiver (rx) is often calculable from equation (1), rssi = −10 n log 10 (d) + a (1) where n is that the path loss exponent (which range from four in indoor environments to two in free space), and a is that the rssi at a reference range from the receiver. whereas the rss primarily based approach is uncomplicated, cheap and energy efficient, it suffers from poor positioning accuracy thanks to further signal attenuation ensuing from transmission through walls and severe rss fluctuation alternative massive obstacles and indoor noise.[11,14] completely different filters or averaging mechanisms will be accustomed mitigate these effects. however, to get high positioning, accuracy will be used with algorithms. channel state information (csi) in ieee 802.11 and uwb, and several wireless systems, the channel frequency is selective frequencies exhibit different amplitude and part behavior due to the coherence information measure of the wireless channel is smaller than the information measure of the signal. nowadays, several ieee 802.11 nics cards will offer subcarrier-level channel measurements for orthogonal frequency division multiplexing systems more stable measurements and better positioning accuracy. fingerprinting fingerprinting: primarily, the main positioning techniques need to be scanned for environmental features of the environment.[15,16] initially, rss is measured in two phases; then the measurements are compared in the online phase with the offline phase to determine the location of the person. there are a number of accessible algorithms that can be discussed below: knn these algorithms utilized if the distances are adopted as weights in the signal space, and based on the rssi utilized root mean square error.[17] neural networks neural networks are used in many prediction and classification situations. when using a neural network in an indoor positioning system, neural network is trained on rss values and coordinates obtained in the offline phase.[18-20] the table 1: a list of abbreviation used in this paper aoa angle of arrival rssi received signal strength indicator poa phase-of-arrival gps global positioning system dbm decibel-milliwatts los line-of-sight rfid radio-frequency identification csi channel state information lbs location-based services rf radio frequency rn reference node knn k-nearest neighbors rx receiver tof time-of-flight s distance uwb ultra-wideband mbl mobile-based localization tx transmitters svm support vector machine dbl device-based localization ble bluetooth low energy ns nanosecond mac medium access control ofdm orthogonal frequency-division multiplexing figure 1: received signal strength indicator based positioning[11] aq1 ali, et al.: survey on wireless indoor positioning systems 44 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 42-47 perceptron multilayer network (multilayer perceptrons) with a hidden node layer is one among the remarkably used ann positioning.[17] support vector machine (svm) it is one of the most common approaches to regression and data classification. svm, they are used extensively in the field of machine learning with great accuracy. the angle of arrival (aoa) primarily, aoa approaches to estimate the angle at that the transmitted signal impact on the receiver utilize antennae arrays.[9] although aoa will offer associate correct estimation once the transmitter-receiver distance is small, whereas its accuracy deteriorates with an increase within the transmitterreceiver distance. moreover, because of multipath effects in indoor environments, the aoa in terms of line of sight (los) is commonly exhausting to get. table 2 provides an outline of the techniques mentioned. indoor positioning techniques in this section, we will present and discuss the different existing technologies that have been utilized to provide indoor positioning services. wi-fi ieee 802.11, called wi-fi, works within the scientific, medical, and industrial (industrial scientific medical [ism]) domain and is originally accustomed to giving internet property and networking for multiple devices publicly, non-public and business environments. in the first place, the wi-fi network had a reception vary of regarding 100 m[17] that have currently inflated to regarding 1 km[21] in ieee 802.11 ah. wi-fi is enabled, most laptops, smartphones, and other portable user devices, creating wi-fi, a perfect candidate for indoor positioning and one in every of the foremost specific positioning technologies in literature.[14,22-24] since wi-fi access points can also be used as reference points,[25] basic positioning systems can be built while not the requirement for extra infrastructure. in addition, uncontrolled interference in the ism domain has been shown the affect on positioning accuracy.[22] the preceding rss, csi, and aoa technologies or any combination of them can be defined as mixed methods. recent wi-fi-based positioning systems[22,23,25] have achieved an average positioning accuracy of up to 23 cm.[23] for elaborate info regarding wi-fi, we direct the reader to go.[26,27] bluetooth bluetooth (ieee 802.15.1) is a formation medium access that control between layers and physical specifications in order to link several mobile or multiple wireless devices among a selected personal space. the most recent version of bluetooth, such as bluetooth smart additionally called bluetooth low energy (ble), as will give a higher power potency and improved data rate of 24 mbps with a coverage range of 70–100 m with. apple proclaimed the ibeacons program at the world software developers conference,[28] whereas it is utilized with different positioning techniques such as rssi and aoa.[29,30] the protocol permits the device that supports ble to send signals or signals at periodic intervals. the most disadvantages of ibeacons are table 2: outline of the most common indoor positioning techniques techniques advantages disadvantages rssi easy to implement, is used with a variety of techniques, cost-effective prone to multipath weakening and lower positioning accuracy, environmental noise, will want fingerprinting csi more powerful for multi-track and indoor noise not readily available on ready nics aoa can provide high positioning, does not require any print, accuracy might need directional antennas, and the comparatively sophisticated algorithms and perhaps needs intricacy hardware fingerprinting partly easy during use the new fingerprinting is needed even if there is a slight difference in space rssi: received signal strength indicator, csi: channel state information, aoa: angle of arrival figure 2: orthogonal frequency division multiplexing (a) transmitter and (b) receiver block diagram[9]aq1 a b ali, et al.: survey on wireless indoor positioning systems 45 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 42-47 that the delayed reporting and reporting on rss may require an important period of time positioning challenges. whereas ibeacons were intended to produce early detection, it was conjointly used for indoor positioning. radio-frequency identification device (rfid) rfid is the transmission and storage of data using electromagnetic transmitters from the transmitter to any appropriate radio frequency circuit.[31] rfid consists of a reader who can communicate with rfid tags. there are two basic types of rfid: active rfid and passive rfid systems. active rfid active rfid operates in the microwave and ultra-high frequency band. active rfid can be used for positioning. passive rfid passive rfid is limited in communication range (1–2 m). they are lighter, smaller, and less expensive than active devices, but their limited range makes them unsuitable for indoor positioning. table 3 provides an outline of the many wireless technologies from a positioning perspective. applications of indoor positioning system 4 indoor positioning technology has recently seen a major increase in usage around the world. some applications within the following: marketing and customer assistance marketing is an essential part of any business because it makes it easier for you to improve your product and brand image. positioning based marketing is fundamentally revolutionary in a world of marketing that improves profits and promotes sales. major airports such as london heathrow airport, tokyo haneda airport in japan, miami international, john f. kennedy in new york and many other improve customer experience proximity-based services and use ibeacons.[32,33] health sector the health sector can profit significantly from the positioning system because it will facilitate save valuable lives, will support hospital workers, patients, and guests.[17] the indoor positioning of doctors can also allow many patients to be traced and track their movements to make sure patient safety.[34] so as, in the event of an emergency, the positioning identifies the employee who is nearby and has the necessary qualifications to deal with the emergency situation. the indoor positioning of doctors can also allow many patients to be traced and track their movements to make sure patient safety. security indoor positioning can greatly improve the security situation of affairs round the world. so as to can be utilizing positioning system to definition potential threats that may cause security risks.[15] using locations, central command can design plans and higher methods that give to troopers within the field. challenges this section, will proposed the most common key challenges that affect the accuracy of the person position.figure 4: model architecture for ibeacon systems[28]aq1 table 3: synopsis of various wireless technologies for positioning technique max. range advantages disadvantages energy consumption ieee 802.11 n[32] 802.11 ac 802.11 ad (250) m outdoor (35) m indoor couple of meters widely available, high accuracy, does not need complex extra hardware prone to noise, needs complex processing algorithms moderate rfid[33] (200) m it has a wide range positioning accuracy is low. low figure 3: angle of arrival positioning[9]aq1 ali, et al.: survey on wireless indoor positioning systems 46 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 42-47 multipath and noise effects multipath effects are a major challenge for positioning. due to the signals intrinsic nature, the signal may be suffering from absorption, refraction, dispersion, reflection from metals, walls, nature of climate, and humans. so as, to get a single signal possibility does not exist. therefore, for accurate site estimation, complex signal processing techniques that can identify the los signal and eliminate the consequences of multiple path signals are needed. however, there is a necessity for improved and efficient multiple noise and noise control algorithms that facilitate to use positioning signals accurately. privacy and security privacy is one of the important challenges facing the positioning system for human. due to not all people have a desire to share their current positions. until now, current positioning systems do not care much about the problem of privacy, but focus more on the accuracy and efficiency of the system in detecting the person’s positioning. however, how the user can trust the system or the positioning service provider? therefore, it is important to consider privacy and security when working with a person’s positioning system. cost cost is one of the fundamental challenges facing a positioning service. sometimes need additional infrastructure in indoor environments and link nodes. in addition, some proprietary software, dedicated servers, and databases are require. despite that, this can be overcome using cellular networks or wi-fi or mixing of them. conclusions this survey, we provided a description of many indoor positioning technologies (wifi, uwb, etc.) and techniques (aoa, rssi, csi, etc.). the default methodology for internal mobility depends on the wireless local area network. over the years, there has been clear progress in attempts to increase the accuracy of time to enhance accuracy. the positioning accuracy depends on the density of the access points and standardization points and can be appropriate for several in typical public buildings such as shopping malls, universities, and airports. however, under non-los multipath or conditions, the resolution deteriorates dramatically. it is also, this survey discuss a number of internal challenges to the positioning and presented some solutions that could help address these challenges. references 1. a. zelenkauskaite, n. bessis, s. sotiriadis and e. asimakopoulou. “interconnectedness of complex systems of internet of things throughsocial network analysis for disaster management”. in fourth international conference on intelligent networking and collaborative systems, ieee, 2012, pp. 503-508. 2. n. marques, f. meneses and a. moreira. “combining similarity functions and majority rules for multi-building, multi-floor, wifi positioning”. international conference on indoor positioning and indoor navigation, ieee, 13-15th nov 2012. 3. y. kim, h. shin and h. cha. “smartphone-based wi-fi pedestrian-tracking system tolerating the rss variance problem”. international conference on pervasive computing and communications, ieee, 19-23 mar 2012. 4. l. chen, b. li, k. zhao, c. rizos and z. zheng. “an improved algorithm to generate a wi-fi fingerprint database for indoor positioning”. sensors, vol. 13, no. 8, pp. 1186-1194, 2013. 5. y. shu, c. bo, g. shen, c. zhao, l. li and f. zhao. “magicol: indoor localization using pervasivemagnetic field and opportunistic wifi sensing”. ieee journal on selected areas in communications, vol. 33, pp. 1443-1457, 2015. 6. p. davidson and r. piche. “a survey of selected indoor positioning methods for smartphones”. ieee communications surveys and tutorials, vol. 19, pp. 1347-1370, 2016. 7. m. nowicki and j. wietrzykowski. “low-effort place recognition with wifi fingerprints using deep learning”. switzerland: springer, 2017. 8. y. e. dari and s. pranowo. “capture: a mobile based indoor positioning system using wireless indoor positioning system”. international journal of interactive mobile technologies, vol. 12, no. 1, p. 61, 2018. 9. f. zafari, a. gkelias and k. k. leung. “a survey of indoor localization systems and technologies”. ieee intelligent vehicles symposium iv, 2018. 10. m. abbas, h. rizk, m. elhamshary and m. torki. “wideep: wifibased accurate and robust indoor localization system using deep learning”. research gate, conference paper, 2018. 11. z. yang, z. zhou and y. liu. “from rssi to csi: indoor localization via channel response”. computing surveys, vol. 46, no. 2, p. 25, 2013. 12. a. m. ladd, k. e. bekris, a. rudys, l. e. kavraki and d. s. wallach. “robotics-based location sensing using wireless ether net”. wireless networks, vol. 11, pp. 189-204, 2005. 13. s. nadler, v. soroka, o. fuchs, r. korenshtein and e. sonsino. “presence zones for contextual location based services”. in innovations in clouds, internet and networks, 2008. 14. j. xiao, k. wu, y. yi, l. wang and l. m. ni. “pilot: passive device free indoor localization using channel state information”. in distributed computing systems, 3rd international conference on, ieee, 2013, pp. 236-245. 15. m. youssef and a. agrawala. “the horus wlan location determination system”. in proceedings of the 3rd international conference on mobile systems, applications, and services, 2005, pp. 205-218. 16. f. zafari, i. papapanagiotou and k. christidis. “micro location for internet-of-things-equipped smart buildings”. ieee internet of things journal, vol. 3, pp. 96-112, 2016. 17. h. liu, h. darabi, p. banerjee and j. liu. “survey of wireless indoor positioning techniques and systems.” ieee transactions on systems, man, and cybernetics-part c: applications and reviews, vol. 37, no. 6, 2007. 18. m. altini, d. brunelli, e. farella and l. benini. “bluetooth indoor localization with multiple neural networks”. 5th ieee international symposium, 2010, pp. 295-300. 19. s. kumar, s. gil, d. katabi and d. rus. “accurate indoor localization with zero start-up cost”. in proceedings of the 20th annual international conference on mobile computing and networking, 2014. 20. m. scherhaufl, m. pichler, d. muller, a. ziroff and a. stelzer. “phaseof-arrival-based localization of passive uhf rfid tags”. ieee mtt-s international microwave symposium 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system”. master’s thesis, purdue university, usa, 2016. 26. b. g. lee and s. choi. “broadband wireless access and local networks: mobile wimax and wi-fi”. usa: artech house publisher, 2008. 27. ibeacon. available from: https://www.decawave.com/sites/ default/files/resources/aps003_dw1000_rtls_introduction.pdf. [last accessed on 2017 dec 11]. 28. s. holm. “hybrid ultrasound-rfid indoor positioning: combining the best of both worlds”. ieee international conference, 2009, pp. 155-162. 29. decawave. “real time-location: an introduction”. available from: https://www.developer.apple.com/ibeacon. [last accessed on 2016 nov 01]. 30. f. ijaz, h. k. yang, a. w. ahmad and c. lee. “indoor positioning: a review of indoor ultrasonic positioning systems”. 15th international conference on, ieee, 2013, pp. 1146-1150. 31. n. b. priyantha, a. chakraborty and h. balakrishnan. “the cricket location-support system”. in proceedings of the 6th annual international conference on mobile computing and networking, 2000, pp. 32-43. 32. p. babu. “10 airports using beacons to take passenger experience to the next level”. available from: http://www.blog.beaconstac. com/2016/03. [last accessed on 2016 oct 08]. 33. intel. “wireless lan standards study”. available from: http://www. intel.com/content/dam/www/public/us/en/documents/casestudies/802-11-wireless-lan-standards-study.pdf. [last accessed 2016 nov 02]. 34. m. shaik. “ultra wide-band vs. wi-fi a study and comparisonof the two technologies”. available from: http://www.academia. edu/4810093/ultra. [last accessed on 2016 nov 03]. author query??? aq1: kindly cite figure 1-4 in text part (example like as table 1 and table 2) tx_1~abs:at/add:tx_2~abs:at 34 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 34-35 review article sand clock and the concept of time in einstein’s theory of relativity waria o. amin* cihan university-erbil, kurdistan region, iraq abstract the sand clock is a device used to measure time, consisting of two conical glass bulbs, which are connected vertically by a small opening neck. the top contains grains of fine dry sand. the upper part symbolizes the future, sand passes through the neck (which indicates present) to the lower part which turned past. in 1907, german scientist hermann minkowski (1864–1909) interpreted the concept of the four-dimensional space-time of einstein’s theory in a two-dimensional cone diagram, in which the upper cone symbolizes the future, the neck the present, and the lower part the past. the researcher has noticed a great likeness, to the point of congruence between the shape and function of both, the sand clock and the diagram. this resemblance aroused speculations that hermann minkowski had been inspired by the sand hour in drawing his diagram. it cannot be attributed to a merely chance. keywords: sand clock, einstein’s theory, minkowski, time introduction the sand clock is a device used to measure time, consisting of two conical glass bulbs, which are connected vertically to a small opening (the neck). the top contains grains of fine dry sand. the duration of the time scales is determined by factors that include the amount of material, its roughness, the size of the bulb, and the width of the neck. there are several accounts of who invented the sand clock. some attribute it to the pharaohs, some to the palmyras, and arab historians say that they were the ones who invented it.[1]. images of the sand clock, as a symbol of time, have been found in art, and on tombstones and other monuments since ancient times in many parts of the world are a template for microsoft word versions 2010 or later, figure 1. the upper part symbolizes the future, because sand will descend in the future, the sand passes through the neck (which symbolizes present), to the lower part and it turns into the past. einstein’s theory einstein (1) published his theory of relativity (2) in 1905, but he did not finish it. he proposed the concept of a four-dimensional space-time.[2] in 1907, hermann minkowski (3) (1864–1909) contributed to the theory of his former student albert einstein, formulated in 1905. his most famous achievement was his development of the special theory of relativity. as a result of minkowski’s contributions, relativity became geometrically possible to the theory of four-dimensional space-time, which is known as minkowski’s space-time.[3] minkowski explained the theory in a two-dimensional cone diagram in figure 2: as indicated in the picture, the upper part symbolizes the future, the neck symbolizes the present tense, and the lower part symbolizes the past. comparison by comparing the sand clock with the diagram, it becomes clear that: 1. both symbolize time 2. both consist of two conical symmetrical shapes connected vertically by a narrow neck 3. the upper part in both symbolizes the future 4. the lower part in both symbolizes the past 5. the neck in both symbolizes the present time corresponding author: waria o. amin, cihan university-erbil, kurdistan region, iraq. e-mail: wariaamin@gmail.com received: may 3, 2021 accepted: june 14, 2021 published: june 20, 2021 doi: 10.24086/cuesj.v5n1y2021.34-35 copyright © 2021 waria o. amin. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) amin: time in einstein’s theory of relativity 35 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 34-35 it appears from the comparison that there is great match to the point of congruence between the shape and function of the sand clock and the graphic image of minkowski. conclusions the conclusion of this article shows that: 1. hermann minkowski quoted his diagrammatic idea in his interpretation of einstein’s theory of relativity from the sand clock 2. the similarity between the sand clock shape and function, and the diagram reaches the point of congruence, which cannot be attributed to sheer coincidence. footnotes 1. albert einstein (1879-1955) was a german-born, swiss, and american physicist. he is famous for the father of relativity theory, for being the author of special relativity and general relativity, which were the first building blocks of modern theoretical physics. 2. relativity or the theory of relativity, one of the most famous theories of modern physics, which was developed by albert einstein at the beginning of the 20th century. there are two theories of relativity, the first is special relativity (1905) and the second is general relativity (1915). 3. hermann minkowski (1864–1909) is a german-born german mathematician and physicist, who worked as a professor of mathematics in konigsberg, zurich, and gottingen. he developed number geometry and used engineering methods to solve problems in number theory, mathematical physics, and the theory of relativity. he is considered one of the founders of convex geometry. minkowski’s most famous achievement was his development of the special theory of relativity in 1907ad, a theory that his former student albert einstein formulated in 1905. as a result of minkowski’s contributions, relativity became geometrically possible to the theory of four-dimensional space-time, which is known as minkowski’s space-time. references 1. j. t. anthony. time and measurement: studies in the history of horology and fine technology. new york: ashgate publishing company, 1993. 2. a. einstein. relativity: the special and general theory (translation 1920). new york: h. holt and company, 1916. 3. p. l. galison. minkowski’s space time: from visual thinking to the absolute world. historical studies in the physical sciences. baltimore, maryland: johns hopkins university press, 1979. 4. j. norton, space-time. einstein for everyone, 2015. available from: https://www.pitt.edu/~jdnorton/teaching/hps_0410/ chapters/spacetime/index.html. [last avccessed on 2015 feb 09]. figure 1: minkowski diagrams. source: norton, 2015 figure 2: minkowski diagrams. source: norton, 2015 . 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 1-4 research article evaluation of the effect of follower jammer on the mobile bluetooth network abdulqadir ismail abdullah1*, adil hussein m. al-dalawie2 1department of computer science, college of science, knowledge university, erbil, iraq, 2department of communication and computer engineering, college of engineering cihan university-erbil, erbil, iraq abstract bluetooth wireless communication technology has spread rapidly in the past 20 years. it has made life much easier with all the devices that use bluetooth technology around us such as mobile phones, laptops, and watches. with all the good and useful applications of bluetooth technology, it has been used in harmful and bad ways such as cheating in an examination. in this article, parameter design considerations have been discussed for a follower jammer to interrupt a bluetooth network, especially when it is used in unauthorized way. the effects of the follower jammer were studied and tested. the effects were calculated for time and jamming-over-signal ratio for three different distances (1, 5, and 10 m). the testing showed that the follower jammer designed was successful in disrupting the bluetooth signal. keyword: bluetooth, follower jammer, mobile jammer introduction bluetooth wireless technology since its development by ericsson in sweden in 1994 has become an essential tool of everyday life. bluetooth wireless technology (bwt) has been used by many devices such as laptops, mobile phones, and car stereo systems. it has made everyday tasks very easy and efficient.[1] bluetooth-enabled devices use ism frequency band range between 2.4 gh and 2.48 gh. it uses a technique called frequency hopping (fh). this technique is useful for reducing interference. there are three classes of bluetooth networks: class 1 is 100-m range, class 2 is 50-m range, and class 3 is 10-m range.[1] bluetooth wireless networks are subject to interference such as jamming which could be intentional, and this disrupts the service. not all disruptions are bad thing because some people nowadays use bluetooth networks for some purposes which are not allowed.[2] there are deferent types of jamming techniques such as overall band noise jamming or spot jamming and swept jamming. research into the evaluation of the effects of these three deferent methods on the mobile bluetooth network provides good motivation to make a comparison between these techniques. the authors in a research article[3] focused on overall noise jamming techniques, spot noise with bandwidth 5 mhz and 20 mhz, and sweep jammer. the conclusions of the research article[3] were that the block jammer signal (noise powered with different bandwidths 5 mhz and 20 mhz) does not affect the bluetooth network, and it needs high power to overcome the processing gain and the path loss attenuator. the effect appears within 1-m distance by increasing the power density of the jamming signal; the bandwidth of the jamming signal was set to 5 mhz and uses very slow swept noise in order to affect the channels sequentially and keep the channel out of service according to adaptive frequency hopping (afh) and that will reduce the processing gain with time. due to the power problems in the previous work, a new method had to be followed in order to reach better results that, in mid, a new technology was tested and studied in the research article such as the follower jammer technique, which is effected on fh.[3] follower jammer technique fh spread spectrum is particularly useful to combat jamming primarily because it is relatively easy to operate corresponding author: abdulqadir ismail abdullah, department of computer science, college of science, knowledge university, erbil, iraq. e-mail: abdulqadir.abdullah@knowledge.edu.krd received: mar 27, 2019 accepted: apr 09, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp1-4 copyright © 2019 abdulqadir ismail abdullah, adil hussein m. al-dalawie. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) abdullah and al-dalawie: evaluation of the effect of follower jammer on the mobile bluetooth network 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 1-4 over very large spread bands. however, fh can be efficiently jammed by follower jammers (also called “repeat back”) under certain conditions. in follower jamming, the jammer intercepts the transmitted signal, tries to determine the frequency of the hop, and then generates jamming in a narrow range about this frequency. it is the purpose of this article to discuss design considerations to account for such jammers.[4] figure 1 shows the block diagram of a simple follower jammer design. design consideration of the follower jammer bluetooth wireless network uses the fh techniques which operates in the 2.4 ghz ism band over 79 mhz bandwidth. this bandwidth is divided into 79 channels, and each channel has 1 mbps capacity; the time for transmission is separated among 625 µs slots, where each slot uses a new hop frequency technique, as shown in table 1.[1,3] according to these specifications, the design of the follower jamming must cover the band of frequency between 2402 mhz and 2482 mhz with sensitivity <−70 dbm and an output between 0.5 watt and 1 watt. in order to achieve this, the antenna that must be used must be an omnidirectional antenna covering 360° in elevation and 60° in horizontal, as shown in figure 2.[5] the band-pass filter must be passing the frequencies between 2402 and 2482 mhz with low attenuation around 2 db and reject rest frequencies out that range with attention −80 db (fourth-order filter), as shown in figure 3. the low-noise amplifier is designed to receive the bluetooth frequencies and amplified for suitable level, as shown in table 2. table 1: specification of bluetooth network items specification power o/p 1 mw channel b.w. 1 mhz operation bandwidth 79 mhz (2402–2482) mhz processing gain 19 db receiver sensitivity −70 dbm transmitting time 625 µs equipment distance (0.30–1.2) m figure 1: follower jammer block diagram figure 2: x-y plane omnidirectional antenna figure 3: band-pass filter characteristic table 2: low-noise amplifiers characteristic parameters min. – typ. – max. units frequency range 2400–2480 mhz gain 20 db noise figure 3.5 db rf input 1 db compression point 18 dbm input third-order intercept point 23 dbm table 3: modulator specification parameters min. – typ. – max. units rf frequency range 2400–2480 mhz if input bandwidth range 0.5 mhz lo input frequency range 2400–2480 mhz table 4: the calculation of the delay and attenuation for the different distances distance delay attenuating 1 m 6.66 ηsec 40.19 db 5 m 33.3 ηsec 54.17 db 10 m 133.2 ηsec 60.17 db abdullah and al-dalawie: evaluation of the effect of follower jammer on the mobile bluetooth network 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 1-4 the modulator should consider the specification, as shown in table 3. the noise generator can use pin diode (pn) noise generator with clock 500 khz or pin diode or a zener diode with amplifier and low-pass filter 500 khz. the power amplifier is going to amplify the modulated bluetooth frequencies by the noise. the output power was around 1 watt or 30 dbm to satisfy the jamming-over-signal ratio (j/s) >3 db. the filter and antenna are considered the same as mentioned before. the critical point in the follower jammer the time response of the jammer and time wave traveling from bluetooth sender to the jammer and from the jammer to bluetooth receiver must reduce the delay as much as possible to follow transmitting time of the bluetooth network. if we consider the wave speed at 3×108 m/sec, meaning the wave traveling 1 m during 3.33 ηsec and this time is very short comparing with the time of the transmission time which is 625 µsec and that gives the confidence in the method. operation consideration an important consideration is the distance of the jammer from the bluetooth network. for the design parameters in this research work we considered the distances where the jammer is located at 1, 5, and 10 m from the bluetooth network, as shown in figure 4. for this purpose, the distances d2 and d3 which represents the distances of the jammer from the bluetooth units as d2 and d3 are equalt to (1, 5, or 10 m). the delay and the attenuation based on the different distances d2 and d3 were calculated. the calculations are shown in the table 4. the estimation of the timing coverage on the frequency slot according to the distance can be seen in table 5 which shows the percentage time coverage on frequency hop, as we notice that the effect of delay from the distance is almost negligible. in order to do a scientific comparison with bluetooth wireless network specification and the follower jammer, we must calculate the link budget for all cases to find the effect from the power view and j/s ratio (table 6). with the follower jammer technique, the gain processing will be neglected because it uses the same frequencies at the same time, so that the gain margin against this technique will change from 19 db and will become 23.81 db. then, the bluetooth power at 1 m will be=−114+46.81=−67.19 dbm. table 7 shows the calculation of the j/s with respect to distance. from the results shown here, it is clear that this technique will affect the bluetooth wireless network and blocks it. conclusion many jamming techniques have been used to disrupt bluetooth wireless networks for many purposes. one useful way of jamming bw networks is when it is used by unauthorized persons. in this article, an approach was studied using the follower jammer in order to evaluate its effects on the network. the design considerations were followed in order to come up with the best results. from the test results, we can determine that the jammer follower can disrupt successfully a bluetooth network when it can receive the bluetooth signal. it was noticed that in order to avoid the circulation path from transmitter of the jammer to the jammer receiver, the antenna of the jammer must be isolated between them with using rf switch single pole single throw (spst) controlled by signal output of the power amplifier. table 5: percentage time coverage on frequency hop distance delay percentage ratio 1 m 6.66 ηsec 99.999 5 m 33.3 ηsec 99.994 10 m 133.2 ηsec 99.978 table 7: calculations of the j/s with respect to distance distance j dbm j/s db 1 m −9.81 57.38 5 m −24.17 43.02 10 m −30.17 37.02 j/s: jamming-over-signal ratio table 6: bluetooth network link budget item value units output power 0 dbm path attenuation @1 m 40.19 db sensitivity −70 dbm processing gain 19 gain margin 46.81 db figure 4: bluetooth network with jammer references 1. d. s. kushwaha. “application of blutooth wireless technology”. international journal of mathematics and computer research, vol. 1, no. 3, pp. 79-82, 2013. 2. p. patel, a. merchant, n. tailor and c. trivedi. “bluetooth security issues”. international journal of computer science and information technologies, vol. 6, no. 6, pp. 5295-5299, 2015. 3. a. h. m. al-dalowi and g. qasmarrogy. “performance evaluation of noise power jammer on the bluetooth network”. international journal of computer networks and communications, vol. 6, no. 5, pp. 167-174, 2014. 4. e. b. felstead. “follower jammer considerations for frequence hopped spread spectrum”. ieee military communications 5. p. d. vita. “antenna selection guidelines”. st microelectronics, switzerland, 2012. . 82 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 82-84 research article detection and evaluation of effective of digital communication of drug on human body zakaria k. jalal1*, abdulrazak a. mohammed2, adil h. mohammed2 1english department, astar education institute, vasteras, sweden, 2department of communications and computer engineering, cihan university, erbil, kurdistan region, iraq abstract nowadays, there is a wide using of mobile and internet from various ages, due to some bad systems which they work to send signals (sound wave frequency) within the music or will be spread to affect the brain of the humankind. therefore, this paper expresses the detection and evaluation of (the signal of sound frequency) which influence the human brain, it has been measured some frequency by programming in the mobile and found a trend to detect such frequencies and a way to produce a human being from it. keywords: digital communication, digital drug, electromagnetic radiation introduction the updated technology of communication (mobile and network) makes our social life more easy for communication and it will be active, but at the same time, it causes our lives many dangerous issues (such as electromagnetic radiation and sound pressure, especially ultrasonic waves) for all ages as well. meanwhile, we need to point to specify the negative side of this new technique equipment and define all the waves with their bad usages. as we know the ear is a sound pressure receptor (or sensor) that influences our behavior (physically or physiology) because these techniques are getting inside houses without any our permission or observations. whereas, the sense of hearing counts on the sensory receptors of the inner ear known as hair cells and the hair cells are extremely vulnerable and can be affected by disease, ageing and over-exposure to loud noise. once destroyed, they do not regenerate. sound properties and characteristics[1] it is the compression wave traveling down material bar. it is a physiological sensation of hearing and it looks like mechanical motion which is considered as longitudinal waves (number of consecutive rarefaction and consecutive compression as shown in figure 1). characteristics of sound amplitude, pitch, frequency (n) =1/t, period (t) =1/f, wavelength (λ). speed (v), quality, tone, and a note. amplitude (a): it is a maximum displacement of particles vibration from their meaning poison and it is two types: (a) louder sound (higher energy) and (b) soft sound. (it counts on wave distribution). wavelength (λ): it is a distance between two consecutions compression and two consecutions rarefaction in the longitudinal frequency (n): it is a number of waves produced by a unit time and its unit is (hertz) n=1/t. pitch: it is an interpretation of the frequency of a sound by the brain. it affects on the wave shape where the high pitch that means high frequency and amplitude and short wavelength (which gives a loud sound [or pitch]). period (t): it is a time duration of the wave (frequency) t=1/n or it is sound propagate on an intervals between two successive consecutions compression and two consecutions rarefaction. quality (timbre) of the sound: it is a character helps us to make a different between two sound different sources but same pitch or loudness. tone, it is a sound wave of single frequency. a note, it is a sound wave of the number of frequencies. speed (v): it is the ratio between distance (wavelength) and the time. cihan university-erbil scientific journal (cuesj) corresponding author: zakaria k. jalal, english department, astar education institute, vasteras, sweden. e-mail: zkarim53@yahoo.com received: apr 04, 2019 accepted: apr 25, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp82-84 copyright © 2020 zakaria k. jalal, abdulrazak a. mohammed, adil h. mohammed. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ jalal, et al.: detection of digital communication of drug on human body 83 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 82-84 speed of sound v =dis./time = λ/t but n=1/t, then v=n λ wavelength: the distance between two consecutive compressions and two consecutive rarefactions in the long of the wave moving. the speed of the sound depends on type of the medium (material). there are several effectives of the sound (depending on sound pressure level as shown in table 1) on humankind such as physic scaly, physiology, and behaviors. sound effects digital communication affects humankind’s body in many various methods such as drug detection, wireless communications, and various channels including several quantities of e-posts, email newsletters, social, targeted outreach to media, and health-care professionals in a bad way. these messages and communications announcement among people can be changed from new or appear risks related to concurrent conditions (for example, pregnancy or existing liver or kidney disease) or attentions about potential medication errors. moreover, digital communication including issues influencing a great deal of patients, potentially serious or life-threatening injuries events, or medication errors that may result in serious or life-threatening conflicting answers. whereas the digital communication influences directly on the behaviorism, mental and psychology of the person without feeling it because he/she will be detected automatically and this will affect humankind’s mentality and it may lead to commit suicide as well. the arriving narrative forms of communication increase information processing and increase the persuasiveness of messages and people become transported into a situation that can enhance emotions, attitudes, and behaviors. as a result, it will be not easy to get rid of this detection which became part of his/her life everywhere and anywhere. digital drugs[2] more accurately called binaural beats which are sounds that can be capable of changing brain wave patterns and inducing an altered state of consciousness similar to that affected by taking drugs or achieving a deep state of meditation. binaural beats occur when two tones with slightly different frequencies are played together. without headphones, the slight difference in the two frequencies is perceived by the listener as a single tone that wavers slightly. hence, headphones, however, the two tones are isolated and the listener hears each frequency clearly in a different ear. as the brain processes the two tones, it must take into account that the slight difference among the frequencies. as for the listener, this difference is perceived as rhythmic beats inside the head. the brain processes rhythmic stimulus as electrical impulses. the goal of digital drugs is to purposely overcome the electrical impulses and encourage the listener’s brain to synchronize its brain waves with the binaural beats. this synchronization which is achieved by selecting binaural tones within a particular frequency level is called frequency following response and is a part of a concept called entrainment. entrainment, the synchronization of one biological rhythm to another is not a new concept. it forms the basis for many sorts of meditation and medical biofeedback. properties of digital drugs[3,4] frequencies are measured in units called hertz (hz) by listening to two tones in difference falls within a particular hz level, the listener hopes to achieve a particular mood or change in energy. for instance, if the listener wants to be very relaxed, he might choose to listen to a tone with 140 hz in one ear and 145 hz in the other. the listener’s brain would perceive the viciousness between the two frequencies (5 hz) and adjust the listener’s brain waves accordingly. if the listener wants to be energized, however, he might choose to listen to 130 hz in one ear and 150 hz in the other. a difference of 20 hz in frequencies would produce a various state of mind as shown in table 2. sound spectrum measuring[5] using a special sound meter program (within mobile application) which has been measured by some frequency spectrum sound for ultrasonic to supersonic (where the range of humane hear from 20 hz to 20,000 hz), some frequencies are out of the hearing range, as shown in figure 2, starting from 2a for frequency before 20 hz to 2 d after 20,000 hz. table 1: effective of sound level sound pressure level permissible exposure time 115 db 0.468 min (≅30 s) 112 db 0.937 min (≅1 min) 109 db 1.875 min (<2 min) 106 db 3.75 min (<4 min) 103 db 7.8 min 100 db 15 min 97 db 30 min 94 db 1 h 88 db 4 h 82 db 16 h figure 1: characteristics of sound wave jalal, et al.: detection of digital communication of drug on human body 84 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 82-84 that means there are a lot of frequencies affect on human’s body without hearing it. results and discussion[6,7] 1. using mobile has a side effect which is the microwave radiation as well as the effect of ultrasound waves 2. the effective of microwaves is physically, but the effective of ultrasound is physiological, physical, and behavior 3. the output of microwave effective will appear after a long period while the effective of ultrasound will appear during a short period 4. digital well-being considerations. when optimizing digital health, multiple health-relatedcomponents can be taken into consideration.[6] 5. the effective is usually different from person to other because it counts on the followingfactors: a. the time period that the person stay under the frequency of sound wave as shown in table 3 b. the behavior and situation of the person during the effective c. the power and pressure level of the sound. table 1 shows the level and time of affecting (exposure time) d. using headphone or without (because using headphone is effected more). conclusion digital well-being is a term used by health professionals, researchers, and device manufacturers to describe the concept of humans interact with technology. the experience should support mental and/or physical health in a measurable way. the goal of improving digital well-being is to design technology in such a way that it promotes healthy use and proactively assists the user to maintain a healthy lifestyle.[4,6] using a digital communication (mobile, laptop, and ipad) with internet, the user certainly will receive and affect the unwanted sound frequency wave; of course, the user effective counted on the factors that mentioned above. we should observe the kids and children to be sure; they are using the digital instruments on right ways by: a. they should use free beneficial programs of any digital drug b. control and check the children not to join or purchasing materials or playing games witha bad group c. observe the children’s behavior in any serious forward motion (such as dancing, doing something, or chatting) and you should know in which programming he is busy with it. references 1. c. j. mosees and c. a. moyer. “modern physics serway”. saunders college publishing, usa, 1989. 2. ryan singel pokes fun at the media frenzy over i-dosing on wired.com. available from: https://www.whatis.techtarget.com›definition›digital. 3. ron doyle blogs about i-dosing: digital drugs and binaural beats for psychology today. available from: https://www.whatis. techtarget.com›definition›digital. 4. available from: https://www.de.wikipedia.org/wiki/digitaldruck. [last accessed on 2019 mar 25]. 5. visual analyzer; 2011. available from: https://www.reviewjournal. com›life›healthon. [last accessed on 2019 dec 26]. 6. available from: https://www goglecom/search2 q=digta druck&rlz= 1c2rucy_entq797iq797. [last accessed on 2019 dec 26]. 7. h. m. michael and n. t. peter. “acoustical imaging”. kluwer academic publishersm, london, 2000. table 2: state of sound frequency type frequency range (hz) state of mind delta 0.5 hz – 4 deep sleep theta 4 hz – 8 drowsiness (also first stage of sleep) alpha 8 hz – 14 relaxed but alert beta 14 hz – 30 highly alert and focused table 3: noises can cause damage to wear hearing type noise value (db) period a 85 db regular and prolonged exposures to noise at or above 85 db (averaged 8 h/day) b 100 db regular and prolonged unprotected exposures of more than 15 min/day risk permanent hearing loss c 110 db regular and prolonged unprotected exposures of more than 1.5 min/day risk permanent hearing loss figure 2: (a-d) sound frequencies spectrum dc ba . 75 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 75-79 research article design and implementation a non-uniform helical antenna in frequency range of 450–850 mhz for ultra-high-frequency television application firas muhammad zeki mahmood*, adil h. mohammed al-dalawie department of communication and computer, college of engineering, cihan university-erbil, iraq abstract in a domestic region, the people are living inside cities, they complaining from the weak of local tv broadcasting signals in frequency range of 450–850 mhz, in this paper we got a solution by increase the gain antenna by design new antenna have a wideband and high gain compare with antennas are available in the local market. a non-uniform helical antenna designed using helical antenna calculator according to equations then implemented and measured the radiation pattern and gain of antenna, then compare with commercial loop antenna gain found increase in received power about 10–12 db more than loop antenna gain. the results of measurement applied in matlab program to draw the pattern of the antenna. keywords: measurements and television, non-uniform helical antenna, ultra-high-frequency antenna introduction the ieee standard definitions of terms for antennas (ieee std 145–1983) defines the antenna or aerial as “a means for radiating or receiving radio waves.”[1] a convenient element for providing circular polarization is the wire helix. the helical antenna consists of a metal conductor wound in the form of helix and backed at one end by a metal ground plane. it conveniently fed from a coaxial cable with the center conductor connected to the helix and the outer sheath connected to a circular ground plane of the radius of half-wavelength approximately. the helical geometry characterized by its diameter of turn spacing, pitch angle, uncoiled length of turn, and number of turns.[2] the people are living our side cities, they complain from weak of the local tv broadcasting signals, so we got a solution by increase the gain of antenna by design new helical antenna has wideband and high gain comparing what available with commercial antenna. link budget equation for simple formula for link budget equation can be state as, pr pt gt gr l= + + − (1) where, pr is received power (db), pt is transmitted power (db), gt transmission gain (db), gr is receiver gain (db), and l is total losses (db). the item gain will contain the two main antennas gain for transmitting and receiving, the focus on receiving gain which is in our control hand and we success to give a way to boost the received power if increase the gain of receiver antenna.[3] non-uniform helical antenna according to exemplary embodiments of the present invention, non-uniform helical antenna is described for use in two or more frequencies tuning to both resonance frequencies can be accomplished by varying parameters of the helical antenna including, for example, the pitch angle, coil diameter, length, and number and spacing of the coil turns; figure 1 shows an example for non-uniform helical antenna.[4] cihan university-erbil scientific journal (cuesj) corresponding author: firas muhammad zeki mahmood, department of communication and computer, college of engineering, cihan university-erbil, iraq. e-mail: firas.zeki@cihanuniversity.edu.iq received: apr 10, 2019 accepted: apr 24, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp75-79 copyright © 2019 firas muhammad zeki mahmood, adil hussein mohammed al-dalawie. al-aubi. this is an open-access article distributed under the creative commons attribution license. mahmood and al-dalawie: design and implementation a non-uniform helical antenna in frequency range of 450–850 mhz for ultra-high-frequency television application 76 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 75-79 main parameters of helical antenna pitch angle pitch angle is the angle between a line tangent to the helix wire and plane normal to the helix axis. α π = arctan s d (2) where, α is the pitch angle, s is the turn spacing (center to center), and d is the diameter of helical. and, h = ns (3) where, h is total height of helix antenna, n is number of turns on the helix antenna, and s is the turn spacing (center to center). the helix antenna functions for pitch angles (α) between 12° and 14°. typically, the pitch angle taken as 13°.[3] the antenna in figure 2 is a left-handed helix antenna because if you curl your fingers on your left hand around the helix your thumb would point up (also, the waves emitted from this helix antenna are left hand circularly polarized). if the helix antenna wounded in another way, it would be a righthanded helical antenna. the radiation pattern will be maximum in the +z direction (along the helical axis in figure 1). the design of helical antenna primarily based on empirical results, and the fundamental equations represented here. helix antennas of at least 3 turns will have close to circular polarization in the +z direction when the circumference c is close to a wavelength: 3 4    4 3 c   ≤ ≤ (4) once the circumference c chosen, the inequalities above roughly determine the operating bandwidth (bw) of the helix antenna.[2,5] for instance, if c = 0.5 m, then the highest frequency of operation will be given by the smallest wavelength that fits into the above equation, or λ = 0.75 c = 0.375 m (5) which corresponds to a frequency of 850 mhz, the lowest frequency of operation will be given by the largest wavelength that fits into the above equation, or λ = 1.33 c = 0.667 m (6) which corresponds to a frequency of 450 mhz. hence, the fractional bw is 56%, which is true of axial helical antennas in general.[6] input impedance the helix antenna is a traveling wave antenna, which means the current travels along with the antenna and the phase varies continuously. in addition, the input impedance is primarily real and can be approximate in ohms[7] by zin c = 140  (7) radiation pattern of the helical antenna the helix antenna functions well for pitch angles (α) between 12° and 14°. typically, the pitch angle taken as 13°. the normalized radiation pattern for the e-field components given by: ε ε ω ω θ φ π θ∝ ∝ sin cos � sin sin2 2 2 n n (8) ω = −( ) − +    ks n �cosθ π1 2 1 � � (9) for circular polarization, the orthogonal components of the e-field must be 90° out of phase. this occurs in directions near the axis (z-axis in figure 2) of the helix.[5] the axial ratio for helix antennas decreases as the number of loops (n) added and can be approximated by: figure 1: non-uniform helical antenna figure 2: main parameters of helical antenna mahmood and al-dalawie: design and implementation a non-uniform helical antenna in frequency range of 450–850 mhz for ultra-high-frequency television application 77 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 75-79 ar n n = +2 1 2 (10) where, ar is axial ratio. gain of the antenna the gain of the helix antenna can be approximated by, g c ns c nfs c = = 6 2 6 22 3 2 3 3 . � � . �  (11) n the above, c is the speed of light. note that for a given helix geometry (specified in terms of c, s, and n), the gain increases with frequency. for an n = 10 turns helix that has a 0.5 m circumference as above, and a pitch angle of 13° (giving s = 0.13 m), the gain is 8.3 (9.2 db).[8] for the same example helix antenna, the pattern is shown in figure 3. the half-power beamwidth for helical antennas can be approximate (in degrees)[3] by: hpbw c ns = 65� /   (12) polarization linear polarization occurs when electromagnetic waves broadcast on a single plane (either vertical or horizontal). compared to linear polarized antennas of the same gain, circular polarized antennas will have a shorter read range because they lose about 3 db splitting their power across two separate planes.[9,10] design specifications it is considered the helical antenna design specifications which are as follows: • frequency range: 450–850 mhz • gain: 10 dbi ± 0.5 db • axial ratio: <1.5. using online antenna calculator program to calculate the parameters that are required to the helical antenna design as shown in figures 4 and 5, respectively.[11] the main factors of helical antenna were entire such as wavelength, number of turns, and the space between each coil to calculate the antenna gain and wire diameter, half-power beamwidth, beamwidth first nulls, and effective aperture. table 1 shows the important factor for implemented the nonuniform helical antenna. figure 3: normalized radiation pattern for helical antenna (db) figure 4: helical antenna design with 450 mhz figure 5: helical antenna design with 850 mhz table 1: main parameters of non-uniform antenna designed parameters at 850 mhz at 450 mhz number of turns 3 5 spacing between coils 8.82 cm 16.7 cm d (diameter) 11.2 cm 21.2 cm g (antenna gain) 9.55 dbi 11.8 dbi z (characteristic impedance) 150 ω 150 ω mahmood and al-dalawie: design and implementation a non-uniform helical antenna in frequency range of 450–850 mhz for ultra-high-frequency television application 78 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 75-79 the implementation of non-helical antenna figure 6 shows the antenna which is designed using helical antenna calculator according to equations then implemented from 5 turns for 450 mhz and 3 turns for 850 mhz with circular plate ground and other parameters as shown in table 1 to cover ultra-high-frequency band for tv application then tested in the laboratory of antenna and microwave/communication and computer department/cihan university – erbil. measurements method the far-field patterns measured on the surface of a sphere of constant radius. any position on the sphere identified by the directional angles θ and φ of the spherical coordinate system. a principal plane must contain the direction of maximum radiation. a simplified block diagram of a pattern measurement system is given in figure 7. the method used a commercial figure 7: set of antenna test[2] figure 8: implemented non-uniform helical antenna under test figure 9: commercial loop antenna radiation pattern figure 6: non-uniform helical antenna manufactured table 2: measurements of commercial loop antenna step no. angle (degree) power (dbm) step no. angle (degree) power (dbm) 1 −180 −16 20 10 −19 2 −170 −19 21 20 −20 3 −160 −22 22 30 −22 4 −150 −25 23 40 −28 5 −140 −28 24 50 −29 6 −130 −29 25 60 −30 7 −120 −30 26 70 −32 8 −110 −31 27 80 −32 9 −100 −32 28 90 −32 10 −90 −32 29 100 −32 11 −80 −32 30 110 −31 12 −70 −32 31 120 −30 13 −60 −30 32 130 −29 14 −50 −29 33 140 −28 15 −40 −28 34 150 −25 16 −30 −22 35 160 −22 17 −20 −20 36 170 −19 18 −10 −19 37 180 −16 19 0 −16 mahmood and al-dalawie: design and implementation a non-uniform helical antenna in frequency range of 450–850 mhz for ultra-high-frequency television application 79 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 75-79 figure 10: non-uniform helical antenna radiation pattern table 3: measurements of non-uniform helical antenna implemented step no. angle (degree) power (dbm) step no. angle (degree) power (dbm) 1 −180 −22 20 10 −9 2 −170 −21 21 20 −10 3 −160 −20 22 30 −12 4 −150 −19 23 40 −15 5 −140 −18 24 50 −17 6 −130 −15 25 60 −18 7 −120 −18 26 70 −19 8 −110 −20 27 80 −20 9 −100 −22 28 90 −21 10 −90 −21 29 100 −22 11 −80 −20 30 110 −20 12 −70 −19 31 120 −18 13 −60 −18 32 130 −15 14 −50 −17 33 140 −18 15 −40 −15 34 150 −19 16 −30 −12 35 160 −20 17 −20 −10 36 170 −21 18 −10 −9 37 180 −22 19 0 −6 radiation pattern of non-uniform helical antenna under test, respectively, these results show the deferent received power about 10–12 db more with non-uniform helical antenna gain. the results of measurements applied in matlab program to draw the pattern of the antenna; figure 9 shows loop antenna radiation pattern and figure 10 shows non-uniform helical antenna radiation pattern, respectively. conclusion in this paper, designed and implemented a non-uniform helical antenna by using helical antenna calculator and equations then tested with frequency range 450 to 850 mhz for tv receiver and compare the gain measurements with commercial loop antenna in local market. the results of measurement show the deferent received power about 10–12 db more with nonuniform helical antenna gain but with circular polarization that meaning the antenna will lost 3 db because the tv broadcasting transmitted linear polarization; still, the gain of antenna manufactured improved about 8 db. references 1. a. c. balanis. “antenna theory analysis and design”. 3rd ed. hoboken, new jersey: john wiley and sons, inc., 2005. 2. f. r. connor. “antennas: introductory topics in electronics and telecommunication”. 2nd ed. london: edward arnold, 1989. 3. j. d. kraus. the helical antenna. in: “antennas”. ch. 7. new york, usa: mcgraw-hill, 1988, p. 265-339. 4. available from: https://www.patentimages.storage.googleapis. com/us6112102. 2019. 5. w. l. stutzman. “antenna theory and design”. 3rd ed. new york: mcgraw-hill, 1998. 6. e. p. ogherohwo and b. barnabas. “design, construction and performance analysis of helical antenna operating at 5.8ghz”. international journal of advanced research in physical science, vol. 2, no. 11, pp. 29-38, nov 2015. 7. m. m. ilić, s. v. savić and p. djondović. “internal matching of uhf helical antenna exciters for magnetic resonance imaging”. api 1.1. proceeding of 3rd international conference on electrical, electronic and computing engineering, zlatibor, serbia, 2016, pp. 1-5. 8. y. liang, j. zhang, q. liu, and x. li. “high power dual branch helical antenna”. ieee antennas and wireless propagation letters, vol. 17, no. 3, pp. 472-475, 2018. 9. j. l. dinkić, d. i. olćan, a. g. zajić and a. r. djordjević. “comparison of optimization approaches for designing non uniform helical antennas,” in 2018 ieee international symposium on antennas and propagation and usnc/ursi national radio science meeting, boston, ma, usa, 2018, pp. 1581-1582. 10. available from: https://www.blog.atlasrfidstore.com/circular -polarization-vs-linear-polarization. 2018. 11. available from: http://www.daycounter.com/calculators/helical antenna-design-calculator.phtml. 2018. loop antenna used in local market and implemented non-uniform helical antenna; figure 8 shows implemented non-uniform helical antenna under test. tables 2 and 3 show the radiation pattern of loop antenna and measurements of . 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 21-24 technical article design and implementation a novel research management system omar talal hamid* department of computer science, cihan university-erbil, kurdistan region, iraq abstract publications and researches are one of the important activities of higher education institutions (heis) worldwide. many heis, especially in iraq, are facing different types of problems in managing publication information. this paper presents the design and implementation of researches management system (rms) to be adopted in one of the state universities in iraq. rms is a web-based application to manage the publications and researches information in heis. the main goals of this application are collecting all submission publication information in a central database, generating different types of publication reports, generating real-time different statistics, saving all researchers information in the hei, and facilitating checking and tracking the submitted researches. keywords: component, database, management information system, web application introduction the researches management system (rms) is aimed to manage researches information in higher education institutions (heis) to improve the efficiency and effectiveness of the research information management processes. scientific researches and publications are crucial activity for heis. the increase in the number of publications, with many types and quantities, has caused many problems in the management, storage, and organization of this information.[1] another important issue is waiting. this problem occurs because a lot of time is required to deliver this publication information to the scientific affairs department, as the data are still manually recording and stored in many files (paper based).[2] furthermore, lack of software availability results in a lot of daily work activity in scientific researches departments, as the clerks must collect publication information, statistics of publications, and reporting by hand (manually).[3] at present, the development of computer system and internet technologies has helped humans to do their work as best as possible.[4] one of these computer systems is management information systems (miss), which is an application used to help and support the human to do his activities such as in a private or public sector institution.[5] the management and monitoring of publications in heis are an extensive and complex undertaking. there are many problems in the current paper-based system. these problems include collecting information from the colleges, organizing the classification of publications, generating publication reports, compiling publication statistics such as local, international, indexed, and no indexed, and displaying all details for each publication such as author names, publisher name, date of publishing, and link for research. the last problem is that there is no database for all researchers’ information. all these issues take a lot of time to do by hand. rms is a web application that is easy to use and work with the different size of screens. rms was developed using various web technologies such as php, html5, css3, and bootstrap. php is used to implement the server-side of the system. html5, css3, and bootstrap are used to develop the graphical user interface. the widespread uses of database technologies have become an important part of information management systems.[6] rms used mysql server as a back-end database because it is free, open source, has strong functions, high speed, access to data by multiple users in real time and handling big size of data.[7] system architecture the general structure of rms is shown in figure 1. there are three types of users (system admin, college admin, and corresponding author: omar talal hamid, cihan university-erbil, kurdistan region, iraq. e-mail: omar_talal20022003@yahoo.com received: apr 11, 2019 accepted: apr 22, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp21-24 copyright © 2019 omar talal hamid. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) hamid: design and implementation a novel research management system 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 21-24 researchers). all users can access the system through any web browser or smartphone. there are three levels of management authentication in rms and each authentication has own specific roles. the first level is for researchers to submit all their publication information to the scientific affairs in the college (college admin). the second level is for college administrators. each college has an administration person to check all publication information that is submitted by his/her college researchers. after checking this information to ensure that everything is correct, the publication is approved. when he/she has approved this information, it will be available for the third level (system admin) or scientific affairs department in the university. the third level is for system admin, and he/she can see all college information separately along with real-time statistics for each college and the entire university. figure 2 shows the use case diagram for rms which illustrates the interaction between users and system functionalities. as depicted in figure 2, the system admin possesses full authentication such as employee management and can generate any kind of report, see all publication information, and change the user roles in the system. college admin has full authentication only inside his/her college. he/she can see all his/her college information such as college reports, college publication information, and college publication statistics and can add/edit/delete the researcher’s information and accounts in the system. the last one is the researcher; he/she can submit his/her publication information to his/her college and see his/ her personal publication statistics. rms database mainly includes the tables of publication_ information, employees_information, departments, colleges, and jobs. the database schema for rms is shown in figure 3. implementation of rms rms is accessible from any smart device with any web browser to allow the staff and researchers to use the system. rms consists of different modules: researchers management module, publications management module, and reports. each module is related to one authentication. the researchers management module is provided to the administration system staff for adding, updating, deleting, and viewing all researchers information. publications management module deals with all functionalities that are related to the management of researches such as adding new article information, viewing article information, editing article information, or deleting article information. reports module is responsible for generating many different types of reports. this module is accessible only for administration staff as they are the ones responsible for reporting on research activities. rms is a multi-authentication system. it means that there are many different roles/authentications in the system (system admin, college admin, and researchers). each user has a username and password to login to the system. if this information is correct, then the system checks his/her role (system admin, figure 1: structure of researches management system figure 2: use case diagrams for researches management system figure 3: database schema for researches management system hamid: design and implementation a novel research management system 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 21-24 college admin, or researcher) and opens the home page as shown in figure 4. the details of this page are different from one user to another depending on his/her role in the system. the main home page consists of three dynamic parts: general statistics, college details, and main menu. general statistics is the current total statistic. these statistics differ from one user to another depending on the user role. system admin shows all approved, waiting (checking) papers, total number of researchers, and total number of college admin and system admin in the university. college admin shows the statistics for his/her college. researcher shows the researcher’s own statistics. figure 4 shows the general statistics part. college details display the detailed statistics of publications depending on publication classification such as local, international, scopus, nature, science, clarivate, sjr, arabic journals, and not indexed. figure 5 shows the college details part of home page. main menu is a dynamic menu which displays different submenu depending on the user role. for example, system admin shows all subpages, college admin shows only publications management, college reports, college researchers management, and my profile, while researcher displays only my publications and my profile. figure 6 shows the main menu part of home page. my publication submenu includes a list of pages (add new publication, displays all my waiting publications, and displays all my approved publications). when the researcher submits a new publication, the rms sends a request to his/her college admin as a waiting publication to check this information. the college admin receives this request and starts to check this information. after checking, he/she can approve this information and will display this information for the system admin as an approved publication. in this case, the researcher figure 4: general statistics part figure 5: college details part figure 6: main menu part figure 7: add new publication page hamid: design and implementation a novel research management system 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 21-24 cannot update this publication information. figure 7 shows how to add new publication information. another important submenu is reports, which include a list of pages such as waiting report to display all waiting publications for college or all university depending on the user role, approved report to display all approved publications. the last and most important one is a custom report, which is a page used to generate different kinds of publication reports, for example, publication within two different dates or specific classification such as only scopus, ieee, local, international or clarivate, college, department and so on. the results can be downloaded as an excel sheet. figure 8 shows how to generate custom report. researchers management submenu is including a list of pages such as add new researcher, display all researchers, edit researcher information, change researcher role, and display all researchers information. figure 9 shows how to display all researchers information. setting submenu also includes a list of pages (add new college and add new department). this submenu is only for system admin to do the system configuration. the last submenu is my profile. in this page, the user can edit personal information such as name, email, password, and phone. conclusions rms was successfully developed and implemented as a web-based application for scientific affairs department. this system was used and tested by one of the iraqi governmental universities. this testing was done by real users for usability and it showed an acceptable performance and ease of use in solving real problems of publication information management at the university. rms is used to improve efficiency and effectiveness of scientific affairs in the university. furthermore, it is used to reduce the time and effort to create publication reports and reduce the time to check the publication information. moreover, rms is an electronic archiving for all researchers information in the university. rms connects all colleges in real time with the scientific affairs department. the evaluation of scientific affairs department for rms was positive. it is easy to use, offers high accuracy statistics, makes it easy to search for any information about the publications, and is faster than paper process and suitable for managing publication and researches information at heis. finally, the rms has achieved all objectives and fulfilled its purpose to develop a novel web-based application to collect and manage all publication information. this could be a low cost, high-efficiency solution, with high accuracy, and faster for managing publication information in heis. references 1. k. liu, j. jiang, x. ding and h. sun. “design and development of management information system for research project process based on front-end and back-end separation,” 2017. 2. y. liu, y. yao, x. zhang, b. liu and c. ding. “design of research management system based on workflow and rapid development platform technology”. international conference on estimation, detection and information fusion, 2015, pp. 329-334. 3. k. i. satoto. “design of management information systems research, publications and community service”. international conference on information technology, computer, and electrical engineering, 2015, pp. 117-122. 4. a. s. perbangsa and j. suhartono. “research grants management information systems model”. conference paper, nov 2016, pp. 247-252. 5. s. shinde, s. nalawade and a. nalawade. “green computing: go green and save energy”. international journal of advanced research in computer science and software engineering, vol. 3, no. 7, pp. 1033-1037, 2013. 6. “developing a web-based research administration and management system: case study of a south african”. theses and dissertations, jun 2017. 7. k. i. satoto, r. r. isnanto, r. kridalukmana and k. t. martono. “optimizing mysql database system on information systems research, publications and community”. 3rd international conference on information technology, computer, and electrical engineering, 2016, pp. 1-5. figure 8: custom report page figure 9: researchers information page . 80 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 80-84 research article exploiting wavelet transform, principal component analysis, support vector machine, and k-nearest neighbors for partial face recognition mustafa zuhaer nayef al-dabagh1, mustafa h. mohammed alhabib2*, firas h. al-mukhtar3, hussein ibrahim hussein4 1department of computer science, knowledge university, kurdistan region, iraq, 2department of communications and computer engineering, cihan university-erbil, kurdistan region, iraq, 3department of information technology, catholic university in erbil, kurdistan region, iraq, 4school of computer and communication engineering, universiti malaysia perlis, malaysia abstract facial analysis has evolved to be a process of considerable importance due to its consequence on the safety and security, either individually or generally on the society level, especially in personal identification. the paper in hand applies facial identification on a facial image dataset by examining partial facial images before allocating a set of distinctive characteristics to them. extracting the desired features from the input image is achieved by means of wavelet transform. principal component analysis is used for feature selection, which specifies several aspects in the input image; these features are fed to two stages of classification using a support vector machine and k-nearest neighborhood to classify the face. the images used to test the strength of the suggested method are taken from the well-known (yale) database. test results showed the eligibility of the system when it comes to identify images and assign the correct face and name. keywords: k-nearest neighborhood, partial face recognition, principal component analysis, support vector machine, wavelet transform introduction smart face recognition systems are now widely used in diverse locations such as universities, laboratories, airports, and many other public service facilities that attempt to identify and prevent the intrusion of undesirable individuals into such institutions.[1] many researches and researchers brought this subject under the spotlight, trying to aid in finding ultimate practical solutions for the final facial identification of suspected or undesirable personnel. the majority of researches aim at the exploration of new approaches where human qualities are its center of interest, which leads to a facial recognition process that requires a relatively small execution time with easy implementation in many applications.[2,3] when using wavelet transform for feature extraction, one of the most common and popular methods for extracting facial features are called “eigenface” technique.[4,5] eigenface is used by the well-known principal component analysis (pca) algorithm, which transforms the facial image and reduces its dimensionality into low-dimensional space. pca converts the image matrix into a reduced single orthogonal vector with some special values called the eigenface. applying pca to more than one image as a training set will align many vectors next to each other, leading to a linear matrix that has some data variances used for later steps.[6] the next step of the presented automatic recognition system is classification. there are many methods that can be used as classifiers to categorize the input image according to the matrix of data variances resulted from the pca step. one of these methods is support vector machine (svm),[7-9] which is basically a binary classifier that is considered a supervised learning algorithm, which aims at finding what is called the optimal hyperplane in the n-dimensional classification space that gives the highest possible margin between classes. svm achieves this by converting the training set obtained in the cihan university-erbil scientific journal (cuesj) corresponding author: mustafa h. mohammed alhabib, department of communications and computer engineering, cihan university-erbil, kurdistan region, iraq. e-mail: mustafa.alhabib1986@gmail.com received: apr 13, 2019 accepted: apr 24, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp80-84 copyright © 2019 mustafa zuhaer nayef al-dabagh, mustafa h. mohammed alhabi, firas h. al-mukhtar, hussein ibrahim hussein. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ al-dabagh, et al.: exploiting wavelet transform, pca, svm and k-nn for partial face recognition 81 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 80-84 case of complex nonlinear data into a higher dimensional space where it is possible to separate the data by a linear hyperplane. svm is often reported to achieve better results than other classifiers.[6,10,11] another classification method is the nearest neighbor (nn) algorithm.[12-14] according to nn, one set of samples is handled such that these samples can be classified by minimizing the distance among them. then, the samples can be represented in different classes where the nn method classifies the testing samples by prediction as it calculates the best linear representation for each class. k-nn is applied primarily due to the fact that it is simple and capable of attaining an outcome that resembles the output of more complicated methods as well as its capability of being used in various applications.[15] many previous researches have been done in the same field with similar methodology. some of these works focused on face identification methods that depend on the concept of cloud computing.[1] the main issue that these methods are trying to address is the unformulated aspect of the human face. here, the authors had suggested a procedure to facilitate the operation of face identification and matching. then, the intent was to design a structure that adheres to the concept of cloud computing side by side with parallel selection process. the authors claimed that their algorithm qualifies as a capable face identification and resolution method according to the practical results they obtained from the system. other works aimed at improving the features gained from facial images by developing methods for this purpose. one of these promising methods is pca and discrete wavelet transform (dwt),[4] in which dwt is utilized for image compression while pca is applied for extracting and identifying features from the images. here, time is to be considered as criteria for measuring the algorithm’s performance. the authors declared that their suggested algorithm required less access time than using pca alone, while achieving up to 93% recognition rate. safety and security are of the main goals of face recognition, which can be applied on personal level such as houses,[4] which presents a system that was set up on a robotic platform used to patrol the area to be covered and give a warning in case of an unidentified personnel. first, the face area was segmented using a method called local binary patterns. afterward, svm is applied for recognizing the face of interest. the authors stated that their real-time robotic architecture had reached 0.59 precision with eigenfaces, 0.81with fisherfaces, and 0.86 with svm. system design the general structure of this research paper consists of four parts. the first part is preprocessing, including image brightness modification using median filtering, followed by contrast enhancement to the extent that qualifies the image for the second step, which involves general implementation of the partial face recognition system. it includes wavelet transform, pca, svm, and k-nn algorithms. in the third part, the recognition system results are reviewed after being applied on a set of training images and compared with the partial face images that are to be distinguished. part four contains the research conclusions. figure 1 demonstrates the steps of the proposed system as below: preprocessing step this step sets the focus on the intensity and brightness features of the image throughout the use of median filtering for enhancing these qualities. the filter acts as a window that sweeps throughout the image pixels, arranging the pixels beneath this window in an ascending order before taking the midpoint and assume it to be the output pixel.[16] this will result in spreading the light evenly across the image. median filtering can be mathematically expressed as follows:[17] y (x, y) = median {g [s, t], (s, t) ∈ w} where, g [s, t] is the degraded image and w is the neighborhood defined by user as a rectangular sub-image window of size m and n. another algorithm is used depending on the output of median filtering, called contrast enhancement method, which is the method that can be applied to enhance the image appearance. contrast is identified as the variation between the highest and lowest pixel intensity levels. the equation for adjusting the image histogram for contrast increment is as follows:[18] g x y f x y f f f bbp, , *min max min ( ) = ( ) − − 2 here, the lowest and highest pixel intensity levels are used in the equation and multiplied by the available range of gray levels. after applying the equation, the quality of the image is improved by enhancing the contrast as the difference among the image histogram bins is increased. read input svm feature k-nn database preprocessi face authorized ? y n end start display the informa�on using gui figure 1: organization of the proposed system al-dabagh, et al.: exploiting wavelet transform, pca, svm and k-nn for partial face recognition 82 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 80-84 feature extraction using wavelet transform a well-known application of wavelet transform in image processing is object detection and classification. dwt aims at providing a representation of time-frequency relevance. in this approach, a group of shifted and expanded wavelet functions ψlh, ψhl, and ψhh is used to portrait the image being applied for two-dimensional (2d) dwt, with a scaling function φll that creates an orthonormal basis for l2(r2). provided a j-scale dwt, an image x (s, t) with a size of n × n can be simplified as follows:[19] x s t u j k i s t w j k i n j k i ll b b j k i n b j , , , , , , , , , ( ) = ( ) + = = = −− ∑ ∑∑∑ 0 1 0 11 φ ∫ kk i j k i s tb, , , ,ψ ( ) with  ll j j jj k i s t s k t i, , , ,( ) ≡ − −( ) − − −2 2 22  b bj k i s t j k i s t. . , , . . ,( ) ( ) ≡ − −( ) − − −2 2 22 j b j js k t i b b bψ ∫, ,� , . where, n j = n/2j. the coefficients lh, hl, and hh are called dwt or wavelet sub-bands. feature extraction techniques using wavelet transform aim at investigating the characteristics of the coefficient values that are produced through each step of the 2d-dwt transform of the image and produce feature vectors to be examined during this step and in the following step. pca pca[4,13] is a well-known numerical process that can convert an amount of related cases into a less amount of unrelated cases called principal components. the largest possible sum of the data variability is contained in the first principal component and the largest possible sum of the remaining variability is contained in each subsequent component.[4] pca reduces the feature space from a bigger amount of quantities into a less amount of aspects in a “non-dependent” process (the specification of a particular dependent case is not presumed). pca is applied mainly for data compression or dimensionality reduction (i.e., the election of a group of cases from a bigger, more correlated group). therefore, original cases (images in our system) enjoy the greatest amount of correlations with the principal component. mathematical pca can be presented in the points below: 1. the facial pictures to be trained can be represented as r: r 1 , r 2 , r 3 , r 4 ……..r n 2. extracting the average facial image from the set of image scan can be done by: ( )…1 2 n 1 v = r + r + + r n the difference between each instance with the average is:1 nq = r v ……… r v 3. the covariance matrix can be computed as below: m n qqt= − 1 1 here, the matrix m is created by aligning n orthogonal vectors, each of which contains some facial image features called eigenfaces. that is, each vector contributes in the original set by a weight vector w, such that: w u qn n t n= where, un t represents the features of the facial image’s orthogonal vectors. classification classification is the final part of the system. in this part, two stages of classifier are used. in the first one, svm is used for determining whether the person in the image authorize to access or not, while in the second stage, k-nn is applied to find the nears image of the person. classification using svm one famous approach that is occasionally used for classification and regression applications is svm. it is classified as supervised machine learning method that is deeply inspired by statistical learning theory and uses the concept of finding the optimal hyperplane for maximizing the separation between classes.[8,20] hyperplane is realized by computing vectors obtained from training data named support vectors. svm tries to address the constraint optimization issue below:[7] ω ξ ξ, , min || || i b i i w c 1 2 2 +       ∑ subject to: y i (<φ(x i ), w>+b) ≥ 1−ξ i ∀ i = 1,…, n ξ i ≥ 0 ∀ i = 1,…, n., from the previous equation, w represents the vector’s weight and b indicates the bias added to find the orthogonal hyperplane distance from the origin point. slack variables are expressed in ε i in addition to a user-defined complexity factor c. due to the presence of slack variables, it is possible that some instances will not be present on the hyperplane, leading to classification errors. following the existence of an upper limit for the co unt of misclassification measure, this kind of svm is called a soft margin classifier.[20] classification using k-nn this method classifies the given attributes using a set of features obtained from the training database examples. it can be implemented by calculating the distance between the trained vectors obtained from the previous steps with the input partial face image to obtain the relevance with each of the available classes. using the euclidean rule,[13] the algorithm tries to determine how much is the similarity between the input face images with the set of training data.[12] euclidian rule is used to determine the distance between two vectors, x = (x1, x2..,, xn) and y = (y1, y2..,, yn) as the following:[14] d x y x yii n i( , ) ( )= =∑ −1 2 depending on the equation above, the minimum difference is calculated, then the input facial image is given the label of nn class. al-dabagh, et al.: exploiting wavelet transform, pca, svm and k-nn for partial face recognition 83 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 80-84 implementation and results the algorithm presented in this paper is tested versus the well-known (yale) face database, which is used as a training database to train the recognition system against the input image to find the extent of similarity and to recognize the image as one of the face classes available in the dataset. the yale face image database includes intensity images of 15 persons stored as gifs; each person has 11 images to present one facial expression or pattern. the patterns are normal, happy, sad, sleepy, surprised, wink, center-light, left-light, write-light, with glasses, and with no glasses, thus yielding 165 images. for training, 60 images are utilized, leaving the rest of 105 to be applied for testing. the graphical user interface designed for the system is displayed below in two cases, one for authorized face image and the other for an unauthorized image. among the 105 images used for testing, the system resulted in the following parameters: • authorized personnel who are correctly accepted (true positive or tp)=35 figure 2: graphical user interface of the proposed system (accepted image) figure 3: graphical user interface of the proposed system (declined image) al-dabagh, et al.: exploiting wavelet transform, pca, svm and k-nn for partial face recognition 84 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 80-84 • unauthorized personnel who are correctly rejected (true negative or tn)=65 • authorized personnel who are incorrectly rejected (false positive or fp)=1 • unauthorized personnel who are incorrectly accepted (true negative or fn)=35. due to the statistics above, accuracy can be calculated as follows: accuracy tp tn tp tn fp fn = + + + + = + + + + = + � * % � * %100 35 65 35 65 4 1 100 35 65 335 65 1 4 100 95 238 + + + =* % . %. therefore, the given system can reach classification accuracy of 95.238%. conclusions in this paper, an image processing system is built for partial face recognition. it relies on various techniques for preprocessing, feature extraction and selection, and classification. it is inferred that the system is able to recognize the faces of the dataset in different expressions and light source position and intensity. it is also noticed that the system outperforms other methods in the field by giving higher accuracy rate with lower misinterpretation percentage. however, the system is expected to achieve higher accuracy rates by enhancing the approaches currently in use, such as applying adaptive median filtering or using one of the available clustering techniques before classification. references 1. p. hu, h. ning, t. qiu, y. xu, x. luo and a. sangaiah. “unified face identification and resolution scheme using cloud computing in internet of things”. future generation computer systems, vol. 81, pp. 582-592, 2018. 2. a. tolba, a. el-baz and a. el-harby. “face recognition: a literature review”. international journal of signal processing, vol. 2, no. 2, pp. 88-92, 2005. 3. l. paul and a. sumam. “face recognition using principal component analysis method”. international journal of advanced research in computer engineering and technology, vol. 1, no. 9, pp. 135-139, 2012. 4. s. kak, f. mustafa and p. valente. “discrete wavelet transform with eigenface to enhance face recognition rate”. academic journal of nawroz university, vol. 7, no. 4, pp. 10-13, 2018. 5. c. bhimanwar, r. biradar, n. bhole and m. rane. “face identification”. international journal of engineering science and computing, vol. 7, no. 5, pp. 11923-11928, 2017. 6. m. mukhedkar and s. powalkar. “fast face recognition based on wavelet transform on pca”. international conference on energy systems and applications (icesa 2015), india, 2015. 7. g. cavallaro, m. riedel, m. richerzhagen, j. benediktsson and a. plaza. “on understanding big data impacts in remotely sensed image classification using support vector machine methods”. ieee journal of selected topics in applied earth observations and remote sensing, vol. 8, no. 10, pp. 4634-4646, 2015. 8. j. wang, j. zheng, s. zhang, j. he, x. liang and s. feng. “a face recognition system based on local binary patterns and support vector machine for home security service robot”. 9th international symposium on computational intelligence and design. ieee, 2016. 9. y. cha, k. you and w. choi. “vision-based detection of loosened bolts using the hough transform and support vector machines”. automation in construction, vol. 71, no. 2, pp. 181-188. 10. i. kavakiotis, o. tsave, a. salifoglou, n. maglaveras, i. vlahavas and i. chouvarda. “machine learning and data mining methods in diabetes research”. computational and structural biotechnology journal, vol. 15, pp. 104-116, 2017. 11. t. patel and b. shah. “a survey on facial feature extraction techniques for automatic face annotation”. international conference on innovative mechanisms for industry applications (icimia). ieee, 2017. 12. y. song, j. huang, d. zhou, h. zha1 and c. giles. “iknn: informative k-nearest neighbor pattern classification”. springerverlag, berlin heidelberg, 2007. 13. k. weinberger and l. saul. “distance metric learning for large margin nearest neighbor classification”. journal of machine learning research, vol. 10, pp. 207-244, 2009. 14. m. z. n. al-dabagh, m. h. m. alhabib and f. h. al-mukhtar. “face recognition system based on kernel discriminant analysis, k-nearest neighbor and support vector machine”. international journal of research and engineering, vol. 5, no. 3, pp. 335-338, 2018. 15. n. pedrajas, j. castillo and g. garcía. “a proposal for local k values for k-nearest neighbor rule”. ieee transactions on neural networks and learning systems, vol. 28, no. 2, pp. 470475, 2015. 16. g. hemalatha and c. sumathi. “preprocessing techniques of facial image with median and gabor filters”. international conference on information communication and embedded system (icices). ieee, 2016. 17. m. sukassini and t. velmurugan. “noise removal using morphology and median filter methods in mammogram images”. the 3rd international conference on small and medium business, 2016. 18. s. perumal and t. velmurugan. “preprocessing by contrast enhancement techniques for medical images”. international journal of pure and applied mathematics, vol. 118, no. 18, pp. 3681-3688, 2018. 19. k. ghazali, m. mansor, m. mustafa and a. hussain. “feature extraction technique using discrete wavelet transform for image classification”. the 5th student conference on research and development scored. ieee, 2007. 20. j. choi, j. choi, m. ha, t. trinh, t. yoon and h. byun. “towards a generalized toxicity prediction model for oxide nanomaterials using integrated data from different sources”. scientific reports, vol. 8, pp. 6110, 2018. tx_1~abs:at/add:tx_2~abs:at 42 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 42-46 research article exploring investigation of the impact of studying statistics at enhancing mathematical skills during years of faculty marwan t. hasan abstract subject of statistics has always known as field related to mathematics and because statistics is commonly taught in schools as part of the mathematics curriculum, it is regarded as a mathematical. in salahaddin university, entering to the department of statistics is limited to students who graduated from science department at high school. thus, we anticipate students with good knowledge in mathematical level. students from all stages of the department were encouraged to participate in the examination and their scores were recorded. the objective of this study are to explore whether there is substantial difference between male and female as well as to discover whether the more higher stages enhance their mathematical level of knowledge. normality test was done on the dataset and if failed to meet it, nonparametric test such as mann–whitney u- and kruskal–wallis test at significant level 0.05. keywords: statistics, mathematics, normality test, mann–whitney u, kruskal–wallis introduction this is unnecessary to stress the value of statistical literacy for today’s person, and we come to an agreement with[1] that lacking of it, having an educated estimation and participating in social and political discussions about the environment, health, education, and other issues is difficult. many teachers, on the other hand, are uncomfortable with the topic. teachers who have not had specialized instruction obtain their knowledge straight from school textbooks and may be unable to spot mistakes.[2] as a result, teachers are unable to recognize and react to students’ ideas, prejudices, and misunderstandings (russell, ibid.). furthermore, the necessity to teach a topic that is not well-understood causes anxiety. “a sense of uncertainty attributed not so much to a deficiency of training in statistics, but also to an absence of preparation in the teaching of statistics,” according to interviews with italian mathematics students.[3] furthermore, teachers are unaware of the breadth of the statistical material they could teach. furthermore, teachers are unaware of the breadth of the statistical material they are required to teach. descriptive numbers, for example, are thought to be simple and uninteresting. as a result, statistics was relegated to the end of the academic year, if not entirely lost, due to a lack of time.[4] there is a lot of works on gender and theoretical achievement in mathematics, with a lot of diverse perspectives and observations. boys outperformed girls in algebra, according to studies undertaken in northern countries.[5,6] demonstrating that on standardized math exams, boys outperformed children. female students, on the other hand, seem to do higher than male students, according to a growing body of foreign research.[7,8] a large-scale analysis in the united states of america[8] found that girls have caught up to boys in mathematics results, including in high school, where there was a difference in previous decades. they confirmed that girls outperform boys even in tasks including complicated problem solving. the international institute for educational planning[9] conducted the second southern and eastern africa consortium for monitoring education quality survey (2000–2002) and found no major gender gaps among students in south africa. only in seychelles did the same research reveal that girls scored slightly higher than boys. boys, on the other hand, ranked slightly higher than girls in tanzania, kenya, mozambique, zanzibar, and malawi. the gaps were not important in other school systems, like those in south africa. received: may 17, 2021 accepted: june 22, 2021 published: june 30, 2021 copyright © 2021 marwan t. hasan. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) doi: 10.24086/cuesj.v5n1y2021.pp42-46 department of statistics, college of adminstration and economics, salahaddin university-erbil, kurdistan region, iraq corresponding author: marwan t. hasan, department of statistics, salahaddin university-erbil, kurdistan region, iraq. e-mail: marwan.hasan@su.edu.krd https://doi.org/10.24086/cuesj.v5n1y2021.pp42-46 hasan: the impact of studying statistics at enhancing mathematical skills 43 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 42-46 another body of evidence suggests that gender differences in mathematical success are dwindling.[10,11] the divide has been shrinking in the united states of america, according to perie et al.[12] gender gaps in mathematics achievement are shrinking and changing in australia, according to research forgasz et al. and vale.[13,14] many studies performed in australasia between 2000 and 2004 reported no substantial variations in mathematics performance between male and female students, while males were more likely to attain higher mean scores, according to vale.[14] researchers have conducted experiments in a variety of ways around the world to look at the aspects that affect gendered math achievement. many of these findings have concentrated on the causes that contribute to disparities in math achievement between boys and girls.[15-18] feminist scholars have attempted to interpret male-female gender dynamics to make sense of the interactions of girls and boys in mathematics classes.[19,20] girls are often overlooked and assigned inferior status in mathematics classes, according to their findings. teachers’ expectations of girls’ math success are based on rote learning, hard work, and perseverance rather than innate flexibility, ability, and risk taking, which are the learning styles of boys, according to the results. in this article, we will attempt to demonstrate how statistics will help students understand mathematics by encasing certain mathematical terms in a genuine and inspiring way. in reality, plentiful of mathematics is established to explain and model life phenomena, from dayto-day depending to medicine and economics, and statistics is often used to do so. another objective of this study is to show how score of math score varies between male and female, as well as the students between stages of studying at department of statistics. importance of simple data analysis mathematics was created to solve issues that people faced on a daily basis. comparisons, charts, and other numerical communications all use numbers in daily life. as a result, “it is vital to create children’s mental representations of numbers at the same time as they learn to count and calculate.”[20] most of this mathematics is encapsulated in statistics that attempt to explain the universe around us. statistics comprises the majority of the mathematics taught in primary school and a portion of the high school curriculum, according to an examination of some school textbooks and accounts of experiments. when it comes to small surveys, descriptive statistics are normally the first step. moreover, preschoolers may benefit from these basic tasks. the development of graphical representations is also ripe for the picking. allowing the children to build their own images is a vital first step and then bringing them to see the elements that are central and those that are absent, such as names, references, and so on. collecting, organizing, categorizing, storing, and symbolizing data are a basic data handling operation. number principles such as cardinal and ordinal, numbering, and basic arithmetical operations are enabled in this activity. it’s just the start. numbers will gradually take the place of dots, and words will replace the sketches that represent each data type. simple pictograms can pave the way for well-organized tables, with numerical details and periods replacing categories. the definition of a number as a metric, such as height or weight, would emerge. importance of proportionality several experiments[21-23] suggest that relational logic and, in particular, fractions and percentage are not well understood. it was discovered that gymnasium students, as well as teachers, had trouble with this sort of figures.[24,25] the principle of proportion is a fundamental concept in mathematics that encompasses the scope of percentage but is also essential to comprehend the linear function. the principle of proportionality is a fundamental mathematical concept that encompasses the concept of percentage while also being essential to comprehending the linear function. it’s often connected to fractions and decimals, as well as graphical representation.[23] there is a “training to obey such laws” rather than a “gain of experience.”[26] statistics, once again, provides beneficial ways to focus on both of these concepts. many everyday circumstances, such as things and their prices, time, and speed, may be the focus of a mathematical survey, which will deal with the multiplicative systems, which are the basis of the proportionality principle. importance of variability and functions variability and functions are examples of mathematical principles that are often found in statistics. line diagrams depicting the association among two variables, for example, can be used to analyze patterns and make forecasts, as well as another technique to view statistics and, finally, modeling. it also serves as a primer on the analysis of heterogeneity and functions. this holds true with linear relationships and regression as well. scatter plots can be seen right away.[27] students should then search for the straight line (or, finally, the shape) that can best translate the relationship between the two variables. they would use the principle of distance between a point and a line or between two lines, as well as absolute values and absolute value functions, to do so. then, you’ll have to do with numbers of distances or squares of distances. methodology data to find out the effect of statistics at enhancing mathematical level, a 20 math questions were developed and designed for a sample of 262 students at salahaddin university – college of economics and administration, department of statistics. the sample includes students from all four stages and both male and female gender. the results were generated using spss v 25 to explore and implement descriptive statistics as well as inferential statistics hasan: the impact of studying statistics at enhancing mathematical skills 44 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 42-46 like non-parametric comparison tests such as mann–whitney test, kruskal–wallis test. an 0.05 significance level was used as threshold value to report the tests results. results descriptive statistics it is always important, as a first step to do any statistical analysis, to have a tour on some statistical measurements to see how the data are distributed, and for that some tables and graphs have been generated. as shown from table 1, the distribution is based on gender and female participants are way higher than males with 181 and 82, respectively, in terms of their distribution, as first thought we can see that there is no huge difference between male and female with respect to mean and sd values. figure 1 confirms that the score variable is sort of equally distributed with a little bit difference and yet cannot be sure until further investigation is made. we can also hint that the score variable is relatively far to be distributed normally for both male and female [table 2]. referencing to stage variable, it is clear that the number of participants for stages 1 and 2 is quite close to each other with 64 and 55 students, respectively, and only 38 students at stage 4 joined the examination. however, a huge number of students were seen in the test with 106. it is worth mentioning that stages 1 (13.203) and 3 (11.783) were much better than students at stages 2 (9.763) and 4 (9.500) concerning to their mean values as they are above the cut value (>10). there might be rational reason for that as at stages 1 and 3, there are some topics which are purely mathematics. figure 2 shows a very useful information in relation to variable distribution. it does tells us that the students in mathematics examination at all four stages had done differently and as seen students at stage 1 were the best one, followed by stages 3, 4, and 2. no normality of the data can be stated. conducting any statistical test to do some comparison requires some assumption to be met. the most common requirement is whether the data are normally distributed. before implementing the normality test, it is better to see the nature of the data. thus, we created the histogram plots for the data and none of the dimensions refer our thoughts to normal distribution. thus, none of the groups strongly suggested to have a perfect normal distribution. however, to be sure about the normality distribution or homogeneity, first let’s see what the shapiro–wilk test leads us. table 3 provides enough evidence to go ahead with nonparametric test to know whether the categorical variables are significantly difference between the levels in response to score achievements. test comparison the time has come to go further into the investigation to discover whether gender has any effect on students to be knowledge with mathematical subjects in statistical department regardless to their stages. since we ended up with not normally distributed and hence, we implemented mann– whitney u-test to compare two groups (in our case gender) and kruskal–wallis test for more than 2 groups (in our case stages). table 4 demonstrates that there is no statistical difference between male and female regarding to their level of knowledge in mathematics since the p-value (0.270) is much larger than the significance level (0.05). referencing to stages as we expected that somehow there might be some discrepancies in their performances at the examination. we thought the more studying in statistics, the more involvement with mathematics is happened. the kruskal table 1: mean and sd exploration for gender gender score n mean sd male 82 11.024 4.128 female 181 11.508 3.769 table 3: normality test for both independent variables in response to score achievement shapiro–wilk statistic df sig. gender male 0.968 82 0.036 female 0.983 181 0.027 score 1 0.976 64 0.250 2 0.933 55 0.004 3 0.977 106 0.057 4 0.960 38 0.190 table 2: mean and sd exploration for stages stages score n mean sd one 64 13.203 3.941 two 55 9.673 3.283 three 106 11.783 3.602 four 38 9.500 3.689 figure 1: boxplot for both male and female and score achievement hasan: the impact of studying statistics at enhancing mathematical skills 45 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 42-46 test confirmed that there were statistical differences in achieving scores in the examination mathematics since the p-value (0.000) is less than 0.05. nevertheless, we cannot know exactly between which stages the difference was occurred, and we needed to compute the multiple comparison tests among each pair. for this purpose, mann–whitney u-test was applied and ended up that there was no difference between stages 2 and 4 only. conclusion as we reach to conclusion, the study reported that there were no differences among males and females in achieving higher or lower mark in mathematical examination at the department of statistics. this result matches with other previous studies. although, female participants were much higher than male participants with about nearly 100 students and still the descriptive result showed no statistical significance result. on the other hand, stages as expected unlike gender had shown that students might increase their level of mathematics while studying statistics and in our study interesting result came up. students in stages 1 and 3 performed better than student at stages 2 and 4 in the examination with reference to descriptive statistics. similar to gender, again non-parametric test was used and consequently showed significant outcome. kruskal– wallis test which works just like anova was calculated and indicated significant conclusion. furthermore, to dive into the detail, mann–whitney u-test was applied again between each pair of stages and reached to report that there was only difference in student’s performance at stages 2 and 4. references 1. c. konold, a. pollastsek and a. gagnon. students analyzing data: research of critical barriers. in: j. b. garfield and g. burrill, (eds.), research on the role of technology in teaching and learning statistics: 1996 proceedings of the 1996 iase round table conference. voorburg, the netherlands: international statistical institute, pp. 151-167. 2. a. hawkins, f. jolliffe and l. glickman. teaching statistical concepts. the effective teachers series. london: longman, 1992. 3. l. gattuso, and m. a. pannone, teacher’s training in a statistic table 4: mann–whitney u and kruskal–wallis test result for the case study mean rank chi-square p-value pairwise comparison gender mann–whitney male 124.341 1.216 0.270 female 135.470 stage kruskal–wallis mann–whitney 1 167.125 35.779 0.000 no difference only occurred between stages 2 and 4 2 96.900 3 141.840 4 96.197 figure 2: boxplot and histograms for detecting how the variables are distributed hasan: the impact of studying statistics at enhancing mathematical skills 46 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 42-46 teaching experimentation. in: b. phillips, (ed.), proceedings of the sixth 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61-66, 1986. 22. s. rené de cotret. étude de l’influence des variables indice de proportionnalité du theme et nombre de couples de données sur la reconnaissance, le traitement et la compréhension de problèmes de proportionnalité chez des élèves de 13-14 ans. thèse présentée à l’université joseph fourier, grenoble i, 1991. 23. m. pézard. à propos de l’enseignement de la proportionnalité les cahiers de didactique. num. 20. irem de paris7, paris, 1986. 24. f. carayol. comportements d’élèves et de futures maîtres de l’école élémentaire face à des questions de mathématiques. thèse de 3e cycle. irem de toulouse, 1983. 25. p. buisson. pourcentages: appropriation transfert. irem de rouen, 1981. 26. f. pluvinage and c. dupuis. la proportionnalité et son utilisation. recherche en didactique des mathématiques, vol. 2, no. 2, 1981. 27. j. mighton. jump at home grade 6: math worksheets for the elementary curriculum. toronto: house of anansi press inc., 2005. . 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 31-36 research article solving isolated nodes problem in zigbee pro for wireless sensor networks nabaa abdul-khudhur hasan*, f. alaa kadhim department of computer science, university of technology, baghdad, iraq abstract wireless sensor network based on the zigbee protocol consists of many sensor devices. in some cases, the sensor nodes may turn to isolated node because random distribution, particularly when creating the network. in this research was suggested two cases to overcome on the isolated node problem, the first case had able to overcome this problem by distributing the isolated nodes on the router nodes that carry the least number of sensor nodes, it helps to minimize the computational overhead on router nodes too, while the second one is able to overcome this problem by calculating the distance between the isolated nodes and the routers and then adds these nodes to the nearest routers. subsequently, this method helps to minimize the energy consumption. the results show our approach able to solve the problem of isolated nodes using these two methods and when compared between them turns out the second method is better in terms of energy consumption. in addition, we are able to make the network larger scale. keywords: isolated nodes, wireless sensor networks, zigbee pro introduction nowadays, wireless sensor network (wsn) plays an important role because it is used in many applications such as military monitoring, fire control, health observation in patient case, and monitoring of traffic and also used in soil to measure humidity.[1] the demand for the construction of large-scale wsns has been increasing rapidly; the use of conventional wsns encounters a number of critical technical challenges such as the network scalability and energy consumption.[2,3] especially, it must apply a network joining technique that is able to link every node in a big scale wsn with reduced power consumption.[4] many of network joining techniques have been suggested to support trusty connection in internet applications (e.g., dynamic host configuration protocol and duplicate address detection).[5-7] even so, because of the broadcast storm issue, may not be applied this technique and the energy may become not efficient and this will be wasted it,[8] appropriate protocols must be applied to solve the problem of rejoining isolated nodes with conserving energy. to reduce power consumption, zigbee has applied as communication protocol. although some papers have been published concerning the problem of communication in zigbee protocol, but has not been enough and did not cover the problem in complete way when compared with other issues such as routing algorithms[9-11] or data transmission.[12,13] in most of the papers, clouqueur et al.,[14] dhillon et al.,[15] kumar et al.,[16] rahman et al.[17] concerned with the coverage of the sensor network, it assumed that communication in the network is good, but communication in the network is an important and will affect performance, it must focus on. in this paper, we will focus on connectivity improving by solving isolate nodes problem so the network will be large scale. moreover, we will solve the problem in two ways. the first method is called load distribution where the isolated nodes are distributed on the router nodes that carry the least number of sensor nodes. the second method is to add the isolated node to the nearest routing node to it. this will help to reduce power consumption. the remainder of the essay arranged in this way. part ii shows relation works. part iii shows the proposed mechanism where part iv displays environment and part v discussed the results of proposed to estimate the efficacy of our suggested. finally, we deduce this essay in part vi. corresponding author: nabaa abdul-khudhur hasan, department of computer science, university of technology, baghdad, iraq. e-mail: nabaa_alkhafagi@yahoo.com received: mar 25, 2019 accepted: apr 22, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp31-36 copyright © 2019 nabaa abdul-khudhur hasan, f. alaa kadhim. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) hasan and kadhim: solving isolated nodes problem in zigbee pro for wsns 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 31-36 relation works zigbee pro zigbee is a protocol based on ieee standard 802.15.4. it is used in wsn because it has the appropriate characteristics of this network such as low-power consumption and low cost. zigbee is considered a worldwide open standard for wireless radio networks in the monitoring and control fields. the standard was developed by the zigbee alliance (a group of international companies). the zigbee protocol consists of three devices:[18] zigbee coordinator: the network should include only one and be responsible for several tasks such as network establishment, the allocation of addresses, and the decisionmaker regarding the admission or departure of the nodes, in addition to the transfer and processing of messages. it should always be powered on. zigbee router: could be missing in a network, the network may include just one or more based on the size and topology of the network. it is not found in a star topology. it is often used to expand the zigbee network (in the tree and mesh). basically, it executes all tasks of the coordinator, except network establishment (start-up). zigbee end devices: it is located in all topologies the purpose of end devices for sending and receiving the packets. zigbee consists of three topologies: star, tree, and mesh topology. in this paper will focus on tree topology, because the isolated nodes problem happens in this topology. a connectivity improving mechanism for zigbee wsns in 2008, tian-wen song and chu-sing yang,[19] they used the tree topology and modified the connection of nodes, and the joining which succeeded by of a shifting is called extended joining. this operation is specified by three elements nwkmaxchildren, nwkmaxrouters, and nwkmaxdepth which determine if an isolated node is able to add to the network or not this enhancement helps not only the reduction of isolated sensors and wasted costs but also the performance growth of the zigbee sensor network. in 2008, enhancing node connectivity by utilizing rssi for zigbee-based wsn in 2016, chung et al.[20] they could overcome the isolated node problem, they proposed two methods, gaussian filtering with averaging and median method based on rssi samples. the first way is objective to get high accuracy, while the another way is objective to reduce the computational cost of node connection. the result of the proposed is more effective than the existing approach. proposed system first, we will apply our proposed on the tree topology. an example of connectivity problem in a zigbee network is shown in figure 1. there are a number of isolated nodes such as node d; the isolated nodes will cause the expected network operations unreached and waste the deployment costs. hence, we proposed two methods to solve this problem and we will discuss in the next sections. however, before that, we must now discover if there are isolated nodes in the network. hence, we assumed that there is table saved in the coordinator, this table includes the number of routers, end devices, and the links between this device, and shown in table 1. after network establishment, it will start many operations such as send and receive the packets. we will determine the specific time (15 min), for example, if one node will not do any action, this node will be considered as isolate node and added to the table as isolated. in this way, we will know if there is any isolated node in wsn. our proposed system is shown in figure 2. figure 1: illustration of zigbee connectivity issue table 1: the table saved in the coordinator (z) zigbee router a, b, c, e, f, g, r, l, t zigbee end devices w, h, k, u isolated nodes p, d, m, n links z a, g, t a b, e, z b a, c, f, u c b e a, r, k f b g l, z h l l g, h, w t z w l r e k e u b hasan and kadhim: solving isolated nodes problem in zigbee pro for wsns 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 31-36 case 1 in this case, we will distribute the isolated nodes on the routers that carry least number of end devices. moreover, this operation starts after the coordinator will know that there are isolated nodes in the network. the coordinator will open the table and search for the least router that carries end devices. for example, z will send two messages, one of t to tell it that there is new node will join into it, the second message will tell p that t it is the router appropriate for it. after that will start the communication between p and t to join. if there is more than one isolated node, it will follow the same method. in figure 3, the node p will be connected to the router t, and the nodes m, n, and d will be connected to f, c, and r, respectively. the protocol clarifying our proposal for the first case is as follows: • the coordinator (z) opens the table stored in it and searches for a router with fewer end devices. • then, z sends a message to that router tells him that there is an isolated node wants to join him by sending the id of that node. • and z sends a message to that node and tells it that there is a router appropriate to it by sending the id of that router. • after that, isolated node will send his id to the router. • finally, the router will compare if the id transmitted by the node matches the id transmitted by z, if they match, the router will accept the node, and otherwise, it will ignore the request. case 2 in this case, the distance will measure between all isolated nodes and every router, and select, the less distance between them to join. we will use the distance equation (1) to calculate the distance between them.[21] 2 2di = (x1 x2) + (y1 + y2) (1) the protocol clarifying our proposal for the second case is as follows: • the coordinator (z) opens the table stored in it and searches for the number of routers and isolated nodes. • then, z will calculate the distance between the isolated nodes and all routers. • after that, z will compare the distance and select least distance between them (node will be added to the router closest to it). • these nodes are added by sending z two messages; one of them to the router containing the id of isolated node and the other one is sent to the node and contains the id of the router. • finally, the router will compare if the id transmitted by the node matches the id transmitted by z, if they match, the router will accept the node, and otherwise, it will ignore the request. figure 2: the flow chart of the proposed system figure 3: illustration to resolve the connection problem in the zigbee protocol by case 1 hasan and kadhim: solving isolated nodes problem in zigbee pro for wsns 34 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 31-36 figure 4 displays the coordinates of nodes. applying the distance formula for finding the every node distance value with the help of sensors, nodes coordinate as showing in figure 5, for example, distance between z (coordinator) and t node. z=(5, 4) t=(2, 4) 2 2d1 = (5 2) + (4 4)  2 2d1 = (3) + (0) d1 = 9 d1=3 similarly, the value of all remaining distance is also calculated, which is also shown in figure 4, that is, d2=2, d3=1, d4=2, d5=2, d6=1, d7=2, d8=4, d9=5, d10=4, d11=5, d12=1, and d13=2, using sensor nodes coordinate and distance measurement formula we able to measure the distance between each node in a wsn area. after calculating the distance between the all nodes within the network, now we will calculate the distance between the isolated nodes and the routers only, table 2 explain that, for example, the distance between node p and router a is 8.1 and between node p and router b is 6. moreover, on this basis, we will compare and choose the nearest router to this isolated node to join, show that in figure 6. to measure the energy consumption[22] and compared it in the two cases used. we will use equations (2, 3): etx (k, d)=eelec*k+εamp*k*d2 (2) erx (k)=eelec*k (3) where, eelec is the energy to run circuitry=50 nj/bit, εamp is the energy for radio transmission=100 pj/bit/m2, etx energy for sending k bits over distance d, and erx energy for receiving k bits. programming environment the proposed system was written in c# language version 2013 on windows10. performance evaluations we suggested two cases to solve the problem of isolated nodes, and during the execution turns out, the method two is better than method one. the first method depends on added the isolated nodes to the routers that carry the least number of sensor nodes. the second method depends on the distance measure way, where the distance of the path between the isolated nodes and the routers is measured. then, the appropriate router is then selected for the node to join based on the least distance between the router and the isolated node. the second method was chosen as the best after measuring and analyzing the results for energy consumption; figure 7 shows the energy consumption in the first and second method. figure 4: show nodes coordinate figure 5: display the distance measurement for the nodes table 2: explain the distance between isolated nodes and routers isolate routers a b c e f g l t r p 8.1 6 4 9.2 9.8 8.9 8.2 6 10.2 m 7.1 9.8 9.2 5.8 8.6 7.8 9.2 8.9 10.6 n 7 7.2 5.7 7.3 9 8 8.2 6.7 10.2 d 7.3 6.3 4.5 8.1 9.2 8.2 8 6.1 10 the results show that there is a significant difference in energy consumption between the two methods when the isolated nodes added to the network where the amount of energy consumed in the first method when joining the node m to the router r is 181.4 mjoule, and node n to f hasan and kadhim: solving isolated nodes problem in zigbee pro for wsns 35 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 31-36 is 131.2 mjoule, node p to t is 59.2 mjoule, and finally, the amount consumed in node d to c is 34 mjoule. as for the amount of energy consumed in the second method when add the node m to the router e is 55.4 mjoule, and node n to router c is 35.7 mjoule, node p to router c is 27.2 mjoule, and finally, the amount of energy that consume for add node d to router c is 34 mjoule. conclusion this work proposed an enhanced the connectivity of zigbee using two ways for solving the problem of isolated nodes in wsn. we have applied the first way by making the isolated nodes join to router who carries the least number of end devices and as for the second way by calculating the distance and added this node to nearest routers. we proved our scheme could overcome the problem of isolated nodes. after that, we compared between the two ways where the second method provides efficiency by achieving less energy consuming compared to the first method. furthermore, our scheme applied to make the network larger scale. references 1. j. sen. “a survey on wireless sensor network security”. international journal of communication networks and information security, vol. 1, no. 2, pp. 59-82, aug 2009. 2. m. s. pan, c. h. tsai and y. c. tseng. “the orphan problem in zigbee wireless networks”. ieee transactions mobile computing, vol. 8, no. 11, pp. 1573-1584, nov. 2009. 3. s. hur, j. kim, j. choi and y. park. “an efficient addressing scheme and its routing algorithm for a large-scale wireless sensor network”. eurasip journal wireless communications and networking, vol. 2008, pp. 765803, sep. 2008. 4. f. ye and r. pan. “a survey of addressing algorithms for wireless sensor networks”. in proceeding ieee wicom, sep. 2009. 5. r. droms. “dynamic host configuration protocol”. internet rfc 2131, mar 1997. 6. r. droms, j. bound, b. volz, t. lemon, c. perkins and m. carney. “dynamic host configuration protocol for ipv6 (dhcpv6)”. internet rfc 3315, jul. 2003. 7. n. h. vaidya. “weak duplicate address detection in mobile ad hoc networks”. in proceeding of acm mobihoc, jun. 2002. 8. j. l. lu, f. valois, d. barthel and m. dohler. “low-energy address allocation scheme for wireless sensor networks”. in proceeding of ieee pimrc, sep. 2007. 9. t. kim, d. kim, n. park, s. yoo and t. s. lopez. “shortcut tree routing in zigbee networks”. 2nd international symposium on wireless pervasive computing (iswpc’07), san juan, puerto rico, feb. 5-7, 2007, pp. 4247. 10. p. ran, m. h. sun and y. m. zou. “zigbee routing selection strategy based on data services and energy balanced zigbee routing”. ieee asia-pacific conference on services computing (apscc’06), guangzhou, china, dec. 2006, pp. 400-404. 11. k. k. lee, s. h. kim and h. s. park. “cluster labelbased zigbee routing protocol with high scalability”. 2nd international conference on systems and networks communications (icsnc’07), cap esterel, france, aug. 2007, p. 12. 12. g. ding, z. sahinoglu, p. orlik, j. zhang and b. bhargava. “tree-based data broadcast in ieee 802.15.4 and zigbee networks”. ieee transactions on mobile computing, vol. 5, no. 11, pp. 1561-1574, 2006. 13. g. ding, z. sahinoglu, b. bhargava, p. orlik and j. zhang. “reliable broadcast in zigbee networks”. 2nd annual ieee communications society conference on sensor and ad hoc communications and networks (secon’05), santa clara, usa, sep. 2006, p. 510. 14. t. clouqueur, v. phipatanasuphorn, p. ramanathan and k. k. saluja. “sensor deployment strategy for target detection”. 1st acm international workshop on wireless sensor networks and applications (wsna’02), atlanta, usa, sep. 2002, pp. 42-48. 15. s.s. dhillon, k. chakrabarty and s. s. iyengar. “sensor placement for grid coverage under imprecise detections”. 5th international conference on information fusion (fusion’02), annapolis, usa, jul. 2002, pp. 1581-1587. 16. s. kumar, t.h. lai and j. balogh. “on k-coverage in a mostly sleeping sensor network”. 10th annual international conference on mobile computing and networking (mobicom’04), philadelphia, usa, sep-oct, 2004, pp. 144-158. 17. o. rahman, a. razzaque and c. s. hong. “probabilistic sensor deployment in wireless sensor network: a new approach”. 9th international conference on advanced communication technology (icact’07), phoenix park, south korea, feb. 2007, pp. 1419-1422. 18. o. g. aju. “a survey of zigbee wireless sensor network technology: topology, applications and challenges”. figure 6: illustrates the way of connection to the nodes by case two (minimum distance) figure 7: illustration the energy consumed in the two methods it used hasan and kadhim: solving isolated nodes problem in zigbee pro for wsns 36 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 31-36 international journal of computer applications, vol. 130, no.9, pp. 975-8887, nov 2015. 19. t. w. song and c. s. yang. a connectivity improving mechanism for zigbee wireless sensor networks”. ieee/ifip international conference on embedded and ubiquitous computing, 2008. 20. s. m. chung, k. t. kim, j. song and y. youn. “enhancing node connectivity by utilizing rssi for zigbee-based wsn”. international conference on information and communication technology convergence, 2016. 21. v. v madhav and j. nigam. “ensuring uniform energy consumption in non-deterministic wireless sensor network to protract networks lifetime”. international journal of electronics and communication engineering, vol. 11, no. 9, pp. 1005-1009, 2017. 22. t. kumar, j. davinder, s. saini and s. v. bhooshan. “cluster head selection in a homogeneous wireless sensor network ensuring full connectivity with minimum isolated nodes”. hindawi publishing corporation journal of sensors, vol. 2014, p. 8, 2014. . 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 12-17 research article improving the level of service of a portion of 60-m ring-road in erbil city dlzar b. qadr*, shorsh a. mohammed, ibrahim a. hasan, rawand f. mohammed ali abstract this study aims to implement the factors that can improve the level of service of 60-m ring-road in erbil, kurdistan region of iraq. these factors are minimizing the traffic volume, providing a higher quality pavement, widening the road carriageway, and constructing bridges. the main objective of this paper is to increase the los of a specific road known as 60-m ring road from new city mall to par hospital in erbil city. for this purpose, the data on speed and traffic flow are collected using video camera to collect data to be reasonably analyzed with respect to the speed of the vehicles and set the los for the proposed road. the result shows that average speed in this study is 19 km/h, and this confirms that the los in this particular road segment is at f category. in addition, the calculated peak hour factor is 0.97 which is more than a typical value in urban area. finally, the study revealed that the traffic volume on this specified road is too high. keywords: bridge ramp, congestion, highway capacity manual, level of service, peak hour factor, space mean speed, urban introduction over the past decade, the number of private cars has been significantly increased at erbil city, where the number tends to be more than a vehicle per family. this is due to the absence of the required public transport in the city such as buses, light rapid traffic system (lrts), railway networks, and underground routes. based on the last statistics in 2011, erbil city as a capital of kurdistan region, iraq, is considered to be one of the major cities in the region, with population about 1.6 million people. human settlement at erbil can be traced back to possibly 5000 bc, and it is one of the oldest continuously inhabited areas in the entire world. at the heart of the city, there is the ancient citadel. apart from its strategic geographical position, commercial development, and tourism of attraction resorts, traffic congestion is one of the challenges that has been affected the movement of vehicles in the city.[1] urban areas can be varied as greatly as the variety of activities performed there: the means of production and the types of goods, trades, transportation, and delivery services. the third definition states that urban areas are those locations where there is a chance for a diversified living environment and various lifestyles.[2] in recent years, traffic congestions have significantly increased in erbil city. there are several reasons which directly and indirectly influence the traffic distribution inside erbil city. the presence of public transport tends to encourage road users to reduce utilizing private cars which does not exist as required in erbil city. the main ring road known as 60 m street around the center of erbil is the most strategic road which has the advantage of its wide carriageway, in which the wide is 18 m per each side, divided into five lanes. geometrically, roads with this geometry should provide a reasonably high level of service (los). in reality, due to the existence of several branch roads from the outer and inner of the ring road, the los has been noticeably decreased. los in urban area the term los is used to qualitatively define the operating conditions of a roadway depending on several factors such as speed, travel time, maneuverability, delay, and safety. the los of a facility is designated with a letter, a to f, with a representing the best operating conditions and f the worst as shown in table 1.[3] corresponding author: dlzar b. qadr, department of civil engineering, cihan university, erbil, iraq. phone: 00964 750 472 77 50, e-mail: dlzar.qadr@cihanuniversity.edu.iq doi: 10.24086/cuesj.v3n1y2019.pp12-17 copyright © 2019 dlzar b. qadr, shorsh a. mohammed, ibrahim a. hasan, rawand f. mohammed ali. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) received: apr 03, 2018 accepted: sep 03, 2018 published: may 13, 2019 department of civil engineering, cihan university-erbil, kurdistan region, iraq https://creativecommons.org/licenses/by-nc-nd/4.0/ qadr, et al.: improving the level of ring-road services 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 12-17 the los is simply one of the pillars of geometric design which is a measurement of any particular road to assess the case of traffic movement depending on the average traffic speed. the los is not smoothly passable defined for heterogeneous traffic flow with different operational features.[4] the los is considered to be a measure of traffic condition on any particular road which has been clarified in the hcm, 2000. depending on the operation characteristics of the vehicles, the los can be differentiated relying on the traffic speed and distribution of the vehicle represented as traffic volume. ideally, the worst los tends to be equal volume to capacity for the traffic condition. in other word, when the volume of capacity ratio is close to one, it indicates poor traffic condition. as a result, the obstacles on the pathway of both sides can significantly drops the traffic speed despite of the multi-lane road.[5] the factors listed below may influence the evaluation of the los:[6] • factors affecting the speed such as traffic restriction. • operating speed and travel time. • quality of traffic condition represented as driving comfort and convenience. • freedom of maneuver with the similar speed required for operation. objective the fundamental aim of this paper is to improve the los for a specified road in which proper solutions are proposed to be constructed in the locations where the vehicle movement becomes reasonably more efficient. this can be achieved using data collection in monitoring system designed for the determination of the vehicle speeds and traffic volume. furthermore, the traffic condition is aimed to be converted from the worst condition to a reasonably better. this may be achieved by proposing proper solutions for that particular road after evaluating the problem. study area the real problem associated with the study area is the branch roads existing on the 60-m ring road. they have a great impact to create traffic jam due to vehicles passing the branch roads to the left turns as shown in figure 1. this possibly generates vehicle conflict minimizing the design speed of the road which is 60 km/h. after inspection stage of 60-m wide ring road of erbil city, the study area was found to be a particular road to conduct the study satisfactorily. the proposed area is located between eskan intersection and koya intersection on the 60-m wide ring road. this portion appeared as the busiest part exposed to traffic congestion. this is due to the branch roads located on the path of the selected road, where the branch roads create traffic conflict as a result of traffic movement passing the branches to the left turns on the medians as shown in figure 1. methodology the following procedure will be undertaken in the field. to set an efficient plan, a map of the proposed area was sketched properly, in a way that the points of two cameras were fixed with the distance of 150 m between them. the ideal position of the cameras has been chosen based on the entering vehicles from the branches to the left turns which exist from the medians of the road. figure 1 shows the opposite side of the 60-m wide ring road, that collect data from two branch roads. consequently, the monitoring process was designed to record the license number of any passing car through the two cameras at two different times. this time is considered to be a travel time to determine the speed of the vehicles based on the actual distance between the two cameras.[7] to guarantee recording the time more precisely, the direction of both cameras was set on the same vision angle. space mean speed (v s ) is the average speed of all the vehicles along a given road segment at a typical instant in time (ortuzar and willumsen, 2011). after the monitoring process, the analysis of the collected data and processing were started as shown in table 2. the determination of the speed for any vehicle is calculated utilizing the following formula: actual distance (d) speed (v)= travel time (t)  (1) where; speed (v): the real speed passing through the portion of table 1: level of service in urban street classes (highway capacity manual, 2000) urban street class i ii iii iv range of speeds 70–90 km/h 55–70 km/h 50–55 km/h 40–55 km/h typical speed 80 km/h 65 km/h 55 km/h 45 km/h los average travel speed (km/h) a >72 >59 >50 >41 b >56−72 >46−59 >39−50 >32−41 c >40−56 >33−46 >28−39 >23−32 d >32−40 >26−33 >22−28 >18−23 e >26−32 >21−26 >17−22 >14−18 f ≤26 ≤21 ≤17 ≤14 los: level of service qadr, et al.: improving the level of ring-road services 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 12-17 the studied road (km/h) actual distance (d): the distance between two fixed cameras in km travel time (t): the time spent by any particular vehicle on the study road can be found by subtracting t 1 and t 0 t 0 : the initial recorded time from the first camera in seconds t 1 : last recorded time spent by a vehicle in seconds. to locate a sufficient approach, two cameras were used per day of monitoring during peak hours (8:30 am and 5:30 pm). the operation of monitoring was done triple for each side of the road. the recording process was designed for peak hours. the average speed of 3 days based on the collected traffic condition was calculated as the final result to be a realistic representation of condition and this is shown in table 2. results and discussion the observations undertaken on the study area generate diverse interpretations regarding the traffic condition. the calculated traffic speed required for the determination of los confirms that the traffic condition varies from e to f. according to table 3, the average travel speed for the vehicles in a specific time is 19 km/h which locates at the f category representing the worst los. this confirms that the influence of the branch roads is considerably high. fi * fi ú ui = ∑ ∑ (2) fi*ui=1376  fi 72 ú= = km/hr 1376 72 19� s fi ui ú n     *( )2 1 (3) fi * ( )2ui ú   664  n-1 = fi-1=71   s2= 664 71 �=9.35 s=±3.05�km / hr→ phf= total�volume�for�1�h 4*maximum�volume�of�15�min (4) phf= 2045 4*525 =0.97 ú is the arithmetic mean speed. s is the standard deviation. phf is the peak hour factor typically between 0.8 and 0.95. tables 3 and 4 illustrate a part of data collection and statistical analysis. the results show that the arithmetic mean speed of the vehicles is 19 km/h and the standard deviation of the data is approximately ±3.05 km/h. this gives a decent indicator of the differences in distribution of the speed vehicles which is not so high in the range of 16–22 km/h. the typical value of the peak hour factor (phf) is between 0.8 and 0.95 indicating a lower phf characteristic. in this study the (phf) is higher than the typical value (0.97). this proves the worst los on this road segment. figure 2 demonstrates the frequency of observed vehicles for the vehicles traveling this particular road section in which the speed of 17 km/h has the highest frequency, and this is statistically called as a modal speed. figure 1: the location of the fixed cameras in the study site qadr, et al.: improving the level of ring-road services 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 12-17 the median speed is obtained from the cumulative frequency distribution curve [figure 3] approximately as 16 km/h, the 50th percentile speed, and 20 km/h, the 85th percentile speed. solution from geometric design points of view since the most problematic points to highlight are the branch roads which are causing traffic conflict, it is fair to state that the 60-m wide ring road has the advantage of its wideness from geometrical viewpoints. this can be of great importance to design an efficient network system of transport to reorganize table 2: typical illustration of data processing and calculation sample/vehicle no. t 0 (s) t 1 (s) actual travel time (h) distance (km) speed (km/h) 1 t 1 –t o speed�(v)�= actual�distance�(d) travel�time�(t)� 2 3 : n-1 n average space mean speed (v s ) (km/h) table 3: sample of data collection and calculation car no. t 0 (s) t 1 (s) t actual (h) distance (km) speed (km/h) 1 2 28 0.0072 0.15 21 2 6 42 0.0100 0.15 15 3 20 48 0.0070 0.15 19 4 14 50 0.0100 0.15 15 5 24 54 0.0083 0.15 18 6 30 97 0.01861 0.15 8 : 0.15 6200 0.15 average travel speed (km/h) 19 table 4: sample of vehicle speed data and statistical analysis speed class km/h class median, ui class frequency (number of observations in class), fi fi*ui percentage of observations in class cumulative percentage of all observations f (ui-ú) 2 11−14.9 0 0 0 0 0 0 15−18.9 17 47 799 65 65 4 19−22.9 21 19 399 26 91 4 23−26.9 25 3 75 4 95 36 27−30.9 29 1 29 1 97 100 31−34.9 33 0 0 0 97 196 35−38.9 37 2 74 3 100 324 totals ∑= 72 1376 664 the movement of traffic. ideally, to fix the traffic conflict at this particular area, it is fair to say that designing bridge ramps would be a helpful option for improving the los, specifically for the vehicles traveling from the branch roads to the left turns on the 60-m ring road. the essential considerations that should be taken into account for designing bridge ramps are as follows:[8] • design speed • number of lanes and its width • grades • curve geometry • effect of alignment skew • crossroad median consideration. qadr, et al.: improving the level of ring-road services 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 12-17 in line with the above-mentioned interpretations, it is highly recommended that the proposed ramp should apply the three fixed terms such as safety, economy, and comfort in the design process. however, the restrictions which exist on the studied area may cause some difficulty in design speed, number of lanes, and radius of the curve which may lead to slightly uncomforted traffic condition for the road users. since there are a limited number of solutions such as changing the position of the u-turns or perhaps adding them as well as changing into intersections, therefore the only logical solution is remained to construct bridge ramps to hold the traffic flow which makes the real problem of this study. conclusions it is obvious that the los has been significantly influencing toward its worst condition on the study area due to the numerous vehicles moving straight foreword conflicting with the vehicles coming from the branch roads. despite the real traffic condition, it has been found that the absence of public transport has negatively affected in minimizing the los as well as the traffic awareness of the road users. the results showed that the average travel speed of vehicles was noticeably less than the design speed of the road (19 km/h), while the design speed specified to be 60 km/h. this drop of speed is considered to be a great difference. since the geometrical advantages of the study area are reasonably appropriate for extra changes more than the restrictions, it is highly recommended that the design and construction of bridge ramps may lead to solve the traffic congestion on that particular road. recommendations the most striking point to emerge is that a tremendous number of private cars are used by the road users due to the inefficient public transport. on a larger scale, the existence figure 2: histogram of observed speed vehicles during peak hour figure 3: cumulative distribution qadr, et al.: improving the level of ring-road services 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 12-17 of a sustainable transport system can be of a favorable support including different pillars of sustainability which are efficiency, economically, and socially. from sustainability perspective, increasing awareness is considered to be vital to encourage people using efficient public transport, for example, establishing a variety of lanes for buses, motorists, cyclists, and tramways (lrts). based on evaluating the gathered data taken from monitoring process at the study area, it was found that the los may be significantly improved toward a reasonably better. this could be achieved by designing proper infrastructures which geometrically are of great support. it is highly recommended that construction of bridge ramps may increase the space mean speed (v s ) of vehicles traveling on the road segment, avoiding conflicts between vehicles moving from branch roads to the left turns against straight forward moving vehicles located on 60-m wide ring road. references 1. o. k. akram, s. ismail and d. j. franco. “the significant of urban form of erbil city, iraq”. international journal of engineering technology, management and applied sciences, vol. 2, no. 4, pp. 96-100, 2016. 2. p. k. bhuyan and k. v. krishna. “defining level of service criteria of urban streets in indian context”. european transport, vol. 1, no. 49, pp. 38-52, 2011. 3. transportation research board. “highway capacity manual. special rep. 209”. washington, d.c: transportation research board, 2000. 4. k. p. patnaik. “level of service criteria of roads in urban indian context”. m.a. thesis, deparment of civil engineering national institute of technology rourkela, rourkela, 2013. 5. r. babit, v. sharma and a. k. duggal. “level of service concept in urban roads”. international journal of engineering science invention research and development, vol. 3, no. 1, pp. 44-48, 2016. 6. s. m. madanat, m. j. cassidy and w. h. w. ibrahim. “transportation research record 1457, trb”. in: methodology for determining level of service categories using attitudinal data”. washington, d.c: national research council, 1994, pp. 59-62. 7. j. o. ensley. “application of highway capacity manual 2010 levelof-service methodologies for planning deficiency analysis”. m.a. thesis, university of tennessee, knoxville, 2012. 8. a. swaroopa and l. n. venkata. “proposed alternative system to existing traffic signal system”. international journal of civil, environmental, structural, construction and architectural engineering, vol. 9, no. 5, pp. 554-557, 2015. tx_1~abs:at/add:tx_2~abs:at 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 13-18 research article educational data mining to improve the academic performance in higher education alyaa a. mahdi* department of computer science, cihan university-erbil, erbil, kurdistan region f.r. iraq abstract globalization and innovation are unlimited interest for the public sector and private business in the world, especially in the higher education institutions. educational data mining (edm) is mainly one of the business processes nowadays that attempt to bring the global innovation through improving and enhancing their processes and procedures to fulfill all the requirements and needs of the students as well as the institutions. the edm considered mostly concern with any research concerning the applications of the dm and developing innovative techniques for dm in the educational sector. this study mainly combined the use of the powerful online e-learning management system (moodle) with dm tools to improve the performance and effectiveness of the learning and teaching manners using the innovative daily data that collected from the educational institutions. learning analytical enhance rubric (lae-r) is the main tool that can retrieve the data from the activities of the students on the moodle. the use of the lae-r can lead for many benefits for both of the educators and students, for example, support the struggling students, a better way for students’ advising, enhance the administration processes and enhance the performance of the students to learn from each other. keywords: data mining, educational data mining, moodle, learning analytical enhance rubric introduction data mining (dm) playing a paramount role in many sectors such as industry, agriculture, retail, banks, health care, and education. due to the globalization and the innovation in the past few decades, the educational institutions faced great challenges and to improve the administrational processes as well as to enhance and develop the academic activities. the international educational dm society defined the educational dm (edm): “an emerging discipline, concerned with developing methods for exploring the unique types of data that come from the educational settings. as well as using those methods to better understand students, and the settings which they learn in.” edm is principally concerned with analyzing the data that mainly generated from the process educational setup using different systems. therefore, edm’s aim is to develop models that enhance and improve the teaching and learning process as well as the effectiveness of the institutions.[1] dm is playing an important role in higher education, which is mainly, improves the level of academicals processes. in every society, higher education institutions are mainly considered the center of knowledge and innovations; it is very important to use the huge data that available in these institutions to improve the level of their operations to prepare the new generations of students that equipped with the new skills to run in different sectors of the society. hence, it is necessary to develop the systems of dm that able to use the available educational data in the higher education institutions that will lead to offer high-quality educational processes to students as well as to enhance the management and research processes that will lead to the innovations and discoveries.[2] collaborative learning (edm and lae) is usually used to deal with the issues that are related to providing instructional strategies that support the collaboration of the students that are working together by groups. the main key indicator was the interaction to measure the effectiveness of the collaboration of the users’ activity logs from their learning platform, which was used as the main key tool for the activities of the learners inferring to fit the demanded behaviors.[3] edm is considered as an emerging field in the area of dm. however, the educational setting is mainly used dm tools to corresponding author: alyaa a. mahdi, department of computer science, cihan university-erbil, erbil, kurdistan region f.r. iraq. e-mail: alyaa.asaad@cihanuniversity.edu.iq received: aug 24, 2020 accepted: sep 12, 2020 published: dec 20, 2020 doi: 10.24086/cuesj.v4n2y2020.pp13-18 copyright © 2020 alyaa a. mahdi. this is an open-access article distributed under the creative commons attribution license cihan university-erbil scientific journal (cuesj) mahdi: educational data mining to improve the academic performance 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 13-18 discover and analyze students’ performance, to predict their results, and to avoid dropout also emphasis on the good and poor academic performers then provide feedback for the instructors and faculties as well as visualization of data is to improve the assessment process. the modern educational institutions mainly need dm techniques to set strategies and plans for the future. mainly, the top-level educational authorities of the institutions may apply the outcomes of the edm results to realize the main key trends and behaviors of the students’ performance, which may lead to design the innovative pedagogical strategies.[4] backround edm mainly the edm is considered as a developed research field to collect big amounts of students’ data then analyze their activities to understand their behavior, which is the base of creating intelligent edm. the analysis of learning and knowledge of the students’ performance is the platform used in a learning environment to measure, scrutinize, as well as report the patterns of learning behavior to improve teaching and learning processes.[5] edm is concerning developing unique methods as well as discovering the educational environment for a better understanding of the students’ behavior. in another word, edm uses a large repository of students’ data from the system such as the moodle with a view to give information regarding how a student is learning process. as well as, the internet and client/service interface utilization that provides a very big repository of the information about students learning process and of learning materials availability and teaching methods.[6] recently, edm garnered the significance for improving and personalizing students’ learning process. many researchers focused on predicting the performance of the learners mainly such a task is not simple to achieve, especially during the course delivery time.[7] learning analytics and edm have been efficient and effective really within a short time. however, the estimates of (student-at-risk) systems are mainly employed at the scale of higher education as well as the adaption of new learning techniques and systems that are used widely in higher education. as well as there is a steady stream of models and discoveries in a very wide range of once challenging areas to the study: the collaborative learning classroom participation, as well as online connections, students’ engagement, and motivation, the meta-cognition as well as self-regulated learning, and many other areas.[8] edm provides appropriate opportunities to evaluate and assess students in an enhanced technological learning system that is considered an easy and efficient platform if we compare it to the traditional systems. to evaluate the students, many factors have to be taken into consideration like their login and logout on the learning system, the time that they spent on each page, participation in the classwork, and the discussion form.[9] edm tools are embedded as a plugin tool in the moodle and the use of these tools is easy even for the non-experts.[10] edm adoption requires a structure and steps for an efficient design that can be upgradable to the system. the methodological structure of this study is mainly based on using data called the cross-industry standard process for data mining (crisp-dm). figure 1 shows the steps and the processes of implementing crisp-dm. crisp-dm technique is mainly recorded as one of the successful guidelines or ways of implementing a dm system. according to sahay and mehta,[11] the foundation of the crisp-dm method was by the european community research fund then the statistical companies such as spss and ncr developed it. the following guidelines discussed briefly the implementation of the dm in the educational sector:[2] 1. understanding the business: the main aim is to understand the objectives and structure of the educational institutions to fulfill the requirements. 2. understanding the data: the main aim is how to deal with and represent the collected data. 3. data preparation: to transfer and clean the collected data to finalize the data of moodle. 4. modeling: the main aim is to apply the technique to represent the crisp algorithm. 5. evaluation: to measure if the model fulfills the requirements of the educational institutions. 6. deployment: implementing and starting the work in the project. educational data mining has significant importance in the higher educational sector. various studies showed that the application of edm can enhance the teaching and learning processes in the universities they studied. for example, at ajman university, different colleges of science and technology in the united arab emirates significantly after applying the edm mentioned that the performance of the students is not very dependent on the academic efforts, but some other factors that may have equal to greater influences. edm motivating and helping the universities to perform edm on the data of their students’ to find out important results figure 1: steps of data mining mahdi: educational data mining to improve the academic performance 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 13-18 that can help both the university, educators, and students in several ways.[12] as well as another edm application, the case was in from vbs purvanchal university, jaunpur (uttar pradesh), the department of computer applications courses, master of computer applications to predict their students’ performance by the end of each semester. significantly, the use of edm supports of both the educators and students to enhance the division their student to identify those students that mainly needed special attention to reducing the ration of fail as well as to take suitable action for the examination of the next semester.[13] higher education institutes in iraq the iraqis have a wide intel lectual history from the ancient of the mesopotamian civilizations the remarkable accomplishments period throughout the arab-islamic in the 9th century. the higher education in iraq dates back to the last century in the year 1908 that law college in baghdad was established, then academies college was established in the 1950s. a number of the other colleges as well as the academies later all these colleges combined as baghdad university in the 1960s. the flourishing of the higher education sectors was between the 1960s and the 1970s. the iraqi college of sciences was the nexus for the research and the literature. medicine and science colleges attracted students from many countries to baghdad university. the iraqi universities according to surveys of the unesco in the year 2004 were about 200 colleges also about 800 departments and 28 specialized institutes, in addition to the american style private universities in the sulaimaniya and koya. however, only missan and muthanna governorates were without universities. then, in 2006, the public universities included in iraq sulaimaniya, dahuk, koya, tikrit, kirkuk, mosul, anbar, baghdad, irbil, islamic studies, al-nah rain, al-mustansiriyya, diyala, technology, qadisiyya, karbala, wasit, kufa, thi qar, babil, and basra. in addition, the number of private universities was (7) in baghdad and (2) in kurdistan region.[14] according to the unesco survey in the year 2003, the number of students in the non-profit colleges was about 400,000 students, as shown in details of figure 2: • the ratios of the students were 58% of male students and 42% of female students. • approximately 50% of the students attended baghdad universities (baghdad, al-nahrain, al-mustansiriyya, technology, and islamic studies). • near to 32% of the students studied the education, 28% were in engineering and sciences colleges, 15% in the law and social sciences colleges, 13% in the medical college, and 12% were in humanities college. the total number of the academic staff was 19,000 member, 36% from them had doctorate, 58% of them had master degree, and also only 6% of them had bachelor degree. most of the iraqi universities did not have the system of data collection, but mainly in the last years, some of the universities, especially private universities, were started to utilize student information management systems with a view to collect and manage information of the students. some universities mainly used systems like moodle for the academic purposes. however, these systems have produced data and information regarding the students in the universities. the generated data were used to produce some reports to improve the operations of learning, teaching, and research at these universities. data collection system in this study, the data collection was mainly suggested to be used by the iraqi universities and the kurdistan region. this system will mainly contain the modules for uploading and downloading lectures and assignments with the sign-in from figure 2: the main actions in the moodle learning system mahdi: educational data mining to improve the academic performance 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 13-18 the students and the lecturers. in addition, it includes the modules for scheduling academic activities. mainly that will provide the use time and resources efficiently and effectively for the university. moodle is a good example of such a system. this system if it’s deployed by the university then used by the lecturers and students will mainly provide an efficient tool to enhance the academic environment. moodle as an academic tool moodle is basically a free online learning management system that provides for administrators, educators, and students by a robust and dynamic platform for all the academic processes. moodle system aims are to help the instructors to present the knowledge better and to support in acquiring skills, knowledge, and experiences.[15] this online system supports the academic staff to enhance their performance and for better assessment methods for their students. furthermore, this system can be used by many other business processes such as the schools, the university, and the firms. the moodle is a free download system, configure, and also an open-source system that can serve various business aspects.[16] moodle system is robust with some training that can be modified and they are available to the user for any higher education institute. the most important activities in moodle are shown in figure 2. dm tool based on moodle learning analytical enhance rubric (lae-r) is considered as one of the main tools that can retrieve the data intelligently from the activities of the students on the moodle to predict the behaviors of the learners to produce clear patterns or trends. mainly this tool works on collecting data from the online activities of the students such as their login and logout from the system, their participation in the discussions, and many other activities for a better understanding of their behaviors that will lead to a better assessment for the students. rubric is an assessment tool developed installed and imbedded as a plug-in on computers of the users that can extracts the interaction students within the moodle system. figure 3 shows the mentioned learning system environment. analysis and visualization an efficient visualization tool called gismo that shows the end result of the lae-r for the educators using graphs of any other descriptive tool. for instance, figure 4 shows the students’ representation login activities in moodle; using this analysis, the teachers can easily assess their dormant students. the analysis includes the pedagogical coherent system assessment aspects, which analyze how the students are accessing the lectures for example as a pdf and/or word formats on the moodle, also studying time per each page of the lecture. the excel pivot tables’ provide the statistics of learning to show the data on spreadsheets report. from a big volume of data available on the moodle using, the mentioned tools will provide the ability for the educators to extract the valuable information.[17] furthermore, this tool helps the students to access their assessment and to get a comparative analysis and efficient recommendation. the platforms of online technological aim to facilitate scalable peer-to-peer training and instructor instructors’ learning dominant channels.[18] combining moodle (online learning management system) with the powerful dm tools like lae-r provides an efficient and effective system for exploring a huge amount of educational data that available to the educational institute also this will provide many improvements to the educational process. figure 3: framework of learning analytical rubric mahdi: educational data mining to improve the academic performance 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 13-18 conclusion and future work the study illustrates how the combination of combining between the online learning management system with the dm applications provides an effective and efficient tool for the higher education institutions using the tools that have been mentioned in this study that enhances the evaluation for the students’ performance from their online activities on the moodle system. from the educator’s perspective, the professor, or educators are able to support the struggling students, especially by providing a better way to advise students. also, it is profitable for the marketing of the universities and supports and enhance the administration processes. for that reason, it will provide the capability to improve and enhance efficiency in the educational process. on the other hand, from the student’s perspective, it encourage and help them to enhance their performance and to learn from each other, leading them to be creative. acknowledgment gratitude and appreciations to my family for giving me the support from various aspects and encouraging me to develop and complete my work. special thanks for ministry of higher education and scientific research in north iraq that provides me by the adequate information. references 1. a. dutt, m. a. ismail and t. herawan. a systematic review on educational data mining. united states: ieee acess, 2017. 2. m. chalaris, s. gritzalis, m. maragoudakis, c. sgouropoulou and a. tsolakidis. improving quality of educational processes providing new knowledge using data mining techniques. procedia-social and behavioral sciences, vol. 147, pp. 390-397, 2014. 3. h. aldowah, h. al-samarraie and w. m. fauzy. educational data mining and learning analytics for 21st century higher education: a review and synthesis. telematics and informatics, vol. 37, pp. 13-49, 2019. 4. s. hussain, r. najoua and n. a. dahan. educational data mining and analysis of students’ academic performance using weka. berlin, germany: researchgate, 2018. 5. z. papamitsiou, a. economides and i. n. sneddon. learning analytics with excel pivot tables. berlin, germany: moodle research conference beuth university of applied sciences, pp. 115-121, 2012. 6. c. romrto, s. ventura, and e. garcıa. data mining in course management systems: moodle case study and tutorial. computers and education, vol. 51, no. 1, pp. 368-384, 2008. 7. m. n. injadat, a. moubayed, a. b. nassif and a. shami. systematic ensemble model selection approach for educational data mining. netherlands: elesiver, 2020. 8. r. s. baker, challenges for the future of educational data mining: the baker learning analytics prizes. journal of educational data mining, vol. 11, no. 1, pp. 1-17, 2019. 9. m. petropoulou, k. kasimatis, i. dimopoulos and s. retalis. lae-r: a new learning analytics tool in moodle for assessing students’ performance. bulletin of the technical committee on learning technology, vol. 16, no. 1, pp. 2-5, 2014. 10. j. hung, k. rice, and a. saba. an educational data mining model for online teaching and learning. journal of educational technology development and exchange, vol. 5, no. 2, pp. 77-94, 2012. 11. a. sahay, and k. mehta. assisting higher education in assessing, predicting, and managing issues related to student success: a web-based software using data mining and quality function deployment. las vegas: academic and business research institute conference, 2010. 12. a. saa. educational data mining and students’ performance prediction. international journal of advanced computer science and applications, vol. 7, no. 5, p. 070531, 2016. 13. b. k. baradwaj and s. pal. mining educational data to analyze students performance. international journal of advanced computer science and applications, vol. 2, no. 6, pp. 63-69, 2011. 14. i. harb. higher education and the future of iraq. united states: institute of peace, 2008. 15. h. f. hasan, m. nat and v. z. vanduhe. gamified collaborative environment in moodle. united states: ieee access, 2019. 16. b. ngom, i. niang, and h. guillermet. enhancing moodle for offline learning in a degraded connectivity environment. figure 4: information about students login in full semester mahdi: educational data mining to improve the academic performance 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 13-18 tangier, morocco: ieee, 2012. 17. h. dierenfeld, and a. merceron. learning analytics with excel pivot tables. berlin, germany: moodle research conference, beuth university of applied sciences, pp. 115-121, 2012. 18. v. z. vanduhe, m. nat and h. f. hasan. continuance intentions to use gamification for training in higher education: integrating the technology acceptance model (tam), social motivation, and task technology fit (ttf). united states: ieee, 2020. . 48 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 48-52 research article energy management in wireless sensor networks for internet of things applications reem jafar ismail1*, samar jaafar ismael2, wisam h. ali2 1department of computer science, cihan university-erbil, kurdistan region, iraq, 2department of electromechanical engineering, university of technology, baghdad, iraq abstract internet of things (iot) aims to develop a smart world based on sensing environment. the energy management of wireless sensor networks (wsns) is a big challenge in iot since sensor nodes have limited energy and they need to have long life for collecting data and information. the aim of this paper is to propose an efficient energy routing algorithm in wsn and infrastructure based on construction an adaptive energy map of sensor nodes. the results show improvement in overall system performance and lifetime of wsn compared to traditional scenario. keywords: shortest energy path, energy map, cluster region, multihop routing introduction wireless sensor networks (wsns) consist of number of sensor nodes which are distributed over the environment to monitor various physical characteristics of the real world, especially in internet of things (iot) applications such as health care, education, transportation, or food industry.[1,2] the life of wsn depends on energy of sensor node.[3,4] some proposal saves energy of nodes itself by energy power management of battery;[5] other proposals save energy by proposing efficient routing algorithm in wsn to maximize lifetime of wsn. many researches had proposed for routing protocol to collect data from sensor nodes depending on central or hierarchical routing. in salim and osamy[6] proposed a routing protocol using sink for data gathering from sensor nodes. in khan et al.,[3] heinzelman et al.,[7] biradar et al.,[8] mahmood et al.,[9] kumar and pal,[10] khan and sampalli,[11] shi et al.[12] researches proposed a cluster-based routing algorithm for data gathering in wsns. in hierarchical cluster-based wsn, the network is divided into clusters, and hierarchy of different nodes is defined. each cluster has a cluster head (ch). sensor nodes in each cluster region get the information and send it to ch.[3] in this research, we propose an efficient optimization algorithm that efficiently manages the energy of the cluster region of hierarchical cluster-based wsn. the main contribution of this work is as follows: • the design and implementation of routing algorithm that saves energy in wsns to maximize lifetime of wsn. • the development and construction of energy map of sensor nodes that are used to manage energy of cluster region when routing algorithm is implemented. the remainder of this research described the following: we present works relate to wsn that applied in iot applications for energy saving; then, the propose algorithm is presented and evaluated; and finally, we conclude the work. literature review routing in hierarchical wsn architecture has advantage over centralized architecture since the route from source to destination is controlled over cluster regions that are led to manage the power of sensor nodes efficiently. chauhan et al. proposed a routing algorithm to consume power using minimum number of nodes from source to destination with a static shortest path using dijkstra’s algorithm.[13] the problem with their proposed algorithm is that they did not consider the energy level of the selected nodes of shortest path, where their measure was the shortest distance from source to destination. corresponding author: reem jafar ismail, cihan university-erbil, kurdistan region, iraq. e-mail: reem_aljanabe@yahoo.com received: apr 13, 2019 accepted: apr 17, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp48-52 copyright © 2019 reem jafar ismail, samar jaafar ismael, wisam h. ali. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) ismail et al.: energy management in wsns for iot applications 49 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 48-52 in, khan et al.[3] a multistage routing protocol is proposed for energy consuming in wsn, where the data are collected from sensor nodes by transferring it from stage to next stage through ch and forwarder to base station by assuming that the ch is usually at the center of the cluster region and the path is direct from sensor node to the ch in the cluster region. table 1 presents an overview for some of routing protocols, where most of them assume that the sensor nodes are distributed randomly in the propose region. proposed routing algorithm for efficient energy management our proposal saves energy by proposing efficient routing algorithm in wsn to maximize lifetime of wsn. in this research, we developed an efficient energy management algorithm that has the following prosperities: • nodes are distributed according to boualem et al.[14] and not randomly, which will affect the overall energy consumption of sensor nodes compared to other protocols • the shortest path between source and destination is not static when same source node sends to same destination node since the path will be updated according to the energy level of intermediate sensor nodes • the controller (ch) will construct an energy map and update it with time since the energy of sensor node is changing with time. in this research, we propose a novel optimization algorithm that efficiently manages the energy of the cluster region of hierarchical cluster-based wsn. in cluster region, we try to collect information from sensor nodes and send it to ch with consuming energy. in our proposal routing, shortest energy path is chosen in cluster region depending on the sensor nodes that have maximum energy. selecting sensor nodes with maximum energy and avoiding sensor nodes with minimum energy will maximize the lifetime of wsn. when sensor nodes send their information to ch, the proposed shortest energy path is used which will maximize the lifetime of wsn. in our proposed algorithm, the ch is responsible on finding shortest energy path and construction energy map of cluster region. energy map of cluster region is constructed when each sensor node in cluster region will send message to ch periodically telling the remaining energy of sensor node. the sensor nodes location is static, but shortest energy path is chosen according to the highest battery charge of sensor nodes not according to distance between them. the shortest energy path from source to destination is not static since in proposed routing algorithm, the shortest energy path will be dynamic according to the energy that will be received from sensor nodes since power level of battery for sensor node is decreasing within time. the dijkstra’s algorithm is an algorithm that finds shortest distance in a given path between the source points to the destination point. dijkstra’s uses these nodes in routing and saves the points where the cost of distance is low.[15] in our proposed energy routing algorithm, the dijkstra’s algorithm is used to construct and find the shortest energy path of sensor nodes. the overall properties of the proposed energy routing algorithm will save energy and increase network lifetime by the following assumptions: • multistage data transmission mechanism where an energyefficient routing protocol for wsns is proposed. it consists of a routing algorithm for data transmission, ch selection algorithm, and a scheme for the formation of clusters[3] also if ch within cluster region died due to energy consumption another head will be chosen for the same cluster region sensor node wants to send data to cluster head (ch) send data with single hop yes no does sensor node have direct link with ch? send data with multiple hop using proposed energy routing algorithm figure 1: sensor nodes sending data to cluster head within cluster region table 1: overview of related works for energy management in wsns algorithms topology organization distribution of sensor nodes in region control node (cluster head) energy map route from s to d within cluster region number of hops chauhan et al., 2015[13] centralized randomly none not constructed static single kayhan et al., 2018[3] hierarchical routing randomly available not constructed static single hop within cluster region since cluster head is centralized proposed energy routing algorithm hierarchical routing enough monitoring for region and not random available is constructed dynamic multihop within cluster region ismail et al.: energy management in wsns for iot applications 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 48-52 • in cluster region, the sensor nodes are not disturbed randomly since the topology of sensor nodes are optimized using semi-random deployment protocol proposed by boualem et al.[14] where the area of interest is covered with a fewer sensor nodes with enough monitoring • if the wireless sensor region is scalable, the proposed algorithm still operate efficiently since the routing is multihop and not single hop in cluster region. this means more sensor nodes could be added within cluster region and the proposed algorithm will still work efficiently. as shown in figure 1, the sensor node will send the data from source to destination using algorithm 1, proposed energy routing algorithm. the objective of algorithm 1 is to maximize lifetime of sensor nodes within the cluster region. algorithm 1. proposed energy routing algorithm input: number of sensor nodes n, graph representation for sensor nodes (node_i, node_j) and energy map vector: [ level_of_node_energy_i,……, level_of_node_energy_n] output: shortest energy path from sensor node to cluster head 1: for each node_i in cluster region specify direct link for all its neighbors as (node_i, node_j) 2: while wsn is life 3: for each node in cluster region send energy level to cluster head periodically 4: cluster head construct an energy map of the cluster region 5: for each node in cluster region 6: find neighbor node which has maximum energy level 7: end for 8: find maximum energy level path from sensor node to cluster head using dijkstra’s algorithm 9: end while. simulation and evaluation in this section, we present the implementation of the proposed energy routing algorithm and then evaluate it, which we did use matlab r2016a (9.0.0.341360). we ran the simulation with 13 nodes, as shown in figure 2; initially, the sensor nodes have full charge so the source node (node no. 13) will reach to destination node (node no. 12), with shortest path using algorithm 1. after the period of time, the energy of the sensor nodes will decrease. as shown in figure 3, which shows the energy map of sensor nodes inside cluster region, the initial shortest path that had been found previously could not be used again since after a period of time, the sensor node energy is decreased, here, all sensor nodes will send a message to the controller about figure 2: proposed wireless sensor network topology figure 3: energy map of sensor nodes inside cluster region where charge is changed overtime where node 13 is source and node 12 is destination node id 1 2 3 4 5 6 7 8 9 10 11 12 13 charge % 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% a: initial charge of nodes node id 1 2 3 4 5 6 7 8 9 10 11 12 13 charge % 100% 100% 99% 100% 100% 99% 100% 100% 99% 100% 100% 99% 99% b: 1st round, send from node 13 to node 12 path is: 13, 3, 6, 9, 12 node id 1 2 3 4 5 6 7 8 9 10 11 12 13 charge % 99% 100% 99% 100% 99% 99% 100% 99% 99% 100% 99% 98% 98% c: 2nd round, send from node 13 to node 12 path is: 13,1,5,8,11,12 node id 1 2 3 4 5 6 7 8 9 10 11 12 13 charge % 99% 99% 99% 100% 99% 98% 100% 99% 99% 99% 99% 97% 97% d: 3rd round, send from node 13 to node 12 path is: 13, 2, 6, 10,12 ismail et al.: energy management in wsns for iot applications 51 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 48-52 the percentage of charge and then the controller will update the energy map and ran the algorithm again according to the received information. figure 5: compare number of dead nodes in each round between traditional algorithm and proposed algorithm figure 4: shortest path from node 13 to node 12 using algorithm in chauhan et al.[13] table 2: compare between traditional algorithm and proposed energy algorithm algorithm round number when the first node is died number of dead node chauhan et al.[13] 500 3 proposed energy algorithm 600 1 after a period of time, the shortest path for routing information will be constructed by choosing the sensor nodes which has maximum energy. we assume that all sensor nodes are homogenous. according to rodrigues et al.,[16] the energy spent in single transmission for sensor node is approximately 0.1 mj when using atmega328p platform. hence, for all sensor nodes in the cluster region, we can apply the proposed equation (1) to find energy of sensor nodes: decrease in power level of sensor node_i new energy node_i = current energy node_i (0.1 x number of transmissions of node_i) value of energy to be decreased with time …(1) where we suppose the value of energy to be decreased with time is constant for all nodes and also in evaluation, the energy of senor node will be calculated as percentage of charge because sensor nodes are homogenous. wsn lifetime performance and energy consumption evaluation consider: scalability, energy level for each sensor node, number of alive nodes, and number of dead nodes as parameters for evaluations per round. • scalability, a property of network which will increase the network size by adding new nodes. • energy level of sensor node, this means sensor node power which will be calculated as charge percentage. • number of alive nodes, these are total nodes that have enough energy to communicate in routing. • number of dead nodes, these are total nodes that did not have energy to communicate in routing. results the proposed energy algorithm is evaluation and compared with chauhan et al. proposed algorithm.[13] [table 2] shows the comparison according to round number when the first node is died and number of dead node. figure 4 shows the cluster region, if we suppose that the source is node number 13 and the destination is node number 12. according to chauhan et al.,[13] the route from source to destination will be static since every time 13 send data to 12 where the algorithm will use the same path which is 13, 3, 6, 9, and 12 because it is the shortest path. if we suppose that each transaction 1% of sensor node charge will decrease; then, after 100 transitions between 13 and 12, three nodes will be died due to energy consuming of same sensor nodes which are node 3, 6, and 9. figure 5 shows number of dead nodes compare to round number since we suppose the source is node 13 and the destination is node 12 , as shown in the figure in round 600 about 23% of sensor nodes are dead in algorithm[13] while in our proposed algorithm, 8% are dead that will increase number of alive nodes approximately to 15% in each round. conclusion energy management in wsn needs to be saved since sensor nodes have limited power. to maximize lifetime of wsn, an efficient routing algorithm is proposed using less number of sensor nodes for transmission information with high energy. ismail et al.: energy management in wsns for iot applications 52 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 48-52 selecting sensor nodes with high energy instead of low energy will keep the topology of wsn stable since the number of alive node will be the same as long time as possible in wsn. each round the energy map is constructed based of sensor nodes power and then the route is found from source to destination with optimized power consuming. the proposed energy routing algorithm considers scalability, where we can add new sensor nodes to the cluster region, especially when some sensor node starts to die. scalability of cluster region will increase number of nodes and then the energy level of the cluster region will also be increased. acknowledgments the authors would like to acknowledge cihan university-erbil, for their support and sponsorship of this project. references 1. x. gu, x. zhou and y. sun. “a data-gathering scheme with joint routing and compressive sensing based on modified diffusion wavelets in wireless sensor networks”. sensors, vol. 18, pp. 724, 2018. 2. w. ejaz, m. naeem, a. shahid, a. anpalagan and m. jo. “efficient energy management for the internet of things in smart cities”. ieee communications magazine, january 2017. 3. m. k. khan, m. shiraz, k. z. ghafoor, s. khan, a. s. sadiq and g. ahmed. “ee-mrp: energy-efficient multistage routing protocol for wireless sensor networks”. wireless communications and mobile computing, vol. 2018, p. 6839671, 2018. 4. y. lu, x. liu and m. li. “study on energy-saving routing algorithm based on wireless sensor network”. journal of computers, vol. 28, no. 4, pp. 227-235, 2017. 5. j. a. khan, h. k. qureshi and a. iqbal. “energy management in wireless sensor networks: a survey”. computers and electrical engineering, vol. 41, pp. 159-176, 2015. 6. a. salim and w. osamy. “distributed multi chain compressive sensing based routing algorithm for wirelesssensor networks”. wireless networks, vol. 21, pp. 1379-1390, 2015. 7. w. r. heinzelman, a. chandrakasan and h. balakrishnan. “energy-efficient communication protocol for wireless microsensor networks”. in: proceedings of the 33rd annual hawaii international conference on system siences (hicss ’00). ieee, vol. 2, 2000. 8. v. biradar, s. r. sawant, r. r. mudholkar and v. c. patil. “multihop routing in self-organizing wireless sensor networks”. international journal of computer science issues, vol. 8, no. 1, pp. 155-164, 2011. 9. d. mahmood, n. javaid, s. mahmood, s. qureshi, a. m. memon and t. zaman. “modleach: a variant of leach for wsns”. in: proceedings of the ieee 8th international conference on broadband, wireless computing, communication and applications (bwcca ’13). compeigne, france, pp. 158-163, 2013. 10. s. v. kumar and a. pal. “assisted-leach (a-leach) energy efficient routing protocol for wireless sensor networks”. international journal of computer and communication engineering, vol. 2, no. 4, pp. 420-424, 2013. 11. z. a. khan and s. sampalli. “azr-leach: an energy efficient routing protocol for wireless sensor networks”. international journal of communications, network and system sciences, vol. 5, no. 11, pp. 785-795, 2012. 12. s. shi, x. liu and x. gu. “an energy-efficiency optimized leach-c for wireless sensor networks”. in: proceedings of the 2012 7th international icst conference on communications and networking in china (chinacom ’12), pp. 487-492, 2012. 13. p. chauhan, a. negi and t, kumar. “power optimization using proposed dijkstra’s algorithm in wireless sensor networks”. international journal of advanced research in computer and communication engineering, vol. 4, no. 7, pp. 326-330, 2015. 14. a. boualem, y. dahmani, a. maatoug and c. de-runz. “area coverage optimization in wireless sensor network by semirandom deployment”. in: proceedings of the 7th international conference on sensor networks (sensornets), pp. 85-90, 2018. 15. l. parungao, f. hein and w. lim. “dijkstra algorithm based intelligent path planning with topological map and wireless communication”. arpn journal of engineering and applied sciences, vol. 13, no. 8, pp. 2753-2763, 2018. 16. l. m. rodrigues, c. montez, g. budke, f. vasques and p. portugal. “estimating the lifetime of wireless sensor network nodes through the use of embedded analytical battery models”. journal of sensor and actuator networks, vol. 6, no. 2, p. 8, 2017. tx_1~abs:at/add:tx_2~abs:at 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 20-23 research article hiv, hbs and hcv in dump site workers of erbil governorate muhsin h. ubeid*, tanya s. salih, ameena s. m. juma department of general biology, cihan university-erbil, kurdistan region, iraq abstract this research targeted eighty-nine males working in kany qrzhala, dumpsite. age and gender comparable apparently healthy subjects were selected as healthy controls, and both of the groups were obliged to fill the study’s questionnaire. further, venous blood samples were collected from each individual for serum collection. the accumulated sera reserved for the seroprevalence for antibodies tests of human immunodeficiency virus, hepatitis b surface (hbs) antigen and hepatitis c virus (hcv), (human immunodeficiency [hiv]), (hbs), (hcv), respectively. the automated immunoassay analyzer cobas e411 facilitated the conducting of the mentioned tests. the serum concentration of hiv and hbs antibodies of dumpsite workers revealed a significant increase when compared to the healthy group, while the hcv antibody serum concentration presented no significant alteration when comparing dumpsite workers to the healthy controls. the antibodies present in the sera that belonged to workers is an indicator of exposure to the viruses due to unsanitary health conditions. this may pose a public health risk to the workers themselves, in addition to the people they are in contact with, including their families. keywords: dumpsite workers, hiv, hbs, hcv introduction the increase in the population of any city will result in increasing production of waste materials. in addition, the poor management systems of domestic wastes and lack of modern technology in the management of wastes, all play a major role in increasing the waste materials.[1] studies have shown that dumpsites do not only affect the dumpsite workers but the surrounding areas and the people living there.[2] it has been shown, that cities in many countries face many health issues owing to inadequate municipal management of solid waste.[3] other studies have shown the link between health and environmental effects due to insanitary dumpsites.[4-6] because dumpsite workers handle solid waste directly, this has been shown to be a consequence to various of diseases; infectious and chronic.[5,7] many of the dumpsites are usually located distant on urban areas borders. they represent a major health problem and many diseases, including infectious diseases, i.e., malaria and cholera, have been recorded in these areas and urban cities around them.[8,9] because of the lack of accurate data on the health status of dumpsite workers in erbil city, this research aimed to discover possible exposure of these workers to specific viral diseases. methodology population selection a dumpsite in erbil governorate named as kany qrzhala. eighty-nine males were chosen for the study in hand. age and gender comparable apparently healthy subjects were selected as healthy controls. the entire dumpsite-working group and the healthy group were asked to respond to the study questionnaire and submit the tests explained in the following sections. samples of the blood collection after applying the tourniquet, disinfecting with 70% ethanol anti-cubital fossa, administering with a 23-guage needle corresponding author: muhsin h. ubeid, department of general biology, cihan university-erbil, kurdistan region, iraq. e-mail: muhsin.hmeadi@cihanuniversity.edu.iq received: june 6, 2021 accepted: september 15, 2021 published: september 30, 2021 doi: 10.24086/cuesj.v5n2y2021. pp 20-23 copyright © 2021 muhsin h. ubeid, tanya s. salih, ameena s. m. juma, this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) ubeid, et al.: hiv, hbs and hcv in dump site workers 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 20-23 attached to a disposable five ml syringe. a 5 ml venous blood samples were accumulated from all the participants. later, the collected blood was retained in plain tubes and was set aside for 60 min at room temperature to clot. then, to collect the serum, tubes for whole 10 min were centrifuged at 4°c at ×450 g, afterwards, by a pasteur pipette the separated serum was aspirated then dispensing it into previously sterilized five ml glass tubes. finally, cooling and storing them at −20°. antibody detection restoring the glass tubes with sera to room temperature seroprevalence tests can be achieved. hence, the research employed the automated cobas e411 immunoassay analyzer for the deduction of human immunodeficiency (hiv), hepatitis b surface (hbs), and hepatitis c virus (hcv) antibodies. results workers’ statistics table 1, illustrates the study’s investigated dumpsite group data, where eighty-nine male workers with age ranging from 12 to 65 years. second, rural areas were the location of 92.13%, while urban areas were the location of 7.87%. third, the table recorded 58.43% smokers and 41.57% non-smokers. fourth, it demonstrated there were 61.80% married and 38.20% single workers. last of all, it has been reported that 37.08% of the workers were not formally educated, 40.45% acquired the education of primary school, and 17.97% earned secondary school certificate, and 2.25% diploma and 2.25% b.sc. degree holders. anti hiv antibody the serum concentration of hiv antibody of investigated dumpsite workers revealed a significant increase when compared to the healthy group (table 2, figure 1). anti hbs antibody the serum concentration of hbs antibody of dumpsite workers showed a significant rise when compared with the healthy controls (table 3, figure 1). anti hcv antibody the serum concentration of hcv antibody presented no significant difference when comparing study’s dumpsite workers to the healthy group (table 4, figure 1). discussion the unsanitary disposal of wastes creates a flourishing habitat for diseases to be defused via insects and rodents. this concurs with the occurrence of the black death in the 14th century epidemic. where closely half of the europe population perished.[10] procedures followed by developing countries for waste management are correlated to employment safety and health issues. it was recorded that, and in many countries, diseases and infections of respiratory, skin, gastro-intestinal, muscular-skeletal complications, along with cutting injuries table 1: kany qrzhala, erbil governorate dumpsite workers information variable no. (n=89) percent workers live in rural areas 82 92.13 workers live in urban areas 7 7.87 smoker workers 52 58.43 non-smoker workers 37 41.57 single workers 34 38.20 married workers 55 61.80 education/illiterate workers 33 37.08 education/primary school 36 40.45 education/secondary school 16 17.97 education/diploma 2 2.25 education/bachelor 2 2.25 0 0.05 0.1 0.15 0.2 0.25 0.3 hiv (coi) hbs (coi) hcv (coi) se ru m c on ce nt ra tio n of a nt ib od ie s virus dumpsite workers healthy group figure 1: hiv, hsb and hcv antibody concentrations in the sera of the workers in comparison to the healthy controls are commonly health ramifications for those who are working in dumpsites.[11] studies have identified the poor working conditions of the dumpsite workers and inadequate sanitation practices making them susceptible to diseases, in addition to their lack of anti-pollution equipment.[12] these studies have identified that these workers are always exposed to injuries with broken glass, for example, exposing these workers to many infectious agents like the viruses in the current study, hiv, and hepatitis viruses. moreover, other studies have found that the landfill sites workers’ respiratory health was poor because of exposure to bioaerosols containing bacteria and fungi.[13] another previous study in africa has indicated that health care waste is dumped with other wastes, hence exposing the dumpsite workers to hiv, being considered as occupational exposure.[14] it has been shown that because health care wastes are also disposed in the dumpsites. as such, the likelihood, workers have to be infected by hepatitis b, hiv, or any other diseases is high.[15,16] in the current study, the significant increase in the antibody levels of hiv and hbs is just another proof that these workers have been exposed to these antigens. cholera, diarrhea, hepatitis a and b, besides typhoid had been recorded among dumpsite workers. besides, disease particularly hbv, hcv, and hiv are to cause infection to healthy people via dumping blood-contaminated tools with ubeid, et al.: hiv, hbs and hcv in dump site workers 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 20-23 infected blood and/or other biomedical waste, i.e., syringes, dressings, needles, body parts, etc. these workers pose a danger to the public because of their possibility of transmitting communicable diseases.[17-19] using bare hands by the workers enrolled in the current study to handle waste could be a risk for injury. it is known that even a small injury carries the risk of hbv, hiv, brucellosis transfers, and other blood-borne diseases.[20,21] hbv has been found to be prevalent among in scavengers rather than those who work at municipals as waste collectors.[22,23] injury with sharp objects might be a source of such infections. in fact, the study in hand showed that workers used their bare hands to handle the wastes, with an evident lack of personal hygiene and safety precautions while handling wastes. it has been shown that most dumpsite workers face health hazards.[24,25] other studies have also documented the high risk of waste scavengers of acquiring hbv infection.[26,27] in pakistan, the same health issues regarding dumpsite workers were recorded. hepatitis was among the infections to be highly prevalent among these workers. this was attributed to, their occupation, insufficient living conditions, absence of sanitation and safety measures, and illiteracy. therefore, training regarding waste handling is required.[28] brazil, despite following safety and health regulations, has been found to face many health problems in what they call recyclable collectors. they were constantly exposed to biological, physical, and more extreme hazards. this was because these workers were not aware of the hazards they will face if not using proper protective materials, hence certainly they will be injured. such injuries are comprised head, hands, arms legs, and feet burns and cuts, lesions of eyes and skin, and even hand amputations.[29] another study in egypt also indicated the prevalence of hbv among dumpsite workers, emphasizing on the necessity for vaccination as a prophylactic measure.[30] studies are still conducted in nigeria, where workers were exposed to hazardous substances, which might cause a variety of diseases. occupational hazards in these individuals might be due to the absence or the lack of appropriate tools and equipment of protection, along with their illiteracy. in conclusion, the authors recommended as a vital response to raise the workers awareness, besides enlightenment training, to survive the health hazards and its risks.[31,32] a periodic, objective and documented assessment and auditing of the dumpsites and their workers is a necessary concern to minimize the health issues that these workers face. vaccination against hbv is an effective measure to control such an infection. in addition, personal hygiene practices and regular training (more than 75% of the individuals enrolled in the current study are either illiterate or had only primary school education) on occupational safety could be a good measure to control risks of viral infections among dumpsite workers. lastly, providing protective clothing and proper equipment as a vital issue to prevent injuries. acknowledments we would like to thank municipal directorate labors of erbil city for providing the help in samples collection. references 1. j. n. fobil, d. carboo and c. clement. defining options for integrated management of municipal solid waste in large cities of low-income economies-the case of the accra metropolis in ghana. journal resource management and technology, vol. 28, no. 2, pp. 106-117, 2002. 2. f. p. sankoh, x. yan and q. tran. environmental and health impact of solid waste disposal in developing cities: a case study of granville brook dumpsite, freetown, sierra leone. journal of environmental protection, vol. 4, no. 7, p. 47076, 2013. 3. n. p. thanh, y. matsui and t. fujiwara. assessment of plastic waste generation and its potential recycling of household solid waste in can tho city, vietnam. environmental monitoring and assessment, vol. 175, no. 1, pp. 23-35, 2011. 4. l. giusti. a review of waste management practices and their table 2: hiv antibody concentration in the sera of the workers when contrasted to the healthy group immunological variable gender dumpsite workers healthy group t-test p value p no. mean±se no. mean±se hiv (coi) m 89 0.251±0.002 40 0.244±0.001 0.000 h.s.** p≥0.05: non significant; *p<0.05: significant; **p<0.01: highly significant, coi=cut‑off index, hiv: human immunodeficiency table 3: comparison of hbs antibody concentration in the sera between dumpsite workers and the healthy group immunological variable gender dumpsite workers healthy group t-test p value p no. mean±se no. mean±se hbs (coi) m 89 0.274±0.001 40 0.261±0.001 0.012 s.* *p<0.05: significant; **p < 0.01: highly significant. coi: cut‑off index table 4: comparison of hcv antibody concentration in the sera between dumpsite workers and the healthy group immunological variable gender dumpsite workers healthy group t-test p value p no. mean±se no. mean±se hcv (coi) m 89 0.080±0.001 40 0.080±0.001 0.325 n.s. p≥0.05: non significant; *p<0.05: significant; **p<0.01: highly significant. coi: cut‑off index, hcv: hepatitis c virus ubeid, et al.: hiv, hbs and hcv in dump site workers 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 20-23 impact on human health. waste management, vol. 29, no. 8, pp. 2227-2239, 2009. 5. j. a. nwanta, j. onunkwo and e. ezenduka. analysis of nsukka metropolitan abattoir solid waste and its bacterial contents in south eastern nigeria: public health implication. archives of environmental and occupational health, vol. 65, no. 1, pp. 21-26, 2010. 6. t. s. salih, s. s. haydar, m. h. ubeid and a. s. m. juma. some immunological and hematological parameters among refugees in kawergosk camp-erbil governorate. cihan university erbil scientific journal, vol. 3, no. 1, pp. 80-84, 2019. 7. s. m. juma, m. h. ubeid and t. s. salih. anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers of erbil governorate. cihan university erbil scientific journal, vol. 3, no. 1, pp. 85-89, 2019. 8. s. abul. environmental and health impact of solid waste disposal at mangwaneni dumpsite in manzini: swaziland. journal of sustainable development in africa, vol. 12, no. 7, pp. 64-78, 2010. 9. m. h. ubeid. prevalence of leukocytes in type 2 diabetic patients in erbil city. medical journal of babylon, vol. 17, no. 1, pp. 19, 2020. 10. britannica encyclopaedia. encyclopaedia britannica almanac 2010. united kingdom: encyclopaedia britannica, inc., 2010. 11. d. bleck and w. wettberg. waste collection in developing countries-tackling occupational safety and health hazards at their source. waste management, vol. 32, no. 11, pp. 2009-2017, 2012. 12. o. i. o. gani and o. h. okojie. environmental audit of a refuse dump site in the niger delta region of nigeria. journal of public health and epidemiology, vol. 5, no. 2, pp. 59-65, 2013. 13. g. f. akpeimeh, l. a. fletcher and b. e. evans. exposure to bioaerosols at open dumpsites: a case study of bioaerosols exposure from activities at olusosun open dumpsite, lagos nigeria. waste management, vol. 89, pp. 37-47, 2019. 14. j. moses. focusing on environmental aspect of hiv/aids prevalence in abuja-nigeria. african health sciences, vol. 6, no. 1, p. 64, 2006. 15. world bank. waste management in china: issues and recommendations. urban development working papers, east asia infrastructure department, world bank-working paper no. 9. united states: world bank, 2005. 16. c. r. n. corrao, a. d. cimmuto, c. marzuillo, e. paparo and g. la torre. association between waste management and hbv among solid municipal waste workers: a systematic review and metaanalysis of observational studies. the scientific world journal, vol. 2013, p. 692083, 2013. 17. h. c. g. nguyen, chalin, t. m. lam, and v. w. maclaren. health and social needs of waste pickers in vietnam. research paper. africa: canadian international development agency (cida) waste-econ progamme south east asia, 2003. 18. s. cointreau. occupational and environmental health issues of solid waste management: special emphasis on middle-and lowerincome countries, urban papers. united states: sage, 2006. 19. m. p. g. mol, d. b. greco, s. cairncross and l. heller. hepatitis b and c in household and health services solid waste workers. cadernos saude publica, vol. 31, pp. 295-300, 2015. 20. l. rushton. health hazards and waste management. british medical bulletin, vol. 68, no. 1, pp. 183-197, 2003. 21. a. adeyemi, o. a. omolade and r. r. raheem-ademola. immunochromatographic testing method for hepatitis b, c in blood donors. journal of antivirals and antiretrovirals, vol. 3, p. 5, 2013. 22. h. o. sawyerr, r. o. yusuf and a. t. adeolu. risk factors and rates of hepatitis b virus infection among municipal waste management workers and scavengers in ilorin, kwara state, nigeria. journal of health and pollution, vol. 6, no. 12, pp. 1-6, 2016. 23. r. o. yusuf, h. o. sawyerr, a. t. adeolu, l. m. habeeb and t. t. abolayo. seroprevalence of hepatitis b virus and compliance to standard safety precautions among scavengers in ilorin metropolis, kwara state, nigeria. journal of health and pollution, vol. 8, no. 19, p. 180914, 2018. 24. f. akinbode. impacts of solid waste on health and the environment. minnesota: walden university, 2011. 25. s. p. kandasamy, a. b. akolkar, a. manoharan and s. paranji. municipal solid waste management at chennai in southern india-an occupational health perspective. international journal of health promotion and education, vol. 51, no. 1, pp. 50-61, 2013. 26. n. o. ariyibi. comparative study of the knowledge and attitude towards hepatitis b virus infection and its seroprevalence among waste scavengers and market porters in lagos state. public health, vol. 2013, pp. 1-5, 2013. 27. ansari-moghaddam, h. ansari, s. khosravi, e. saneimoghaddam, m. mohammadi, s. m. alavian, and h. poustchi. the prevalence of hepatitis b virus among municipal solid waste workers: necessity for immunization of at-risk groups. hepatitis monthly, vol. 16, no. 3, p. e30887, 2016. 28. majeed, s. a. batool, m. n. chaudhry and r. a. siddique. scavenging demeanor in bahawalpur, pakistan: social and health perspective. journal of material cycles and waste management, vol. 19, no. 2, pp. 815-826, 2017. 29. t. r. zolnikov, d. ramirez-ortiz, h. moraes, v. r. n. cruvinel, a. dominguez and d. galato. continued medical waste exposure of recyclable collectors despite dumpsite closures in brazil. journal of health and pollution, vol. 9, no. 23, p. 190905, 2019. 30. abd el-wahab, ekram w., and safaa mohamed eassa. seroprevalence of hbv among egyptian municipal solid waste workers. heliyon, vol. 5, no. 6, p. e01873, 2019. 31. a. adeyi and a. m. adeyemi. potential occupational health effects of municipal solid waste management in nigeria, the case of lagos and ibadan. ife journal of science, vol. 21, no. 2, pp. 417-430, 2019. 32. m. h. homady, juma, ameena sm, ubeid, muhsin h, salih, tanya s, al-jubori, mm. age and gender in relation to colorectal cancer in najef province: a histopathological study. acta scientific pharmaceutical sciences, vol. 5, no. 3, p. 0686, 2021. tx_1~abs:at/add:tx_2~abs:at 46 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 46-51 research article automatic classification of coronavirus disease-19 chest x-ray images using local binary pattern and binary particle swarm optimization for feature selection bazhdar n. mohammed1, firas h. al-mukhtar2, raghad z. yousif1, yazen s. almashhadani3 1department of physics, salahaddin university-erbil, iraq, 2department of information technology and computer science, catholic university in erbil, iraq, 3department of communication and computer engineering, cihan university-erbil, kurdistan region, iraq abstract novel coronavirus disease 2019 (covid-19) is a type of pandemic viruses that causes respiratory tract infection in humans. the clinical imaging of chest x-ray (cxr) by computer-aided diagnosis (cad) plays an essential role in identifying the patients infected by covid-19. the objective of this paper presents a cad method for automatically classifying 110 frontal cxr images of contagious people according to normal and covid-19 infection. the proposed method contains four phases: image enhancement, feature extraction, feature selection, and classification. gaussian filter is performed to de-noise the images and adaptive histogram equalization for image enhancement in the pre-processing step for better decision-making. local binary pattern features set are extracted from the dataset. binary particle swarm optimization is considered to select the clinically relevant features and develop a robust model. the successive features are fed to support vector machine and k-nearest neighbor classifiers. the experimental results show that the system robustness in classification covid-19 from normal images with average accuracy 94.6%, sensitivity 96.2%, and specificity 93%. keywords: computer-aided diagnosis, chest-x-ray, coronavirus disease 2019, local binary pattern, binary particle swarm optimization, classification introduction at the end of december 2019, a new type of worldwide outbreak, coronavirus: the severe acute respiratory syndrome coronavirus 2 (sars-cov-2), is called novel coronavirus (coronavirus disease 2019 [covid-19]) has been announced in wuhan, china. after that, it was declared a global health emergency by the centre of disease control and prevention (cdc) and the world health organization (who).[1] since the past two decades, other known case of a typical pneumonia sars-cov has been discovered originating from a healthcare worker in guangdong province, china.[2] sars-cov was a relatively rare epidemic disease; the incidence killed approximately 800 persons among 8437 infected cases which belongs to 11% case of fatality rate.[3] furthermore, 9 years after guangdong’s plague, another type of this virus is middle east respiratory syndrome (mers) identified in saudi arabia in 2012, also known as camel flu, because it is believed this kind originated from the camel then transported to humans by contacting.[4] during the tests, many differences and similarities in the symptoms, screening features, and management of corona disease types have been identified. the rapid rate in spreading of covid-19 is considered one of the main diversities of nowadays novel coronavirus.[5] nowadays, many radiological images are captured in medical centers and hospitals. still, defects in diagnostic tools are one of the major problems that all countries face during the initial stages of this pandemic disease. for these reasons, fast, reliable screening of covid-19 is very crucial and impressive. besides therapeutic and clinical guides, imaging provides extra help to the physicians, and it helps to control the disease from cross-infection effectively. the american college of radiology suggests that chest x-ray (cxr) may be advised to reduce the risk of spreading.[6] computed tomography (ct)/cxr tool for imaging is one of the key components besides the corresponding author: bazhdar n. mohammed, department of physics, salahaddin university-erbil, iraq. e-mail: bazhdar.sh.mohammed@su.edu.krd received: may 16, 2021 accepted: october 18, 2021 published: november 10, 2021 doi: 10.24086/cuesj.v5n2y2021.pp46-51 copyright © 2021 bazhdar n. mohammed, firas h. al-mukhtar, raghad z. yousif1, yazen s. almashhadani. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) mohammed, et al.: covid-19 chest x-ray images 47 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 46-51 clinical assessments that may help early screening suspected pneumonia cases. moreover, cxr is a convenient and low-cost modality for clinical screening and follow-up many pulmonary diseases.[7] the images of various viral pneumonia are similar and they overlap with other infectious during exposure dose that makes a poor contrast. as a result, it may not be easy for radiologists to differentiate covid-19 from normal chest images in some screening cases that may lead to make a wrong decision about the patient’s situation.[8] throughout this pandemic, many groups and researchers have reported different systems of covid-19 pneumonia detection by using computed aided diagnosis (cad). the cad consists of pre-processing, features extraction and selection, classification, and post-processing techniques. the main purpose of this work-study is to develop a cad program for screening and classifying ct/cxr images according to normal and covid-19 on a small data set that will be useful in decreasing the workload of healthcare personnel. the contributions provided to this study is (1) implementation diagnosis system for enhancement the images using gaussian filter, (2) the usage of local binary pattern (lbp) to extract the texture features from the images, and (3) performing the classification technique using k-nearest neighbor (knn) and support vector machine (svm) classifier for obtaining suitable accuracy, sensitivity, specificity, and precision. during the spreading of covid-19 as a pandemic disease till now, researchers have tried to use different machine learning techniques to classify the patient’s diagnostic tests based on radiological images that assist the doctors for accurate clinical decision which reduced the number of deaths. abbas et al. suggested a method for classifying covid-19 images in xrd diagnostic images by performing a transfer algorithm. the feature vector of datasets is classified based on convolution neural network (cnn) technique.[9] another intelligence system was proposed by purohit et al.[10] to separate covid-19 infected x-ray and ct scan images of chest based on k-mean clustering as an unsupervised method for feature extraction; then the images are classified by cnn machine learning technique. to identify covid-19 cases from normal images narin et al.[11] proposed a method deep cnn. for this purpose, the dataset was divided into two parts: 80% for the training phase and 20% for the testing phase based on 5-fold cross validation. in elasnaoui et al.[12] the authors presented a deep learning model that allowed to detect covid-19 from healthy people using cxr images using recent deep learning methods. method and materials the proposed methodology for the covid_19 cxr image diagnosis system passes through the fundamental steps in machine learning data acquisition, image pre-processing, feature extraction, feature selection, and classification steps. the proposed method is shown in figure 1. data acquisition data acquisition was retrieved from an open dataset of covid-19 cases with ct/cxr. the database is already publicly available in github and it includes complete mailing addresses, telephone numbers, and e-mail addresses. this document represents the stage-two template.[8] this collaborative open data source allows radiologists and researchers to develop a machine learning model for covid-19. for this work, 110 digital radiographic images were used and equally distributed on normal and covid-19 cases. pre-processing in the pre-processing step, the data were prepared to maximize the diversity between the intensity of pixels and improve the accuracy of classification results.[13] here, at first, the images were resized to (576 × 576) to reduce the processing time and a gaussian filter was performed to minimize the noise; then, adaptive histogram equalization was applied to increase the contrast and image qualification improvement. feature extraction feature extraction plays an essential role in any classification task. medical images of different categories usually appeared by three matrices: object, edge, and background, which can be distinguished with their feature characteristics.[14] if the features are precisely selected, the classification result will provide helpful information and pattern to distinguish between the normal and covid-19 images. lbp is an effective set of texture descriptor that thresholds the intensity of neighboring pixels based on the value of the central pixel.[15] having a value equal or greater than the threshold pixel will assign the bit 1 and the remaining others whose values are smaller than the central pixel value will have bit 0. a bit vector is formed by restricting all these binary digits in a clockwise motion. the decimal point of the central pixel can be achieved by converting the binary sequences. mathematically, the lbp for every pixel is represented as: [16] lbr p r f g gp c p p p ( , ) ( )� � � � � 2 0 1 (1) where p denotes the total number of pixels surrounding the central pixel in radius r. furthermore, g p , and g c represent the intensity of central pixel and neighbor pixels, respectively. in the lbp feature vector, each central pixel in (x,y) image is surrounded by eight neighbors if the radius of circulation (r) equals one. at the same time, the central value is equal to the threshold value of any neighbor pixel. feature selection the role of feature selection is to simplify the model by making a vector with relevant features, shorter training time, and reducing the over fitting case because a real dataset contains statistically irrelevant features that may create a problem in classification tasks.[17,18] nowadays, the binary particle swarm optimization (bpso) metaheuristic algorithm is considered as an effective modern technique for feature selection with less running consumption figure 2.[19] it works based on a set of candidate solutions of randomly generated particles. the particles are moving around the search space to find the global best-optimized solution due to mathematical formula over both particle’s position x and velocity v. according to the used objective function, global best (g best ) and local best (l best ) of moving particles will be initiated in regards to the optimization problem. then, after mohammed, et al.: covid-19 chest x-ray images 48 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 46-51 each iteration, the particles try to update their positions and velocities according to the particle’s last destination and best moving, which the best candidate records until the searching process is completed.[20] the next position and velocity for each particle can be determined by: x x vid t id t id t� �� �1 1 (2) v w v c r p x c r p xid t id t id id t gd id t� � � � � �1 1 1 2 2* * ( ) * ( ) (3) where p id and p gd represent l best and g best , respectively, in the dth dimension. c1 and c2 are acceleration constants. r1 and r2 are random change values between 0 and 1. w is inertia weight to set up a balance between previous and current velocity, which refers to exploitation and exploration [table 1]. with defining the construction factor (k), the update of eq. (3) becomes: v k v c r p x c r p xid t id t id id t gd id t� � � � � �1 1 1 2 2* * ( ) * ( ) (4) where k � � � � 2 2 4 2� � and ϕ=c1+c2, c1+c2< 4 (5) under the equation’s balance, mathematically equation w from eq. (3) is equal. once the relevant features were selected, passed features were labeled and split into training and testing sets based on a 10-fold cross-validation strategy. class 0 refers to the normal images and class 1 determines the presence of covid-19 disease. classification image classification is an important step for observing whether the model can classify infected images from normal images or not. two classification methods svm and knn were performed figure 1: flow chart of proposed methodology mohammed, et al.: covid-19 chest x-ray images 49 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 46-51 table 1: binary particle swarm optimization algorithmic parameters for searching the best solution by swarms algorithm parameter name value bpso up (upper band) 10 bpso lb (lower band) −10 bpso cognitive component (c1) 2 bpso social component (c2) 2 bpso maximum inertia (wmax) 0.9 bpso minimum inertia (wmin) 0.4 bpso population size 10 bpso iteration 100 on training and testing datasets. svm classifier determines the best hyperplane to distinguish positive and negative training samples.[21] it can project the low dimensional input data in a higher dimensional feature by choosing the proper kernel function that separates cases of class labels linearly in multidimensional space.[22] knn captures the idea of similarity with some mathematics, we might have learned in calculating the distance between the points on the graph. the selection of a suitable value of distance (k) makes the model’s performance better.[23] the confusion matrix was used to better describe the gained classification models and understand which data classes are most often misplaced when it makes a prediction. each column of the matrix corresponds to a predicted classes and the row represents the actual classes. the number of incorrect and correct predictions is summarized with count values. the system scores perfectly when all the classified rates locate at the diagonal position of the matrix. if any data is misclassified, the values have deviated from diagonal cells.[24,25] finally, the performance of the model was calculated using following performance measurement equations: accuracy= (tp+tn)/(tp+tn+fp+fn) (6) sensitivity= tp/(tp+fn) (7) specificity= tn/(tn+fp) (8) precision= tp/(tp+fp) (9) results and discussion all the analysis was done in matlab 2016 software with intel core i5, 2.4 ghz, and 6 gb ram. the analysis of image preprocessing outcomes is shown in figures 3 and 4. eight neighbor points (p = 8) with central radius (r=1) are applied with uniform patterns; hence, the feature vector length reduces from 256 gray scale to 59 binary patterns. among 59 lbp extracted features, only 20 features were remained by bpso feature selection which is statistically significant in medical imaging and other irrelevant features were canceled from the feature vector. the best solution achieved by the objective function in bpso is 0.87 [figure 5]. in table 2, the performance measurements comparison is represented applying different functions (linear, polynomial, gaussian) for svm and (fine, cubic, weighted) knn. for passed features dataset, the svm (gaussian) classifier achieves the best accuracy 94.6%, sensitivity 96.2%, specificity 93%, and precision 92.7%. the performance measurements are calculated by applying eq. (6) to eq. (9). the confusion matrix of the proposed method is tabulated in table 3. in figure 6, the performance measurements comparison is represented applying different functions (linear, polynomial, and gaussian) for svm and (fine, cubic, weighted) knn. the passed features dataset, the svm (gaussian) classifier achieves the best accuracy 94.6%, sensitivity 96.2%, specificity 93%, and precision 92.7%. the performance measurements are calculated by applying eq. (6) to eq. (9). the confusion matrix of the proposed method is tabulated in table 2 below. figure 6 represents the bar graph to depict the comparison between the performance parameters of average specificity, table 2: confusion matrix representation gaussian support vector machine classification after ten-fold cross-validation actual case predicted case 51 4 55 2 53 55 53 47 110 figure 2: local binary pattern process over gray scale pixels with radius (r=1) and 8 neighbor pixels (a) observed pixel of gray scale image (b) binary encode between the center pixel and its neighbors (c) lbp string value and converted from binary to decimal a b c figure 3: results of affecting pre-processing filters on covid-19 cxr images (a) original resized image (b) image denoising using gaussian filter (c) image enhancement using adaptive histogram equalization b ca mohammed, et al.: covid-19 chest x-ray images 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 46-51 table 3: summarizes of classification results after calculation average accuracy, sensitivity, specificity, and precision from the confusion matrices classifiers type accuracy (%) sensitivity (%) specificity (%) precision (%) knn fine 90.9 86.9 95.9 96.4 cubic 84.5 83.9 93.2 94.5 weighted 93.6 92.9 94.4 94.5 svm linear 91.8 77.9 82.4 76.4 polynomial 92.7 94.3 82.4 94.5 gaussian 94.6 96.2 93 92.7 sensitivity, and precision values achieved by weighted knn and gaussian svm. for sake of a fair comparison between proposed work and the work presented in baity[26] this research did not use any feature selection algorithm whereas in the proposed work an intelligent feature selection strategy has been used which is based on a practical swarm optimization (bio-inspired technique), which is served as effective tool to filter out all irrelevant features before classification stage. in addition, two this work proposed two types of classifiers (svm and k-nn), whereas in baity[26] only one classifier (k-nn) had been suggested. eventually, the maximum accuracy attained is 94.6%for covid -19 virus detection which outperform the system best recognition accuracy attained in baity[26] which was only 93.9%. conclusion since a few days after spreading this outbreak, many cad methods for diagnosing covid-19 have been proposed. there is no doubt that early detection by imaging machines can survive the current rate of contagious patients. till now, many attempts are under progress to improve the performance measurements in data imaging for helping the doctors to diagnose that disease precisely. in this study for automatically classify 110 frontal cxr images according to normal, and covid-19 stages, lbp was used to extract the texture features from the images after pre-processing step. the clinically significant features are selected by bpso. finally, the proposed method is evaluated by performing knn and svm classifiers. the results show that the system robustness in covid-19 detection. our experimental results show that the proposed method can be used as a second purpose to help the radiologists for classifying the frontal cxr images either positive or negative covid-19. references 1. m. y. ng, e. y. p. lee, j. yang, f. yang, x. li, h. wang, m. m. s. lui, c. s. y. lo, b. leung, p. l. khong, c. k. m. hui, k. y. yuen, and m. d. kuo. imaging profile of the covid-19 infection: radiologic findings and literature review. radiology: cardiothoracic imaging, vol. 2, no. 1, p. e200034, 2020. 2. m. thomas, g. ksiazek, d, erdman, c. s. goldsmith, s. r. zaki, t. peret, s. emery, s. tong, and c. urbani. respiratory and enteric virus branch. united states: national center for immunization and respiratory diseases, 2003. 3. m. chan-yeung, and r. xu. sars: epidemiology. respirology, vol. 8, pp. s9-s14, 2003. 4. a. zumla, d. s. hui, and s. perlman. middle east respiratory syndrome. the lancet, vol. 386, no. 9997, pp. 995-1007, 2015. 5. a. j. rodriguez-morales, j. a. cardona-ospina, e. gutiérrezocampo, 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using adaptive histogram equalization b ca mohammed, et al.: covid-19 chest x-ray images 51 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 46-51 e. y. p. lee, e. y. f. wan, i. f. n. hung, t. p. w. lam, m. d. kuo, and m. y. ng. frequency and distribution of chest radiographic findings in covid-19 positive patients. radiology, vol. 296, no. 2, pp. 72-78, 2020. 7. l. fan, d. li, h. xue, l. zhang, z. liu, b. zhang, l. zhang, w. yang, b. xie, x. duan, x. hu, k. cheng, l. peng, n. yu, l. song, h. chen, x. sui, n. zheng, s. liu, and z. jin. progress and prospect on imaging diagnosis of covid-19. chinese journal of academic radiology, vol. 3, no. 1, pp. 4-13, 2020. 8. m. e. h. chowdhury, t. rahman, a. khandakar, r. mazhar, m. a. kadir, z. b. mahbub, k. r. islam, m. s. khan, a. iqbal, n. al-emadi, m. b. i. reaz, and t. i. islam. can ai help in screening viral and covid-19 pneumonia? arxiv, vol. 8, pp. 132665–132676, 2020. 9. a. abbas, m. m. abdelsamea, and m. m. gaber. classification of covid-19 in chest x-ray images using detrac deep convolutional neural network. applied intelligence, vol. 51, no. 2, pp. 854-864, 2021. 10. k. purohit, a. kesarwani, d. r. kisku, and m. dalui. covid-19 detection on chest x-ray and ct scan images using multi-image augmented deep learning model. biorxiv. vol. 1, p. 205567, 2020. 11. a. narin, c. kaya, and z. pamuk. automatic detection of coronavirus disease (covid-19) using x-ray images and deep convolutional neural networks. pattern analysis and applications, vol. 24, pp. 1207-1220, 2021. 12. k. el asnaoui, and y. chawki. using x-ray images and deep learning for automated detection of coronavirus disease. journal of biomolecular structure and dynamics, vol. 39, no. 10, pp. 36153626, 2020. 13. s. h. wady, r. z. yousif, and h. r. hasan. a novel intelligent system for brain tumor diagnosis based on a composite neutrosophic-slantlet transform domain for statistical texture feature extraction. biomed research 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statistical analysis and data mining applications. cambridge, massachusetts: academic press, 2017. 23. s. jang, y. e. jang, y. j. kim, and h. yu. input initialization for inversion of neural networks using k-nearest neighbor approach. information science, vol. 519, pp. 229-242, 2020. 24. e. el-din hemdan, m. a. shouman and m. e. karar. covidx-net: a framework of deep learning classifiers to diagnose covid-19 in x-ray images. arxiv, vol. 2003, p. 11055, 2020. 25. d. r. j. ramteke, and k. y. monali. automatic medical image classification and abnormality detection using k-nearest neighbour. international journal of advanced computer research, vol. 2, no 4, p. 6, 2012. 26. s. baity. covid-19 detection for chest x-ray images using local binary pattern. international journal of emerging trends in engineering research, vol. 8, pp. 78-81, 2020. tx_1~abs:at/add:tx_2~abs:at 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 6-8 research article series solution for single and system of non-linear volterra integral equations narmeen n. nadir* department of petroleum equipment, erbil technology institute, erbil polytechnic university, erbil, iraqi abstract in this paper, taylor series expansion is used for finding the approximate series solution of single and system of non-linear volterra integral equations of the second kind. the method allows us to overcome the difficulty caused by integrals and non-linearity; also, it has more precise and rapidly convergent to the exact solution. four examples are considered to show the accuracy and efficiency of the method keywords: integral equations, non-linear, taylor expansion, approximate solution introduction one of the most useful aspects of numerical analysis is to study approximate or/and numerical solution of mathematical equations, including differential and integral equations. the integral equation appears in many scientific problems, particularly, engineering problems such as fluid and solid mechanics, for example, potential flow theory, dynamics of population, and elasticity of structures.[1-3] at the end of the 18th century, the volterra integral equation (vie) was introduced by volterra, in which a variable appears at the upper boundary of the integrals, and it has first and second kind.[4] there are very few available techniques to handle the solution of integral equation in general, and non-linear integral equation in particular. therefore, we introduce approximation, because we aim to carry out some mathematical calculation involving these knowing functions such as performing integrations and differentiations. taylor’s expansion is a traditional and very important tools in the approximation theory, in which we can replace certain complicated functions by an approximated and simple polynomial.[5] taylor’s expansion[6] and its properties are used to find an approximated solution of different types of equations. some important, efficient, and classical numerical techniques for solving different equations are based on the taylor series expansion, for instance, euler method, modified euler method, and fourth-order runge–kutta method.[7-10] many works for finding a numerical solution of nonlinear vies have been proposed (see[11-13]). kanwel and liu[14] presented an algebraic technique based on the taylor expansion technique to find solution of linear integral equation, while saeed[5] used this expansion to treat system of linear problems, including volterra and fredholm integral equations and integrodifferential equations. in this study, we are concerned with the use of taylor series expansion to find an approximate series solution of the single and system of non-linear vies of the second kind, that is, u x v x k x t u t dt a x ( ) ( ) ( , , ( )) ,� � � (1) where v(x) is known, and it is continuous and differentiable on the given domain (a, b), and, u(x) is undetermined and required function, and k (x, t, u(t)) is called kernel function and continuous with respect to the three variables x, t and u(t) on the domain a≤x≤b, a≤t≤x. solution of equation (1) by taylor expansion expand u(x) in taylor series expansion at x=c, i.e. corresponding author: narmeen n. nadir, department of petroleum equipment, erbil technology institute, erbil polytechnic university, erbil, iraqi e-mail: narmeen1712@gmail.com received: jan 12, 2020 accepted: feb 16, 2020 published: jul 20, 2020 doi: 10.24086/cuesj.v4n2y2020.pp6-8 copyright © 2020 narmeen n. nadir. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) nadir: solution for single and system of non-linear volterra integral equations 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 6-8 u x i x a u ai i i ( ) ! ( ) ( )( )� � � � � 1 0 (2) hereafter, we will attempt to find u (a), u’ (a), u” (a). when we put x=a, in equation (1) we have u (a)=v (a). take the first derivative for equation (1) with respect to x using generalized leibnitz formula[9] and get the different equations such as first-order approximation which is known as euler’s method, second-order approximation which is called predictor-corrector or modified euler method and fourth-order approximation which is called fourth runge–kutta scheme.[7-10] in this paper, we were concerned to use the taylor series expansion (2) to find an approximate solution of the nonlinear vie of the second kind. therefore � � � � � ��u x v x k x t u t dt k x x u x a x ( ) ( ) ( , , ( )) ( , , ( )), (3) when we put x=a, in this equation we have � � � �u a v a k a a u a( ) ( ) ( , , ( )) . again, to find u” (a), differentiate equation (3) with respect to x and get �� � �� � �� � ��u x v x k x t u t dt k x x u x a x ( ) ( ) ( , , ( )) ( , , ( ))2 (4) again, when setting x=a, in equation (4) we get �� � �� � �u a v a k a a u a( ) ( ) ( , , ( ))2 , and so on we repeat this process n-time to obtain u a u a nk a a u an n n( ) ( ) ( )( ) ( ) ( , , ( ))� � �1 , n=1, 2,…. (5) putting the values of equation (5) for n=1, 2,… in the taylor series expansion (2) in the order, we can obtain the finite and approximate series solution of equation (1). numerical examples example 1 consider the problem u x x x e e u t dtt x t t a x ( ) ( )( ( ))( )� � � �� ��1 2 2 2 2 where the exact solution is u x ex( ) .= 2 solution: by using the procedure in section two, where a=0, we obtained u u u( ) , ( ) , ( ) ,( ) ( )0 1 0 2 0 122 4= = = u u( ) ( )( ) , ( ) ,6 80 120 0 1680= =  and u u u u( ) ( ) ( ) ( )( ) ( ) ( ) ( ) ...1 3 5 70 0 0 0 0= = = = = . using all these values in the taylor series expansion (2), we arrive u x x x x x x n n ( ) � � � � � � � � � � � � � � � 1 2 6 24 2 2 2 2 3 2 4 2  this equation is the nth partial sum of the maclaurin series of ex 2 . example 2 consider the non-linear vie u x u t t dt x ( ) ( ) � � �� 1 10 where the actual solution is u(x)=x and v(x)=0. solution: from equation (5), when x=0 we obtain u u u u u n( ) , ( ) , ( ) , ( ) ,..... ( )( ) ( ) ( )0 0 0 1 0 0 0 0 0 02 3� � � � � � using all these obtained values in the taylor series expansion in (2) to arrive, the exact solution u(x)=x. this approach also can be exploited for solving system of non-linear vie by using lebibntz generalized formula[15] which state that: d dx k x t u t dt x k x t u t dt k x b a x b x a x b x ( , , ( )) ( , , ( )) ( , ( ( ) ( ) ( ) ( ) � � � � � � xx u b x db x dx k x a x f a x da x dx ), ( ( ))) ( ) ( , ( ), ( ( ))) ( ) � (6) in this investigation a(x)=a is constant and b(x)=x, therefore (6) reduce to d dx k x t u t dt x k x t u t dt k x x a x b x b x a x ( , , ( )) ( , , ( )) ( , , ( ) ( ) ( ) ( ) � � � � � � uu x( )) (7) we can use this principle to convert initial or boundary value problem into an integral equation. for instance, suppose we the following initial value problem �� � �u x k x u x u x( ) ( , ( ), ( )) (8) with u(a)=u 0 and u’(a)=u 1 are given. take integral for both side of (8), we get �� � � � � � � � �u t dt k t u t u t dt t a t x t a t x ( ) ( , ( ), ( )) (9) use principle of (7) for both side of (9), we obtain � � � � � � �u x u k t u t u t dt t a t x ( ) ( , ( ), ( ))1 (10) take integral again for both side of (10), we can rewrite (10) as, � �� � � �� ��u t u dt k z u z u z dzdx a x a z a x ( ) ( , ( ), ( ))1 (11) u x u a u x a x a k z u z u z dz a x ( ) ( ) ( ) ( ) ( , ( ), ( ))� � � � � ��1 nadir: solution for single and system of non-linear volterra integral equations 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 6-8 thus, we get, u x u a u x a x a k z u z u z dz v x x a k t a x ( ) ( ) ( ) ( ) ( , ( ), ( )) ( ) ( ) ( , � � � � � � � � � �1 uu t u t dt a x ( ), ( ))�� which is non-linear of the second kind and v(x)= v(a)+ u 1 (x−a). in addition, equation (7) can be used for each equation in the system of non-linear vies as illustrated in the following two examples. example 3[16] consider the following system u x e x t u t dt u x xe e t e x x x x x u 1 2 0 2 2 2 0 2 1 4 1 4 2 1 1 ( ) ( ) ( ) ( ) � � � � � � � � � � �� � (( )t dt where actual solution u x x1 1 2 ( ) � � and u x ex2( ) .= u 1 (0)=1 and u 2 (0)=1 � � � � � � �� � � � � � � � � u x e x x t u t dt x x u x u x e x x x 1 2 2 2 0 2 2 2 1 2 ( ) ( ) ( ) ( ) ( ) ( ) �� � � � � � � �� � ��xe e x t e dt x e x x x u t u x2 0 2 21 1( ) ( ) so, � � � � �u u1 20 1 2 0 1( ) ( )and �� � �� �u u1 20 0 0 1( ) ( ) .and by continuing in this process, we can the exact solution. example 4 consider the following system u x x x u t u t dt u x x x u t x x 1 1 2 0 2 2 2 1 2 0 3 ( ) sec( ) ( ) ( ) ( ) tan( ) ( ) � � � � � � � � � �� u t dt22( ) with exact solution u 1 (x)=sec(x) and u 2 (x)=tan(x). u 1 (0)=1 and u 2 (0)=1 � � � � � � �� � � ��u x x x x u t u t dt u x u x x 1 1 2 2 2 0 1 2 2 21( ) sec( ) tan( ) ( ) ( ) ( ) ( )) ( ) sec ( ) ( ) ( ) ( ) ( � � � � � � � � �� � � ��u x x x u t u t dt u x u x x 2 2 1 2 2 2 0 1 2 2 23 1 ))� � so, � � � �u u1 20 0 0 1( ) ( )and by the same manner, we can get the exact solution. conclusion taylor series expansion was applied successfully to find an approximate or exact series solution of single and system of non-linear vies in the second kind. the method was tested by taking four numerical examples and good results were obtained. we also conclude that the solution obtained by taylor series expansion is given by a function and not only at some points and numerical computations of this method are simple and the convergence is satisfactory. references 1. j. l. hess. review of integral-equation techniques for solving potential-flow problems with emphasis on the surface-source method. computer methods in applied mechanics and engineering, vol. 5, no. 2, pp. 145-196, 1975. 2. s. a. edalatpanah and e. abdolmaleki. a new collocation method for systems of nonlinear fredholm integral equations. applied mathematics and physics, vol. 2, no. 1, pp. 15-18, 2014. 3. h. ibrahim, f. attah, g. t. gyegwe. on the solution of volterrafredholm and mixed volterra-fredholm integral equations using the new iterative method. applied mathematics, vol. 6, no. 1, pp. 1-5, 2016. 4. a. m. wazwaz. linear and nonlinear integral equations. berlin: springer, 2011. 5. r. k. saeed. computational methods for solving system of linear volterra integral and integro-differential equations (doctoral dissertation, ph. d. thesis, university of salahaddin/ erbil-collage of science), 2006. 6. p. j. davis. interpolation and approximation. new york: dover publication, inc., 1975. 7. k. e. atkinson. an introduction to numerical analysis. 2nd ed. hoboken, new jersey: john wiley and sons, 2008. 8. s. c. chapra, r. p. canale. numerical methods for engineers. 4th ed. boston: mcgraw-hill higher education, 2010. 9. e. w. cheney, d. r. kincaid. numerical mathematics and computing. monterey county: cengage learning, brooks/cole publishing company, 2012. 10. g. hall, j. m. watt. modern numerical methods for ordinary differential equations. oxford: oxford university press, 1976. 11. c. t. baker, g. f. miller. treatment of integral equations by numerical methods: based on the proceedings of a symposium held in durham from 19-29 july, 1982, organized under auspices of the london mathematical society. london: academic press, 1982. 12. l. m. delves, j. walsh. numerical solution of integral equations. oxford, england: claredon press oxford, 1974. 13. o. a. faour. solving non-linear integral equations using (b2-spline) function. journal of university of babylon for pure and applied sciences, vol. 7, no. 3, pp. 372-377, 2002. 14. r. p. kanwal, k. c. liu. a taylor expansion approach for solving integral equations. international journal of mathematical education in science and technology, vol. 20, no. 3, pp. 411-414, 1989. 15. a. j. jerry. introduction to integral equations with applications, a series of monographs and textbooks. new york: marcel dekker, inc., 1985. 16. a. m. wazwaz. the modified decomposition method for analytic treatment of non-linear integral equations and systems of nonlinear integral equations. international journal of computer mathematics, vol. 82, no. 9, pp. 1107-1115, 2005. tx_1~abs:at/add:tx_2~abs:at 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 1-5 research article occupational exposures and associated health effects among dumpsite workers: a case study of kani qirzhala landfill site, erbil, iraq tanya s. salih, ameena s. m. juma, muhsin h. ubeid department of general biology, cihan university-erbil, kurdistan region, iraq abstract eighty-nine male workers from the landfill site in (kani qirzhala), erbil, iraq, aged from 12 to 65 years were investigated in this study. each one had filled out a questionnaire sheet, in addition to their signed consent to take a blood sample from them. plus, a 7  ml of venous blood samples were collected from them. total ige and syphilis antibodies were detected. thus, cbc was conducted on each sample. the results recorded 92.13% of the workers lived in rural areas, while 7.87% lived in urban areas. further, there were 58.43% smokers and 41.57% non-smokers. furthermore, the research sample contained 38.20% single and 61.80% married people. as education illustrated, 37.08% had no schooling, primary school education 40.45%, less than 20% had secondary school education, 2.25% acquired a diploma, and a b.sc. degree. the serum concentration of total ige of dumpsite workers revealed a significant increase when compared to the healthy group. none of the workers’ sera revealed syphilis antibodies except one case which was positive; however, it shows non-significant differences between both groups. wbc count was soared significantly in dumpsite workers when compared to the healthy individuals, yet, lymphocyte and granulocyte numbers showed non-significant increment, while monocyte numbers showed a statistically insignificant rise in workers as compared to healthy group. the number of rbcs and hb level of the landfill workers exhibited a substantial increase. even though, both groups’ platelets did not show significant variance. the rise in wbc counts and ige levels may be due to the exposure of these workers to allergens at the dumpsites as for allergies is the common consequence when exposed to waste and garbage. keywords: ige and allergy, dumpsite workers, complete blood count, syphilis and hematological factors introduction wastes management using open landfill sites is considered a primitive practice and an inadequate service that is still used in many parts of the globe, in addition to the absence of efficacy and scientific concepts. these sites commonly are located at the brinks of towns. such dumpsites are considered health and environmental hazard,[1] they will affect and infect not only the contacted workers but they also will influence the communities who live in the surrounding adjacent areas.[2,3] as such, those people who work and live in a dirty atmosphere are at risk of illness as of their surroundings. they have a high possibility to be caught by many microbial diseases and exposure to many chemical substances causing them, namely, irritation (allergies) of skin, nose, and eyes, in addition to chest and stomach pains, headaches, and diarrhea.[4-7] in fact, the study had isolated various of pathological species of bacteria from the site’s surroundings; top soils, leachates, and the study group for instance; bacillus, staphylococcus, escherichia coli, proteus, streptococcus, klebsiella, pseudomonas, citrobacter, bacteroides, clostridium, and serratia. all these bacteria are known to cause serious respiratory, skin, gastrointestinal, and even central nervous system infections.[8,9] further, dumpsites are known to emit air pollutants, that is, bioaerosols, which are hazardous to those who work in landfill sites and put them at risk of respiratory diseases. admittedly, cough was the most reported symptom by the investigated literature.[10] furthermore, surveys of community corresponding author: tanya s. salih, department of general biology, cihan universityerbil, kurdistan region, iraq. e-mail: tanya.salam@cihanuniversity.edu.iq received: may 31, 2021 accepted: july 28, 2021 published: august 10, 2021 doi: 10.24086/cuesj.v5n2y2021.pp1-5 copyright © 2021 tanya s. salih, ameena s. m. juma, muhsin h. ubeid. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) salih, et al.: occupational exposures and associated health effects among dumpsite workers 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 1-5 health from africa had documented a vast of well-being issues: psychological disorders, gastrointestinal problems, respiratory symptoms, skin inflammation besides to nose, and eyes.[2] based on the aforementioned statements, and the scarcity of information, this study is subjected to shed the light on the health status of the local landfill workers. because of their continuous exposure to toxic materials and allergens, hence, the investigation on the immune status of the mentioned workers was carried out. research methods location and groups selection from kani qirzhala, erbil, iraq, dumpsite eighty-nine males were investigated for this study in hand. age, gender, and apparently healthy subjects had selected as criteria for healthy group. figure 1 illustrates the questionnaire that was filled by both groups. in addition, they were subjected to all tests mentioned in the next section. sample collection to collect 7 ml of venous blood from each worker, a standard clean venipuncture technique was administered by disinfecting the anti-cubital fossa with 70% ethanol and using a disposable hypodermic syringe and 23-gauge needle after tourniquet application. afterward, 2 ml of venous blood was dispensed; into dipotassium ethylenediaminetetraacetic anticoagulant tube, and evenly stirred, hence, to facilitate the establishment of the hematological criteria (cbc). the remaining, 5 ml were placed in a plain tube and left aside for 60 min at room temperature for blood to clot. then, the tubes were in a centrifuge for 10 min. afterward, tubes cooled at 4°c at 450× g to collect the serum. later, using a pasteur pipette, the serum was aspirated and dispensed into 0.5 ml sterile glass tubes and frozen at –20℃. antibody detection once they are restored to room temperature, the stored sera were used for the seroprevalence tests. the automated cobas table 1: kani qirzhala landfill site workers’ information characteristics frequency percent live in rural areas 82 92.13 live in urban areas 7 7.87 smoker 52 58.43 non-smoker 37 41.57 single 34 38.20 married 55 61.80 illiterate 33 37.08 primary school education 36 40.45 secondary school education 16 17.97 diploma degree 2 2.25 b.sc. degree 2 2.25 figure 1: the study’s questionnaire sheet to investigate worker’s antibodies in the sera, hematological, and other parameters e411 immunoassay analyzer facilitated the total ige and syphilis antibodies detection. hematology the hematology tests were achieved after 1 h of blood samples collection using an automated hematological analyzer coulter machine (mythic 18 hematology analyzer, orphee, switzerland) for 20 parameters. results workers’ data this research investigated 89 males who were 12–65 years old (no females were found in that site). no significant difference (p> 0.05) was found in the age intervals regarding all results. the majority 92.13% lived in rural areas, while urban areas consisted only of 7.87%. results demonstrated that there were 58.43% smokers and 41.57% non-smokers. further, it showed that most of them were married 61.80%, and only 38.20% were single. more into this, the education level illustrated, 37.08% had no schooling, primary school education 40.45%, 17.97% had secondary school education, 2.25% acquired a diploma, and a b.sc. degree [table 1]; these workers supported their families financially by going into this occupation. concentration of total ige table 2 reveals a high and significant increase the serum concentration of total ige of the dumpsite workers when compared to the healthy group. frequency of syphilis antibodies the results show a non-significant difference between both groups, as none of the collected sera demonstrated the availability of syphilis antibodies except one case was positive [table 3]. hematological parameters table 4 and figure 2 indicate that wbc count increased significantly (p< 0.01) in dumpsite workers in comparison salih, et al.: occupational exposures and associated health effects among dumpsite workers 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 1-5 table 2: total ige concentration in the sera of the dumpsite workers compared with healthy individuals immunological characteristics gender dumpsite workers healthy group t-test p-value p-value no. mean±se no. mean±se ige ng/ml m 89 255.62±51.47 40 98.40±9.04 0.003 h.s.** p>0.05: non-significant; *p<0.05: significant; **p<0.01: highly significant table 3: syphilis antibodies in the sera of dumpsite workers compared with healthy individuals immunological parameter gender dumpsite workers healthy group t-test p-value p-value no. mean±se no. mean±se syphilis ng/ml m 89 0.313±0.019 40 0.296±0.004 0.399 n.s. p>0.05: non-significant; *p<0.05: significant; **p<0.01: highly significant table 4: hematological parameters in the sera of dumpsite workers in comparison to healthy individuals immunological and hematological parameters gender dumpsite workers healthy group t-test p-value p-value no. mean±se no. mean±se wbc (103/gl) m 89 7.97±0.62 40 6.19±0.21 0.009 h.s.** lymphocyte (103/gl) m 89 2.70±0.22 40 2.20±0.14 0.058 n.s. granulocyte (103/gl) m 89 4.51±0.31 40 4.05±0.18 0.209 n.s. monocyte (103/^l) m 89 0.77±0.11 40 0.47±0.04 0.014 s.* rbc (106/^l) m 89 5.27±0.07 40 4.95±0.09 0.006 h.s.** hb (g/ml) m 89 16.06±0.24 40 13.86±0.18 0.000 h.s.** platelet (103/^l) m 89 225.49±13.85 40 242.97±4.74 0.235 n.s. p>0.05: non-significant; *p<0.05: significant; **p<0.01: highly significant figure 2: hematological parameters in the sera of dumpsite workers in comparison to healthy individuals to the healthy individuals, while lymphocyte and granulocyte numbers non-significantly increased (p> 0.05), and monocyte number shows a slightly significant rise (p< 0.05) in workers as compared to healthy individuals. the table illustrates also, significant increment in the number of rbc’s and hb level as contrasted to the control group. further, it signifies no substantial variance among platelet of groups. discussion at present, more than 15 million individuals in economically poor countries endure by retrieving domestic waste. shockingly, children are a big category among those people. they wade through households’ garbage or factories, collecting and selling materials of recycling context to earn their income.[11] many of the enrolled individuals in this current study were children. pollution of garbage emissions is one of the reasons for many microbial diseases.a wide variety of diseases and sickness symptoms have been diagnosed in dumpsite workers including cholera, typhoid, malaria, skin diseases gastrointestinal, stomach pain, vomiting, diarrhea, and respiratory allergies. it is should be mentioned that none of the individuals included in this study showed any symptoms of diseases, despite the fact that most of them looked malnourished. insects and specific mosquitoes can also spread various diseases in the area.[4-7,12,13] whereas, the international solid waste association reported a variety of microbial diseases that can be caught from garbage including bacterial, viral, fungal, and parasitic, in addition to various allergic infections.[14] in general, degraded living conditions have a direct relationship with the immunological status of a person, which makes them more liable to infections.[15] landfill wastes are identified of spreading very highrisk diseases. moreover, the major types are the medical and biowastes.[12] in addition, the dumpsite dwellers are likely to inhale poisonous gases, that is, methane, ammonia, carbon monoxide, and hydrogen sulfide. moreover, many other medical conditions have been diagnosed in these workers including cardiovascular degeneration, disorders such as osteoarthritic changes and intervertebral disc herniation, problems of skin and respiratory system, and altered pulmonary function parameters.[16] salih, et al.: occupational exposures and associated health effects among dumpsite workers 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 1-5 the research revealed high levels of ige and wbcs in the dumpsite workers. this could indicate allergic reaction to the substance(s) that these workers exposed to. it is of interest to emphasize on the fact that none of the workers were on any medication, including anti-allergic medications. allergic reactions associated with hypersensitivity to substances that are characterized by excessive activation of certain white blood cells named as mast cells and basophil granulocyte by ige.[17] for instance, hay fever triggers the nose, sneezing, itching, and eyes redness irritation.[18] inhaled allergens can generate asthmatic indications, causing the lung’s airways to narrow, mucus production, dyspnea, as well as to wheezing and coughing.[19] further studies have stated that those who work for long hours in compost are vulnerable to acute and (sub-) chronic non-immune or type iii allergic inflammation.[20] in lebanon, through observation, it was assured the escalation tendency in the acute health symptoms among dumpsite people, such as respiratory, gastrointestinal, constitutional, and dermatological.[21] values of increased wbcs may refer to their involvement in immunological responses as they tend to engage in a descending form of instances resulting in inflammatory and certain sickness conditions, that is, hypersensitivity, immunosuppression, immunomodulation, and autoregression on exposure to allergens or environmental contamination.[22] despite the distinct case of syphilis, which have documented in this research, still it is necessary to it put into consideration. this agrees, where evidence indicated that if threats of biological, chemical, and physical exceeded their limitations, work-related diseases may emerge. many workers, and even retired persons, have been found to have many illnesses including ulcers, bronchitis, in addition to hypertension, heart diseases, arthritis, and diabetes.[23] multitude studies reported that waste collectors risk their health and safety when exposed to significant concentration of biological aerosols,[20,24-26] namely, respiratory and gastrointestinal complaints,[6,27-30] infectious diseases such as hepatitis (a, b, and c), and hiv, in addition to syphilis.[31-33] it is worthy to mention that there have been some studies in the kurdistan region regarding the dumpsites. in sulaymaniyah, poor management and lack of waste treatment have resulted in the severe contamination of soil, water, and air of the surrounding area. besides, urban waste, oil refineries, and cement plants nearby unload production residues into the city dump. around 75% of medical waste from local hospitals is also disposed in the same place. soil and water samples taken in proximity of the landfill showed concentrations of iron (fe), manganese (mn), cadmium (cd), nickel (ni), cobalt (co), and chromium (cr) above permissible levels.[34] life on kani qirzhala has been described elaborately and how it is a family affair and lives of families depend on this occupation.[35] the concern about the protection of the environment has largely increased due to the contaminants mainly with anthropogenic origin and has affected and continues to threaten human resources, especially air quality, surface soil, and groundwater in erbil city where the government is working hard to prevent any health issues, especially with the rapid development of the city.[36] conclusion municipal landfills areas and the workers establish a major public health hazard to themselves, families, in addition to nearby communities. because of relationship between dumpsites as a source of income, and the landfill site administration, the workers’ health is at risk. thus, education of these workers on how to protect themselves and their families is essential. references 1. f. p. sankoh, x. yan and q. tran. environmental and health impact of solid waste disposal in developing cities:a case study of granville brook dumpsite, freetown, sierra leone. journal of environmental protection, vol. 4, no. 7, pp. 665-670, 2013. 2. s. abul. environmental and health impact of solid waste disposal at mangwaneni dumpsite in manzini: swaziland. the journal of sustainable development in africa, vol. 12, no. 7, pp. 64-78, 2010. 3. n. p. thanh, y. matsui and t. fujiwara. assessment of plastic waste generation and its potential recycling of household solid waste in can tho city, vietnam. environmental monitoring and assessment, vol. 175, no. 1, pp. 23-35, 2011. 4. h. issever, h. gul, m. erelel, f. erkan and g. y. gungor. health problems of garbage collectors in istanbul. indoor and built environment, vol. 11, no. 5, pp. 293-301, 2002. 5. p. o. njoku, j. n. edokpayi and j. o. odiyo. health and environmental risks of residents living close to a landfill:a case study of thohoyandou landfill, limpopo province, south africa. international journal of environmental research and public health, vol. 16, no.12, p. 2125, 2019. 6. m. r. ray, s. roychoudhury, g. mukherjee, s. roy and t. lahiri. respiratory and general health impairments of workers employed in a municipal solid waste disposal at an open landfill site in delhi. international journal of hygiene and environmental health, vol. 208, no. 4, pp. 255-262, 2005. 7. r. maheshwari, s. gupta and k. das. impact of landfill waste on health: an overview. iosr journal of environmental science, toxicology and food technology, vol. 1, no.4, pp. 17-23, 2015. 8. f. oviasogie and d agbonlahor. the burden, antibiogram and pathogenicity of bacteria found in municipal waste dumpsites and on waste site workers in benin city. the journal of medicine and biomedical research, vol. 12, no. 2, pp. 115-130, 2013. 9. f. e. oviasogie, c. u. ajuzie and u. g. ighodaro. bacterial analysis of soil from waste dumpsite. archives of applied science research, vol. 2, no. 5, pp. 161-167, 2010. 10. g. f. akpeimeh. bioaerosol emission from msw open dumpsites and the impact on exposure and associated health risks. leeds, england: university of leeds, 2019. 11. m. medina. solid wastes, poverty and the environment in developing country cities: challenges and opportunities. wider working paper, 2010. 12. a. jorgensen. diseases caused by improper waste disposal. geneva: world health organization, 2007. 13. a. s. juma, m. h. ubeid and t. s. salih. anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers of erbil governorate. cihan university-erbil scientific journal, vol. 3, no. 1, pp. 85-89, 2019. 14. a. mavropoulos and d. newman. wasted health the tragic case of dumpsites. vienna: international solid waste association, 2015. 15. t. s. salih, s. s. haydar, m. h. ubeid and a. s. juma. some immunological and hematological parameters among refugees in kawergosk camp-erbil governorate. cihan universityerbil scientific journal, vol. 3, no. 1, pp. 80-84, 2019. 16. r. r. tiwari. occupational health hazards in sewage and sanitary salih, et al.: occupational exposures and associated health effects among dumpsite workers 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 1-5 workers. indian journal of occupational and environmental medicine, vol. 12, no. 3, p. 112, 2008. 17. d. ogundele, v. olayemi, f. folaranmi, o. oludele and a. oladejo. haematological, liver function parameters and heavy metal assessment of kwmc workers at roadside and mixed waste dumpsites in kwara state, nigeria. journal of applied sciences and environmental management, vol. 23, no. 7, pp. 1213-1219, 2019. 18. e. t. bope and r. e. rakel. conn’s current therapy. philadelphia, pa: wb saunders, 2002. 19. d. t. win. chemical allergies/chemical sensitivities. au journal of technologies, vol. 12, no. 4, pp. 245-250, 2009. 20. j. douwes, h. dubbeld, l. van zwieten, i. wouters, g. doekes, d. heederik and p. steerenberg, work related acute and (sub-) chronic airways inflammation assessed by nasal lavage in compost workers. the annals of agricultural and environmental medicine, vol. 4, no. 1, pp. 149-152, 1997. 21. r. z. morsi, r. safa, s. f. baroud, c. n. fawaz, j. i. farha, f. el-jardali and m. chaaya. the protracted waste crisis and physical health of workers in beirut:a comparative cross-sectional study. environmental health, vol. 16, no. 1, p. 39, 2017. 22. b. majkowska-wojciechowska and m. l. kowalski. allergens, air pollutants and immune system function in the era of global warming. in: air pollution-monitoring, modelling, health and control rijeka. london: intech, pp. 221-254, 2012. 23. j. a. cimino and r. mamtani. occupational hazards for new york city sanitation workers. environmental health, vol. 1, no. 1, pp. 8-12, 1987. 24. j. lavoie, c. j. dunkerley, t. kosatsky and a. dufresne. exposure to aerosolized bacteria and fungi among collectors of commercial, mixed residential, recyclable and compostable waste. science of the total environment, vol. 370, no. 1, pp. 23-28, 2006. 25. i. m. wouters, s. spaan, j. douwes, g. doekes and d. heederik. overview of personal occupational exposure levels to inhalable dust, endotoxin, p (1^3)-glucan and fungal extracellular polysaccharides in the waste management chain. annals of occupational hygiene, vol. 50, no. 1, pp. 39-53, 2006. 26. s. widmeier, a. bernard, a. tschopp, s. jeggli, x. dumont, s. hilfiker, a. oppliger and p. hotz. surfactant protein a, exposure to endotoxin, and asthma in garbage collectors and in wastewater workers. inhalation toxicology, vol. 19, no. 4, pp. 351-360, 2007. 27. c. y. yang, w. t. chang, h. y. chuang, s. s. tsai, t. n. wu and f. c. sung. adverse health effects among household waste collectors in taiwan. environmental research, vol. 85, no. 3, pp. 195-199, 2001. 28. k. kheldal, a. halstensen, j. thorn, w. eduard and t. halstensen. airway inflammation in waste handlers exposed to bioaerosols assessed by induced sputum. european respiratory journal, vol. 21, no. 4, pp. 641-645, 2003. 29. g. de meer, d. heederik and i. m. wouters. change in airway responsiveness over a workweek in organic waste loaders. international archives of occupational and environmental health, vol. 80, no. 7, pp. 649-652, 2007. 30. m. r. ray, s. roychoudhury, s. mukherjee s, s. siddique, m. banerjee, a. b. akolkar, b. sengupta and t. lahiri. airway inflammation and upregulation of p2 mac-1 integrin expression on circulating leukocytes of female ragpickers in india. journal of occupational health, vol. 51, no. 3, 2009, pp. 232-238. 31. g. dounias and g. rachiotis. prevalence of hepatitis a virus infection among municipal solid-waste workers. international journal of clinical practice, vol. 60, no. 11, 2006, pp. 1432-1436. 32. m. a. rozman, i. s. alves, m. a. porto, p. o. gomes, n. m. ribeiro, l. a. nogueira, m. m, caseiro, v. a. da silva, e. massad and m. n. burattini. hiv and related infections in a sample of recyclable waste collectors of brazil. international journal of std and aids. vol. 18, no. 9, 2007, pp. 653-654. 33. p. luksamijarulkul, d. sujirarat and p. charupoonphol. risk behaviors, occupational risk and seroprevalence of hepatitis b and a infection among public cleansing workers of bangkok metropolis. hepatitis monthly, vol. 8, no. 1, pp. 35-40, 2008. 34. a. tinti. available from: https://www.ejatlas.org/conflict/dumpsite-in-sulaymaniyah-kurdistan-region-of-iraq. [last accessed 2021 may 20]. 35. e. fitt. life on the kani qirzhala landfill site in iraq, 2017. available from: https://www.recyclingwasteworld.co.uk/ in-depth-article/life-on-the-kani-qirzhala-landfill-site-iniraq/164658. [last accessed 2021 may 20]. 36. s. q. gardi. environmental impact assessment of erbil dumpsite area west of erbil city-iraqi kurdistan region, 2018 available from: https://www.researchgate.net/ publication/324262622environmentalimpactassessmentof_ erbil_dumpsite_area_-_west_of_erbil_city-iraqi_kurdistan_ region. [last accessed 2021 april 01]. tx_1~abs:at/add:tx_2~abs:at 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 research article the prevalence of microorganisms and antibiotics activity tests of natural and industrial products in surgical site infections in referral hospitals in erbil city, kurdistan region – iraq raid d. thanoon1*, yara m. mohammed2 1department of medical biochemical analysis, cihan university-erbil, kurdistan region, iraq, 2department of general biology, cihan university-erbil, kurdistan region, iraq abstract background and objectives: to study the prevalence of microorganisms and antibiotics activity tests of natural and industrial products, surgical site infection was found in referral hospitals in erbil, iraq’s kurdistan province. methods: pus samples were collected from 29 surgical patients using sterile swabs and primed for culture collection. disk diffusion method was used to determine the bacterial activity of three types of plants (cinnamon, ginger, and radish) against three bacterial species including staphylococcus aureus, proteus spp., and staphylococcus epidermidis. results: overall, 29 pus samples were processed for culture testing. one gram-negative bacterium (7.14%) and 13 gram-positive cocci (92.85%) were isolated from the 14 bacteria genera. s. aureus, proteus spp., and s. epidermidis most potent isolates were found to be resistant to the variable of tested antibiotics, while they showed any sensitivity to the tested plant extracts. cinnamon, ginger, and radish extracts were discovered to be among the few variable effective plant extracts against the tested pathogenic bacteria isolates. conclusion: the data collected from this research showed a better way of explanation of the microorganism’s etiology of ssis in hospitals where they might have epidemiological and healing insinuations. keywords: surgical site infections, bacteria isolated from wounds, antimicrobial resistance, pus, plant extract introduction surgical site infection occurs when surgical wounds get infected with germs from a previously diseased location. according to the report, it can result in illnesses, hospitalizations, and even death.[1] the third most often observed nosocomial infection is surgical site infections.[2] the most prevalent causes of these contagions are exogenous and endogenous germs that reach the operating wounds during surgery.[2] the occurrence of infected operational injuries might be affected by such factors as pre-operative therapy, theater, post-operative care, and surgery.[2] staphylococcusaureus is a naturally occurring bacterium that colonizes about 30% of the human population’s nares and a new study indicates that very diverse nasal microbiosis is required if colonizations of s. aureus are to be encouraged or prevented. it spreads easily from person to person, onto health care workers’ hands and clothing, as well as onto surfaces and into the air.[2,3] staphylococcus epidermidis,once considered a comparatively harmless pathogen, is the biggest and best investigated member of the cons group.[4,5] it is considered as an unsafe commensal of the human skin and is becoming known for its useful role in skin immunity and microbiota.[4] the active metabolites of cinnamon bark (cinnamomum verum) belonging to the lauraceae family were cinnamon aldehyde and trans-cinnamic acid. the nutritional content, phenolic profile, and antioxidant activities of cinnamon were studied. according to a compositional study, this species is a great source of protein and minerals. the nutritional content, phenolic profile, and antioxidant activities of cinnamon were studied. according to a compositional study, this species is a great source of protein and minerals.[6] radish (raphanus sativus l.), belonging to the brassicaceae family, is an annual or biennial root crop globally of economic importance. there are many resources of radish germplasm in china, and for conservation and use of the radish resources, cultivars and genetic diversity studies are very important.[7] since antiquity, ginger (zingiber officinale) belonging to the zingiberaceae corresponding author: raid d. thanoon, department of medical biochemical analysis, cihan university-erbil, kurdistan region, iraq. e-mail: raid.duraid@ cihanuniversity.edu.iq received: may 30, 2021 accepted: july 18, 2021 published: august 10, 2021 doi: 10.24086/cuesj.v5n2y2021. pp 6-12 copyright © 2021 raid d. thanoon, yara m. mohammed. this is an openaccess article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) thanoon and mohammed: antibiotics activity tests of natural and industrial products in surgical site infections 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 family for more than 200 years, traditional chinese medicine has been utilized as a spice and medication. ginger is a major herb with numerous nutritional and therapeutic properties that are utilized in traditional asian and chinese medicine.[8] ginger and its general components, such as fe, ca, vitamin c, flavonoids, phenolic compounds, paradols, and so on, are used for a wide variety of symptoms, including vomiting, pain, and other herbal medications.[8] objectives 1. isolation of microorganisms from surgical wounds 2. natural and industrial products were tested for antibiotic activity. materials and procedures population research the research was carried out at cihan university-erbil’s biology department. specimens were collected from four hospitals between november 2019 and march 2020. specimen collection and transport under aseptic conditions, two sterile swabs were used to extract pus samples from each patient. swabs were taken to the department of biology, cihan university-erbil. the specimens can be stored in the sample tube at 4°c for 72 h. types of antimicrobials identification the bacterial growth of the incubated plates was identified on the basis of cultural, morphological, and biochemical characteristics (table 1).[9] production of specimens inoculated specimens of pus have been immediately inoculated on macconkey, chocolate, and mannitol salt agar dishes. they were incubated aerobically at 37°c for 24–48 h with a co2 concentration of 7–10%. after this, the specimens was inoculated on the plates using four flame methods.[10,11] procedure of catalase testing fill a slide with a small amount of bacterial growth. drops of h2o2 were added to the smear and blended with a toothpick.[12] gram staining overflow slide with mineral violet – 60 s by gram – 180 s, iodine floods. prudently decolor with 95% ethanol until the thinnest part of the smear is colorless. flood with safranin (pink color) (10% fuchsine) – 60 s (wash on the water). air dehydrated or spot by permeable paper.[13] slide coagulase test identify where the test strain (t) and the control (c) are placed by marking the slide. set up positive and negative control organisms on the same slide for simultaneous testing. emulsified test strain to obtain a homogeneous thick suspension. observed self-agglutination. within 10 s, there will be a visible cluster. negative result: no visible clustering within 10 s.[14] testing for antimicrobial sensitivity drugs have been chosen on the basis of their impact on a specific organism, as well as hospital antibiotic policies.[20] the diameter of the inhibitor was measured on a scale of 24 h after 18–24 h of incubation. each antimicrobial disk’s area size was used to determine if it was susceptible, intermediate, or immune (clsi, 2006). mueller-hinton agar and antibiotic disks were used.[15] ginger extraction ginger was bought from local marketplaces in erbil city, using ginger for this investigation. in boiling water bath for 30 min, 10 g of ginger powder combined with 40 ml soy oil was heated. before disk diffusion testing, extracts were kept at 4°c to retain their antibacterial activities. the plates were dried in the air flow chamber of the laminar for 15 min. extracts of ginger (20 μl) have been applied individually for 18–24 h on each disk.[16] radish extraction a local supermarket provided raphanus sativus (white radish variety). radish was lyophilized and crushed into a fine powder before ethanol extraction. powder root was extracted for 24 h with 300 ml of ethanol at room temperature and 150 rpm.a filter paper disk diffusion test was used to assess the extract’s antibacterial function.[17] the research bacteria inoculums were prepared according to mcfarland standard 0.5. uniform bacterial lawns were developed, using 100l of inoculum on a mueller-hinton agar plate. the plates were incubated at 37°c for 24 h.[18] cinnamon extraction after cleaning the spices, they were dried in sunlight for 2 days. the dried material was pulverized by a grinder into a table 1: antimicrobials and their concentrations no. name dose manufactures 1 amikacin 10 mcg amikin 2 amoxicillin 25 mcg trimox 3 ampicillin/cloxacillin 25/5 mcg ampoxin 4 ceftazidime 30 mcg ceptaz 5 ceftriaxone 10 mcg rocephin 6 chloramphenicol 10 mcg fenicol 7 ciprofloxacin 10 mcg ciloxan 8 gentamicin 10 mcg gentasol 9 imipenem 10 mcg primaxin 10 netilmicin 30 mcg netmicin 11 oxacillin 10 mcg bactocill 12 rifampin 5 mcg rifadin 13 tetracycline 10 mcg declomycin figure 1: the prevalence rate of surgical site wound infections figure 2: number of isolated bacteria according to gram-positive and gram-negative bacteria figure 4: staphylococcus aureus organism control thanoon and mohammed: antibiotics activity tests of natural and industrial products in surgical site infections 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 fine powdered substance. twenty grams of cinnamon powder (weighted by electric balance) were transferred to 100 ml of conical flask where 40 ml of ethanol was added. the ethanol crude was filtered using whatman no. 1 filter paper. cinnamon bark extract was made using dimethyl sulfoxide as a solvent at concentrations of 2.5%, 5%, and 10%. with a sterile cotton bud, the bacterial solution was spread on the whole surface of mueller-hinton agar. at 37°c, the petri dish was incubated overnight.[19] results specimens of surgical wound infection a total of 29 pus specimens were processed for culture testing. the present study being conducted between november (2019) and march (2020). the overall percentage of infection was (48.27%) while the remaining (51.72%) was negative [figure 1]. two patients from the maternity hospital were cultivated with macconkey agar and the mannitol salt agar had a negative result and a positive result of s. aureus bacteria and specimens were taken from females. ten patients from the cardiac center were sampled, two males and eight females, and nine of the 10 were negative on macconkey agar, with proteus spp. bacteria being the only positive result. eight patients from sardam hospital, two specimens from females and six specimens from males, five of whom were cultivated with mannitol salt agar, and all of whom had a positive result were s. aureus bacteria and three of them cultivated with chocolate agar, had a positive result, two of whom were s. aureus bacteria, and one of them was s. epidermidis bacteria. nine patients from jamhuri hospital, specimens taken from males, cultivated mannitol salt agar, five of whom were negative, and four of whom were positive, were s. aureus and s. epidermidis bacteria [table 2]. out of the 14 isolated bacteria, 1 was gram-negative bacteria (7.14%) and 13 were gram-positive cocci (92.85%). s. aureus was the most common bacteria 10 isolates (71.42%). other isolated bacteria were proteus sp. (7.14%) and s. epidermidis (21.42%) [figure 2]. antimicrobial activity of plant extracts andantibacterial susceptible of s. aureus sensitivity pattern for s. aureus to the following antibiotics; rf (20 mm), te (16 mm), net (27 mm), and the other antibiotics resistance for s. aureus such as c, ox, cro, ipm, caz, apx, and cro [figure 3a and b]. the result presented in figure 4 was s. aureus control bacteria. each extract was tested against s. aureus isolated. the results showed cinnamon resistance to antibacterial activity against s. aureus [figure 5]. antimicrobial activity of plant extracts and antibacterial susceptible of s. epidermidis antibacterial resistant and susceptible of s. epidermidis isolated from patients. the results show that s. epidermidis isolated from patients were susceptible to net (34 mm) and apx (28 mm), and other antibiotic resistant for s. epidermidis such as te, c, and cro [figure 6]. the result presented in figure 7 exhibited s. epidermidis control bacteria. the results given in figure 8 show that ginger, cinnamon, and radish root extracts are not effective against s. epidermidis. figure 3: (a and b) antibacterial inhibition zoneagainst staphylococcus aureus ba figure 5: inhibition zone of plants against staphylococcus aureus figure 6: antibacterial inhibition zone against staphylococcus epidermidis figure 7: staphylococcus epidermidis bacteria control thanoon and mohammed: antibiotics activity tests of natural and industrial products in surgical site infections 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 antimicrobial activity of plant extracts and antimicrobial susceptible of proteus spp. the results show that proteus spp. isolated patients were susceptible to net (17 mm), rf (19 mm), ak (25 mm), te (26 mm), and cn (20 mm). moreover, other antimicrobial is resistant to proteus spp. ox, apx, caz, and ax [figure 9a-c]. results given in figure 10 showed that ginger and radish root extract are not effective against proteus species. the results showed that cinnamon had very little (5 mm) effect on proteus species [figure 10]. discussion surgical site infection is a prevalent kind of disease related to health-care research conducted by the way.[20-22] ssis table 2: the patients number, gender, isolated bacteria, media cultured from the surgical site of infection, and the name of the hospitals serial no. of patient media isolated bacteria hospitals sex 1 macconkey agar negative (–) cardiac center ♀ female 2 macconkey agar negative (–) cardiac center ♀ female 4 macconkey agar negative (–) cardiac center ♂ male 5 macconkey agar positive (+) proteus sp. cardiac center ♀ female 6 macconkey agar negative (–) maternity hospital ♀ female 7 macconkey agar negative (–) cardiac center ♀ female 8 macconkey agar negative (–) cardiac center ♀ female 9 macconkey agar negative (–) cardiac center ♀ female 10 macconkey agar negative (–) cardiac center ♀ male 11 macconkey agar negative (–) cardiac center ♀ female 12 mannitol salt agar positive (+) s. aureus maternity hospital ♀ female 13 mannitol salt agar positive (+) s. aureus sardam hospital ♀ female 14 mannitol salt agar positive (+) s. aureus sardam hospital ♂ male 15 mannitol salt agar positive (+) s. aureus sardam hospital ♂ male 16 mannitol salt agar positive (+) s. aureus sardam hospital ♂ male 17 mannitol salt agar positive (+) s. aureus sardam hospital ♂ male 18 chocolate agar positive (+) s. aureus sardam hospital ♂ male 19 chocolate agar positive (+) s. aureus sardam hospital ♀ female 20 chocolate agar positive (+) s. epidermidis sardam hospital ♂ male 21 mannitol salt agar positive (+) s. aureus jamhuri hospital ♂ male 22 mannitol salt agar positive (+) s. epidermidis jamhuri hospital ♂ male 23 mannitol salt agar negative (–) jamhuri hospital ♂ male 24 mannitol salt agar negative (–) jamhuri hospital ♂ male 25 mannitol salt agar positive (+) s. aureus jamhuri hospital ♂ male 26 mannitol salt agar positive (+) s. epidermidis jamhuri hospital ♂ male 27 mannitol salt agar negative (–) jamhuri hospital ♂ male 28 mannitol salt agar negative (–) jamhuri hospital ♂ male 29 mannitol salt agar negative (–) jamhuri hospital ♂ male s. aureus: staphylococcus aureus, s. epidermidis: staphylococcus epidermidis figure 10: antimicrobial disk diffusion test against proteus spp. figure 8: antimicrobial disk diffusion test against staphylococcus epidermidis thanoon and mohammed: antibiotics activity tests of natural and industrial products in surgical site infections 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 (infections at surgical site) were found to be 48.27 percent prevalent in the present specimens, with s. aureus and s. epidermidis being the most common etiological agents among gram-positive bacteria. gram-positive bacteria while the gram-negative bacteria were proteus spp. respectively.[23] these findings support that previous research s. aureus was the most common bacteria found in post-operative wound infections.[24,25] s. aureus was found in 71.42% of surgical site infections. the most common gram-positive s. aureus isolate (71.42%) followed by s. epidermidis (21.42%) and proteus spp. (7.14 percent of the total). in the current study, a shorter postoperative stay was seen in patients with acquired infection, who had a roughly triple longer patient without operational site infection. in hospitals, neonatal infections, operative care for the elderly and malnutrition, and individuals with diabetes and other chronic diseases, s. aureus is an important source of infection.[26] the minimal number of s. aureus sensitivity found in the results of this investigation against te and c was compatible with studies published in eritrea by certain nigerian employees.[27,28] sensitivity to rf, te, net, and the other antibiotics resistance for s. aureus such as c, ox, cro, ipm, caz, apx, and cro did not match the findings of chowdhury et al.,[29] however, resistance to other antibiotics did. caz and cip were consistent with the report published by the commission.[29] moreover, s. aureus was sensitive to te and resistance to cro and c, which is not in agreement with the report published by the commission.[29] pattern of antibiotic resistance of s. epidermidis isolated from hospital infectious specimens s. epidermidis resistance against: c, cro, and te were consistent with reports published by chabi and momtaz,[30] wijesooriya et al.,[31] and s. epidermidis sensitive to apx and net.[32] results show that proteus spp. isolated from patients were susceptible to net, rf, ak, te, and cn and resistant to proteus spp.;ox, apx, caz, and ax.[33] current results showed that cinnamon, radish, and ginger were resistant to s. aureus. when opposed to the previous findings,[18,34] the present findings show a higher level of activity. raphanus extract had the best activity against a multiresistant strain of p. aeruginosa, while p. mirabilis was the most resistant to the extract’s influence.[35] piper nigrum and zingiber officinale are active in s. aureus and k. pneumoniae. cinnamon essential oil shows maximum activity against e. coli and s. aureus.[36] the results show that ginger extract and root radish extract are not effective against s. epidermidis.[37] the current results showed that cinnamon and ginger extract are not effective against proteus species. the different concentrations of radish root extract had no effect on proteus spp.[18] the antibacterial action of cinnamon was ascribed to the existence or longterm storage of specific active components which have been declined by melting and exposure to air.[38] the negative result of cultured pus specimens may not have been effective due to the transfer of the swabs, and the bacteria may have been dead. it could be the species of bacteria that may not grow in these media. they need special media or anaerobic bacteria could not grow aerobically in the 37°c incubator. conclusion the findings of this research will aid in our understanding of the microbiological etiology of ssis in our hospitals, which could have epidemiological and therapeutic implications. s. aureus was the most common surgical site infection we found (71.42%). this study demonstrates that rf, te, and net are sensitive to s. aureus, and net and apx are sensitive to s. epidermidis and net, rf, te, ak, and cn are affective against proteus spp. and c, ox, cip, ipm, caz, apx, and cro figure 9: (a-c) antimicrobial inhibition zone on proteus spp. c ba thanoon and mohammed: antibiotics activity tests of natural and industrial products in surgical site infections 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 are resistant to s. aureus. te, c, and cro are resistant to s. epidermidis. ox, apx, caz, and ax are resistant against proteus spp. plant extracts have no antimicrobial activity against s. aureus, s. epidermidis, or proteus ssp., which typically cause surgical wound infections, according to the findings of this report. the findings showed that cinnamon, radish root, and ginger are ineffective at inhibiting the growth of wound pathogenic bacteria. recommendation the findings of this study can be used to launch an effort to strengthen hospital antimicrobial policy and antimicrobial prescribing guidelines. it is difficult to choose the best drug for any condition, particularly in infectious diseases. since it is performed in vitro, the susceptibility pattern cannot be used as the sole criterion because it ignores the patient’s immunological status and clinical condition. this research may be useful in determining which plant extracts to use for further isolation of constituents responsible for the studied species’ behavior. acknowledgments i’d want to offer my heartfelt gratitude to dr. raid duriad thanoon, the research supervisor, for assisting me through each step of my research paper it would have been impossible to write such a significant and fascinating work without his help. and a special thanks to ahmed majeed mohammed, who inspired us to achieve many goals during this research. many thanks to biolaboratory for their help and guidance. finally, many thanks to all the other people and teachers for their help and advice. references 1. t. anggrahita, a. wardhana and g. sudjatmiko. chlorhexidinealcohol versus povidone-iodine as preoperative skin preparation to prevent surgical site infection:a meta-analysis. medical journal of indonesia, vol. 26, no. 1, pp. 54-61, 2017. 2. k. s. bhaskarrao, r. vaishali and v. j. katkar. study of bacteriological profile of post-operative wound infections in obstetrics and gynaecological surgeries in a tertiary care hospital. national journal of integrated research in medicine, vol. 9, no. 2, pp. 77-83, 2018. 3. b. krismer, c. weidenmaier, a. zipperer and a. peschel. the commensal lifestyle of staphylococcus aureus and its interactions with the nasal microbiota. nature reviews 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in clean, clean-contaminated and contaminated cases. journal of medical science and clinical research, vol. 4, no. 12, pp. 14981-14986, 2016. 21. s. shrestha, r. shrestha, b. shrestha and a. dongol. incidence and risk factors of surgical site infections in kathmandu university hospital, kavre, nepal. kathmandu university medical journal, vol. 14, no. 54, pp. 107-111, 2016. 22. j. segreti, j. parvizi, e. berbari, p. ricks and s. i. berríos-torres. introduction to the centers for disease control and prevention and healthcare infection control practices advisory committee guideline for prevention of surgical site infection: prosthetic joint arthroplasty section. surgical infections, vol. 18, no. 4, pp. 394400, 2017. 23. h. guan, w. dong, y. lu, m. jiang, d. zhang, y. aobuliaximu, j. thanoon and mohammed: antibiotics activity tests of natural and industrial products in surgical site infections 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 6-12 dong, y. niu, y. liu, b. guan, j. tang and s. lu. distribution and antibiotic resistance patterns of pathogenic bacteria in patients with chronic cutaneous wounds in china. frontiers in medicine, vol. 8, p. 609584, 2021. 24. p. neopane, h. p. nepal, r. shrestha, o. uehara and y. abiko. in vitro biofilm formation by staphylococcus aureus isolated from wounds of hospital-admitted patients and their association with antimicrobial resistance. international journal of general medicine, vol. 11, pp. 25-32, 2018. 25. x. zhu, x. sun, y. zeng, w. feng, j. li, j. zeng and y. zeng. can nasal staphylococcus aureus screening and decolonization prior to elective total joint arthroplasty reduce surgical site and prosthesis-related infections? a systematic review and metaanalysis. journal of orthopaedic surgery and research, vol. 15, no. 1, p. 60, 2020. 26. b. c. g. slingerland, m. c. vos, w. bras, r. f. kornelisse, d. de coninck, a. van belkum, i. k. m. reiss, w. h. f. goessens, c. h. w. klaassen and n. j. verkaik. whole-genome sequencing to explore nosocomial transmission and virulence in neonatal methicillin-susceptible staphylococcus aureus bacteremia. antimicrobial resistance and infection control, vol. 9, no. 1, p. 39, 2020. 27. l. ying, x. zhe, y. zhou, s. juan and m. lin. characterization of community-associated staphylococcus aureus from skin and softtissue infections:a multicenter study in china. emerging microbes and infections, vol. 5, no. 1, pp. 1-11, 2016. 28. l. radlinski, s. e. rowe, l. b. kartchner, r. maile, b. a. cairns, n. p. vitko, c. j. gode, a. m. lachiewicz, m. c. wolfgang and b. p. conlon.pseudomonas aeruginosa exoproducts determine antibiotic efficacy against staphylococcus aureus. plos biology, vol. 15, no. 11, p. e2003981, 2017. 29. m. h. chowdhury, t. debnath, s. bhowmik and t. islam. presence of multidrug-resistant bacteria on mobile phones of healthcare workers accelerates the spread of nosocomial infection and regarded as a threat to public health in bangladesh. journal of microscopy and ultrastructure, vol. 6, no. 3, p. 165, 2018. 30. r. chabi and h. momtaz. virulence factors and antibiotic resistance properties of the staphylococcus epidermidis strains isolated from hospital infections in ahvaz, iran. tropical medicine and health, vol. 47, no. 1, p. 47, 2019. 31. w. r. p. wijesooriya, d. n. kotsanas, t. m. korman and m. graham. teicoplanin non-susceptible coagulase-negative staphylococci in a large australian healthcare network: implications for treatment with vancomycin. sri lankan journal of infectious diseases, vol. 7, no. 1, p. 10, 2017. 32. m. a. mohammed, a. m. j. abd al-abbas and a. m. alsamak. distribution of oata alleles detected by a new designed primer in bacteria isolated from eye infections in basrah governorate/iraq. annals of the romanian society for cell biology, vol. 25, no. 3, pp. 8258-8277, 2021. 33. d. girlich, r. a. bonnin, l. dortet and t. naas. genetics of acquired antibiotic resistance genes in proteus spp. frontiers in microbiology, vol. 11, p. 256, 2020. 34. s. haq, s. dildar, m. b. ali, a. mezni and a. hedfi. antimicrobial and antioxidant properties of biosynthesized of nio nanoparticles using raphanus sativus (r. sativus) extract. materials research express, vol. 8, no. 5, p. 055006, 2021. 35. ç. güceyü, g. goncagül, e. günaydin and p. akpinar. zencefil’in antibakteriyal etkisi (antibacterial effect of zingiber officinale (ginger)). etlik veteriner mikrobiyoloji dergisi, vol. 30, no. 1, pp. 44-50, 2019. 36. y. zhang, x. liu, y. wang, p. jiang and s. quek. antibacterial activity and mechanism of cinnamon essential oil against escherichia coli and staphylococcus aureus. food control, vol. 59, pp. 282-289, 2016. 37. e. julianti, k. k. rajah and i. fidrianny. antibacterial activity of ethanolic extract of cinnamon bark, honey, and their combination effects against acne-causing bacteria. scientia pharmaceutica, vol. 85, no. 2, p. 19, 2017. 38. k. i. al-zubaidy. cinnamon’s antibacterial activity on the bacterial isolates from urinary tract infections. world journal of pharmaceutical research, vol. 6, no. 17, pp. 81-91, 2017. . 85 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 research article a multiband biconical log-periodic antenna for swarm communications bessam al jewad* department of computer and communication engineering, cihan university-erbil, kurdistan region, iraq abstract in this paper, we present a specific multiband antenna design that addresses the problem of communication with unmanned aerial vehicles (uavs). we consider a scenario where multiband single-antenna uav communicates with the rest of the swarm members that are equipped with similar antennas. the key point in the design is that the communication does not require high or low elevation angles in most of the cases. the suggested design has a sufficient degree of freedom to select the desired features for the field pattern while keeping other features such as antenna impedance and gain relatively stable or at least in the acceptable operation region. keywords: biconical antenna, broadband antenna, component, drone-to-drone, drone-to-infrastructure, frequency-independent antennas, log-periodic antenna introduction in recent years, the use of unmanned aerial vehicles (uavs), also known as drones, has witnessed an unprecedented increase for both military and civilian applications. from airborne traffic surveillance to the more recent mail package delivery, the list of interesting applications for uav swarms keeps growing.[1-3] however, the introduction of the swarm concept, communications have always been a challenge.[4,5] in uav swarms, there are two types of communications: droneto-drone (d2d) in which drones can communicate important navigational information between each other, and drone-toinfrastructure (d2i) in which drones can communicate and take general commands from the ground station (gs). while the former can be relatively simple, the latter is much more complicated and challenging for many reasons. first, in most applications, the uav is equipped with a high-resolution camera sending images and video. therefore, each uav requires a high-speed low-latency communication link in the order of tens of mbps of bandwidth and few tens of milliseconds of latency.[6] that is, of course if such images and videos are to be used for any reliable form of navigation or interaction. second due to the dynamic nature of the uav movement in three-dimensional at speeds ranging from 20 m/s to 50 m/s,[6] the communication channel with the gs becomes time varying with non-stationary noise. it is further hampered by the constant loss of the line-of-sight and the change in antenna characteristics (radiation pattern, polarization, etc.) as well as orientation. currently, existing wireless technologies, such as wireless fidelity, zigbee, and xbee-pro as well as massive multiple-input multiple-output (mimo) and ultra-wideband communication were devised for communication with uavs.[6] however, none of these techniques was designed for this particular application making them highly unreliable for many practical situations.[7] in recent artificial intelligence ai studies, it was suggested that navigational information and low-latency interaction or decision-making between the uav and gs can be minimized if some form of ai algorithm like swarm guidance algorithm was implemented locally on each uav causing the group to have a massive collective processing power.[8] in such an application, the general heading command can be sent by the gs and the swarm can follow a stochastic general path in which each uav can communicate the obstacles, constraints and locally-optimized path solution to the rest of the group.[9] collectively, the group will perform a distributed cost-function minimization and will find the optimal path that minimizes the time and energy for the swarm.[10] in such an application, the design of a special low-cost and ergonomically efficient antenna for d2d communication is highly desirable. at a basic level, a drone swarm is a floating dynamic wireless network, commonly known as a wireless mesh corresponding author: bessam al jewad, cihan university-erbil, kurdistan region, iraq. e-mail: bassam.jawad@cihanuniversity.edu.iq.test-google-a.com received: apr 17, 2019 accepted: apr 19, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp85-91 copyright © 2019 bessam al jewad. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) al jewad: a multiband biconical log-periodic antenna for swarm communications 86 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 network. a mesh network is a network topology in which each node relays data for the network. all nodes cooperate in the distribution of data in the network. a mesh network whose nodes are all connected to each other is a fully connected network. many studies suggested the use of multi-carrier, ultra-wideband or multi-band communication. however, one of the limiting factors for such an approach is the antenna. many conventional antennas are highly resonant and only operate over a bandwidth that represents a few percent’s of their center frequency. these narrow bandwidth antennas may be entirely satisfactory or even desirable for single-frequency or narrowband applications where high gain is required. wider bandwidths are required in d2d communication applications, especially as the level of the swarm intelligence and its processing power increase. the more sophisticated the applied ai algorithm gets, the more it becomes necessary to exchange high bandwidth or latency dependent data between swarm members. conventionally, there has been a broad range of work on wideband antennas[11-13] for various airborne vehicle applications. however, d2d rarely will need to communicate at high (or low) elevation angles, especially while flying in a swarm. therefore, a wideband, omnidirectional antenna, with low beam-width is proposed as a solution for such an application. this antenna needs to be effectively small to maintain the weight constraints of the drone design. in addition, it needs to be a broadband antenna to facilitate exchanging high-bandwidth data. in this paper, we present a specific antenna design that addresses the problem of multi-band communications and has a higher degree of freedom in its design to select the desired features of the field pattern while keeping other features such as antenna impedance and gain relatively stable or at least within the acceptable operation region. wideband frequency independent antenna the proposed design is a biconical log-periodic spiral antenna whose conceptual image is shown in figure 1. the goal is to create a broadband antenna with center frequencies of 3.9 and 5.9 ghz and a frequency range of 3.4–10 ghz. the objective radiation pattern is a one with high gain at the horizon and less gain toward the sky or ground. the reason why an array antenna would not work is that arrays are typically narrow band. conventionally, several broadband antennas were utilized in the past. however, biconical log-periodic spiral antennas have an advantage in that they can operate in two main modes: the end-fire axial mode with circular polarization and the broadside normal mode with linear polarization. thus, depending on the operating frequency, they can serve the dual purpose of d2d and d2i communications. this design draws from a range of common antennas including dipoles, arrays, spirals, loops, log-periodic, and conical. the aspects of these antennas and the amount they are used in the design are dependent on the frequency range of operation. in this particular design, the cones are actually present just to support the wire.[14] however, a multi-layered ferrite material can be used inside these cones to adjust the center frequency and change the polarization. as will be shown later each part of a loop will have a different center frequency, and since ferrite material is narrow band, in general, a multi-layer stack of ferrite materials can be utilized depending on the desired characteristics from the antenna. the material around the core can be filled with dielectrics to enhance the capacitive effect of the antenna impedance to adjust the performance. in general, a broadband antenna is defined by two factors: percentage of center frequency and ratio of operation. the percentage of the center frequency is defined by − − = × = ×100% 100% * u l u l p cl u f f f f f ff f (1a) while the frequency ratio of operation is defined as the ratio of the upper and lower frequencies of operation f u and f l . = u l f f f (1b) if the impedance and the pattern do not change significantly over about an octave (𝐹=2) or more, we classify the antenna as a broadband antenna. to meet the frequency independence requirements in a finite structure, the current must attenuate along with the structure and be negligible at the point of the truncation. for radiation and attenuation to occur the charge must be accelerated (or decelerated) and this happens when the conductor is curved or bent normally to the direction, in which the charge is traveling. thus, the curvature of the spiral provides a frequency-independent operation over a wide bandwidth.[12] broadly speaking, each arm of this antenna can be designed independently and placed strategically to provide a set of electrically small loop antenna array elements spaced by half wavelength, and excited at the center. starting from one arm the logarithmic (log) spiral is defined in cylindrical coordinates by r=αθ or ln(r)=θ ln(α) and is shown in detail in figure 2. the radial distance to a point p on the spiral is 𝑟, the angle with respect to the x-axis is 𝜃, and 𝛼 is a constant representing the apex angle. the relationship between 𝜃 and 𝑟 can alternatively be given by θ=tan(β) ln (r). in both relations, the z is not specified. an important control for a log-periodic spiral angle is the relationship for the spacing between each loop.[15] to achieve the broadband nature, the ratio between the 𝑖th loop spacing 𝑠 and the subsequent loop must stay constant. the log-periodic loop array seems a reasonable approximation to the spirally wrapped wire in our design. the loop diameters increase along with the antenna so that the included angle α is a constant and the spacings s of adjacent elements are scaled. this can also be translated in terms of the height of the loop z, which gives: + + − − = = − 1 1 1 i i i i i i s z z k s z z (2) where k is some constant. from this equation, it can be easily seen that z follows a simple difference equation. since the arrangement is logarithmic, it is more convenient to let z=mθ. now to find the height of each loop z i we will assume that completes one full rotation for each discrete loop and is therefore an integer i times 2π. thus, the height of the al jewad: a multiband biconical log-periodic antenna for swarm communications 87 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 ith loop can be written as z i = mi. therefore, z i+1 =m(i+1)=mz i in the same way z i+1 =m2 z i-1 . substituting this in (2) and solving for m, we find that m=k for a non-trivial solution and therefore z=kθ. it is fairly reasonable to assume that only a single loop (or part of the loop) from the spiral will contribute to the major farfield component and will have the major current distribution according to the frequency of interest. this assumption was validated by several simulations. in this sense, the diameter for any element n+1 is approximately kn greater than the first element, or + = ≈1 1 nnd f k d (3) where 𝐹 is the frequency ratio from equation (1); the smallest loop in the antenna has a circumference equal to a tenth of a wavelength to quantify for the small loop antenna definition which gives the constraint for the diameter of the first loop. therefore, the geometric representation for one arm of the biconical antenna is given by q q q= =/ tan    , r e z k (4) equations (3) and (4) constitute the design equations for one arm of the antenna. each antenna arm can be placed in either a frequency selective biconical construct shown in figure 1 or stacked in a broadband uni-conical construct. multi-band antenna matching in the previous section, the antenna was designed to meet several broadband requirements. in general, most selfcomplementary broadband antennas have almost constant impedance. the problem with most wire antennas (like our design) is that the antenna impedance is highly frequency dependent. thus, even though the antenna arm in the previous section may have broadband properties, it can only be used with a single frequency dictated by the matching network. the alternative would be to utilize a broadband matching network and in this case, the matching would not be perfect or easy. in addition, broadband matching in general means a compromise on the antenna gain due to the constant gain-bandwidth product constraint. using the design equations of the previous section, the parameters selected for our design are shown in table 1. the simulation results for the antenna impedance assuming a wire gauge of awg33 are shown in figure 3 for an input port impedance of 50 ohm. the approach that was adopted in our matching is the dual-band impedance matching where the antenna is matched at 3.9 ghz and 5.9 ghz. the same approach can be expanded to match any set of frequencies within the band. in this section, the approach is introduced. analyzing the actual antenna response on the smith chart (from a vector network analyzer) is also performed. the antenna’s frequency response (that is, the reflection coefficient γ) is plotted as a function of frequency on the smith chart, as shown in figure 4. the blue curve in this figure 4 represents the antenna impedance, plotted on the smith chart continuously from 3.4 ghz to 6.2 ghz since the rest of the frequency response is not required for the matching. the black circles indicate the locations on the curve corresponding to 3.9 ghz–5.9 ghz. the corresponding vswr plot for the same impedance above is given in figure 5. the target is to bring both of the above impedance locations to the center of the smith chart with a single matching network. to do this, the low band is matched first. for this step, use is made of only series capacitors and parallel inductors. this is because series capacitors affect lower figure 1: proposed antenna concept table 1: design parameters for the initial antenna matching circuit parameters values circumference ≤λ/10 low frequency 3.481 ghz large frequency 10 ghz β 1.54 radians κ 1.235 radius of wire 0.0899 mm number of turns 5 figure 2: log-periodic loop array structure al jewad: a multiband biconical log-periodic antenna for swarm communications 88 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 frequencies more than higher frequencies. similarly, parallel inductors affect low frequencies more than high frequencies. as a result, if only those components are used, the low band can be matched with minimal effect on the high band. the matching is done by calculating the required series and parallel impedances by moving on a constant x circle on the immittance smith chart. for the lower band matching a series, capacitor will move the impedance to intersect the re[y]=1 circle. by inspection, we find that a 0.738 pf series capacitor will move the 3.9 ghz impedance to intersect the re[y]=1 circle. from there, we complete the low band match with a parallel inductor of approximately a 0.692 nh. the resulting impedance curve for the group (the antenna, the 0.738 pf series capacitor and then the parallel 0.692 nh inductor) is shown in figure 6. from this, we see that the low band (3.9 ghz) is moved to the center of the smith chart, i.e., we have impedance matched in this region. however, the high band (5.9 ghz) region has moved from its original location, even though that was not intentional. the next step is to try to match the 5.9 ghz point without undoing the 3.9 ghz match. this can be done with the use of parallel capacitors and series inductors (both of which affect high frequencies more than low frequencies). to this end, we want to swing the high band point in figure 5 into the center of the smith chart. using the concepts of the parallel capacitor, we can move the 5.9 ghz impedance to intersect the re[z]=1 circle through a 0.483 pf parallel capacitor. note that by doing so, the 3.9 ghz point is moved slightly away from the center but that can be tolerated. the high band is now set up to be matched with a series inductor. the required value can be found to be approximately 1.61 nh of series inductance. the addition of this component to the overall matching network produces the final impedance curve, as shown in figure 7. figure 3: un-matched antenna impedance figure 4: antenna reflection coefficient frequency response figure 5: original vswr for the antenna al jewad: a multiband biconical log-periodic antenna for swarm communications 89 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 from figure 7, the high and low bands may not appear exactly matched. however, figure 8 presents the vswr corresponding to the impedance plot of the above smith chart. at 3.9 ghz and at 5.9 ghz, the vswr is <2.0. this is typically considered a good match. if a better match was desired, the four component values could be further optimized to improve the response. the trick to optimize for the component values is to assume that each value has a correction factor of δ. from the location of each desired point, we can determine the sign and value of each δ. the key factor to remember if lumped elements are to be used is the parasitic of each element. the final step in our matching is to convert the lumped elements found into short-embedded transmission lines, as shown in figure 9. the reason for this step is that lumped passive elements rarely behave as pure elements at very high frequencies. for example, a simple inductor will be hampered by stray capacitance and by wire resistance converting it into a more complicated circuit element, and the same applies to capacitors. in addition, the values we found for the matching network are not very practical. thus, the initial lumped-element values found were converted to distributed elements by utilizing the theory of embedded short transmission line.[16] in general, a shortembedded transmission line can approximate a shunt capacitance or a series inductance. as long as the length of the transmission line is kept very small compared to the minimum wavelength of interest, the implementation is possible. other examples exist in[16] for more complicated matching networks, and the selection between different possible matching networks depends entirely on the practicality of the element dimensions. figure 6: matching of the lower frequency band figure 7: matching of the upper frequency band figure 8: vswr for the matched antenna figure 9: short-embedded transmission line implementation of the matching network al jewad: a multiband biconical log-periodic antenna for swarm communications 90 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 the resulting overall matching network was implemented on a rogers 4350b substrate, as shown in figure 10 for the entire frequency band. simulation results the resulting antenna design from the previous section for a biconical construct is shown in figure 11. this design had been simulated with ansoft hfss. the separation between the two conic sections was selected to give resonance at the 3.9 ghz–5.9 ghz. the 𝑆 11 or reflection coefficient is shown in figure 12 and the radiation pattern for the 5.9 ghz is shown in figure 13 for 5.9 ghz. the reflection coefficient response is <−10 db over the entire band of interest and the center frequencies of 3.9 ghz–5.9 ghz have very apparent resonant peaks. this is due to the wideband nature of the spiral antennas. as was devised previously, only part of each loop contributed to the radiation pattern in each frequency. the rest of the stacked loops aided in increasing the broadside gain to 5 dbi by working as a reflector and helping to limit the beam width at high and low elevation angles at the center frequency by working as a guide. conclusion this paper studied a broadband biconical log-periodic antenna design for uav communications. a geometrical analysis of the design is presented along with an hfss simulation. the results show that this antenna has good performance at the proposed frequency range with center frequencies at 3.9 ghz and 5.9 ghz. the results show that this antenna can achieve a gain of 5 dbi. further work will focus on increasing the gain and miniaturizing the antenna size using ferrite stacks and proper dielectric materials. references 1. s. hayat, e. yanmaz and r. muzaffar. “survey on unmanned aerial vehicle networks for civil applications: a communications viewpoint”. ieee communications surveys and tutorials, vol. 18, no. 4, pp. 2624-2661, 2016. 2. t. andre, k. a. hummel, a. p. schoellig, e. yanmaz, m. asadpour, c. bettstetter, p. grippa, h. hellwagner, s. sand and s. zhang. “application-driven design of aerial communication networks”. ieee communications magazine, vol. 52, no. 5, pp. 129-137, 2014. 3. j. abatti. “small power: the role of micro and small uavs in the future”. air command and staff college, maxwell afb, al, 2005. figure 11: the biconical log-periodic spiral antenna (dimensions are in mm) figure 10: actual implementation of the matching circuit figure 12: simulated reflection coefficient or s 11 (db) figure 13: radiation pattern of antenna (dbi) al jewad: a multiband biconical log-periodic antenna for swarm communications 91 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 85-91 4. p. olsson, j. kvarnstrom, p. doherty, o. burdakov and k. holmberg. “generating uav communication networks for monitoring and surveillance”. in: proceeding international conference on control automation robotics vision, pp. 1070-1077, 2010. 5. e. yanmaz, r. kuschnig and c. bettstetter. “achieving air-ground communications in 802.11 networks with three-dimensional aerial mobility”. in: proceeding ieee infocom, pp. 120-124, 2013. 6. p. chandhar, d. danev and e. g. larsso. “massive mimo for communications with drone swarms”. ieee transactions on wireless communications, vol. 17, no. 3, pp. 1604-1629, 2018. 7. a. pospischil and j. p. rohrer. “multihop routing of telemetry data in drone swarms”. in: proceeding of itc, 2017. 8. m. v. ramesh, p. l. divya, r. v. kulkarni and r. manoj. “a swarm intelligence based distributed localization technique for wireless sensor network”. proceeding international conference on advances in computing, communications and informatics, pp. 367-373, 2012. 9. l. rosenberg. “artificial swarm intelligence vs human experts”. international joint conference on neural networks, 2016. 10. c. papagianni, k. papadopoulos, c. pappas and n. d. tselikas. “communication network design using particle swarm optimization”. proceeding international multiconference on computer science and information technology, vol. 3, pp. 915920, 2008. 11. v. rumsey. “frequency independent antennas”. ire international convention record, vol. 5, pp. 114-118, 1957. 12. j. d. kraus. “helical beam antennas for wide-band applications”. proceeding of the ire, vol. 36, no. 10, pp. 1236-1242, 1948. 13. y. mushiake. “self-complementary antennas”. ieee antennas and propagation magazine, vol. 34, no. 6, 1992. 14. j. dyson. “the unidirectional equiangular spiral antenna”. ieee transactions on antennas and propagation, vol. 7, no. 4, pp. 329334, 1959. 15. j. d. kraus and r. j. marhefka. “antenna for all applications”. mcgraw-hill, new york, 2002. 16. b. c. wadell. “transmission line design handbook”. artech print on demand, boston, 1991. . 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 7-12 research article characterization of lactobacillus isolates from human mouth and feces as probiotics wala’a sh. ali*, aya t. abdulameer reza department of biology, college of science, university of baghdad, baghdad, iraq abstract probiotics are live microbes that give many health benefits to human beings and animals, the most studied and commonly used probiotics are gram-positive bacteria; lactobacilli and bifidobacteria. at nowadays, lactobacillus spp. constitute more than two-thirds of the total numbers of probiotic species. the present study aimed to characterize lactobacillus that locally isolated from human mouth and feces as probiotics. a total of three lactobacillus isolates; lactobacillus fermentum lb2, lactobacillus rhamnosus lb9, and lactobacillus paracasei lb10 were investigated in respect to acid and bile salts tolerance, antibiotics susceptibility, and cell surface hydrophobicity in vitro using bacterial adhesion to hydrocarbons method. in comparison with the other two isolates, the isolate l. fermentum lb2 was able to grow in all ph values and in the presence of different concentrations of bile salts. antibiotics susceptibility profile showed that the tested lactobacillus isolates were sensitive to ampicillin, amoxicillin, and erythromycin, while they were resistant to the other antibiotics that used in this study. l. fermentum lb2 exhibited high surface hydrophobicity (77.26%), while the other tested isolates; l. rhamnosus lb9 and l. paracasei lb10 revealed moderate adhesion abilities, 68.56% and 65%, respectively. l. fermentum lb2 exhibited good probiotic behavior with respect to acid and bile salt tolerance as well as adhesion ability to hydrocarbons. keywords: acid and bile salt tolerance, cell surface hydrophobicity, lactobacillus, probiotic introduction the attention of probiotic as an effective approach to prevent and treat a wide range of diseases is observed in recent years.[1] the term probiotic consists of two words which are pro, means for, and biotic, means life and totally means for life.[2] in 2001, the food and agriculture organization of the united nations and world health organization defined probiotics as “live microorganisms which, when administered in adequate amounts, confer a health benefit on the host.”[3] the us food and drug administration stated that the probiotic is considered as generally recognized as safe.[4] many studies showed that probiotics are used effectively to do many beneficial functions to the host[5-9] when they are taken in adequate quantity, and the recommended dosage of them is ranged usually between 108 and 1010 cfu/day.[10] probiotics can be found in a wide assortment of commercial dairy products[11] as well as they can be prepared as capsules, powder, granules, and fermented or pelleted feed.[12] microorganisms are considered as probiotic when they have several criteria such as the ability to adhesion to gastrointestinal tract, the ability to tolerance acidity and bile salts, and the resistance to antibiotics[13] have health benefits for the host[14] and safety.[15] the most commonly used probiotics are lactobacillus and bifidobacterium species.[16] lactobacilli are the largest group included in lactic acid bacteria[15] and have many functions, such as improvement of growth performance and health of gastrointestinal tract, this answers why they are among the bacteria mostly used as probiotics in animal feeds and human foods.[17,18] the aim of this study was to characterize three locally lactobacillus isolates as probiotics. materials and methods bacterial isolates a total of three locally lactobacillus isolates were used in this study; lactobacillus fermentum lb2 that isolated from the mouth, lactobacillus rhamnosus lb9 and lactobacillus paracasei lb10 that isolated from feces.[19] corresponding author: dr. wala’a sh. ali, department of biology, college of science, university of baghdad, baghdad, iraq. e-mail: dr.walaashali@scbaghdad.edu.iq received: apr 13, 2019 accepted: apr 19, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp7-12 copyright © 2020 wala’a sh. ali, aya t. abdulameer reza. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ ali and reza: lactobacillus isolates from human mouth and feces as probiotics 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 7-12 acid tolerance test fresh broth cultures of lactobacillus isolates (lb2, lb9, and lb10) were used at concentration of 1.5×108 cfu/ml according to mcfarland tube no. 0.5 (hardy diagnostics, santa maria). serial dilutions were made in tubes contain mrs broth (oxoid, germany), then bacteria were transferred individually to mrs broth at various ph values (2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, and 6), and tubes were incubated at different times (1, 1.5, 2, 2.5, and 3 h) h; thereafter, 0.1 ml from each culture was taken and spread on mrs agar (oxoid, germany) with two replicates. plates were incubated anaerobically for 24 h at 35°c. the pronounced colonies were counted, and results were recorded corresponding to control.[20] bile salt tolerance test fresh broth cultures of lactobacillus isolates (lb2, lb9, and lb10) were used at the concentration of 1.5×108 cfu/ml according to mcfarland tube no. 0.5. serial dilutions were made in tubes contain mrs broth, then bacteria were transferred individually to mrs broth supplemented with various concentrations of bile salts (difco, usa) (0.3%, 0.5%, and 0.7%), tubes were incubated at different times (1, 1.5, 2, 2.5, and 3 h); thereafter, 0.1 ml from each culture was taken and spread on mrs agar with two replicates, and then plates were incubated anaerobically for 24 h at 35°c. the pronounced colonies were counted, and results were recorded corresponding to control.[21] the optical density of bacterial isolates growth in each bile salt concentration was measured at wavelength 660 nm after 3 h of incubation. antibiotics susceptibility test fresh broth cultures of lactobacillus isolates (lb2, lb9, and lb10) were used at concentration of 1.5×108 cfu/ml according to mcfarland tube no 0.5; thereafter, 0.1 ml from each culture was taken and spread on mrs agar, and then antibiotics discs (25 µg ampicillin, 25 µg amoxicillin, 10 µg tetracycline, 1.25/23.75 µg trimethoprim-sulfamethoxazole, 10 µg erythromycin, 10 µg gentamycin, 25 µg streptomycin, 10 µg amikacin, 10 µg neomycin, and 10 µg chloramphenicol) (bioanalyse, turkey) were gently placed on mrs agar plates, plates were incubated anaerobically at 35°c for 24 h. the inhibition zones were measured and recorded; the work was done with two replicates.[22] cell surface hydrophobicity test it was determined in vitro using bacterial adhesion to hydrocarbons (bath) method,[23] a fresh cultures of lactobacillus isolates were subjected to centrifugation (6000 rpm, 10 min), cells were washed by phosphate buffer saline (oxoid, germany), resuspended in the same buffer and the optical densities of suspensions were adjusted to 0.8–1 at (od 560), and the data were recorded as a 0.6 ml of lactobacillus suspensions were mixed with 1.2 ml of n-hexadecane (sigma, usa), gently vortexed, and tubes were incubated anaerobically at 35°c for 30min. the mixtures were vortexed rigorously for 2 min and then allowed to stand for 15 min at room temperature to ensure complete separation of the organic and the aqueous phase. the absorbance of aqueous phase was measured at 560 nm and recorded as a, the affinity to solvent was expressed using the formula: hydrophobicity�percentage a0-a a0 �= ( ) ×� 100 a0, before adding n-hexadecane; a, after adding n-hexadecane. statistical analysis statistical analysis system[24] program was used to show the effect of different factors in study parameters. furthermore, least significant difference (lsd) test was used to significant compare between means in this study. results and discussion probiotic strains must be able to survive and colonize in the presence of acidic conditions and bile salt so as to travel through the gastrointestinal tract and reach to digestive tract where they give their therapeutic effect to host.[25] in current study, three lactobacillus isolates were subjected to different ph values with different incubation periods to examine their ability to tolerate acidic conditions. according to the results in table 1, all three isolates were unable to grow at ph 2, while they were able to grow at all times of incubation at ph 4, 4.5, and 5 with a decline in their growth at ph 6 after its rise at ph 5.5. at ph 5, there was no significant difference in the growth of the three isolates except the 1st h of incubation with significant difference in time 3 h for lb9. the gradual decrease in the survival rate of bacterial growth, from ph 5 down to ph 2, as well as the decrease in bacterial growth at ph 6 after its increasing at ph 5.5 can be observed, it seemed there is an affinity of the three isolates to ph 5.5 where their survival percentage reached the highest at this ph value, these results are similar to previous studies that used ph 5.5 in preparation of media as it allows optimum growth for lactobacillus.[26,27] at ph 6, the reduction of the growth of the three isolates is noticeable at the 1st h of incubation while there was no significant difference in their growth in times 1.5 and 2 h for lb2 and in times 2 and 2.5 h for lb10. the significant decrease in the bacterial growth at the 1st h of incubation is due to the sudden change in ph as reported by fernández-calviño and bååth.[28] the ability of three lactobacillus isolates to survive at three different concentrations of bile salts for different incubation periods was examined. the results in table 2 show that all isolates were able to survive at all bile salts concentrations. the results of this study are agreed with previous studies used 0.3% and 0.5% of bile salts concentrations for 2.5 h of incubation.[29,30] the gradual decrease in the survival rate of bacterial growth as a result of the increase of concentration of bile salts can be observed. the significant decrease in bacterial growth at the 1st h of incubation is due to the sudden exposure to bile salts stress while there was no significant decrease in bacterial growth after the 1st h of incubation particularly at high concentration of bile salts. the resistance of lactobacillus to bile salts may be due to different mechanisms such as changes in the composition of cell membrane and cell wall, active efflux of bile acids, and bile salt hydrolysis.[31] figure 1 shows ali and reza: lactobacillus isolates from human mouth and feces as probiotics 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 7-12 the tolerance of lactobacillus to three bile salts concentrations after 3 h of incubation. for probiotic bacteria to be active for a long time in the gastrointestinal tract, they should be able to resist administrated antibiotics. in this study, ten antibiotics were used. the results in figure 2 show that lb2, lb9, and lb10 were highly sensitive to ampicillin, amoxicillin, and erythromycin; this was in accordance with reported by other authors.[32-35] for lb2, the inhibition zone of ampicillin was higher than that of amoxicillin and erythromycin while the inhibition zones of these three antibiotics were the same for lb9 while for lb10, amoxicillin made a higher inhibition zone than other two antibiotics. lb9 was resistant to trimethoprim-sulfamethoxazole, amikacin, and neomycin where there was no inhibition zone; these results are similar to those of coppola et al.[36] table 1: acid tolerance percentages of lactobacillus isolates at different ph values ph h mean of viable count (log10 cfu/ml) survival percentage lb2 lb9 lb10 lb2 lb9 lb10 2.5 1 8.505b 8.612b 2 3 0 3 1 8.748b 8.959b 8.643b 1.5 8.913a 4 25 4 2 8.838a 2.5 8.643b 3.5 1 8.851b 9.017b 8.799b 1.5 8.740a 8.949a 8.505b 2 8.886b 10 33 9 2.5 8.770b 3 8.643b 4 1 9.120b 9.086b 9.004b 1.5 9.110a 9.079a 8.959a 2 9.008b 9.004a 8.869a 46 45 39 2.5 8.954b 8.949b 8.819b 3 8.919b 8.892b 8.698b 4.5 1 9.178b 9.264b 9.093b 1.5 9.152a 9.136b 9.096a 2 9.123a 9.113b 9.079a 55 59 61 2.5 9.079b 9.071b 9.060a 3 8.963b 9.004b 9.045a 5 1 9.274b 9.204b 9.152b 1.5 9.276a 9.198a 9.130a 2 9.247a 9.146a 9.113a 77 64 66 2.5 9.235a 9.130a 9.086a 3 9.227a 9.123b 9.082a 5.5 1 9.298b 9.301a 9.250a 1.5 9.294a 9.278a 9.230a 2 9.271a 9.247a 9.204a 81 80 80 2.5 9.255a 9.230b 9.158b 3 9.247a 9.225b 9.130b 6 1 9.235b 9.206b 9.117b 1.5 9.220a 9.146a 9.082a 2 9.178a 9.127b 9.056a 65 59 58 2.5 9.143b 9.079b 9.045a 3 9.079b 9.075b 8.977b capital letters (a and b) indicate differences between the numbers of 1 h of incubation and control while the small letters indicate differences between the numbers of current hour and 1 h of incubation at p<0.05 based on the lsd test; a,ano significant difference; b,bsignificant difference; -: no viable cells; lb2: l. fermentum; lb9: l. rhamnosus; lb10: l. paracasei; missing ph (2) and hours, no viable cells ali and reza: lactobacillus isolates from human mouth and feces as probiotics 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 7-12 cell surface hydrophobicity of lactobacillus isolates was determined to correlate the data with their ability to adhere to the intestinal epithelium. cell surface hydrophobicity was determined using the method of bath. the percent cell surface hydrophobicity was calculated, as shown in table 3. according to three levels of hydrophobicity (low: 0–35%, medium: 36–70%, and high: 71–100%) that determined by ocana et al.[37] lb2 showed a high hydrophobicity which was 77.26% while lb9 and lb10 showed medium hydrophobicity with percentage of 68.56% and 65%, respectively, the results of lb2 table 2: bile salt tolerance percentages of lactobacillus isolates at different bile salts concentrations bile salt concentrations % h mean of viable count (log10 cfu/ml) survival percentage lb2 lb9 lb10 lb2 lb9 lb10 0.3 1 9.385a 9.372b 9.245b 70 68 58 1.5 9.369a 9.332a 9.212a 2 9.235b 9.278b 9.133b 2.5 9.193b 9.262b 9.130b 3 9.146b 9.260b 9.079b 0.5 1 9.305b 9.324b 9.103b 52 49 43 1.5 9.176b 9.206b 9.089a 2 9.133b 9.146b 9.021a 2.5 9.082b 9.082b 8.924b 3 8.991b 8.968b 9.004a 0.7 1 9.170b 9.060b 9.068b 42 30 44 1.5 9.117a 9.029a 9.060a 2 9.045b 8.954a 9.056a 2.5 9.029b 8.857b 9.037a 3 8.908b 8.851b 9.017a capital letters (a and b) indicate differences between the numbers of 1 h of incubation and control while the small letters indicate differences between the numbers of current hour and 1 h of incubation at p<0.05 based on the lsd test; a,ano significant difference; b,bsignificant difference (p<0.05); lb2: l. fermentum; lb9: l. rhamnosus; lb10: l. paracasei table 3: cell surface hydrophobicity of lactobacillus isolates lactobacillus isolates absorbance hydrophobicity percentage a0 a lactobacillus fermentum lb2 0.8273 0.1881 77.26 (high) lactobacillus rhamnosus lb9 0.9965 0.3132 68.56 (medium) lactobacillus paracasei lb10 0.8206 0.2872 65.00 (medium) a0: before adding n-hexadecane; a: after adding n-hexadecane; low: 0–35%; medium: 36–70%; high: 71–100% figure 1: tolerance of lactobacillus isolates; lb2, lactobacillus fermentum; lb9, lactobacillus rhamnosus; and lb10, lactobacillus paracasei to bile salts concentrations (0.3%, 0.5%, and 0.7%) figure 2: antibiotics susceptibility test of lactobacillus isolates; am: ampicillin; ax: amoxicillin; te: tetracycline; sxt: trimethoprim-sulfamethoxazole; e: erythromycin; cn: gentamycin; s: streptomycin; ak: amikacin; n: neomycin; c: chloramphenicol ali and reza: lactobacillus isolates from human mouth and feces as probiotics 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 7-12 and lb9 are similar to previous study of garcía et al.[35] the bath test quantifies the surface hydrophobicity of bacteria, and it has widely been used to estimate the physicochemical component in adhesion ability of bacterial strains.[38] hence, based on the hydrophobicity or the charge of the bacterial outer layer, the adhesion ability to hydrophobic surfaces such as mucus can be characterized indirectly.[39] however, consumption products that contain probiotic bacteria such as dairy products and fermented vegetables as well as eat healthy foods rich in fibers and prebiotics will contribute to increase the numbers of these useful bacteria in our gastrointestinal tract, and this lead to reestablishment of normal microbiota balance and enhancement their activities. acknowledgment we would like to thank dr. nasr noori al-anbari for his help in statistical analysis. conclusion depending on the results of this study, the bacterial isolate l. fermentum lb2 showed up good probiotic behavior in respect of acid and bile salt tolerance and adhesion ability to hydrocarbons. however, more studies are required about this isolate and thus could be good candidate to use as probiotic. references 1. w. s. ali and n. n. mahmood. “probiotic therapy: review”. iraqi journal of medical sciences, vol. 14, no. 1, pp. 1-6, 2016. 2. s. fijan. “microorganisms with claimed probiotic properties: an overview of recent literature”. international journal of environmental research and public health, vol. 11, no. 5, pp. 4745-4767, 2014. 3. r. frei, m. akdisa and l. o’mahony. “prebiotics, probiotics, synbiotics, and the immune system: experimental data and clinical evidence”. current opinion in gastroenterology, vol. 31, pp. 1-6, 2015. 4. s. doron and d. r. snydman. “risk and safety of probiotics”. clinical 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2018. 10. s. sarkar. “worldwide consumer’s awareness and inclination towards healthful food products”. international journal of food sciences and nutrition, vol. 5, no. 2e, p. 1, 2016. 11. g. gardiner, c. stanton, p. b. lynch, j. k. collins, g. fitzgerald and r. p. ross. “evaluation of cheddar cheese as a food carrier for delivery of a probiotic strain to the gastrointestinal tract”. journal of dairy science, vol. 82, no. 7, pp. 1379-1387, 1999. 12. y. ohashi and k. ushida. “health-beneficial effects of probiotics: its mode of action”. animal science journal, vol. 80, pp. 361-371, 2009. 13. n. farahmand. “characterization of probiotic lactobacillus spp. isolates from commercial fermented milks”, ph.d thesis, london metropolitan university, england, 2015. 14. c. dunne, l. o’mahony, l. murphy, g. thornton, d. morrissey, s. o’halloran, m. feeney, s. flynn, g. fitzgerald, c. daly, b. kiely, g. c. o’sullivan, f. shanahan and j. k. collins. “in vitro selection criteria for probiotic bacteria of human origin: correlation with in vivo findings”. american journal of clinical nutrition, vol. 73, no. 2, pp. 386s-392s, 2001. 15. s. gulel. “molecular identification and properties of ‘lactobacillus acidophilus’ group isolates from turkish kefir,” m.sc.thesis, middle east technical university, turkey, 2014. 16. m. a. ciorba. “a gastroenterologist’s guide to probiotics”. clinical gastroenterology and hepatology, vol. 10, no. 9, pp. 960-968, 2012. 17. v. coeuret, m. gueguen and j. p. vernoux. “numbers and strains of lactobacilli in some probiotic products”. international journal of food microbiology, vol. 97, no. 2, pp. 147-156, 2004. 18. r. dowarah, a. k. verma and n. agarwal. “the use of lactobacillus as an alternative of antibiotic growth promoters in pigs: a review”. animal nutrition, vol. 3, no. 1, pp. 1-6, 2017. 19. a. t. a. reza and w. s. ali. “antibacterial activity of lactobacillus spp. against listeria monocytogenes”. indian journal of public health research and development, in press. 20. w. p. charteris, p. m. kelly, l. morelli and j. k. collins. “development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic lactobacillus and bifidobacterium species in the upper human gastrointestinal tract”. journal of applied microbiology, vol. 84, no. 5, pp. 759-768, 1998. 21. m. f. fernandez, s. boris and c. barbes. “probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract”. journal of applied microbiology, vol. 94, no. 3, pp. 449-455, 2003. 22. w. p. charteris, p. m. kelly, l. morelli and j. k. collins. “antibiotic susceptibility of potentially probiotic lactobacillus species”. journal of food protection, vol. 61, no. 12, pp. 1636-1643, 1998. 23. m. rosenberg, d. gutnick and e. rosenberg. “adherence of bacteria to hydrocarbons: a simple method for measuring cellsurface hydrophobicity”. fems microbiology letters, vol. 9, no. 1, pp. 29-33, 1980. 24. sas. statistical analysis system, user’s guide. statistical. version 9.1th ed. cary. n.c. usa: sas. institute inc., 2012. 25. e. tuomola, r. crittenden, m. playne, e. isolauri and s. salminen. “quality assurance criteria for probiotic bacteria”. american journal of clinical nutrition, vol. 73, no. 2, pp. 393s-398s, 2001. 26. l. samaranayake. “essential microbiology for dentistry e-book”, 4th ed. amsterdam, netherlands: elsevier health sciences, 2011. 27. a. m. grumezescu and a. m. holban. “advances in biotechnology for food industry”. cambridge, usa: academic press, 2018. 28. d. fernández-calviño and e. bååth. “growth response of the bacterial community to ph in soils differing in ph”. fems microbiology ecology, vol. 73, pp. 149-156, 2010. 29. m. mirlohi, s. soleimanian-zad, s. dokhani, m. sheikh-zeinodin and a. abghary. “investigation of acid and bile tolerance of native lactobacilli isolated from fecal samples and commercial probiotics by growth and survival studies”. iranian journal of biotechnology, vol. 7, no. 4, pp. 233-240, 2009. 30. m. tavakoli1, z. hamidi-esfahani, m. a. hejazi, m. h. azizi and s. abbasi. “characterization of probiotic abilities of lactobacilli isolated from iranian koozeh traditional cheese”. polish journal of food and nutrition sciences, vol. 67, no. 1, pp. 41-48, 2017. 31. l. ruiz, a. margolles and b. sánchez. “bile resistance mechanisms ali and reza: lactobacillus isolates from human mouth and feces as probiotics 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 7-12 in lactobacillus and bifidobacterium”. frontiers in microbiology, vol. 4, pp. 1-8, 2013. 32. n. zdolec, i. filipović, ž. c. fleck, a. marić, d. jankuloski, l. kozačinski and b. njari. “antimicrobial susceptibility of lactic acid bacteria isolated from fermented sausages and raw cheese”. veterinarski arhiv, vol. 81, no. 1, pp. 133-141, 2011. 33. s. k. allameh, f. m. yusoff, h. m. daud, e. ringø, a. ideris and c. r. saad. “characterization of a probiotic lactobacillus fermentum isolated from snakehead, channa striatus, stomach”. journal of the world aquaculture society, vol. 44, no. 6, pp. 835-844, 2013. 34. r. georgieva, l. yocheva, l. tserovska, g. zhelezova, n. stefanova, a. atanasova, a. danguleva, g. ivanova, n. karapetkov, n. rumyan and e. karaivanova. “antimicrobial activity and antibiotic susceptibility of lactobacillus and bifidobacterium spp. intended for use as starter and probiotic cultures”. biotechnology and biotechnological equipment, vol. 29, no. 1, pp. 84-91, 2015. 35. a. garcía, k. navarro, e. sanhueza, s. pineda, e. pastene, m. quezada, k. henríquez, a. karlyshev, j. villena and c. gonzález. “characterization of lactobacillus fermentum uco-979c, a probiotic strain with a potent anti-helicobacter pylori activity”. electronic journal of biotechnology, vol. 25, pp. 75-83, 2017. 36. r. coppola, m. succi, p. tremonte, a. reale, g. salzano and e. sorrentino. “antibiotic susceptibility of lactobacillus rhamnosus strains isolated from parmigiano reggiano cheese”. lait, vol. 85, no. 3, pp. 193-204, 2005. 37. v. s. ocana, e. bru, a. a. de ruiz holgado and m. e. nader-macias. “surface characteristics of lactobacilli isolated from human vagina”. journal of general and applied microbiology, vol. 45, no. 5, pp. 203-212, 1999. 38. t. de wouters, c. jans, t. niederberger, p. fischer and p. a. ruhs. “adhesion potential of intestinal microbes predicted by physico-chemical characterization methods”. plos one, vol. 10, no. 8, pp. 1-17, 2015. 39. m. c. van loosdrecht, j. lyklema, w. norde, g. schraa and a. j. zehnder. “the role of bacterial cell wall hydrophobicity in adhesion”. applied and environmental microbiology, vol. 53, no. 8, pp. 1893-1897, 1987. . 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 26-31 research article investigation of the zinc oxide nanoparticles effect on thyroid and testosterone hormones in male rats noori m. luaibi,* noor a. zayed department of biology, college of sciences, university of al-mustansiriyah, baghdad, iraq abstract exposure to zinc oxide nanoparticles (zno nps) has been increasing steadily, causing more attention being paid to their potential toxicity, including cytotoxicity and genotoxicity. hence, this study aimed to investigate the effect of zno nps on thyroid hormone triiodothyronine (t3), thyroxine (t4), and thyroid-stimulating hormone (tsh) as well as testosterone hormone in male adult rats. a total of 54 sprague-dawley albino adult male rats were divided into nine groups each of six rats, daily treated intraperitoneal with zno nps two different doses (30 and 60) mg/kg in three different periods of time (7, 14, and 28) days, as following: control groups (groups 1, 2, and 3): respectively received intraperitoneal injection with distilled water for 7, 14, and 28 days, experimental groups (groups  4, 5, and 6): they were rats, respectively, received intraperitoneal dose (60 mg/kg) of zno nps for (7, 14, and 28) days, and group (7, 8, and 9) experimental groups were rats, respectively, received intraperitoneal dose (30 mg/kg) of zno nps for (7, 14, and 28) days. data showed high significant decrease (p < 0.01) in level of t3, t4, tsh, and level of testosterone also decrease at high and low dose for 7, 14, and 28 days. keywords: t3, t4, testosterone, thyroid-stimulating hormone, zinc oxide nanoparticles introduction zinc oxide nanoparticles (zno nps) are inorganic compounds, called multifunctional material due to unique chemical and physical properties.[1] zinc is an essential metal it is an activator for more than 300 enzymes in the body.[2] zno nps are one of most widely used in cosmetics and sunscreen because of their efficient uv absorption properties, they are used due to the antimicrobial properties in food packaging.[3] they are also being explored for their potential use as fungicides in agriculture.[4] as well zno nanoparticles are used in anticancer drugs and in biomedical applications.[5] commonly considered to be a material with low toxicity because the zno is an essential trace element in human body and is existing in food or added as nutritional complement, so zinc attracts little care during assessment of toxicity of nps.[6] zno nps can induce the formation of reactive oxygen species (ros) that disrupt intracellular metabolic activities and the antioxidant system; these alterations permit generated ros to interact with and damage dna, lipids, carbohydrates, and proteins.[7] thyroid gland is one of major endocrine glands of the body responsible of creating the hormones, thyroxin, and triiodothyronine which are essential for the proper organism development in particular for the nervous system and heart, normal growth, and skeletal maturation.[8] it is considered one of the main regulators of biological processes, during development, and childhood.[9] decrease in the production of thyroid hormones means hypothyroidism.[10] many evidence suggesting the role of zinc in the formation and function of thyroid hormones.[11] zno nps cause higher levels of oxidative stress, resulting in inflammation, and cell toxicity.[12] man testis has two main functions, the production of testosterone and the production of male germ cells, maintenance of spermatogenesis process needs testosterone, the androgen production is regulated by luteinizing hormone, while follicle-stimulating hormone is critical for initiation and maintenance of spermatogenesis.[13,14] testosterone is the main androgenic hormone in males, it is largely produced by the leydig cells of the testes, zno nps were internalized by sertoli cells and leydig cells resulted in cytotoxicity in a time and dose-dependent manner through the induction of apoptosis, caused by increase in ros related with loss of corresponding author: noori m. luaibi, department of biology, college of sciences, university of al-mustansiriyah, baghdad, iraq. e-mail: sznl@uomustansiriyah.edu.iq received: apr 25, 2019 accepted: apr 28, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp26-31 copyright © 2020 noori m. luaibi, noor a. zayed. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ luaibi and zayed: zinc oxide nanoparticles effect on thyroid and testosterone hormones 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 26-31 mitochondrial membrane potential, so injection of zno nps produced structural alterations in the seminiferous epithelium and sperm abnormalities in male rats.[15] materials and methods adult male sprague-dawley albino rats, age about 2.5–3 months, average bodyweight 200–225 g. they were obtained from the national center for drug control and research/ministry of health, then transferred to the animal house of the college of science, mustansyriah university. all animals were allowed to acclimatize to the laboratory conditions for 7 days before starting the study. they were kept in clean separated plastic cages with metal network cover under climate-controlled condition of the animal house with 22–25 temperature, 60% humidity, 12 h light, and darkness period and allowed free access to food ad libitum and water. this study was conducted after obtaining the approval from the ethical consideration to deal with experimental animals that is the ethical committee of the college of science, mustansiriyah university and all ethical behavior with the laboratory animals was taken care and has priority in our work in accordance with the globally adopted guidelines of the animal care and experiments. preparation of zno nps solution zno nps used in this study was obtained from sky spring nanomaterials, they were in white to light yellow colored powder with 99.8% purity, particle size was 10–30 nm in diameter. the stock suspension was prepared by dissolving 1 g of powder zno in 10 ml of distilled water and then mixed by vortex for 10 min to prevent agglomeration, then distributed in to following groups: group of 60 mg/kg of zno nps (high dose) 120 µl of stock +880 µl of distal water. group of 30 mg/kg of zno nps (low dose) 60 µl of stock +940 µl of distal water. experimental design to study the effect of zno nps, animals were divided into nine groups with six rats each as follows: • groups 1, 2, and 3 (control group), respectively, received intraperitoneal injection of distilled water for (7, 14, 28) days. • groups 4, 5, and 6 (the experimental groups); rats, respectively, received intraperitoneal dose (60 mg/kg) of zno nps for 7, 14, and 28 days. • groups 7, 8, and 9: (the experimental groups); rats, respectively, received intraperitoneal dose (30 mg/kg) of zno nps for 7, 14, and 28 days. measurement of the levels of hormones concentration it was represented by the enzyme immunoassay tests (tosoh) for the quantitative determination of concentrations of thyroid gland hormones t3 according to braverman et al.,[16] t4 according to ormston et al.,[17] and thyroid-stimulating hormone (tsh) according to fisher.[18] furthermore, the reproductive hormone testosterone was measured according to sigberg.[19] collection of blood samples the end of each experiment animals were completely anesthetized by diethyl ether for several minutes, and blood samples were obtained by heart puncture collected in to non-heparinized tubes used in biochemical examination, 4 ml of blood collected from each rat was used to obtain sera (0.5–1.0) ml separated by centrifugation 3000 rpm for 5 min, then they were kept in −20°c until analysis for the measurement of the level of tsh, measurement of the level of t3, measurement of the level of t4, and measurement of the level of testosterone. results thyroid hormone function statistical analysis for the effect of zno nps on t3, t4, and tsh serum levels is shown in figures 1-3. t4 (µg/dl) serum level displayed high significant decrease (p < 0.01) in both treated groups (30, 60) mg/kg (3.10 ± 0.01), (2.90 ± 0.02) (µg dl), respectively, at day 7 compared to control groups (3.34 ± 0.01) (µg/dl), at day 14 also showed high significant decrease (p < 0.01) at the same concentrations (2.71 ± 0.01), (2.21 ± 0.02) (µg/dl) when compared with control group (3.35 ± 0.01) (µg/dl), in addition to 28 period of time exposing to zno nps observed high significant decrease (p < 0.01) of the level of t4 in different concentration (30, 60) mg/kg (1.91 ± 0.02), (1.66 ± 0.01) (µg/dl) compared with control groups (3.36 ± 0.01) (µg/dl), as shown in figure 1. statistical analysis of the level of t3 (ng/ml) showed high significant decrease (p < 0.01) in both treated groups (30, 60) mg/kg (0.685 ± 0.007), (0.600 ± 0.010) (ng/ ml), respectively, exposed to zno nps for 7 days compared to the control groups (0.786 ± 0.008) (ng/ml), also there was high significant decrease (p < 0.01) in the level of t3 at both concentrations (30, 60) mg/kg at 14 days (0.581 ± 0.01), (0.438 ± 0.019) (ng/ml) in comparison to control groups (0.786 ± 0.008), and at 28 days (0.440 ± 0.01), (0.335 ± 0.01) (ng/ml) in comparison to control groups (0.795 ± 0.009) (ng/ml), as shown in figure 2. while serum level of tsh (µlu/ml) showed high significant decrease (p < 0.01) at different treatment durations exposing to zno nps in both treated groups (30 and 60) mg/kg (0.186 ± 0.007), (0.190 ± 0.010) (ng/ml), respectively, for 7 days compared to the control groups (175 ± 0.008) (ng/ml), also there was high significant decrease (p < 0.01) in the level of (tsh) at both concentrations (30 and 60) mg/kg in 14 days (0.188 ± 0.01), (0.195 ± 0.019) (ng/ml), respectively, in comparison to control group (0.175 ± 0.008), and at 28 days (0.191 ± 0.01), (0.198 ± 0.01) (ng ml) in comparison to control group (0.174 ± 0.009) (ng/ml), as shown in figure 3. testosterone statistical analysis of the present study of zno nps on testis function is observed in figure 4 there was a high significant decrease (p < 0.01) in testosterone level at 30 and 60 mg/kg for 7 days (160.83 ± 1.16), (149.00 ± 1.29) ng/dl, respectively, compared to control group (184.67 ± 1.05) ng/dl. testosterone level exhibited significant decrease (p < 0.01) at day14 luaibi and zayed: zinc oxide nanoparticles effect on thyroid and testosterone hormones 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 26-31 when treated with zno nps (30 and 60) mg/kg (138.50 ± 0.99), (121.50 ± 0.99) ng/dl when compared with control groups (181.00 ± 1.31) ng/dl. there was significant decrease (p < 0.01) showed in testosterone level at a period of 28 days treatment (30 and 60) mg/kg (117.00 ± 1.52), (104.17 ± 1.42) ng/dl in comparison to control groups (184.17 ± 1.24) ng/dl. discussion results of the present study are in agreement with previous study of zno nps by cheric and rafieirad[20] showed that injecting zno nps with three different doses (1.25, 2.5, and 5) mg/kg to 42 male rats divided into 7 groups, the blood samples of experimental with different doses were taken on the 1st, 3rd, 4th, and 15th days after receiving nano zno intraperitoneally (ip) to measure the amount of t3 and t4 hormone levels, results observed that acute injection of nano zno reduced the amount of thyroid hormone t3 and t4, the effect occurred in different doses, but is more significant in the medium and long term, even in small amounts can cause negative effects on the activity of the thyroid gland and disrupt creation of thyroid hormones. another study by espanani et al.[21] who treated 48 male rats by zno nps ip (5, 10, 20, and 40 mg/kg), after a 21 days period results showed significant increase in tsh hormone level in animals that were treated by high dose of zno np, but those who treated by 5, 10, and 20 mg/kg do not show this effect. dean et al.[22] found a reduction in thyroid function t3, t4 when he fed chicken with zn in diet at 73 ppm or 5280 ppm for 1–2 weeks, have demanded that high zn intake change the production and secretion of thyroid hormones, they suggested that decrease in thyroid hormone function was due to reduced effects of thyroid gland might be induced by the reduced regulatory effect of pituitary gland on thyroid gland, and decreased circulating thyroid hormones figure 2: effect of different concentrations of zinc oxide nanoparticles (30 and 60) mg/kg on t3 levels of rats with different periods of time (7, 14, and 28) days in comparison with control groups and between treated groups themselves. **high significant decrease (≤0.01). a,b,crepresents the significant difference between groups with days as a fixed factor and concentrations as a variable factor. a,b,crepresents the significant difference between groups with concentrations as a fixed factor and days as a variable factor figure 1: effect of different concentrations of zno nps (30 and 60) mg/kg on t4 levels of rats with different periods of time (7, 14, and 28) days in comparison with control groups and between treated groups themselves. **high significant decrease (≤0.01). a,b,crepresents the significant difference between groups with days as a fixed factor and concentrations as a variable factor. a,b,crepresents the significant difference between groups with concentrations as a fixed factor and days as a variable factor figure 3: effect of different concentrations of zinc oxide nanoparticles (30 and 60) mg/kg on thyroid-stimulating hormone levels of rats with different periods of time (7, 14, and 28) days in comparison with control groups and between treated groups themselves. **high significant decrease (≤0.01). a,b,crepresents the significant difference between groups with days as a fixed factor and concentrations as a variable factor. a,b,crepresents the significant difference between groups with concentrations as a fixed factor and days as a variable factor figure 4: effect of different concentrations of zinc oxide nanoparticles (30 and 60) mg/kg on testosterone levels of rats with different periods of time (7, 14, and 28) days in comparison with control groups and between treated groups themselves. **high significant decrease (≤0.01). a,b,crepresents the significant difference between groups with days as a fixed factor and concentrations as a variable factor. a,b,crepresents the significant difference between groups with concentrations as a fixed factor and days as a variable factor luaibi and zayed: zinc oxide nanoparticles effect on thyroid and testosterone hormones 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 26-31 may be indicative of hypothyroidism due to zinc toxicity. lu et al.[23] stated that zn decrease the binding of t3 to its receptor in several preparations in vitro. kaya et al.[24] conducted the experiment using 130 hisex brown laying hens from 56 weeks to 68 weeks of age, then they were divided into five zinc treatment groups (0, 25, 50, 100, and 200) mg/zinc/kg diet respectively, the values of 100 and 200 mg/zn/kg decreased plasma level of t4 compared to control, while plasma level of t3 was reduced by 100 mg/zn/ kg compared to groups fed less zn, these data might suggest that a high intake of zn changes production or secretion of the thyroid hormone.[25] used 12 lambs and 12 goats that were divided into two equal groups as control and zn groups in separate experiments, both species of animals in the zn groups were fed a basal ration supplemented with zinc sulfate adjusted to 250 mg zn/kg diet showed significant decrease in hormone level t3 and t4.[26] randomly distributed 120 male rabbits into four groups, the control groups were fed on a basal diet with zinc-free premix, while experimental groups received the basal diet supplemented, group 1 with 60 mg/kg nano zno/kg diet, group 2 60/kg mg nano zno/kg diet, and group 3 30/kg mg nano zno/kg diet, respectively, results observed that rabbits showed no significant changes among the treated groups in respect to serum tsh concentration. in contrast, baweja et al.[27] demonstrated that the levels of tsh were raised after 8 weeks of zinc supplementation in female wistar rats. another study that disagree with present report by shirband et al.[28] that administered iron oxide nps with three different dose (20 µg/kg, 50 µg/kg, and 150 µg/kg) for 15 days to male rats, results showed significant increase in t4 level in groups receiving a dose 50 µg/kg and caused significant decrease in tsh in the group receiving a dose 50 µg/kg and 150 µg/kg. it is likely that np effects can be applied through the inhibition of endocrine pituitary axis hypothalamus which affects the hypothalamus, and probably due to decreased tsh levels. nps affect hypothalamic pituitary thyroid axis and therefore affect the level of thyroid hormones that were demonstrated in this study. in a study by[29] used 32 adult male mice treated daily for 35 days: 5, 50, and 300 mg/kg zno nps, respectively, while control group received distilled water orally for 35 consecutive days, results observed significantly changed in 50 and 300 mg/kg zno nps treated mice in epididymal sperm parameters including sperm number, motility, and percentage of abnormality, while significant decrease in seminiferous tubule diameter, seminiferous epithelium height, and maturation arrest was observed at 50 and 300 mg/kg zno nps, this study established that znp has cytotoxic actions on testicular germ cells in a dose-dependent manner, nps might also affect sertoli cell functions.[30] who treated 40 female wistar rats with zirconium oxide nps with a dose of 100, 200, and 400 ppm injected intraperitoneally showed a significant decrease in level of testosterone at high doses. reports of a study by mohammadi et al.[31] injected titanium dioxide tio nps intraperitoneally to wistar rats weighing 150–250 g, 1 ml tio2 nps in doses (30 and 50) mg/kg, injection repeated every other day, results showed decrease in testosterone level, could be caused by adverse effects of nps in the leydig cells, as leydig cells are testosterone production factories. another study by fuse et al.[32] showed that zinc nps disorder leads to atrophy to the seminiferous tubules and impaired spermatogenesis in male rats. in a study by shirvani et al.[33] was intraperitoneal injected of different doses (25, 50, and 100 mg/kg) of zno nps (25 nm) to male wistar rats showed decrease but no significant in level of testosterone, synthesis of testosterone is inhibited as a result of responding to the inflammation which is caused by zno nps. [34] treated rats with zno nps with dose (5, 10, 20, and 40 mg/ kg) results revealed significant increase in level of testosterone in blood serum of rats at high dose of zno nps (40 mg/kg), results assured exposing to zno nps damaged to public health and reduced fertility potential. zno nps are related to (ros), which results in an increase in dna double-strand breakage and a decrease in sperm motility.[33] kolesarova et al.[35]reported that metal nps induce changes in reproductive organs, histology of laboratory animals and cause disruption in reproductive cells production and hormones. thyroid hormone deficiency affects all tissues of the body including multiple endocrine changes that alter growth hormone, corticotrophin, gonadal function, and glucocorticoids, as primary hypothyroidism is associated with hypogonadotropic hypogonadism, so hypothyroidism decrease free testosterone concentration as thyroid hormone affects in sex hormone-binding globulin (shbg).[36] alterations in gonadal steroid genesis and pituitary functions have been stated in hypothyroid males, hypothyroidism was found to be associated with an increase in level of total cholesterol and reduction in the levels of testosterone and progesterone without any alteration in the levels of gonadotropins and estradiol, the decline in level of testosterone could be explained by reduction in serum triiodothyronine, a higher rate of alteration of testosterone to estradiol or the further decline in the rate of alteration of progesterone to testosterone.[37] increasing in tsh level that was caused by zno nps decreased gonadotropinreleasing hormone secretion by negative feedback inhibition that led to lh and fsh reduction, while inhibin hormone that is usually released by sertoli cells can affect fsh hormone level.[21] considering the results of the above mentioned studies and the alterations in results from the present study, these outcomes could be related to the dosage of zno nps used, animal species diversity, route of administration of nps and different durations of exposing to nps, nps showed cytotoxic effect on testosterone and thyroid hormone levels in dose and time-dependent manner. conclusions the results of this study showed that the size, doses, route of administration, and time depended can be a factor that effects thyroid hormone level and testosterone, the decrement in t3 andt4 can be caused by dose and duration of zno nps, decrease in levels of thyroid hormones due to toxic effect of zno nps that effects on the function of thyroid gland or on release of tsh. nps affects the reproductive system of male by complex and varied mechanisms; results indicate that thyroid hormone deficiency has effects on testosterone level, as hypothyroidism decrease free testosterone concentration, also thyroid hormone has effects on shbg. acknowledgments the authors would like to thank mustansiriyah university (www.uomustansiriyah.edu.iq) baghdad, iraq, for its support in the present work. 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pp. 17-25, 2004. 37. a. kumar, b. p. mohanty and l. rani. “secretion of testicular steroids and gonadotrophins in hypothyroidism”. andrologia, vol. 39, pp. 253-260, 2007. tx_1~abs:at/add:tx_2~abs:at 36 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 36-41 research article antibiotic susceptibility pattern, molecular characterization of virulence genes among pseudomonas aeruginosa isolated from burn patients mustafa d. younis1*, omar f. bahjat1, sirwan a. rashid2 1department of general biology, college of science, cihan university-erbil, kurdistan region, iraq, 2department of biomedical sciences, college of science, cihan university-erbil, kurdistan region, iraq. abstract in this research, a total of 150  samples were obtained from burn and wound patients admitted to the west erbil emergence hospital during period from september 2020 to january 2021. through cultural, morphological features, biochemical testing, and vitek 2 compact systems, 40 isolates of p. aeruginosa have been identified. p. aeruginosa produced various pigments, including blue/green and yellow/green. the isolates of p. aeruginosa were subjected to 14 different antibiotics. imipenem was the most effective antimicrobial agents against all p. aeruginosa isolates, and most of isolates showed high resistance degree to ampicillin 100%, chloramphenicol 100%, amoxicillin-clavulanic acid 100%, cefotaxime 100%, and penicillin 100% while for aztreonam 32.5%, meropenem 42.5%, tobramycin 45%, gentamycin 45%, amikacin 45%, ciprofloxacin 62.5%, ceftazidime 67.5%, and tetracycline 80%. all pseudomonas aeruginosa isolates were screened using multiplex polymerase chain reaction (pcr) to check for the presence of pvda, lasb, protease, exoa, exot, exou, and plch on its genomic dna. the findings have shown that pvda was 55%, lasb 75%, protease 65%, exoa 60%, exot 75%, exou 60%, and plch 55% of isolates harbored these genes as a virulence genes. keywords: pseudomonas aeruginosa, burn patients, antibiotic resistance, virulence genes, and multiplex pcr introduction pseudomonas aeruginosa is a gram-negative aerobic non-spore rod with remarkable capacity to survive and persist under many environmental circumstances.[1] in both hospitals and communities, p. aeruginosa is a common, opportunistic human pathogen.[2] burning and wound infections are a challenge because they slow down the healing process, promote cicatrix, and can lead to bacteremia, sepsis (or organ failure) syndrome, however, organ from several systems cannot regulate homeostasis on its own and need immediate treatment.[3] the most severe pathogenic burn injuries are bacteria and fungi. multiple species biofilms are formed on burning injuries in 48–72 h of the wound injury.[4] organisms are acquired by the patient’s own skin, digestive, and respiratory flora, as well as association with contaminated environments and health-care providers.[5] the human skin is considered the principal protective layer of the body’s tissues and may contribute to damage and destruction of bacteria transmitted to the internal blood tissue, which is rich in proteins.[6] isolation and laboratory diagnosis is used to diagnose p. aeruginosa infection. this aerobic bacterium is needed and thrives in the majority of laboratory culture media. on pseudomonas agar (selective media) and cetrimide agar, bacteria can be isolated, warmth, no spores, flagella morphology, positive, exercise catalase, lactose intolerance (positive oxidase reaction), fruit odor (grape flavor), and ability to grow at 42°c are used to detect bacteria.[7] p. aeruginosa is a ubiquitous microorganism that can quickly develop resistance to various antibiotics of broad spectrum.[2] moreover, in recent years, resistance to a broad range of antibiotics by these microorganisms has made it difficult to treat infections caused and leads to higher death rates.[8] the development of soluble pyocyanin pigment, a water-soluble blue-green compound formed in large amounts, is one of p. aeruginosa characteristics. pyocyanin acts as an antibiotic against a variety of bacteria and fungi.[9] p. aeruginosa has a variety of virulence factors in corresponding author: mustafa d. younis, department of general biology, college of science, cihan universityerbil, kurdistan region, iraq. e-mail: mustafa.thanoon@cihanuniversity.edu.iq received: may 5, 2021 accepted: june 15, 2021 published: june 30, 2021 doi: 10.24086/cuesj.v5n1y2021.pp36-41 copyright © 2021 mustafa d. y. ali, omar f. bahjat, sirwan a. rashid. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) younis, et al.: antibiotic susceptibility, molecular characterization of virulence genes among p. aeruginosa 37 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 36-41 host defenses and infection. these factors include hemolysin production, pyocyanin production, gelatinase, and biofilm formation, which act by enhancing the damage of tissue and helping bacteria to avoid the action of antibiotics.[10] another virulence factor such as exotoxin a, exoenzyme s, elastase, and sialidase, which are powerfully controlled by cell-to-cell signals. exotoxin1a (eta) plays1a major role in the pathogenesis1of infections caused by1this organism as the primary virulence factor provided by most p. aeruginosa isolates. such exotoxins may contribute to leukopenia, acidosis, and blood circulation, necrosis of the liver, pulmonary edema, bleeding, and kidney tubular necrosis[11] so that the aim of this study is screening of antimicrobial sensitivity profile of p. aeruginosa and detection of certain virulence genes through pcr technique. materials and methods patients and samples collection one hundred and fifty samples were collected 1from patients admitted to west erbil emergency hospital, during the period from september 2020 to january 2021. following collection, each sample was cultured on different culture media and p. aeruginosa was identified by cultural characters, biochemical methods, and vitek 2 compact system. antibiotic sensitivity pattern the isolates were examined for antibiotic sensitivity in accordance with the national committee for clinica1 laboratory guidelines 1and the antimicrobial susceptibility testing protocols by disc diffusion method on muellerhinton agar.[12] adjustment of the bacterial inoculates to the clinical and laboratory standards institute of 0.5 mcfarland standards.[12] a sterile cotton swab was used to disperse the sample inoculum to mueller-hinton agar. the antimicrobial products tested, including: imipenem (ipm), ceftazidime (caz), ampicillin (am), aztreonam (atm), chloramphenicol (c), amoxicillin-clavulanic acid (amc), amikacin (ak), cefotaxime (ctx), gentamycin (cn), ciprofloxacin (cip), tetracycline (te), penicillin (p), meropenem (mem), and tobramycin (tob) were placed aseptically and incubated overnight. the zones of inhibition were interpreted and measured.[12] color production by p. aeruginosa isolates all isolates had been inoculated on cetrimide agar, incubated for 18–24 h by streaking method at 37°c, and then, the pigment production was examined.[13] dna extraction protocol two hundred microliters of overnight growth were centrifuged for 30 s at 13,000 rpm, after that 1.5 ml was separated from supernatant in 2 ml microcentrifugal tube. the pellet has been dissolved in 200 μl tl buffer, then removed and fully mixed with 20 ul proteinase k solutions to achieve a uniform suspension. the sample has been incubated in the water bath at 56°c for 10 min until the cells have been completely lysed. two hundred microliters of gb buffer applied to the specimens, then by vortexing mixed thickly for approximately 15 s up to a uniform mixture and then incubated for 10 min at 56°c. then, 200 μl of absolute ethanol is added and pipetted or vortexes. the lysate transferred carefully without wetting the rim into the spin column reservoir for 1 min at 10,000 rpm, and the column 1centrifuged the collection tube then discharged containing the flow-through solution a new 2 ml tube has been placed with the genet bio genomic dna purification column. five hundred microliters of gw1 buffer were added0 then centrifuged for 1 min at 10,000 rpm, the flow-through discarded and the purification column placed back into the collection tube, 500 μl of gw2 was added to the genet bio genomic dna purification column, centrifuged for 1 min at 10,000 rpm. then after centrifuging the tube, remove the flow-through and reassemble the spin column with its collection tube, again, centrifuge at 12,000 rpm for 12 min to extract ethanol completely and check that the droplet is not attached at the bottom of the tube. then, 1.5 ml of the spin column moved to a new tube to do the elution. two hundred microliters of the elution buffer were added to the center of the genet bio genomic dna purification kit column membrane. the genomic dna elution kept a side at room temperature for 1 min and centrifuged for 1 min at 10,0000 rpm. then, the purified dna was immediately removed and stored at −20°c for further applications (primeprep genomic dna extraction kit, genet bio, korea) protocol of pcr technique pcr conducted for all genes was performed in a 25 μl of reaction volume. master mix tube contains l2.5 μl, forward and reverse primers with 1 μl for each primer, dna template 1 μl, and finally sterile (d. w) deionized water 9.5 μl.[14] detection of pvda, lasb, protease, exoa, exot, exou, and plch virulence genes in p. aeruginosa multiplex pcr also was used for the detection of pvda, lasb, protease, exoa, exot, exou and plch genes in pseudomonas aeruginosa genome as shown in table 1: protocol of agarose gel electrophoresis to perform gel electrophoresis, a method of judelson[15] was followed with minor modifications. adding 1.2 g agarose to 100 ml 1x tbe buffer was used as an agarose gel, the mixture melted for 1–2 min in the microwave oven or until it was apparent and fully dissolved. left to cool at 50°c, 10 μl of primary safe dye was carefully added to the agarose solution then thoroughly mixed with a gentle stirring. the tray borders are sealed with the tape and inserted into the tray the right comb. then, the agarose gradually poured in the tray and any bubbles were removed with a disposable tip, then kept away to the side at room temperature, the agarose solidified (15–30 min). the tape was removed from the tray and then the tray was placed in the electrophoresis tank. the tank was filled with more tbe buffers so that the ge1 is completely under buffer. the pcr product loaded into the wells (15 μl) with loading buffer. depending on the size of the pcr sample, the first well (5 ul) (1 kb or 100 bp) was used. the gel runs for 50 min at 100 v. finally, the uv transilluminator and gel photographed. younis, et al.: antibiotic susceptibility, molecular characterization of virulence genes among p. aeruginosa 38 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 36-41 table l: primers sequences, target genes, amplicon sizes and cycling conditions for conventional and multiplex pcr[14] target gene primer sequences amplified segment (bp) initial denaturation denaturation annealing extension final extension pvda f-gactcaggcaac tgcaac l28l 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 min r-ttcaggtgctgg tacagg lasb f-ggaatgaacgaagcg ttctc 300 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 min r-ggtccagtagtagcg gttgg protease fatttcgccgactcc ctgta 752 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 min r-gaatagacgccgctg aaatc exoa f-aaccagctcagccac atgtc 207 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 min rgctggcccattcgctccagcgct exot f-aatcgccgtccaactgca tgcg l52 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 minr-tgttcgccgaggtac tgctc exou f-ccgttgtggtgccgt tgaag l34 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 min r-ccagatgttcaccga ctcgc plch f-gaagccatgggctac ttcaa 307 96°c 5 min 96°c 1 min 55°c 1 min 72°c 2 min 72°c 10 min r-agagtgacgaggagc ggtag results and discussion collection of p. aeruginosa isolates a series of confirming tests were conducted to verify that out of 150 bacterial isolates only 40 belong to species of p. aeruginosa. these smear preparations of bacterial cells were gram-negative rods, non-spore forming, arranged in single or short chains. the colonies were thin, rough, or smooth on solid media with flat edges and high appearance, but some were mucoid in aspect. these isolates were found non-lactose ferment creating negative pale yellow colonies on macconkey agar and on blood agar shows β-hemolytic colonies. because of the production of the soluble pyocyanin and pyoverdin which are water soluble, the colonies were surrounded by bluish color on nutrient agar. the colonies pigments in selective media (cetrimide agar) are more apparent yellow-green pigment. biochemical tests confirmed p. aeruginosa burn contamination confines, biochemical testing was negative for indole, tsi, positive for oxidase and catalase, positive for citrate, positive for urease (slowly hydrolysis the urea), all p. aeruginosa 40 isolates had been also confirmed using vitek 2’s compact system bacterium id method. antimicrobial sensitivity screening test for p. aeruginosa forty p. aeruginosa isolates were screened for their resistance to (14) widely used antibiotics including amikacin, amoxicillin-clavulanic acid, ampicillin, cefotaxime, penicillin, ciprofloxacin, chloramphenicol, gentamycin, imipenem, meropenem, tetracycline, ceftazidime, aztreonam, and tobramycin. the results of antibiotic resistance pattern for the bacteria1 isolates understudy are shown in table 2. olayinka[16] reported that 20% of p. aeruginosa isolated from clinical samples obtained from the surgical units of ahmadu bello university teaching hospital in nigeria were sensitive for imipenem which disagreed with our results, imipenem and meronem are ßlactam antibiotics that they have broad-spectrum activity against both gram-negative and gram-positive bacteria.[17] all bacterial isolates displayed a low resistance and the majority of enterobacteriaceae isolates younis, et al.: antibiotic susceptibility, molecular characterization of virulence genes among p. aeruginosa 39 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 36-41 showed no resistance. it might be because they are reserve medicines and they are used as the last option in our hospital environment for multidrug-resistant bacteria which agreed with our result. ebrahimpour[18] reported that all p. aeruginosa isolated from burn patients were sensitive to imp. this may be attributed to the inability of p. aeruginosa to produce enzymes that degrade or inactivate the antibiotic. therefore, imp is the most effective drug for the treatment of infections caused by p. aeruginosa. in the case of fattma,[19] 98% of p. aeruginosa isolates resist amikacin, 96% for cefotaxime, 80% for rifampicin, 70% for ampicillin, 70% for augment, and 60% for doxycycline, which is near with our performance. resistance by p. aeruginosa can both be due to inducible of beta-lactamases, which can make cephalosporin of broad-spectrum inactive and to beta-lactamases mediated by plasmid, which can lead to several penicillin’s and ancient cephalosporin becoming resistant.[20] mechanisms of aminoglycoside resistance in clinical isolates are usually controlled by enzymatic antibiotic inactivation since nine different enzymes that are capable of catalyzing phosphorylation, acetylation, and aminoglycosides coradenylylation in bacteria had been described.[21] the development of p. aeruginosa multiresistant and its antibiotics mechanisms involves decreased cell permeability, efflux pumps, and changes in target enzymes and antibiotics inactivation.[22] detection of pvda, lasb, protease, exoa, exot, exou, and plch virulence genes by multiplex pcr in p. aeruginosa multiplex polymerase chain reaction (pcr) is a variant of pcr in which two or more loci are simultaneously amplified in the same reaction. since its first description in l988,[23] this method has been successfully applied in many areas of dna testing, including analyses of deletions,[24] mutations[25] and polymorphisms,[26] or quantitative assays[27] and reverse transcription pcr.[28] the role of various reagents in pcr has been discussed,[29] and protocols for multiplex pcr have been described by a number of groups. however, few studies[30] have presented an extensive discussion of some of the factors (e.g. primer concentration and cycling profile) that can influence the results of multiplex analysis. in the present study, 40 isolates of p. aeruginosa were tested for the detection of some virulence genes using polymerase chain reaction (multiplex). in our study, detection of virulence genes results showed that pvda was 55%, lasb 75%, protease 65%, exoa 60%, exot 75%, exou 60%, and plch 55% among tested strains, as shown in figures 1 and 2. other findings also showed that in 100 strains of p. aeruginosa, all the virulence genes studied were detected. therefore, the virulence genes studied might carry strains isolated from bovine meat, fresh fish, and smoked fish. the analysis revealed that the lasb genes are most frequently detected (89.0%) and exos genes (84.0%) which could be explained by the fact that p. aeruginosa, secrets elastase (lasb).[31] the previous studies showed a high lasb prevalence in p. aeruginosa despite its isolated origin.[32] another studies obtained by holban[33] also agreed with our results, who reported that lasb 55%, protease 75%, exot 95%, and plch 55%, these virulence genes were detected using multiplex pcr, in p. aeruginosa which isolated from wound secretions. mitov[34] also agrees with our results, who found that 100% for lasb and 71% for plch, the protease and lasb both genes encode for proteases activity, and they are found in the majority of tested strains, lending support to the phenotypic data demonstrating that isolates obtained from burned patients can undergo hemolysis. exos, exot, and exoa related exotoxins were distributed differently amongst the genes codified for the type iii secretion system (t3ss). the most positive for t3ss exotoxins which encode genes in isolates from burn patients were also followed by tracheobronchial isolates. exou, codified for a major enzyme involved in pyoverdine synthesis, codifies for a highly cytotoxic exoenzyme exou and pvda gene.[34] p. aeruginosa has been estimated to be involved in between 10% and 22.5% of hai both in adults and in children,[13] leading to increased costs for health care and prolonged hospital admission, respectively.[35] the clinical results of an infection with a combination of bacteria-related factors (intrinsic and table 2: percentage of resistance bacterial isolates to different antibiotics antibiotics symbo1 total no. of isolates no. of resistant isolates % of resistant amikacin ak 40 18 45 ampicillin am 40 40 100 amoxicillin-clavulanic acid amc 40 40 100 aztreonam atm 40 13 32.5 chloramphenicol c 40 40 100 ceftazidime caz 40 27 67.5 ciprofloxacin cip 40 25 62.5 gentamycin cn 40 18 45 cefotaxime ctx 40 40 100 imipenem imp 40 0 0 meropenem mem 40 17 42.5 penicillin p 40 40 100 tetracycline te 40 32 80 tobramycin tob 40 18 45 younis, et al.: antibiotic susceptibility, molecular characterization of virulence genes among p. aeruginosa 40 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 36-41 antimicrobial resistance, prevalence and persistence in the hospital environment, and cocktail expression of a virulence) and individual differences in host susceptibility. in favorable environmental conditions, bacterial virulence is reduced and greatly increased if stressful conditions arise.[36] conclusion p. aeruginosa showed resistance to most antibiotics, and imipenem was the most effective antibiotic against p. aeruginosa isolated from burn patients. seven virulence genes were detected through the amplification of pvda, lasb, protease, exoa, exot, exou, and plch by multiplex pcr. references 1. s. santajit and n. j. b. indrawattana. mechanisms of antimicrobial resistance in eskape pathogens. biomed research international, vol. 2016, p. 2475067, 2016. 2. k. singh, m. panghal, s. kadyan and u. chaudhary, j. p. j. yadav. antibacterial activity of synthesized silver nanoparticles from tinospora cordifolia against multi drug resistant strains of pseudomonas aeruginosa isolated from burn patients. journal of nanomedicine and nanotechnology, vol. 5, no. 2, p. 1, 2014. 3. g. héry-arnaud, e. nowak, j. caillon, v. david, a. dirou, k. revert, m. r. munck, i. frachon, a. haloun, d. horeau-langlard, j. le bihan, i. danner-boucher, s. ramel, m. p. pelletier, s. rosec, s. gouriou, e. poulhazan, c. payan, c. férec, g. 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and b. a. onile. antibiotic susceptibility and plasmid pattern of pseudomonas aeruginosa from the surgical unit of a university teaching hospital in north central nigeria. international journal of medicine and medical sciences, vol. 1, no. 3, pp. 79-83, 2009. figure 2: pcr amplification of plch, pvda, and protease virulence genes of p. aeruginosa through multiplex pcr, lanes 6 negative control, lanes 7, 8, 9, and 10 represent amplified genes with product size amplicon size (307 bp, 1281 bp, and 752 bp) of p. aeruginosa isolates, m representing the ladder 100 bp figure 1: pcr amplification of exot, exou, exoa, plch, pvda, lasb, and protease virulence genes of p. aeruginosa through multiplex pcr, lanes (l, 2, 3, 4, and 5) represent amplified genes with product size amplicon size (152 bp, 134 bp, 207 bp, 307 bp, 1281 bp, 300 bp, and 752 bp) of p. aeruginosa isolates, m representing the ladder 100 bp younis, et al.: antibiotic susceptibility, molecular characterization of virulence genes 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chifiriuc, a. i. cotar and c. bleotu. virulence markers in pseudomonas aeruginosa isolates from hospital acquired infections occurred in patients with underlying cardiovascular disease. vol. 18, no. 6, pp. 8843-8854, 2013. 34. i. mitov, t. strateva and b. markova. prevalence of virulence genes among bulgarian nosocomial and cystic fibrosis isolates of pseudomonas aeruginosa. the brazilian journal of microbiology, vol. 41, no. 3, pp. 588-595, 2010. 35. d. jr. hayes, s. e. west, m. j. rock, z. li, m. l. splaingard and p. farrell. pseudomonas aeruginosa in children with cystic fibrosis diagnosed through newborn screening: assessment of clinic exposures and microbial genotypes. pediatric pulmonology, vol. 45, no. 7, pp. 708-716, 2010. 36. m. ledizet, t. s. murray, s. puttagunta, m. d. slade, v. j. quagliarello and b. kazmierczak. the ability of virulence factor expression by pseudomonas aeruginosa to predict clinical disease in hospitalized patients. plos one, vol. 7, no. 11, p. e49578, 2012. . 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 57-61 research article radish juice promote kidney stone deposition in ethylene glycol-induced urolithiasis in rats falah m. aziz1*, dlshad h. hassan2 1department of biology, college of science, salahaddin university-erbil, erbil, iraq, 2department of biology, faculty of science, soran university, soran, iraq abstract urolithiasis is a well-known problem that stones could form in various parts of the urinary system and it is the most common disease of the urinary tract. the current study was planned to investigate the effect of radish juice on ethylene glycol (eg)-induced urolithiasis. twenty-one rats randomly divided into three groups. the first group was the control group was received normal standard diet and drinking water and the second group represented the model group received 0.75% eg in drinking water ad libitum. the third group received radish juice (2 ml/kg of body weight) by gavage plus eg (0.75%) in drinking water ad libitum. the experiment was conducted for 28  days. the light microscope examination revealed a disturbed histological architecture of the kidney tissues, including dilated renal tubules, aggregation of infiltrated leukocytes inflammatory cells, and crystal deposition in the model group. the eg plus radish juice treated rats showed higher crystal density with improved renal tubule structure and alleviated inflammation. both treated groups showed various biochemical alterations compared to control group, but the most interest biochemical result was the significant decrease of malondialdehyde, a lipid peroxidation marker, and in the radish plus eg group compared to the eg group. scanning electron microscopy showed clear structural detail about calcium oxalate crystals in which radish-treated group showed higher crystal deposition and calcified tissue compared to eg group. the present study concluded that radish juice promotes stone deposition but exerted an antioxidant effect. keywords: ethylene glycol, kidneystone, radis, urolithiasis introduction urolithiasis is a well-known problem that stones could form in various parts of the urinary system.[1] mankind has been afflicted by urinary stones for centuries dating back to 4000 b.c. and it is the most common disease of the urinary tract.[2] urolithiasis affects about 12% of the world population at some stage in their lifetime.[3] urolithiasis is a complex process that occurs due to imbalance between promoters and inhibitors.[4] the chemicals that can promote stone formation are calcium, sodium, oxalate, urate, and cystine, while magnesium, citrate, and pyrophosphate can inhibit the urolithiasis process.[5] calcium containing stones are the most common renal stones with a percentage of 80% of cases.[6] the process of stone formation begins with supersaturation that is the driving force for crystallization in solutions like urine.[7] as a result of supersaturation, solutes precipitate in urine leads to nucleation and then crystal growth. crystallization happens when chemical concentrations exceed their point of saturation.[8] then after, small hard mass of crystals sticks to each other to form a larger stone that is called crystal aggregation.[7] plants such as oxalis corniculata,[9] phyllanthus niruri,[10] bergenia ligulate,[11] corresponding author: falah m. aziz, department of biology, college of science, salahaddin university-erbil, erbil, iraq. e-mail falah.aziz@su.edu.krd received: apr 23, 2019 accepted: apr 25, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp57-61 copyright © 2020 falah m. aziz, dlshad h. hassan. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) and nigella sativa[12] have been used as a remedy for renal calculi. radish (raphanus sativus) is a member of the family cruciferae. this family consists of annual, biennial, or perennial herbs with pungent oils in the sap. there are about 380c genera and about 3000 species.[13] varieties of radish are now broadly distributed around the world, but there are almost no archeological records available to determine its early history and domestication.[14] radish has been shown medicinal benefits such as gastroprotective effects[15] as a remedy for diabetes treatment[16] and antimicrobial activity.[17] the present study has been designed to evaluation of protective effect of radish juice on renal stone. https://creativecommons.org/licenses/by-nc-nd/4.0/ aziz and hassan: radish juice promote kidney stone deposition 58 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 57-61 materials and methods experimental animals and radish juice preparation for conducting this experiment, 21 mature male rats were bred under standard animal house conditions. for radish juice preparation, radish obtained from local market in erbil city, systematic identity confirmed in college of science/ salahaddin university, washed, cut to small pieces, and homogenized then filtered by gauze. the rats were divided randomly into three groups; each group consisted of seven rats per cage group 1, control, the rats of this group were given standard rat chow and tap water for 28 days. group 2, ethylene glycol (eg)-induced urolithiasis model. the rats of this group were given standard rat chow and eg (0.75%) in drinking water ad libitum for 28 days. group 3, radish, the rats received standard rat chaw, radish juice (2 ml/kg of body weight) by gavage, and 0.75% eg in drinking water ad libitum for 28 days. the animals were anesthetized by intraperitoneal injection of combination ketamine hydrochloride 80 mg/kg (trittau, germany) and xylazine 12 mg/kg (interchem, holland). after sacrification, kidneys removed then fixed in desired fixative according to the type of microscopical preparation. kidney weight recording both kidneys have removed, weighted, and expressed as gram per 100 g of body weight. urine flow animals remained in urine collector and urine was collected for 24 h then urine flow measured as ml/h/kg. fluid intake fluid intake recorded every 4 days during the experiment. blood collection at the end of the treatment period, blood samples were collected from all anesthetized rats of four groups through cardiac puncture in which the collected blood samples were immediately placed into gel tube for serum collection, later were centrifuged (hettich d-78532/germany) at 3000 rpm for 15 min. the sera were stored at −80°c (sanyo – ultra-low temperature, japan) until assayed. serum urea determination urine and serum urea was determined by urea kit (biolabo, france). enzymatic and colorimetric method of urea determination is based on hydrolysis of urea to ammonium and carbon dioxide by urease. ammonium reacts with chloride and salicylate and makes a blue-green complex that is proportional to urea concentration and measured at 600 nm. estimation of creatinine and glomerular filtration rate serum-urine creatinine was determined spectrophotometrically using biolabo kit (france). figure 1: left (a) and right (b) kidney weight per 100 g of body weight. different letters on bars mean significant change and the same letters means no significant change figure 2: fluid intake (a) and urine flow (b) of control and ethylene glycol treated groups. different letters on bars mean significant change and the same letters means no significant change a b a b aziz and hassan: radish juice promote kidney stone deposition 59 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 57-61 figure 4: level of tissue malondialdehyde in control, model, and ethylene glycol +radish treated groups. different letters on bars mean significant change and the same letters means no significant change creatinine reacts with picrate to form a colored complex in alkaline solution. absorbency of samples was read at 500 nm. determination of kidney tissue malondialdehyde (mda) the right kidney of each rat was washed in ice-cold normal saline solution then homogenized in 20 mm phosphate buffer (ph = 7.4; tissue/buffer ratio, 1/10 w/v) by handheld glass homogenizer (chowdhury et al., 2013). homogenates were centrifuged at 4000 g at 4°c for 10 min (beckman j2-21). the supernatants were collected and stored at −8°c until assayed. mda level was estimated according to method of kartha and krishnamurthy (kartha and krishnamurthy, 1978). 1 ml of tissue homogenate was added to 20% trichloroacetic acid. after centrifugation for 10 min, 2 ml of supernatant was taken in a test tube and 2 ml of 0.7% thiobarbituric acid was added to each tube and kept it in the boiling water bath for 20 min. the development of pink color was measured at 535 nm and mda concentration was calculated by the following equation: mda nmol/g of tissue = absorbance at 535 nm×d/ (l×eo) l: lightpath (1 cm) e o: extinction coefficient 1.56×105 m−1.cm−1 d: dilution factor. light microscopy kidneys were removed from the anesthetized animals, immediately fixed in bouin’s solution for 24 h followed by dehydration using a series of ethanol in ascending concentrations (50%, 70%, 95%, and 100%), then immersed in xylene for clearing process, infiltrated with paraffin wax, and embedded in paraffin wax. 5 µm thick paraffin sections were obtained using rotary microtome (bright, mic) and stained by hematoxylin and eosin. the specimens were examined and photographed under light microscope (digital binocular compound microscope ×40–×2000, built-in 3mp usb camera). figure 3: serum creatinine (a) and serum urea (b) of control and ethylene glycol treated groups. different letters on bars mean significant change and the same letters means no significant change a b figure 5: kidney sections of control group (a) show normal appearance(g): glomerulus.×400. hematoxylin and eosin (h&e). (b) kidney sections of model group (b) show a lot of crystal deposition (c) ×400. h&e. kidney section of radish treated group (c and d) shown inflammatory cells and crystal deposition (c) ×400. h&e a b c d aziz and hassan: radish juice promote kidney stone deposition 60 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 57-61 radish group leading to more kidney damage, this may explain the higher serum urea noticed in this group in comparison to the control group. furthermore, urea is by product of protein metabolism and this rising in serum urea may return to the fact that radish itself contain total protein of 6.5%.[22] as shown in figure 4, the level of mda in model group raised when compared with control group while there was no significant difference between control and radish treated group. mda is by product of lipid peroxidation and is used as marker for this process and oxidative stress induced by eg through elevating mda level confirmed previous findings.[23,24] under hyperoxaluria, oxalate reacts with polyunsaturated fatty acid in cell membrane[25] and make damage. calcium oxalate which induced oxidative stress can help crystal attachment to epithelial cells of nephrons (cellcrystal attachment).[26] radish juice succeeded in lowering the mda level compared with eg group and it may be due to the antioxidant contents of radish since it has been found to contain phenolic compounds and ascorbic acid.[27] paraffin sections of kidney belong to control group (figure 5a) show normal and healthy appearance. kidney sections of model group have shown crystal deposition and dilated tubules [figure 5b] that is match with finding of the previous study.[28] inflammatory cells (ic) accumulation also observed that is maybe due to ability of calcium oxalate to triggering immune system.[29] oxalate, the major stone-forming constituent, has been reported to induce lipid peroxidation and causes tissue damage by reacting with polyunsaturated fatty acids in cell membrane.[23] on the other hand, some investigations say that lipid peroxidation is not the underlying cause of renal injury in hyperoxaluric rats.[30] paraffin section through the kidney of eg + radish juice treated rats has shown more quantity of crystal deposited in lumen of kidney tubules [figure 5d] and a lot of inflammatory leucocytes were observed in kidney tissues [figure 5c]. radish has been found to contain four major oxalic, malic, malonic, and erythroic acid.[31] oxalic acid itself is one of the promoters of renal stone formation. despite promoter activity of oxalic acid, total acidity of these four acids may decrease urine ph and makes a favor environment for calcium oxalate crystals. scanning electron microscopy showed the three-dimensional image of crystals more clearly in comparison to other techniques. the kidneys in the control group were appeared with normal and healthy structure having no crystals [figure 6a]. clear large electron microscopy scanning electron microscopy was done at the university of niece, france. kidneys were fixed in 2.5% glutaraldehyde in 0.1m cacodylate buffer ph 7.2–7.4. after washing dehydrated in ethanol (50%, 70%, 85%, 100%, and 100%), the samples were put in desiccator for air drying; after mounting, they were coated by coater machine with gold and then examined by scanning electron microscope (sem). statistical analysis data entry was done by microsoft excel 2010 and analyzed statistically by one-way analysis of variance using graphpad prism version 6.01 and spss version 20 and data expressed as mean ± standard error. newman–keuls tests were chosen as post hoc tests. results and discussion as shown in figure 1, both the left and right kidneys weight in model and radish-treated group increased significantly when compared with control group. although many cells may degenerate in response to eg toxicity or to the formation and presence of crystals in the kidney, the increase in kidney weight in eg and eg-treated plant juice was expected at least due to the presence of renal crystals.[18] fluid intake [figure 2a] in eg and radish-treated groups elevated in compare of control group. increasing fluid intake in eg-induced group maybe return to sweet taste of eg[19] or nephron damage that could not reabsorb glomerular filtrate again. as illustrated in figure 2b, urine flow in model group increased significantly when compared to control group; however, there is no statistical difference between model and radish-treated group. with respect of serum creatinine, figure 3a shows significant raising in eg and radish-treated groups when compared to control group, while there is no statistical difference between both eg-treated groups. the elevation of serum creatinine may be due to renal tubules obstructions.[20] figure 3b revealed that there was no statistical difference of serum urea between control and model group and between model and radish treated group but serum urea elevated in radish group when compared to control group. increasing of serum urea is related to the level of nephron injury[21] and since more crystals were deposited in the kidney of eg plus figure 6: scanning electron microscopy of kidney (a) control group showing normal appearance (b) ethylene glycol (eg)-treated group showing crystal deposition (white arrow) and calcified tissue (blue arrow). (c) eg + radish treated group showing more crystal deposition (white arrow) and calcified tissue (elbow arrow) a b c aziz and hassan: radish juice promote kidney stone deposition 61 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 57-61 crystals were revealed in the kidney tissue of eg-treated rats [figure 6b]. sem of rat kidneys belong to eg + radish juice treated group showed crystals deposition in wide distribution and in the most area, the tissue appeared calcified [figure 6c]. conclusion radish treated group has shown more crystal deposition, damage, and ic in compare to model; however, radish juice decreased mda level significantly. radish juice could not improve the state of experimental rats in kidney weight, renal function tests, and fluid intake. acknowledgment the authors would like to appreciate mr. sarwan barzani for his supports and the assistance of miss chnar najmaddin in light and electron microscopy preparation. references 1. k. kaczmarek, a. gołąb, m. soczawa and m. słojewski. “urethral stone of unexpected size: case report and short literature review”. open medicine, vol. 11, pp. 7-10, 2016. 2. m. lópez and b. hoppe. “history, epidemiology and regional diversities of urolithiasis”. pediatric nephrology, vol. 25, p. 49, 2010. 3. c. chauhan, m. joshi and a. vaidya. “growth inhibition of struvite crystals in the presence of herbal extract commiphora wightii”. journal of materials science: materials in medicine, vol. 20, p. 85, 2009. 4. k. g. ingale, p. a. thakurdesai and n. s. vyawahare. “effect of hygrophila spinosa in ethylene glycol induced nephrolithiasis in rats”. indian journal of pharmacology, vol. 44, p. 639, 2012. 5. d. r. basavaraj, c. s. biyani, a. j. browning and j. j. cartledge. “the role of urinary kidney stone inhibitors and promoters in the pathogenesis of calcium containing renal stones”. eau-ebu update series, vol. 5, pp. 126-136, 2007. 6. f. l. coe, a. evan and e. worcester. “kidney stone disease”. the 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sharma, w. khan, r. parveen, m. j. alam, i. ahmad, m. h. ansari and s. ahmad. “antiurolithiasis activity of bioactivity guided fraction of bergenia ligulata against ethylene glycol induced renal calculi in rat”. biomed research international, vol. 2017, p. 1-11, 2017. 12. p. hayatdavoudi, a. k. rad, z. rajaei and m. a. r. hadjzadeh. “renal injury, nephrolithiasis and nigella sativa: a mini review”. avicenna journal of phytomedicine, vol. 6, p. 1, 2016. 13. j. glimn-lacy and p. b. kaufman. “botany illustrated: introduction to plants, major groups, flowering plant families”. new york: springer science and business media, 2006. 14. d. zohary, m. hopf and e. weiss. “domestication of plants in the old world: the origin and spread of domesticated plants in southwest asia, europe, and the mediterranean basin”. oxford: university press on demand, 2012. 15. s. alqasoumi, m. al-yahya, t. al-howiriny and s. rafatullah. “gastroprotective effect of radish raphanus sativus l. on experimental gastric ulcer models in rats”. farmacia, vol. 56, p. 204, 2008. 16. s. a. banihani. “radish (raphanus sativus) and diabetes”. nutrients, vol. 9, p. 1014, 2017. 17. s. s. beevi, l. n. mangamoori, v. dhand and d. s. ramakrishna. “isothiocyanate profile and selective antibacterial activity of root, stem, and leaf extracts derived from raphanus sativus l”. foodborne pathogens and disease, vol. 6, pp. 129-136, 2009. 18. l. m. besenhofer, m. c. cain, c. dunning and k. e. mcmartin. “aluminum citrate prevents renal injury from calcium oxalate crystal deposition”. journal of the american society of nephrology, vol. 23, pp. 2024-2033, 2012. 19. j. brent. “current management of ethylene glycol poisoning”. drugs, vol. 61, pp. 979-988, 2001. 20. h. han, a. m. segal, j. l. seifter and j. t. dwyer. “nutritional management of kidney stones (nephrolithiasis)”. clinical nutrition research, vol. 4, pp. 137-152, 2015. 21. j. ran, j. ma, y. liu, r. tan, h. liu and g. lao. “low protein diet inhibits uric acid 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park, m. linnes, h. j. han, j. h. kim, j. c. lieske and s. h. kim. “gallotannin suppresses calcium oxalate crystal binding and oxalate-induced oxidative stress in renal epithelial cells”. biological and pharmaceutical bulletin, vol. 35, pp. 539-544, 2012. 27. r. goyeneche, s. roura, a. ponce, a. vega-gálvez, i. quispefuentes, e. uribe and k. di scala. “chemical characterization and antioxidant capacity of red radish (raphanus sativus l.) leaves and roots”. journal of functional foods, vol. 16, pp. 256-264, 2015. 28. d. h. hassan and f. m. aziz. “histological and ultrastructural study on the effect of celery juice on ethylene glycol induced urolithiasis in male albino rats”. zanco journal of pure and applied sciences, vol. 29, pp. 1-12, 2017. 29. s. r. mulay, o. p. kulkarni, k. v. rupanagudi, a. migliorini, m. n. darisipudi, a. vilaysane, d. muruve, y. shi, f. munro and h. liapis. “calcium oxalate crystals induce renal inflammation by nlrp3-mediated il-1β secretion”. the journal of clinical investigation, vol. 123, pp. 236-246, 2012. 30. m. l. green, r. w. freel and m. hatch. “lipid peroxidation is not the underlying cause of renal injury in hyperoxaluric rats”. kidney international, vol. 68, pp. 2629-2638, 2005. 31. r. m. p. gutiérrez and r. l. perez. “raphanus sativus (radish): their chemistry and biology”. the scientific world journal, vol. 4, pp. 811-837, 2004. . 64 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 64-69 research article simulation-based evaluation of mobile ad hoc network routing protocols: ad hoc on-demand distance vector, fisheye state routing, and zone routing protocol mohamad t. sultan*, khaled n. yasen, ali q. saeed department of computer science, cihan university-erbil, kurdistan region, iraq abstract mobile ad hoc network (manet) is an infrastructure-less and decentralized network without any physical connections. nodes are mobile, free to move, and independent of each other which makes routing a difficult task. hence, a dynamic routing protocol is needed to make manet reliable and function properly. several routing protocols have been proposed with different working mechanisms and performance levels. therefore, the performance study of those protocols is needed. this paper evaluates the performance of manet routing protocols using simulation based experiments to observe the behavior of the network as the density of the nodes increases. the paper evaluates the performance of proactive (fisheye state routing), reactive (ad hoc on-demand distance vector), and hybrid (zone routing protocol) routing protocols in terms of the packet delivery fraction, average throughput, and average end-to-end delay. the simulations of protocols to analyze their performance in different conditions were performed using the network simulator 2 (ns 2). keywords: mobile ad hoc network, routing protocols, wireless networks introduction today, mobile wireless networking is one of the most innovative topics in computer technologies.[1,2] there are two types of the wireless network. the first type is called an infrastructure network. in an infrastructure network, there exists a device called a base station and all mobile devices within the area will communicate with the base station. the base station, in turn, is connected to the wired network. the second type of wireless network is called the ad hoc network.[3] mobile ad hoc network (manet) is a set of nodes, which are able to link on a wireless medium forming an arbitrary and dynamic network. the ad hoc network which is also called infrastructure-less network is deployed in places where there is no availability of network infrastructure. the ad hoc network typically consists of nodes that are portable devices operating in a dynamically changing topology. the base station in such a network is not available. all the mobile devices connect and communicate with each other dynamically without the assistance of fixed devices. the wireless links are the means by which the mobile nodes get connected with each other in an arbitrary manner. in such network, all nodes play an important role in route discovery and maintenance, and at the same time, each mobile node acts as a router where it has the ability to forward data packets as necessary from the source to the destination. in wireless ad hoc network, the mobile nodes are free to leave and join the network at any time without any kind of restriction on their behavior. this is results in a rapid change in the network’s interconnections and topology.[4] manet has many features, and due to its flexibility, it has become suitable to be used in emergency rescue operations, military operations, and surveillance tasks. however, at the same time, it is not easy to handle the operation in the ad hoc networks because of the irregular change in the network’s structure. given this dynamically changing environment, links can become unreliable which creates further challenges for those ad hoc networks. this warrant the need for efficient routing protocol specially designed to handle the dynamic topology of manet. the main issue of routing protocols in manet is how to send data packets from one node to another when there is no direct link exists between the source and destination nodes. this resulted in many routing protocols being developed for manet; these protocols are implemented with some basic objectives. some of these protocols are suitable for limited network’s nodes, and some of them deal with high power consumption or low bandwidth or high error rate.[5] corresponding author: mohamad t. sultan, cihan university-erbil, kurdistan region, iraq. e-mail: mohamad.taha@cihanuniversity.edu.iq received: mar 31, 2019 accepted: apr 21, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp64-69 copyright © 2019 mohamad t. sultan, khaled n. yasen, ali q. saeed. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ sultan, et al.: simulation-based evaluation of mobile ad hoc network routing protocols: ad hoc on-demand distance vector, fisheye state routing, and zone routing protocol 65 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 64-69 the rest of the paper is organized as follows: section 2 describes the studied routing protocols, and section 3 gives the details of the simulation environment and the implementation of the routing protocols and the simulation setup used in this research. the simulation results are presented in section 4 and finally, in section 5, we conclude the paper and describe our future scope. the routing protocols in manet routing is a process to choose or to find a path from the source to the destination within the network.[6] the network routing is being used in numerous types of networks, such as electronic data, internet, and telephone networks. in the ad hoc networks, the concept of routing refers to the technique of choosing the correct path to deliver packets from the source to the destination. throughout the process of routing, there will be at least one intermediate node within the network is encountered. the two main activities that have to be mentioned about routing are how to determine the optimal path for routing and how to transfer the data packets throughout the network. moreover, manet routing protocols have to preserve the processing and communication time spent to the lowest possible, and these protocols have to adapt and get used to the unpredictable sudden changes in the network topology. routing protocols in manet can generally be categorized into three approaches, namely, (a) table-driven routing protocols (proactive), (b) on-demand routing protocols (reactive), and (c) hybrid routing protocols.[7] this is shown in figure 2. table 1 presents a categorization of manet routing protocols studied in this research. in the following sub-sections, the three different routing approaches will be discussed. proactive routing protocols in the proactive routing protocols, each node maintains routing information for every other node in the network. the routing information is often sheltered in a number of different routing tables.[7] the tables must be updated as the network topology is changing dynamically. the routing protocol adapts to the sudden changes in topology by broadcasting network updates whenever changes occur. fisheye state routing (fsr), destination sequence distance vector, and optimized link state routing are examples of proactive routing protocols.[8] fsr the fsr protocol[9] is one of the table-driven routing protocols based on link state routing algorithm and uses a hierarchical routing scheme. the mobile nodes refresh the information about the destinations by repeatedly update their routing tables. the main aim of the fisheye approach is to decrease the volume of the information required to represent data using the fisheye technique. the link state packets in the fsr are not flooded. instead, there will be a link state table maintained by the mobile nodes based on the updated information received from neighboring nodes, and this table is exchanged periodically only with their local neighbors without any flooding.[8,9] the fisheye routing is able to see objects when they are close to its focal point in a better way rather than seeing these objects when they are far which means each node keeps precise data about near neighborhood nodes and not so precise data (less detail) about the nodes which are not near or distant. the scope of fisheye is described as a group of nodes which might be reached within a specified number of hops. the fsr has the ability to minimize the bandwidth used to transmit link state update packets between surrounding nodes, and it attempts to decrease the size of the message belongs to topology information because of the exclusion of topology information about far nodes.[9] figure 3 below illustrates how the fisheye technique is applied to a manet. in figure 3, three scopes of fisheye routing are defined with regard to node number 11 which is the focal point. a group of nodes for each scope is defined which can be reached using a particular number of hops. the fsr has the ability to minimize the bandwidth used to transmit link state update packets between surrounding nodes, and it attempts to decrease the size of the message belongs to topology information because of the exclusion of topology information about far nodes. figure 1: example of mobile ad hoc network figure 2: mobile ad hoc network protocols table 1: categorization of manet routing protocols selected manet routing protocols table-driven routing protocols (proactive) on-demand routing protocols (reactive) hybrid routing protocols fisheye state routing ad hoc on-demand distance vector zone routing protocol manet: mobile ad hoc network sultan, et al.: simulation-based evaluation of mobile ad hoc network routing protocols: ad hoc on-demand distance vector, fisheye state routing, and zone routing protocol 66 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 64-69 reactive routing protocols the on-demand or reactive routing approach is the second and most prominent category of routing protocols in manet. these routing protocols choose routes to other nodes only when they are needed. when a node in the network wants to communicate with another node, a route discovery process is launched. the route discovery process helps the communicating nodes to collect routing information to find the best possible path between the source and destination inside the network. the ad hoc on-demand distance vector (aodv), dynamic source routing, and the temporally ordered routing algorithm are the examples of reactive routing protocols.[8] aodv aodv is a reactive protocol.[10] it uses the hop-by-hop methodology for routing the data. nodes in aodv maintain a route which includes only the next hop routing information for the destination nodes. in aodv, only the active network routes are maintained. when there is a route needed in the network, aodv will execute a route discovery process to find the route to the desired destination. as soon as a route is generated in the network, it is maintained as long as it is still needed using a route maintenance process. the routing information in aodv routing protocol is preserved in routing tables at the mobile nodes. one characteristic of aodv is that it aids nodes in the network by responding quickly to any change in topology and to link failure, and this protocol works in loop-free manner. there are three different types of messages used in aodv which are the route request (rreq), route reply (rrep), and route error (rerr).[11] using those main routing messages, aodv organizes its work as an on-demand routing protocol in ad hoc networks. in case a mobile node in the ad hoc network demands to communicate with another node, it first searches through its existing routing information for a valid route to the other node. if one route is present there the node uses that existing route for communicating with the destination node. otherwise, the node broadcasts a rreq message to its neighbor’s nodes throughout the network to discover a route to the destination. this rreq message contains information such as the ip address of the source and destination nodes, current sequence number, broadcast id, and latest sequence number for the destination known to the source node. then, the intermediate nodes start forwarding the rreq. when the nodes which are either a destination node or one of the intermediate nodes receive the request, it establishes a rrep which includes information that the source node needs with the valid route. if in case any problem happens in the network or link failure is discovered, the protocol will initiate a rerr message. this message will be sent throughout the network with a list of all unreachable destinations.[12] figure 4 represents a rreq broadcast in aodv routing protocol. hybrid routing protocols the hybrid approach in manet is very popular and over time has gained a wide-spread acceptance and applied in various ad hoc networks.[13] in the hybrid approach, the route starts with a simple proactive mode but later uses the reactive flooding to support the request from the mobile nodes in the network. hybrid routing tries to offer a way to minimize the inefficiencies of manet routing. however, the performance evaluation and the implementation in practical conditions of the hybrid routing are still an on-going process. the network in the hybrid routing approach is initially divided into different zones to reduce overhead. the proactive approach is applied when routing to another node in the same zone or when nodes communicate with their neighbors while the use of reactive routing approach is mainly for the purpose of routing to far distance nodes which are located outside the zone area. the hybrid protocols are designed to conform and adapt to any random arbitrary ad hoc network. the most popular protocol that falls under this category is the zone routing protocol (zrp).[14,15] zrp zrp is a proactive or reactive protocol (hybrid routing) that divides the network into zones.[15] zrp tries to address the problems of the proactive and reactive routing by exploiting the preferable properties and eliminate the weaknesses of both approaches. zrp takes advantage of proactive detection within a node’s local neighborhood and applying a reactive protocol for the contact between these neighborhoods. the proactive one that controls and monitors the local zones which are used within routing zones is called intra-zrp (iarp), and the reactive one that controls and monitors the global zones which are used between the routing zones is called inter-zrp (ierp). in the case when the source and destination happen to be in the same zone, the data packets can be transferred directly. this happens because the establishment of a path to a figure 3: fisheye scope figure 4: route request broadcast sultan, et al.: simulation-based evaluation of mobile ad hoc network routing protocols: ad hoc on-demand distance vector, fisheye state routing, and zone routing protocol 67 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 64-69 target node inside the local zone can be done using the stored routing table of the source by iarp. nearly, all of the current proactive systems can be applied as iarp in zrp. the process of determining and finding a route occurs reactively for routes which may be located out of the range of a specific local zone. this happens when the source node initiates a request message to the border nodes of the zone. the message carries the source node’s address, the destination node’s address, and a special sequence number. the borders investigate their local zone for the target to check its availability. if the destination is not available or not found, the node tags to the request packet the current address that it possesses and sends that packet to its borders again as this target is not a member of the current local zone. if it happens that the target node is available and found, it sends back a rrep on the reverse direction to the source as this target is a member of the current local zone. the origin node benefits from the path kept in the reply packet to transfer data packets to the target. simulation setup mobility model the mobility model must attempt to mimic the actions and movements of real mobile nodes. for this research study, the selected mobility model is the random waypoint mobility model which is one of the most widely used mobility models among the research community.[16] this mobility model can highly represent the actions and movements of real mobile nodes in real conditions. simulation parameters in the simulation, various parameters have been defined to analyze the comparative performance of proactive fsr, reactive aodv, and hybrid zrp routing protocols. the configured simulations parameters are shown in table 2. ns-2 simulator the network simulator (ns2) is a discrete-event driven simulation software developed as part of the vint project at the university of california in berkeley and targeted for network simulation.[17] the ns2 simulation project is mainly funded and supported by darpa in cooperation with xerox palo alto research center and lawrence berkeley national laboratory. among networking community, the performance of ns2 as a simulator is greatly trusted and it is an effective standard in simulations. performance metrics to study the routing protocols (aodv, fsr, and zrp), we selected three performance metrics for evaluation which are, packet delivery fraction, average end-to-end delay, and average throughput. packet delivery fraction the packet delivery fraction is known as the ratio of successfully delivered data packets to destination nodes over the total number of data packets produced for those destinations. the packet loss ratio is defined as the packet delivery fraction. average end-to-end delay the average delay in transmission of a data packet between two nodes from a source to destination in the network is known as the average end-to-end delay. this indicates the time that a data packet utilizes to move from the source node to the application layer of the destination node. average throughput the throughput of the network denotes the average rate of successful message delivery over a communication channel. it is the average number of packets successfully obtained their destinations per unit time. the throughput is usually measured in bits per second (bit/s or bps). simulation results this research consists of three main experiments that have been conducted for the aim of an effective evaluation study regarding three different protocols of manet networks under different environmental conditions by the varying the number of nodes. the results after simulation are viewed in figures. packet delivery fraction figure 5 shows the comparison between the simulated routing protocols (fsr, aodv, and zrp) on the basis of packet delivery fraction as an evaluation metric. from the results in figure 5, it can simply be noticed that almost all routing protocols performed relatively well in small mnaet, less number of nodes (20 nodes). this due to the fact that only a few hops need to be taken by the packets to reach the destination node. it can also be observed that as the manet size grows more and the scenario become more stressful, all routing protocols experience lower performance. however, the reactive routing protocol (aodv) has performed better and table 2: simulations parameters simulation parameters parameters values platform linux (ubuntu) 10.04 simulation tool network simulator 2 routing protocols fsr, aodv, zone routing protocol pause time 10 s experiment duration 200 s number of nodes 20, 40, 60 traffic model constant bit rate packet size 512 bytes packet rate 2 packets/s area 500 m×500 m maximum speed 20 m/s mobility model random waypoint mac layer protocol ieee 802.11b antenna type antenna/omni antenna sultan, et al.: simulation-based evaluation of mobile ad hoc network routing protocols: ad hoc on-demand distance vector, fisheye state routing, and zone routing protocol 68 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 64-69 delivered more packets than the proactive (fsr) and hybrid (zrp) counterparts. it can be noticed that the performance of the aodv routing protocol decreases only slightly with an increase in manet size. this is due to the behavior of reactive protocols which always try to guarantee that the packets will be delivered to the destination even if that may cause some delays. therefore, it is the most appropriate consideration for a network if data delivery is of the highest consideration. average end-to-end delay figure 6 shows the simulation results of the average end-to-end delay in milliseconds. the simulation results indicate that all routing protocols experience higher delay values as the number of wireless nodes or network size grows. this is due to the fact that when manet size grows; there will be more broken routes. as those routes break, nodes have to discover new routes which make it possible to have longer end-to-end delays. according to simulation results, the proactive fsr routing protocol has performed well having the lowest end-to-end delay compared to other protocols in this simulation scenario. the reason behind that could be due to the proactive nature of the fsr where the routes for all the destinations are preserved in routing tables and there is no need to initiate the rreq procedure more frequently. therefore, it has less network delay. the performance of the reactive routing protocol (aodv) is almost uniform in this simulation scenario. the results also indicate that the zrp routing protocol did not perform well. it has higher values of average end-to-end delay, especially with a higher number of nodes. average throughput figure 7 illustrates the average throughput results of the comparison between the three simulated routing protocols (fsr, aodv, and zrp). in this scenario, a significant difference in throughput can be observed. this is because of the increase in the number of mobile nodes which results in higher load in the network. the on-demand reactive aodv routing protocol has achieved a higher throughput compared to other protocols. the throughput of aodv seems to be more stable and consistent as the number of nodes increases. among the three routing protocols, fsr has the worst performance. the performance of fsr degrades with a higher number of nodes and it can be noticed that fsr is nowhere near the other protocols in terms of consistency. overall, it can be claimed that the best performance achieved is by aodv closely followed by zrp routing protocol. conclusion manet is an infrastructure-less network with a dynamic nature. nodes are mobile and independent of each which triggers the need for a reliable routing protocol to make the network function properly. several routing protocols have been developed by a researcher, yet choosing a reliable protocol could be a challenging task. this research, study the performance of proactive routing mechanism in manet and compare it with the reactive and hybrid counterparts, to evaluate the strengths and weaknesses of each of these mechanisms. this study indicates that the routing protocols play a prominent role in developing better communication between nodes in the network. it observed generally that increasing number of nodes results in decline in performance for all routing protocols. according to our simulation results, the reactive routing protocol (aodv) has demonstrated the first best performance and outperformed fsr and zrp in terms of packet delivery fraction and average throughput, figure 5: packet delivery fraction versus number of nodes figure 6: average end to end delay versus number of nodes figure 7: average throughput versus number of nodes sultan, et al.: simulation-based evaluation of mobile ad hoc network routing protocols: ad hoc on-demand distance vector, fisheye state routing, and zone routing protocol 69 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 64-69 while the hybrid routing protocol zrp has presented an overall average performance except it has suffered from delays, especially under a high number of nodes scenario. fsr has shown the lowest end-to-end delay as a proactive routing protocol. thus, in cases where the delay is a major concern in the ad hoc network, this table-driven protocol could be a good compromise. however, fsr performance in terms of packet delivery fraction and throughput was poor. for future works, other aspects of routing protocols in manet such as privacy and security will be considered. references 1. n. sze-yao, t. yu-chee, c. yuh-shyan and s. jang-ping. “the broadcast storm problem in a mobile ad hoc network”. proceedings of the 5th annual acm/ieee international conference on mobile computing and networking. acm, 1999. 2. s. t. mohamad and k. n. yasen. “homomorphic encryption implementation to ensure data security in cloud computing.” journal of theoretical and applied information technology, vol. 96, no. 7, pp. 1826-1836, 2018. 3. k. young-bae and n. h. vaidya. “location-aided routing (lar) in mobile ad hoc networks.” wireless networks vol. 6, no. 4, pp. 307-321, 2000. 4. u. muhammad, m.a. jan, x. he and p. nanda. “qasec: a secured data communication scheme for mobile ad-hoc networks”. future generation computer systems, vol. 93, pp. 737-750, 2018. 5. a. mehran, t. wysocki and e. dutkiewicz. “a review of routing protocols for mobile ad hoc networks.” ad hoc networks, vol. 2, no. 1, 1-22, 2004. 6. r. stefano, k. krużelecki, g. heitz, d. floreano and b. rimoldi. “dynamic routing for flying ad hoc networks.” ieee transactions on vehicular technology, vol. 65, no. 3, pp. 1690-1700, 2016. 7. h. alex, m. ngulube, s. zhu and h. al-aqrab. “a review of routing protocols for mobile ad-hoc networks (manet)”. international journal of information and education technology, vol. 3, no. 1, p. 1, 2013. 8. m. shima, r. hassan, a. patel and r. razal. “comparative review study of reactive and proactive routing protocols in manets”. 4th ieee international conference on digital ecosystems and technologies, 2010. 9. p. guangyu, m. gerla and t.w. chen. “fisheye state routing: a routing scheme for ad hoc wireless networks.” vol. 1. 2000 ieee international conference on communications. icc 2000. global convergence through communications. conference record, 2000. 10. p. charles, e. belding-royer and s. das. “ad hoc on-demand distance vector (aodv) routing”. no. rfc 3561. document, 2003. 11. g. prakhar, p. gole, p. varshney and n. tyagi. “reliability factor based aodv protocol: prevention of black hole attack in manet”. smart innovations in communication and computational sciences. singapore: springer, 2019, pp. 271-279. 12. s. a. rashid, m. s. alkatheiri, s. anamalamudi and j. liu. “cognitive aodv routing protocol with novel channel-route failure detection”. multimedia tools and applications, vol. 2019, pp. 1-18, 2019. 13. b. m. lubdha, r. l. jain and g. d. patil. “study of various routing protocols in mobile ad-hoc networks”. international journal of scientific research in network security and communication, vol. 6, no. 1, pp. 1-5, 2018. 14. y. xueqin, q. chen, c. chen and j. zhao. “improved zrp routing protocol based on clustering”. procedia computer science, vol. 131, pp. 992-1000, 2018. 15. h. j. zygmunt, m. r. pearlman and p. samar. “the zone routing protocol (zrp) for ad hoc networks”. working group, 2002. 16. b. christian, g. resta and p. santi. “the node distribution of the random waypoint mobility model for wireless ad hoc networks”. ieee transactions on mobile computing vol. 2, no. 3, pp. 257-269, 2003. 17. i. teerawat and e. hossain. “introduction to network simulator 2 (ns2)”. introduction to network simulator ns2. boston, ma: springer, 2009, pp. 1-18. tx_1~abs:at/add:tx_2~abs:at 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 research article improving coronavirus disease tracking in malaysian health system mohammed h. shukur* department of computer science, cihan university-erbil, kurdistan region, erbil, iraq abstract this paper proposes coronavirus disease tracing system acts as a portal for the health center to update and track their patients’ contacts and collect data for further analysis. all of the contents are handpicked, filtered, and selected to the best of our ability by assigned individuals (mostly medical staff ) to ensure that sources are credible and free of hoaxes for the public’s benefit. this application likewise means to assemble information for the top to the bottom investigation (for example, time arrangement to screen the development, in-house patients) and synchronization with a contact following application, for instance, mysejahtera app that was created and set up by the malaysian government and utilized from one side of the country to the other it assumes that all patients have internet access and smartphone, to handle such delinquent, sms driven application with ability to update status through using patient credentials through system website, and to add simplicity for clients’ understanding using graphic visualizations and dashboards are incorporated. keywords: coronavirus disease, e-government, e-health, tracking system introduction coronavirus disease (covid-19) is a newfound virus causing severe, intense respiratory disorder (coronavirus sickness covid-19)[1] rose in wuhan, hubei province, china, and quickly spread to different nations worldwide. at present, the malaysian government and health center are trying their best to learn and stop the spread. this virus is perceived to spread through human-to-human contact, particularly those with close contact and respiratory droplets from an infected person sneezing or coughing. with the number of cases expanding each day, government departments are looking to cooperate with health centers to track down patients effectively. the health center needs a system to store patient’s details, trace the patient visited places, and send the report to government agencies whenever required. hence, this covid-19 tracing system (cts) was intended to assist clinical experts and medical care communities in keeping track of their covid patients and updating the government agency regularly. this system allows the health cares to store patients’ details, their last visited locations, and linked cases if there are any. these patients’ profiles are then updated to the government. the problem statement for this project is that the medical center cannot store, track, and update case details with government agencies as the number of cases grow every day. moreover, it a challenging to trace the linkage infections between two or more cases. furthermore, public locations attended by the infected person are unknown. the system’s scope is separated into two parts: employee scope and system scope. the employee scope enables the employee to login to the system using their employee id and password. the system also enables the employee to add new case details and link them to any existing past cases. the employee can put in a location to see if there were any reported cases. at the same time, the system scope shows the latest case updates of the covid-19 cases. moreover, it can store the new case details and link to any existing past cases. it also shows the background of the infected person, details such as last 14 days visited places, how to contact the virus, and sent the reports to government agencies through email. this project aims to help health centers store, trace, and update the government agencies on covid-19 case details. a webbased application to store the cloud data will be easy to integrate with many government applications such as mysejahtera. it is expected to reduce the work on government agencies to trace the contacts linked to the cases. simultaneously, the project works with an internet connection and location enabled through the web browser, yet it collects patients information through sms or through qr code scan by nearest smartphone. it is *corresponding author: mohammed h. shukur, department of computer science, cihan university-erbil, kurdistan region, erbil, iraq mohammed.shukur@cihanuniversity.edu.iq received: march 22, 2021 accepted: april 30 2021 published: may 20, 2021 doi: 10.24086/cuesj.v5n1y2021.pp11-19 copyright © 2021. mohammed h. shukur. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) shukur: improving coronavirus disease tracking in malaysian health system 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 anticipated that the government will approve it and integrate it with government-linked applications to allow for seamless data transfer. the default email services for health care will be used. literature review health centre app acts as a portal for the health center staff to update and track their patients’ contacts and collect data for further analysis. all of the content are handpicked, filtered, and selected by assigned medical staff to ensure that sources are credible and free of hoaxes for the public’s benefit. this app also aims to collect information for the in-depth analysis, such as time series to monitor in-house patients and monitor the growth and synchronization with a government’s contact tracing. it is conveyed using graphic visualizations and dashboards to make it easier for users to learn. review of foreign countries tracing applications new york h1n1 surveillance system on april 23, 2009, a medical attendant from a secondary school in new york city (nyc) called the department of health and mental hygiene (dohmh) to report a flare-up of respiratory disease[2]. the reason for the episode was quickly affirmed to be flu a pandemic (h1n1) 2009 infection. only a few days after initial reports of mild illness caused by pandemic (h1n1) 2009 infection in california and texas[3,4], this episode was identified as a flare-up of a significant respiratory condition linked to pandemic (h1n1) 2009 infection in mexico[5]. the clinical severity and transmission characteristics of this new flu infection were kept a secret. dohmh launched a wideranging public health response in response to initial media reports about the mexican flare-up and concerns that nyc could also be hit by unending severe infection. before the spring of 2009, dohmh routine observation frameworks for flu included (1) syndromic reconnaissance for medicine deals, school non-attendance, and crisis office visits for flu-like ailment (ili)[5,6]; (2) electronic research facility announcing of affirmed cases from business and emergency clinic laboratories; (3) dynamic observation of all nyc virology laboratories to decide the week after week number of examples submitted for flu testing and the level of those positive; (4) composing tests of flu disconnects acquired from patients in nyc emergency clinics at the dohmh public health laboratory; (5) upgraded latent observation for pediatric flu passing; (6) checking patterns in flu and pneumonia-related mortality through the dohmh vital registry; and (7) observing outpatient ili through the centers for disease control and prevention (cdc; atlanta, ga, usa) influenza-like illness surveillance network [6], a sentinel arrange through which suppliers revealed week after week on the extent of ili in their works on during flu season. the dohmh had also arranged a nearby reaction to a potential flu pandemic, remembering upgraded observation to control general well-being authorities to decide how to organize the utilization of antiviral specialists and antibodies[7]. reconnaissance information could likewise advise network control measures, for example, school terminations. proposed observation methodologies in this arrangement concentrated on systems for checking patterns in hospitalizations and passing, yet not attempting to tally each severe case. techniques were additionally proposed for acquiring more definite clinical and epidemiologic information for an example of cases. the dohmh also has an incident command system (ics), which is different from the standard dohmh structure in that it is an office wide structure for dealing with and responding to crises. the ics is divided into 10 sectors and is led by an episode officer who reports directly to the commissioner of health[1]. dohmh workers are allocated to an area inside the ics and can be approached to help their endless supply of the framework. in a general well-being crisis, the surveillance and epidemiology section builds up and directs observation to evaluate the sickness and passing related to the occasion and leads any required epidemiologic examinations to manage the general well-being reaction. ics actuation gives flood limit by expanding the workforce accessible to lead observation or epidemiologic exercises past the staff individuals who are regularly liable for the particular infection or general medical problems associated with the crisis. healthmap – h1n1 surveillance system. h1n1 swine flu pandemic: 2009–2010. the 2009 swine flu pandemic was caused by a new h1n1 strain that started in mexico in the spring of 2009 and quickly spread worldwide. the virus infected up to 1.4 billion people worldwide in a year, killing between 151,700 and 575,400 people. according to the cdc, the 2009 flu pandemic primarily affected children and young adults, with 80% of deaths occurring in people under 65. considering that most flu virus strains, including those that cause seasonal flu, kill people aged 65 and up, this was important. in the case of the swine flu, however, older individuals seemed to have built up enough immunity to the virus family that h1n1 belongs to, so they were less affected. the annual flu vaccine also includes vaccination for the h1n1 virus that caused swine flu. healthmap is an example of a new initiative in public health surveillance that is open and global. the healthmap system integrates automated, around-theclock data collection and processing with expert review, and analysis to aggregate reports by type of illness and geographic location. through a freely accessible web site, healthmap sifts through vast amounts of event information acquired from various online sources in multiple languages to provide a detailed view of ongoing global disease activity. healthmap collaborated with the journal to develop the h1n1 interactive map (as part of the journal’s h1n1 influenza center) to improve situational awareness of public health practitioners, clinicians, and the general public regarding the worldwide spread of 2009 h1n1 influenza infection. the healthmap shukur: improving coronavirus disease tracking in malaysian health system 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 infrastructure was used to provide information about outbreaks on this interactive map. it used informal sources (e.g., the news media, mailing lists, and contributions from individual users) and formal announcements to compile the data. (mainly from the world health organization [who], the cdc, and the canadian public health agency) visitors to the site might sort reports by alleged or confirmed cases or deaths and view reports over time to see how the disease spread. during the two most giant waves of the h1n1 pandemic, healthmap gathered over 87,000 reports from both informal and official sources (43,738 reported during the first wave of infection, from april 1 to august 29, 2009, and 43,366 reported during the second wave, from september 1 to december 31, 2009) (between august 30 and december 31, 2009, during the second wave of infection). all reports submitted into the healthmap system had their geographic location documented, allowing for easy monitoring of h1n1 influenza’s regional and global spread. according to the who regions and pandemic stages, healthmap also detected a growing number of countries with informal reports of suspected or confirmed cases overtime. there had already been reports of suspected or confirmed cases in 32 countries by the end of the who pandemic phase 4 (april 28). the spread of the virus to new countries was most rapid in europe and the americas during the pandemic’s early stages. because of the implementation of containment measures and the early involvement of countries with a high amount of air travel, this spreading steadily slowed to about one country per day by the end of phase 5 (june 10). it further decreased to one country per 2 days in phase 6, potentially due to the implementation of containment measures and countries’ early involvement with a high volume of air travel. although air travel may have aided the virus’s intercontinental spread, other migration patterns are likely to have shaped the virus’s spread within continents. regions that are foreign travel hubs (e.g., france and the united kingdom) in europe, for example, identified infections earlier than areas with fewer commercial flights (e.g., eastern european nations). public health capability and ability to report cases influenced this trajectory as well. johns hopkins university csse dashboard when a disease can spread so quickly, data must move at a much faster pace; thus, map-based dashboards become extremely useful. seven coronavirus dashboards are among the top 10 cited applications from esri arcgis online assistance at the time of this writing in mid-february 2020, with more than 160 million views. in light of the rising pandemic, this was first issued on january 22, 2020[4]. the johns hopkins university’s center for systems science and engineering (jhu csse) dashboard stands out in late january 2020, gathering 140 million viewpoints. lauren gardner (a disease transmission specialist) and her team from the jhu csse came up with the idea. the dashboard was shared widely on the internet, along with numerous news stories and offers. this enthusiastic reaction to the jhu csse and other dashboards reveals the public’s interest in health risks. anyone with internet access will learn a massive amount of information about the covid-19 infection in a few short clicks from these resources. the intuitive guide on the jhu csse dashboard detects and counts confirmed contaminations, deaths, and recoveries. the diagrams show the progression of the infection overtime. the day and season of the most recent information update and the information sources are visible to viewers. the who, the us centers for disease control and prevention, the national health commission of the people’s republic of china, the european center for disease prevention and control, and the chinese online clinical asset dxy.cn are among the dashboard’s five valid information sources. the dashboard provides links to these and other sources. this work is subtly described in a blog entry. the related data are stored in github as google sheets. web administrations enable gis customers to use and display various information contributions without the need for centralized facilitating or handling, facilitating information sharing, and speeding data collection. the hopkins group physically refreshed information twice a day from january 22 to january 31, 2020, as part of the dashboard focusing. shukur: improving coronavirus disease tracking in malaysian health system 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 esri arcgis living atlas team helped them adopt a semicomputerized living information stream approach to refresh the dashboard in february 2020. it is primarily reliant on the dxy.cn information asset, which updates case reports at the standard and national levels every 15 min. nonetheless, lauren gardner’s blog points out that other information assets were faster to refresh than dxy.cn for countries outside of china, so those case checks are physically renewed for the day. the arcgis living atlas currently has no restrictions on element layers. who dashboard the who coordinates and organizes global health, combating infectious diseases through surveillance, preparation, response, and incorporating gis technology into its work. on january 26, 2020, the who released its arcgis operations dashboard for covid-19, which also tracks and records coronavirus cases as well as the total number of cases in each country and chinese territory, as well as educational boards about the guide and its data assets. the who and jhu csse dashboards made them curious comparisons before february 18, 2020. as shown in figures 1 and 2, each had an endlessly particular all-out case (both taken on february 16, 2020). the who dashboard only included laboratory-confirmed cases, while the jhu csse included cases that were examined using a side effect exhibit and chest imaging (representing exactly 18,000 extra reports). in any case, beginning february 19, 2020, the two dashboards are in sync, displaying comparative all-out case tallies. the who dashboard includes a pandemic forecast, which shows cases by date of disclosure. placing the epi bend perception over the combined cases chart provides helpful information about episode movement. it may reduce anxiety, as the absolute number of new cases has consistently decreased every day since february 4, 2020. (except for a spike on february 13, 2020, more than 15 k cases were added after china started to include “clinically analyzed” cases in its figures rather than just laboratory affirmed cases). a “hamburger” icon in the upper right corner of the who dashboard leads to additional covid-19 data and an intuitive web map that places covid-19 in context with other who identified health crises. dengue fever, rift valley fever, and west nile fever are just a few examples. as a result, the who is updating its covid-19 dashboard using arcgis geoevent server to push updates to a single location. every day, component management takes place on several occasions. review of malaysian systems: my trace mytrace is a mobile application that aids in the management of covid-19 outbreaks by the health authority. when an app identifies another nearby device with mytrace installed, it uses a community-driven approach to exchange proximity information. the app allows people who have been near an infected person to be identified. shukur: improving coronavirus disease tracking in malaysian health system 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 review of malaysian systems: mysejahtera sejahtera is a mobile application designed by the malaysian government to aid in managing covid-19 outbreaks in the country[3]. it helps people to self-assess their health and that of their family members. during the covid-19 epidemic, people can also track their health progress. sejahtera also allows the malaysian ministry of health to keep track of users’ health and respond quickly with the necessary treatments, yet it assumes that all patients are using smartphone with wifi connection. comparison of coronavirus disease tracing system with malaysian products feature/product mytrace mysejahtera coronavirus disease tracing system login security    store patient/user details    trace locations    health-care use    send data to the government    needs for internet to communicate    collect information when no smartphone under use    methodology to develop this project, we have used the extreme programming methodology. extreme programming is one of the agile methods. the main reason is allowing changes to the project according to the current situation or customer requirement even late in the life cycle. feedback and communication are taken as essential aspects to improve better software development. we can communicate with users by giving them a chance to test using the software and gain feedback to change or add to the software (if any). these criteria help developers to change requirements and technology. extreme programming methodology involves five phases and below has been explained the progress thoroughly in each stage: planning the problems for cts were identified and listed down. a research study was conducted to gather related information on covid-19 tracings apps in malaysia, a reliable application for health centers to store and trace their patients’ details. i have searched for associated scenarios and cases that explain and support well the problem statements. informal interview sessions were held to gather information from the health center employees. finally, the objectives and scopes are drawn to get a picture of the system’s functionality. designing in this phase, generally, the website’s design is created. first of all, i have used a prototyping tool to design the interface. next, i develop a finalized user-friendly interface that can make it easy to learn and use. user experience is ensured for the employees to feel at ease when using the website. at this phase, the functionality is not added yet. in figure 1, it illustrates entity relationship, and figure 2 shows the relation table. coding in this phase, the entire step involves coding to develop each objective is set into a practical and working application. each shukur: improving coronavirus disease tracking in malaysian health system 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 button will be added with functionality. certain functionalities were integrated with a government-linked application such as mysejahtera to load the places the patient has visited. adding the ability to generate a distinguished code with unique credentials to be printed out as qr code, define the mechanism to read sms from patient identify their status, and written location, that will after done, the program is compiled and build. testing each class of code is tested to detect any bugs and errors. those detected bugs and errors should be fixed to produce an error/bug-free application. besides, system testing and user acceptance testing were done, and the data were documented. managing management is vital throughout the development process. it is crucial to managing time in developing the software within the figure 1: entity relationship diagram figure 2: rational table figure 3: login screen figure 4: dashboard shukur: improving coronavirus disease tracking in malaysian health system 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 personas. employees from the health-care center can update patient information such as patient id, ic, name, address, visited locations, gender, and age. employees would have qualities such as their name, employee id, and password. furthermore, when entering the patients’ locations, a new entity called zone will be created, which includes attributes such as location id and address. the most basic requirement is that employees must associate a patient with a room with qualities such as room id and room type. with the help of erdplus, the relational table was generated, with minor tweaks to maintain the precision of data explained between the erd diagram and relational table. we then developed the sql from erdplus, did minor tweaks, and ran through a phpmyadmin web-based tool. we used an apache server to run the code and get the output. attached below are the erd diagram, relational table, and sql code samples used. below are the erd diagram and relational table. results and analysis cts is developed as a web-based system. the front end is built using an angular framework with bootstrap styling. whereas the database is created using sql. before the development, a simple design brainstorming is done to ensure all the scenarios are catered to address the health-care need while storing the patients’ details and send the updates to the government every often. the employee can log in using their employee id [figure 3]. an employee can also use their id name generated figure 5: patients list figure 6: patients details given time. the works should be documented and presented clearly on the fixed dates. when carrying out research methods such as surveys or asking for feedback, good ethics and proper communication should respect the user. it is vital to keep the application up to date and obtain input from the supervisor or user who may assist in its development. er diagram and sql codes to make the erd diagram drawing precise and efficient, the team has used the erdplus web tool. we started defining the shukur: improving coronavirus disease tracking in malaysian health system 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 figure 7: location list figure 8: locations details uniquely for them by the administrator, which will detect the employee id in the backend. the home shows the country’s entire cases [figure 4], the whole point within the health center (currently assumed it is a hospital), with daily case breakdowns, filter able to see new only, recovered only, and death only. patients in treatment at the hospital and overall growth of several cases also shown. (for illustration purpose, dummy data shown) [figure 5]. this screen will reveal the high-level details of patients on clicking the patient id and patient name. it will take you to the patient’s information screen. the button update new government queries will submit the attachment of further patient information through email. for now, the email of government authority is embedded within the system. figure 6 shows that all the details will be stored; here, the past visited locations will also be synced through mysejahtera. the dashboard will be helpful for the government agencies in the contact tracing work. (for illustration purpose. dummy data shown) figure 7 shows the list of places visited by patients that are admitted or diagnosed within the health center. on clicking the location id, it will show the patients who visited the location and their time as its shown in figure. 8. conclusions at the point when disease numbers begin to ascend in a state, general well-being authorities and inhabitants stress over whether there are sufficient medical clinic beds for genuinely sick patients. a zone’s medical clinic limit is a gauge, took care of by the information that special emergency clinics report. general well-being authorities depend on the assessments to see whether emergency clinics are in danger of being overpowered. clinics should report day by day to the government what number of beds they have, the number involved, and the accessibility of escalated care beds. under the new framework, the department of health and human services totals the data at a state level and offers a simple spreadsheet of the data accounted for – holes whatnot. to aid the health center in keeping track of their covid19 patients, cts featured to store the patients details, with added vital functionality such as communicate with patients through sms when wifi is down or non-smartphone being used, qr code scan in case of phone switched off, or out of charges, or even no wifi connection, to link cases based on visited place and time and locations visited, it helps the government agency track and traces the contacted people based on comparing patients location and distance. shukur: improving coronavirus disease tracking in malaysian health system 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 11-19 this website can be enhanced with built-in bluetooth functionality where multiple phones can be keyed in the future (auto-communication). the system security can also be enhanced using blockchain technology[7]. references 1 m. cattalini, s. d. paolera, f. zunica, c. bracaglia, m. giangreco, l. verdoni, a. meini, r. sottile, r. caorsi, g. zuccotti, m. fabi, d. montin, a. meneghel, a. consolaro, r. m. dellepiane, m. c. maggio, f. la torre, a. marchesi, g. simonini, a. villani, r. cimaz, a. ravelli, a. taddio and rheumatology study group of the italian pediatric society. defining kawasaki disease and pediatric inflammatory multisystem syndrome-temporally associated to sars-cov-2 infection during sars-cov-2 epidemic in italy: results from anational, multicenter survey. pediatric rheumatology, vol. 19, no. 1, 2021. 2 h. awang, e. l. yaacob, s. n. s. aluawi, m. f. mahmood, f. h. hamzah, a. wahab, n. abdul rashid, r. razali, g. s. ning, k. embong and n. a. yunus. a case control study of determinants for covid-19 infection based on contact tracing in dungun district, terengganu state of malaysia. infectious diseases, vol. 53, no. 3, pp. 222-225, 2020. 3 s. pillai, n. siddika, e. hoque apu and r. kabir. covid-19: situation of european countries. archives of medical research, vol. 51, no. 7, pp. 723-725, 2020. 4 contact tracing in the context of covid-19, 2021. available from: https://www.who.int/publications/i/item/contact-tracing-inthe-context-of-covid-19. [last accessed on 2002 dec 01]. 5 contact transmission of covid-19 in south korea: novel investigation techniques for tracing contacts. osong public health and research perspectives, vol. 11, no. 1, pp. 60-63, 2020. 6 j. abeler, m. bäcker, u. buermeyer and h. zillessen. covid19 contact tracing and data protection can go together. jmir mhealth and uhealth, vol. 8, no. 4, p. e19359, 2020. 7 m. turki, f. laith, n. abdullah. secure electronic healthcare system based on block chain technology with wireless body area network. technology reports of kansai university, vol. 62, no. 8, 2020. . 37 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 37-41 research article measuring the score matching of the pairwise deoxyribonucleic acid sequencing using neuro-fuzzy safa a. hameed*, raed i. hamed department of computer science, college of engineering and science, university of bayan, erbil, iraq abstract the proposed model for getting the score matching of the deoxyribonucleic acid (dna) sequence is introduced; the neuro-fuzzy procedure is the strategy actualized in this paper; it is used the collection of biological information of the dna sequence performing with global and local calculations so as to advance the ideal arrangement; we utilize the pairwise dna sequence alignment to gauge the score of the likeness, which depend on information gathering from the pairwise dna series to be embedded into the implicit framework; an adaptive neuro-fuzzy inference system model is reasonable for foreseeing the matching score through the preparation and testing in neural system and the induction fuzzy system in fuzzy logic that accomplishes the outcome in elite execution. keywords: component, dynamic programming, matching, neuro-fuzzy, sequence alignment introduction deoxyribonucleic acid (dna) sequence matching is an essential area and more approaching nearby in computational biological data.[1] dna sequence analysis is an imperative exploration topic in bioinformatics. assessing the similarity between sequences, this is important for sequence analysis, because similarity proves congruence.[2] the dna atom contains biological, physical, and chemical data; it has turned out to be essential to examine dna sequences statistically.[3] string matching is a strategy to find a design from the predefined info string.[4] similarities between dna sequences may emerge due to the functional, structural, or transformative relationship among them.[5] sequence alignment of two biological sequences may be called pairwise sequence alignment, also in the event, more than two sequences are involved; it may be called multiple sequence alignment.[6] the dynamic programming is the method to implement the dna alignment using the needleman– wunsch[7] and smith–waterman algorithms.[8] here, in this article, we use the pairwise sequence alignment in a global and local algorithms and examined the measure of the matching based on the collected data for dna alignment. the neuro-fuzzy model[9] is used in the matlab tool that implemented by the data set files of measure score matching of dna sequences that deal with the set of biological data. this tool is efficient and fast to evaluate the scoring measure of matching the dna sequences. literature review the study of a biological sequence has been growing exponentially, while the applications of the sequence alignment cover the wide range in bioinformatics. the previous research work has been studied to provide new algorithms with the main purpose for the requirements of matching sequences; the techniques have been used all the latest with providing fast and efficient sequence alignment algorithms. bhukya and somayajulu[1] suggested a new pattern for matching technique defined as exact multiple pattern-matching algorithms that utilize dna sequence. the current method is used to avoid unneeded comparisons in the dna sequence. gill and singh[6] proposed a multiple sequence alignment algorithm which performs fuzzy logic to measure the similarity of sequences based on the fuzzy parameters. nasser et al.[10] suggested the fuzzy logic model for approximate matching of dna subsequences. kim et al.[11] suggested a dna sequence alignment, which uses quality information and a fuzzy inference implementation developed based on the features of dna parts and a fuzzy logic system. chai et al.[12] explained how to perform pairwise sequence alignments utilizing the biostrings bundle using the pairwise alignment function. hameed and hamed[13] discussed how to cihan university-erbil scientific journal (cuesj) corresponding author: safa a. hameed, department of computer science, college of engineering and science, university of bayan, erbil, iraq. e-mail:safa.hamid@bnu.edu.iq received: mar 21, 2019 accepted: apr 24, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp37-41 copyright © 2019 safa a. hameed, raed i. hamed. this is an open-access article distributed under the creative commons attribution license. hameed and hamed: measuring the score matching of dna using nf 38 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 37-41 implement the pairwise alignment technique to get the score of similarity for a pair of characters. in our work, we use the neuro-fuzzy model that utilizes the biological dataset files for matching dna and measures the score of matching the dna sequences with global and local alignment. sequence alignment dna matching is a significant venture in the sequence alignment. since sequence alignment is a discretionary matching process, there is a need for better algorithms.[10] dna sequence alignment algorithms over computational biological science have been enhanced eventually by different techniques:[11] the (needleman-wunsch) global, the (smithwaterman) local, and (ends-free) cover pairwise sequence alignment issues.[12] pairwise alignment is a technique for scoring the similarity of a pair of characters. it decides the correspondences between the substrings in the sequences like the similarity score is amplified.[13] for it is a large portion basic form, known as pairwise sequence alignment, we provided for two sequences a and b and discover their best alignment (either global or local).[12] aligned sequences represented as rows in a grid. gaps (“−“) need aid embedded between the characters with the goal.[6] the ways we use it to perform the alignment are global and local alignment, these algorithms uses the proposed matrix to measure the similarity of bases in the two sequences. for the needleman– wunsch algorithm, a scoring matrix is ascertained for those two provided for sequences a and b, by setting one sequence along column side, furthermore on the turn sequence side. it is additionally frequently referred as optimal matching algorithm and the global alignment technique.[7] the smith– waterman algorithm, which is the method used to perform the local sequence alignment, local alignment algorithms find the sections of the highest similarity between two sequences and create the alignment to abroad from there, that is, identify the most similar portion comparable subregion imparted between two successions.[8] the proposed method we use the nero-fuzzy technique in matlab tool. the neuro-fuzzy model is very well established approach and has a tremendous potentiality to outcome results with high accuracy ratio and the efficiency with biological data to determine the measure score of matching dna sequencing. those recommended sequence-matching algorithm utilize the three input variables – match score (match), mismatch score (mismatch), and gaps, as shown in “figure 1.” these three inputs would then fuzzified utilizing the following membership functions (mfs) equations and giving the calculated resulting score: matching = 0 if there is no similarity 1 if there is highest simmilarity 100% 1,0 matching score / lenseq          (1) mismatch = 0 if there is no mismatch 1 if there is no similarityy 1,0 mismatching score / lenseq          (2) gap = 0 if there is no need to put a gap 1, 0 gaps score / lens  eeq      (3) figure 1: the three input variables and the output figure 2: the membership function and the training testing phase for the lowest possible error. (a) input membership function. (b) the testing data ba hameed and hamed: measuring the score matching of dna using nf 39 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 37-41 score = 0 if the resulting score £ 0 1 if the resulting perfect 11, 0 resulting score / perfect score          (4) the variable “lenseq” mean the entire length of the sequence. score = match + (− mismatch) + (− gabs) (5) the simulation results in our work, we perform the neuro-fuzzy model by an adaptive neuro-fuzzy inference system (anfis) tool in matlab, using the data set about 600 samples for a matching measure score of dna sequencing; these data divide into two data files for the training and testing, for the training step, we use the data set about 450 samples, and for the testing step, we use the data about 150 samples.[14] we use these dataset files in anfis system in the range value in equation 1, 2, 3, and 4 and output the result according to the equation 5. we use different processing systems to implement the matching process, and each system has different results with convergent values, and that for choosing the most suitable one with less error percentage and depends on it to calculate the matching score. “figure 2” shows the chosen attempt that gives the results with high accuracy. “figure 2” explains the most suitable anfis system with the lowest average testing error; table 1 shows the different processing systems; we implement it with the details. in this table the most suitable system which has chosen is the system that has the following: trapezoidal mfs that has three mfs, constant mfs output, backpropegation train fuzzy inference system method and the number of epochs which are 500, this system has the lowest average training and testing error which are 0.016572 and 0.01657 respectively, and gives the result with high performance, thus we use it to get the score matching of the numeric data from the dna sequence alignment, show tables 2 and 3. in table 3, we aligned the sequence using the local algorithm; in this method, the algorithm takes the most similar part of the pair sequence, not must in the order and not need to input the gap; the resulting score is the perfect, there are no mismatch and no gaps, and is 100% identical. discussion sequence alignment is a necessary condition for analyzing dna sequencing; in our method, we use the pairwise sequence alignment; it is applied using the global and local alignment algorithm method. in this method, we use the numeric biological data for sequence alignment using anfis system in matlab; this system implements several processing systems to get the most suitable results as shown in table 1; the most suitable system is the trapezoidal with three mf for each input and 500 epochs; it has the lowest average testing error. we use several different sequences to be aligned, as shown in table 2; we aligned the sequences in the global alignment algorithm. in this method, we insert the gaps when the base in the sequence is not similar with the other in the same order in this pair, and shifted the character and input the gap in order to be identical; we use the number of times for (matching), mismatching and gaps as the input in the anfis view tool, and output the score matching, as shown in “figure 3” we can compute the percentage similarity of the alignment by using this code in matlab [score, alignment] = nwalign(‘s1’,’s2’); showalignment(alignment); in order to display a pairwise sequence alignment, as shown in “figure 4.” this use the table 1: the various anfis testing results sequences global alignment identities (%) score aggttgc aggttgc 7-may 0.149 aggtc aggt--c −71% gtaggcttaaggtta gtaggcttaaggtta 15-may 0 tagatc at-ctag −33% agtcca a – gtcca 7-may 0.149 atgtcc atgtcc−71% cggga cggga6-feb 0 attgac attgac 33% ctatccg ctatc cg 7-may 0.425 ctagtcg ctagtcg −71% anfis: adaptive neuro-fuzzy inference system table 2: the score matching for global alignment of anfis tool sequences local alignment identities (%) score aggttgc aggt 100 1 aggtc aggt gtaggcttaaggtta tag 100 1 tagatc tag agtcca gtcc 100 1 atgtcc gtcc cggga ga 100 1 attgac ga ctatccg cta 100 1 ctagtcg cta anfis: adaptive neuro-fuzzy inference system hameed and hamed: measuring the score matching of dna using nf 40 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 37-41 number of times similar in the sequence alignment divided on the entire length of the alignment sequence. in table 3, we aligned the sequence using the local algorithm, in this method the algorithm take the most similar part of the pair sequence, not must in the order and not need to input the gap, the resulting score is the perfect there is no mismatch and no gaps, and have 100% identical. conclusion the proposed technique, here, is used to obtain the similarity measure of the pairwise dna sequence alignment; the pattern matching is an essential task of example the disclosure process in this day and age for finding the basic and utilitarian conduct in the dna sequencing. in spite of the fact that, the example of matching is commonly utilized as a part of computer science and information processing. in this paper the proposed algorithms are used that are global and local alignment to measure the score matching, which are utilized; it as a method to be align the two dna sequences. the neuro-fuzzy model is used to evaluate the score similarity by the anfis tool in table 3: the score matching for local alignment of anfis tool mf type input the number of mf for the inputs the mf output train fis method the number of epochs the average training error the average testing error triangular mf 3 3 3 constant backpropagation 50 0.04657 0.06078 triangular mf 5 5 5 constant backpropagation 250 0.01109 0.04306 trapezoidal mf 3 3 3 constant backpropagation 100 0.01898 0.0453 trapezoidal mf 3 3 3 constant backpropagation 500 0.01657 0.01657 gaussian mf 5 5 5 constant backpropagation 350 0.01436 0.04436 gaussian2 mf 5 5 5 constant backpropagation 500 0.02804 0.05415 mf: membership functions, anfis: adaptive neuro-fuzzy inference system, fis: fuzzy inference system figure 3: view rules adaptive neuro-fuzzy inference system figure 4: the identical alignment hameed and hamed: measuring the score matching of dna using nf 41 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 37-41 matlab; we implement the method in several processing systems and depend on the most suitable system with the lowest average testing error; we obtain the score matching result for several patterns of dna sequencing; this model presented the matching implementation in fast and efficient. references 1. r. bhukya and d. v. l. somayajulu. “exact multiple pattern matching algorithm using dna sequence and pattern pair”. international journal of computer applications, vol. 17, no. 8, pp. 32-38, 2011. 2. x. xie, j. guan and s. zhou. “similarity evaluation of dna sequences based on frequent patterns and entropy”. bmc genomics, vol. 16, no. 3, p. s5, 2015. 3. w. deng and y. luan. “analysis of similarity/dissimilarity of dna sequences based on chaos game representation”. abstract and applied analysis, vol. 2013, p. 926519, 2013. 4. p. pandiselvam, t. marimuthu and r. lawrance. “a comparative study on string matching algorithms of biological sequences”. in: international conference on intelligent computing, pp. 1-5, 2014. 5. t. chakrabarti, s. saha and d. sinha. “dna multiple sequence alignment by a hidden markov model and fuzzy levenshtein distance based genetic algorithm”. international journal of computer applications, vol. 73, no. 16, pp. 26-30, 2013. 6. n. gill and s. singh. “biological sequence matching using fuzzy logic”. international journal of scientific and engineering research, vol. 2, no. 7, pp. 1-5, 2011. 7. s. b. needleman and c. d. wunsch. “a general method applicable to the search for similarities in the amino acid sequence of two proteins”. journal of molecular biology, vol. 48, no. 3, pp. 443453, 1970. 8. t. f. smith and m. s. waterman. “identification of common molecular subsequences”. journal of molecular biology, vol. 147, no. 1, pp. 195-197, 1981. 9. d. nauck, f. klawonn and r. kruse. “foundations of neuro-fuzzy systems”. john wiley and sons, inc., new york, 1997. 10. s. nasser, g. l. vert, m. nicolescu and a. murray. “multiple sequence alignment using fuzzy logic. in: 2007 ieee symposium on computational intelligence and bioinformatics and computational biology, ieee, pp. 304-311, 2007. 11. k. kim, m. kim and y. woo. “a dna sequence alignment algorithm using quality information and a fuzzy inference method”. progress in natural science, vol. 18, no. 5, pp. 595-602, 2008. 12. n. chai, l. r. swem, m. reichelt, h. chen-harris, e. luis, s. park and j. mcbride. “two escape mechanisms of influenza a virus to a broadly neutralizing stalk-binding antibody”. plos pathogens, vol. 12, no. 6, p. e1005702, 2016. 13. s. a. hameed and r. i. hamed. “analysing the score matching of dna sequencing using an expert system of neurofuzzy”. journal of theoretical and applied information technology, vol. 95, no. 6, pp. 1255-1262, 2017. 14. dna matching data base-ncbi”. https://www.ncbi.nlm.nih. gov/nucleotide. . 77 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 77-81 research article ad hoc on-demand distance vector inherent techniques comparison for detecting and eliminating the black hole attack nodes in mobile ad hoc network ghassan a. qasmarrogy*, yazen s. almashhadani department of communication and computer engineering, cihan university, erbil, kurdistan region, iraq abstract it is important to connect wirelessly a group of moving mobile nodes together in a static or dynamic form, to transfer digital data between them, this form known as a mobile ad hoc network. this private network can be used in different essential situations where it depends on each connected mobile node to deliver and pass the data between them, without any fixed access point or router. unfortunately, there are different types of attacks that can affect these nodes, and steal or corrupt the data inside, one of these attacks called the black hole attack. in this paper, a compared study will be done between two major innovative techniques derived from the ad hoc on-demand distance vector routing protocol to avoid the black hole attack; the paper will compare the two techniques in delay, throughput and packet dropping efficacy. keywords: black hole attack, extended data routing information technique, mobile ad hoc network, secure ad hoc on-demand distance vector routing protocol, trusted ad hoc on-demand distance vector introduction some scenarios or situation needs a group of moving nodes wirelessly connected together in a static or dynamic infrastructure form; this can be known as mobile ad hoc network (manets).[1] this form can broadcast or unicast data between each wireless mobile node separately or together. manet can change its form dynamically depending on the moving nodes speed as shown in figure 1; therefore, it shows better moving infrastructure to be used for different types of situations such as military, search and rescue, wireless sensor networks, and many more.[2] where each node can move separately or with its private group to form the new infrastructure without the need of fixed access point or routers. each node in manet is responsible for cooperating with other nodes to send and receive the data smoothly using a different type of routing protocols (rps) that are designed, especially for manet. there are three types of rps for networking transmission operations and routing management, namely, proactive, reactive, and hybrid rps.[3] each protocol is depending on the size and the dynamic nature of the topology of manet.[4] the proactive protocols basically are table driven, meaning. it depends on a specific table that used for routing decisions,[5] while reactive protocols are depending on an on-demand decision where it calculates the path needed for sending the data between the nodes on the same time that they need to transmit the data.[6] hybrid protocols use both reactive and proactive techniques to deliver the data between the nodes.[7] figure 2 shows the types of manets rps. it is essential to secure the data transmitted between the moving nodes to provide maximum security; unfortunately, wireless nodes have many security issues to be attacked.[8] one of these attacks called the black hole attack. in this paper, two major annotative techniques, namely, the trusted ad hoc on-demand distance vector (taodv) rp technique and the extended data routing information (edri)-based technique will be compared and evaluated to prevent and detect the black hole attack. this paper will be organized as follows: in section 1, an explanation will be shown for both black hole attack and it’s two preventing techniques; section 3 will demonstrate a comprising between the two techniques; and finally, section 4 will conclude the paper. cihan university-erbil scientific journal (cuesj) corresponding author: ghassan a. qasmarrogy, cihan university, erbil, kurdistan region, iraq. e-mail: ghassan.qasmarrogy@cihanuniversity.edu.iq received: apr 04, 2019 accepted: apr 25, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp77-81 copyright © 2020 ghassan a. qasmarrogy, yazen s. almashhadani. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ qasmarrogy and almashhadani: ad hoc on-demand distance vector inherent techniques comparison for detecting and eliminating the black hole attack nodes in mobile ad hoc network 78 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 77-81 the black hole attack and aodv rp in manets, all rps have security issues, therefore, creating a robust rp to transfer data between the nodes without any security issues is difficult, due to many problems such as share link broadcast wireless channel, node’s environment insecurity, the absence of center administration, the limitation of resources, and many more.[9] the essential requirements in any communication system to transfer data are integrity, confidentiality, and availability.[10] these requirements insurance the data will be intact without any eavesdroppers that can intercept or replace the transferred data. there are different types of attacks depending on the involvement of the attacker which can be classified into the passive attack and active attack. moreover, each one of these attacks can be classified into external and internal attacks.[11] the black hole attack one of the active attacks in manet is the black hole attack, in this attack, one communicating node in the private network, after receiving the rreq message from the sender node to form a path to the destination and start to transmit the data, replies with a rrep message with a highest sequence number compared to other nodes, forcing the sender to form a path through this node to the destination node, and transmits all the data through this path to that suspicious node, in the end when the suspicious node receives all the data, it will drop it all, without forwarding them to the neighbour nodes,[11] figure 3 shows the fake rrep from the suspicious node. this attack can be considered a serious threat in any wireless private network because all the transmitted data are dropped without any succeed end to end delivery; therefore, it will increase the threshold of successful packet delivery to 100%. aodv inherent black hole preventing techniques aodv is a reactive rp that starts a discovery route from source to destination when manet topology changes and the source node want to transfer data, in aodv, each node in manet stores and uses a routing table, to find the next neighbor node, which has the highest sequence number. in general, before the sender wants to transmit the data, it checks its routing table, therefore if there is a recent route to destination, it will send the data through that path, otherwise it will start a discovery route phase to find more recent route, where the sender will broadcast a request message rreq to their neighbour nodes and then each node will reply with rrep message, and the highest sequence number node will participate in the path formed.[12] trusted aodv routing technique this technique is built on the original aodv trusted protocol. when new manet topology is established and a node wants to transmit information to a destination node, it will send a hello message to all other nodes to find the destination, each node after receiving the rreq message will reply and respond to the sender node with their unique id; after that, the sender node will be built a neighboring list consist of all nodes around its wireless range, which is stored in the node’s memory. this table stores each node’s id in the range, also each node has a trust factor value stored in the table. when the value is small, it means that the node is suspicious, unreliable, and new to the network, and when the value is high, it means that the node is trusted and it was participating in old transmission path figure 1: mobile ad hoc network topology figure 2: the types of mobile ad hoc networks routing protocols qasmarrogy and almashhadani: ad hoc on-demand distance vector inherent techniques comparison for detecting and eliminating the black hole attack nodes in mobile ad hoc network 79 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 77-81 before. therefore, when the sender node broadcasts the rreq message, the trusted value function will be used as a feature found in aodv rp to compute the level of trustiness for each node, and when each node receives the rreq message, they will reply back directly to the sender, so it will be giving a trusting value. after that, when the node involved in a root establishment mechanism, the sender will verify this node by the forward route request rreq to check its integrity and if the integrity is ok, the trusted value will be increased. after each node stores the trust value factor for intermediate nodes, it would become easier to detect the black hole nodes, so when the sender node will send the rreq and establishes the path to the destination node, it will make sure that each intermediate node in the path has a higher trust factor, and other nodes with small trust factor will not participate in the future path for a definite time.[13] figure 4 shows the trusting technique of aodv, where the nodes with higher trusting value are most reliable and with lower trusting value are unreliable. edri technique the second technique also is an extension from aodv rp; this approach uses a data routing information table (dri) and an extended data routing information table (edri); it also uses a data control packet to detect and discover the suspicious nodes in manet. using the edri table, each node dynamically updates and stores their current and previous neighbors node’s id, the dri table including two columns (from and through) and the black hole number (bhn), so when the middle nodes receive a packet from any neighbors’ node, it writes 1 in the from column, but when the same node sends the packet to the neighbor note, it writes in the through column 1. when a neighbour node is detected as a black hole the flag of bhn number will be set to 1 and it will be set to 0 when there is no detection. figure 5 shows the edri table.[14] as known, any suspicious black hole node drops partially or fully all received packets; therefore, control data packets will be sent to check all the middle nodes in the path from source to destination. this packet contains node’s id, next node hop nhn, and a random number that are constant for all data packets in that path, this control’s data packet cannot be forwarded by a suspicious node. another data control packet will be also used which can be transmittable by the suspicious node, to check which node did not respond to the previous control data packet. figure 6 shows the composition of the data control packet. after receiving all the rrep messages, the sender node will analyze and select the best-secured path to the destination, for transmitting the data packets and detect any suspicious black hole nodes in the path and bandit from future mapping. comparison and results to evaluate the two techniques to prevent the black hole attack and isolate the suspicious nodes from manet, a scenario was made to simulate the prevention of the attack; the same metric where used to evaluate the two algorithms, packet overhead, throughput, and delay was measured. when the number of suspicious black hole nodes increases, the end-to-end delay will also be increased, due to the fact of packet dropping, renewal of path formation, and resending of the data message. all of these causes force the sender node to retransmit the packets until it gets acknowledgment from the receiving destination node. in the trusted aodv (taodv) technique, when the sender node requests a fresh route, a path to the destination node will be formed, and only the trusted node with higher trusting value will participate in the new path, the trusted nodes figure 3: mobile ad hoc network’s black hole attack figure 4: ad hoc on-demand distance vector trusting technique figure 5: example of extended data routing information table figure 6: the component of the data control packet in extended data routing information technique qasmarrogy and almashhadani: ad hoc on-demand distance vector inherent techniques comparison for detecting and eliminating the black hole attack nodes in mobile ad hoc network 80 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 77-81 number in total are small; therefore, the end-to-end delay will increase. as a result, from the first running transmission, the number of trusted nodes cannot be increased, which causes the sender to use only the available trusted ones, so the delay will not be decreased rapidly overtime. moreover, when the delay of the message increases, the packet dropping will also increase. furthermore, the packet overhead will increase and the packet delivery ratio will be decreased due to the previous causes. figure 7 shows how the number of trusted nodes values increase over multiple transmissions overtime. in edri technique, the sender node sends an rreq packets to generate a safe path, by sending to the neighbor nodes the data control packet, the delay will increase at the beginning of transmission, after that, when all suspicious nodes detected, overtime the delay will be decreased rapidly, due to the fact that all the suspicious nodes were detected in the first running safe path, and no more rreq packet needs to be broadcasted. figure 8 shows the decreasing number of rreq packets overtime. moreover, overtime when the number of trusted nodes increases due to edri table, the overall delay will be decreased. moreover, the packet overhead will also be decreased overtime, due to the fact that the sender will reach more trustable nodes so connection packet will not be needed anymore between the neighbor nodes. figure 9 shows the main comparison of the delay between the two techniques. the throughput of the network was also compared, in the black hole attack, every single suspicious node sends an rrep to a source node and receives all the data and drops it from that sender, when the number of suspicious node increases, the number of dropping packet increases; therefore, the throughput will decrease. moreover, when the number of suspicious nodes becomes equal or more than the sender nodes, the throughput will be equal to zero. in the taodv technique, the suspicious nodes will not participate in the safe path to the destination, due to the trust value factor, therefore, only the trusted ones will join the safe path, still, the trusted nodes number starting is lower than the normal ones, as a result less throughput and higher packet overhead. in edri technique, all the suspicious nodes will be detected by each sender node in the first run; therefore, the number of figure 8: rreq number evaluation 0 2 4 6 8 10 12 14 16 18 5 10 15 20 25 30 e nd to e nd d el ay (m s) no. of suspicious nodes taodv edri figure 9: delay comparison between trusted ad hoc on-demand distance vector and extended data routing information figure 7: graph represents the trusted nodes overtime qasmarrogy and almashhadani: ad hoc on-demand distance vector inherent techniques comparison for detecting and eliminating the black hole attack nodes in mobile ad hoc network 81 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 77-81 figure 10: throughput comparison between trusted ad hoc on-demand distance vector and extended data routing information dropped packets will decrease, throughput will be higher in general, also by time, all the suspicious nodes will be detected, and they will be eliminated, therefore, the throughput will increase through time rapidly. figure 10 shows the throughput for the two techniques. conclusion in this paper, two annotative techniques were compared to detect and eliminate the black hole node attack in manet, the paper compared three metrics; delay, packet overhead, and throughput. as a conclusion, the taodv technique shows a lower delay at the beginning of the transmission, due to the fact that no data control packet was used to detect the suspicious nodes, after that, the delay increases slowly, due to no elimination of suspicious nodes. while in the edri technique, the delay higher in the beginning then it decreases rapidly during the time. in throughput, taodv shows lower packet delivery compared to edri, while edri shows better throughput due to the elimination of suspicious nodes overtime, which increases the packet successful delivery. finally, both techniques show good delay and throughput to detect the problem of black hole attack nodes, but these techniques work only in one run transmission, new protocols needed to detect multiple runs from the 1st time. references 1. s. b. sharma and n. chauhan. “security issues and their solutions in manet”. ieee international conference on futuristic trends on computational analysis and knowledge management (ablaze), pp. 289-294, 2015. 2. r. sheikh, m. s. chande and d. k. mishra. “security issues in manet: a review”. ieee 7th international conference on wireless and optical communications networks, pp. 1-4, 2010. 3. m. n. alslaim, h. a. alaqel and s. s. zaghloul. “a comparative study of manet routing protocols”. ieee 3rd international conference on e-technologies and networks for development, 2014, pp. 178-182. 4. m. chitkara and m. w. ahmad. “review on manet: characteristics, challenges, imperatives and routing protocols”. international journal of computer science and mobile computing, vol. 3, no. 2, pp. 432-437, 2014. 5. r. shenbagapriya and n. kumar. “a survey on proactive routing protocols in manets”. ieee 2014 international conference on science engineering and management research, pp. 1-7, 2014. 6. d. n. patel, s. b. patel, h. r. kothadiya, p. d. jethwa and r. h. jhaveri. “a survey of reactive routing protocols in manet”. ieee international conference on information communication and embedded systems, pp. 1-6, 2014. 7. r. dilli and p. c. s. reddy. “trade-off between length of the hash code and performance of hybrid routing protocols in manets”. ieee 2nd international conference on applied and theoretical computing and communication technology, pp. 732-735, 2016. 8. a. dorri, s. r. kamel and e. kheirkhah. “security challenges in mobile ad hoc networks: a survey”. international journal of computer science and engineering survey, vol. 6, no. 1, 15-29, 2015. 9. m. m. alani. “manet security: a survey”. ieee international conference on control system, computing and engineering, pp. 559-564, 2014. 10. m. kuzlu, m. pipattanasomporn and s. rahman. “communication network requirements for major smart grid applications in han, nan and wan”. computer networks, vol. 67, pp. 74-88, 2014. 11. r. ranjan, n. k. singh and a. singh. “security issues of black hole attacks in manet. ieee international conference on computing”. communication and automation, pp. 452-457, 2015. 12. m. s. el-azhari, o. a. al-amoudi, m. woodward and i. awan. “performance analysis in aodv based protocols for manets”. ieee international conference on advanced information networking and applications workshops, pp. 187-192, 2009. 13. m. b. m. kamel, i. alameri and a. n. onaizah. “staodv: a secure and trust based approach to mitigate black hole attack on aodv based manet”. ieee 2nd advanced information technology, electronic and automation control conference, pp. 1278-1282, 2017. 14. a. dorri. “an edri-based approach for detecting and eliminating cooperative black hole nodes in manet”. wireless networks, vol. 23, no. 6, 2017, pp. 1767-1778. tx_1~abs:at/add:tx_2~abs:at 52 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 52-55 review article enhancing wireless sensor networks routing protocols based on cross layer interaction reem j. ismail1*, khalid f. jasim1, samar j. ismael2, soma a. m. solaimanzadeh1 1department of computer science, college of science, cihan university-erbil, kurdistan region, iraq, 2department of electromechanical engineering, university of technology, baghdad, iraq abstract wireless sensor networks (wsn) aim to develop a smart city based on sensing environment. the routing protocols of wsn are important to transfer the data in smart cities since sensor nodes have limited power and transmission range. the aim of this research is to enhance wsn routing protocols based on proposed cross-layer interaction between physical layer and network layer also a proposed routing table information of wireless sensor nodes is developed to consider the transmission power of neighbor’s nodes to determine the next hop. cross-layer interaction provides a useful information and effective adaptation for wsn routing protocols. as a result, the proposed routing protocol shows an improvement in network performance when number of intermediate nodes is minimized. keywords: transmission power, wireless sensor node, routing protocol, cross-layer interaction introduction wireless sensor networks (wsns) are under growth and revolution technology in wireless communication since it is the most widely used in smart cities to monitor or to sense the environment.[1,2] there are two types of wireless sensor nodes: fixed wireless sensor nodes and mobile wireless sensor nodes. in fixed wireless sensor nodes, the nodes are placed in strategic and suitable positions while, in mobile wireless sensor nodes, the nodes are moving and sensing for data collection.[3,4] wireless sensor nodes have different applications that are used on land, underground or under water or even in human body and each one of them face different problems and challenges.[5,6] wsns can be classified into homogeneous (identical mobile sensor nodes) and heterogeneous (different abilities in node property).[7,8] the traditional layered approach concerning osi layered model is not suitable to be used in the mobile wireless sensor nodes design due to the unstable conditions of mobile nodes and wireless links reliability changing. the dynamic environment performance can be improved by considering cross interaction between the layers. these cross-layer interactions provide an exchange information to control the dynamic environment and will allow improvement of network performance.[9] in this research, an interaction of the physical layer with the network layer is used with different parameters that result in a new physical-network cross-layer design, the new physicalnetwork cross-layer design is elaborated to implement the proposed wsns routing protocol that will effect and improve network performance. the main contributions of this research are as follows: 1. design an efficient routing protocol for wsns with reliable intermediate nodes that participate in the routing process. 2. the development of a new structure for wireless sensor nodes routing table to consider cross-layer interaction parameters. 3. minimize the flooding problem when sending information from source to destination. the remainder of this research will be as follows. we present some of the work related to wsns concepts; we present the wsn routing protocols based on cross-layer interaction; we evaluate the proposed routing protocol in an urban scenario; and finally, we conclude this work. literature review in the literature, wsns use routing protocols with cross-layer interaction to exchange information over wireless sensor nodes to find the path from source to destination. corresponding author: reem j. ismail, department of computer science, college of science, cihan university-erbil, kurdistan region, iraq. e-mail: reem.jafar@cihanuniversity.edu.iq received: june 26, 2021 accepted: november 12, 2021 published: november 30, 2021 doi: 10.24086/cuesj.v5n2y2021.pp 52-55 copyright © 2021 reem j. ismail, khalid f. jasim, samar j. ismael, soma a. m. solaimanzadeh. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) ismail, et al.: enhancing wsn routing protocols based on cross layer interaction 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 52-55 urmila et al.[9] proposed a cross-layer design with fewer intermediate sensor nodes and keep others in the sleep mode to maximize the lifetime of sensor nodes and to manage sensor node energy efficiently. azlan et al.[10] proposed a cross layer design of medium access control mac and routing, to deliver sensing data from wireless sensor nodes to the sink. when wireless sensor node has data to transmit, it broadcasts the packets to all its neighbors, and this will affect the efficiency of the routing algorithm when flooding problem will happen. nabil et al.[11] used cross-layer design parameters and exchanged between different layers to ensure efficient use of energy. the stored energy system is used to take decisions for the wireless sensor node state and then in the routing protocol. mobile wireless sensor node architecture in general, wireless sensor node architecture includes processing unit, sensing unit, power unit, and transceiver unit while mobile wireless sensor nodes have additional unit relate to positioning, mobility, and power generation, as shown in figure 1.[12,13] in jung et al.,[14] they developed a sensor node hardware that has processor of 8 mhz, ram of 8 kb, and storage of 116 kb, the nodes are limited in its specification due to their small size. understanding transmit power of wireless sensor node in wsns, use the radio frequency (rf) signals for transceiver unit in wireless sensor nodes. wireless sensor nodes transceiver operates at the high frequencies and high sampling rates to have specific rf signal processing.[15,16] the distance that a rf signal can be transmitted depends on several hardware factors of wireless sensor nodes, includes: 1. transmitter power: the power of the signal is measured in dbm (decibels) or mw (mill watts).[17] 2. cable losses between the transmitter and its antenna and cable losses between the receiver and its antenna.[18,19] 3. antenna gain of the transmitter and receiving antenna gain.[18,20] 4. localization of the two antennas: the distance between sender and receiver is important and a better data transfer will be gain when there are no obstacles between both antennas and when both antennas are in line of sight. free space path loss problem refers to power lost, it happened when energy of rf disperses into the air.[21] 5. receiver sensitivity: receiver sensitivity is the minimum signal power level that is necessary for the receiver to accurately decode a given signal it is measured in dbm or mw.[18] proposed wsn routing protocols based on cross layer interaction flooding in network is the process of broadcasting packets form one node to all other nodes. in wsn routing protocols, the wireless sensor node broadcasts the receiving packet to discover the route to all neighbors, in which each intermediate wireless sensor node will rebroadcast the packet again that will cause a broadcasting storm and a flooding problem.[22] based on these analyzes, we propose a new protocol which interacts between the different layers: physical layer and network layer, at the aim of: improving the routing protocol and minimizing the broadcasting storm problem for intermediate nodes. the transmit power range of the sensor node is a parameter that will be considered from the physical layer to be crossed to network layer. this parameter will affect the performance of the network layer and the proposed routing protocol, as shown in figure 2. a new routing algorithm is proposed and implemented to send information from source to destination with minimum number of intermediate nodes. communication link in wireless sensor node will has unreliable wireless link, unreliable links need to prevent from the use.[19,23] the wireless sensor node will detect all the neighbor nodes to find next hop to send the information, as shown in figure 3, where s 0 coverage area and its neighbors s 1 , s 2 , and s 3 are detected (s 4 is outside its range). the routing table will be updated whenever the wireless sensor node is moving from point to point or every t-time and the next hop will be chosen according to the maximum transmit power range and transmission direction to reach to the destination. the proposed routing table structure needs to be developed to consider the physical layer parameter (transmit power range of wireless sensor node), as shown in table 1: network layer physical layer the proposed routing protocol for wireless sensor networks considering physical layer parameter for selective flooding the power range of the wireless sensor nodes figure 2: the proposed physical-network cross-layer parametersfigure 1: architecture of the mobile wireless sensor node ismail, et al.: enhancing wsn routing protocols based on cross layer interaction 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 52-55 according to the proposed routing table for wireless sensor node s 0 that is shown in table 1, the next hop to be selected will be the wireless sensor nodes: s 1 or s 3 since they have high power range relate to s 0 . algorithm 1: proposed wsn routing protocols input: source wireless sensor node (s), destination wireless sensor node (d), and transmit power range of (t) wireless sensor nodes (n) output: path from (s) node to (d) node and updated routing table begin for each wireless sensor node in the region n discover neighbor wireless sensor nodes get transmit power range for each neighbor wireless sensor node by exchanges information every 3 s if d = neighbor wireless sensor nodes send data from s to d else find next hop with the higher transmit power range t from the neighbor wireless sensor nodes to send data update the routing table with next hop and new transmit power range from the neighbor wireless sensor nodes end implementation and results in this section, we present the implementation of the proposed wsn routing protocols using matlab r2016a (9.0.0.341360). we proposed a heterogeneous mobile wireless sensor node, where the sensor nodes are mobile and have a different hardware specification that relates to transmit power and energy management. the evaluation of the results is shown as line plots using pyplot module of matplotlib library in python programming language. for this purpose, python 3.8 was used. figure 4 shows neighbors’ discovery of sensor nodes. we observed that proposed selective flooding based on crosslayer interaction shows better performance compared to broadcasting to all neighbors since fewer number of neighbors for sensor nodes will deliver packets over routing protocols because only high power transmit wireless sensor node will be considered in routing protocols. table 1: proposed routing table for wireless sensor node s0 wireless sensor node (wsn) transmit power range of wireless sensor node next hop s1 high s1 s2 medium s3 high s3 s4 low s3 s0 s1 s2 s4 coverage areas of s0 figure 3: coverage area of wireless sensor node s 0 and neighbor nodes. wireless sensor node, wireless link figure 4: compares neighbors’ discovery of sensor nodes of broadcasting flooding with proposed selective flooding based on cross-layer interaction s0 ismail, et al.: enhancing wsn routing protocols based on cross layer interaction 55 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 52-55 conclusion in this research, we propose a new routing table is updated with physical layer parameter relate to power range of wireless sensor nodes that aimed to enhance the reliability of wsn routing protocol. the proposed cross-layer interaction of wsn routing protocols will minimize the number of sensor nodes that will deliver the packets in routing protocol when selective forwarding neighbors with the high power range will participate in routing compared to broadcasting flooding of packets to all neighbors. furthermore, reliable transmission in routing protocol will improve the performance of wsns when weak power range of wireless sensor nodes will not be considered to be as an intermediate node in routing. references 1. k. z. ghafoor, m. guizani, l. kong, h. s. maghdid, and k. f. jasim. enabling efficient coexistence of dsrc and c-v2x in vehicular networks. ieee wireless communications, vol. 27, no. 2, pp. 134-140, 2020. 2. m. k. faaeq, n. k. n. mat, a. k. faieq, m. m. rasheed, and q. h. al-salami. towards of smart cities based on the sustainability of digital services. international journal of engineering and technology, vol. 7, no. 4, p. 28, 2018. 3. m. carlos-mancilla, e. lópez-mellado, and m. siller. wireless sensor networks formation: approaches and techniques. journal of sensors, vol. 2016, p. 2081902, 2016. 4. h. f. hasan, a. a. mahdi, and v. zira. cost and energy efficient hybrid wireless optical networks: fiwi access network. massachusetts: lap lambert academic publishing, p. 978, 2019. 5. l. chelouah, f. semchedine, and l. bouallouche-medjkoune. localization protocols for mobile wireless sensor networks: a survey. computers and electrical engineering, vol. 6, no. 4, pp. 1-19, 2017. 6. a. n. abdulfattah, c. c. tsimenidis, and a. yakovlev. ultralow power m-sequence code generator for body sensor node applications integration. vlsi journal, vol. 12, no. 3, pp. 89-93, 2019. 7. c. h. wu, and y. c. chung. heterogeneous wireless sensor network deployment and topology control based on irregular sensor model. in: c. cérin, and k. c. li, (eds.), second international conference on grid and pervasive computing. gpc 2007. lecture notes in computer science. paris, france: springer berlin, heidelberg, pp. 78-88, 2007. 8. s. sankar, h. ranganathan, and s. venkatasubramanian. a study on next generations heterogeneous sensor networks. in: 2009 5th ieee gcc conference and exhibition; 17-19 march 2009. kuwait: ieee, pp. 1-4, 2009. 9. u. patil, a. v. kulkarni, r. menon, and m. venkatesan. a novel aeb-aodv based aadithya cross layer design hibernation algorithm for energy optimization in wsn. wireless personal communication, vol. 117, pp. 1419-1439, 2020. 10. a. awang, x. lagrange, and d. r. sanchez. a cross-layer medium access control and routing protocol for wireless sensor networks. journées doctorales en informatique et réseaux, vol. 4, p. 02161417, 2009. 11. n. a. alrajeh, s. khan, j. lloret, and j. loo. secure routing protocol using cross-layer design and energy harvesting in wireless sensor networks. international journal of distributed sensor networks, vol. 9, no. 1, p. 374796, 2013. 12. v. ramasamy. mobile wireless sensor networks: an overview. india: intechopen, 2017. 13. n. sabor, s. sasaki, m. abo-zahhad, and s. m. ahmed. a comprehensive survey on hierarchical-based routing protocols for mobile wireless sensor networks: review, taxonomy, and future directions. wireless communications and mobile computing, vol. 2017, p. 2818542, 2017. 14. w. jung, s. hong, m. ha, y. j. kim, and d. kim. ssl-based lightweight security of ip-based wireless sensor networks. in: international conference on advanced information networking and applications workshops, toronto, canada: leonard barolli laboratory, 2009. 15. á. lédeczi, and m. maróti. wireless sensor node localization. philosophical transactions of the royal society a, vol. 370, pp. 85-99, 2012. 16. l. r. flaih, and s. a. b. al-deen. machine to machine measurement (m3) framework for web of things. journal of xi’an university of architecture and technology, vol. 11, no. 11, pp. 118-125, 2019. 17. g. a. qasmarrogy. improving voip transmission for ieee 802.11 n 5ghz manet. zanco journal of pure and applied sciences, vol. 33, no. 1, pp. 157-162, 2021. 18. rf power values. united states: cisco systems inc., 2008. available form: http://www.cisco.com/en/us/tech/tk722/ tk809/technologies_tech_note09186a00800e90fe.shtml. [last accessed on 2021 jun 01]. 19. d. son1, b. krishnamachari, and j. heidemann. experimental study of the effects of transmission power control and blacklisting in wireless sensor networks. in: 1st annual ieee communications society conference on sensor and ad hoc communications and networks. united states: ieee, 2004. 20. g. a. qasmarrogy. optimizing video transmission performance in 5ghz manet, journal of duhok university, vol. 23, no. 2, pp. 402-411, 2020. 21. a. h. m. al-dalawie, and f. m. zeki. design and implementation a non-uniform helical antenna in frequency range of 450-850 mhz for ultra-high-frequency television application. cihan universityerbil scientific journal, vol. 3, no. 2, pp. 75-79, 2019. 22. s. yamazaki, y. abiko, and h. mizuno. a simple and energyefficient flooding scheme for wireless routing. wireless communications and mobile computing, vol. 2020, p. 8832602, 2020. 23. g. a. qasmarrogy, and y. s. mahmood. capacity analysis of multiple-input-multiple-output system over rayleigh and rician fading channel. cihan university-erbil scientific journal, vol. 3, no. 2, pp. 70-74, 2019. . 70 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 70-74 research article capacity analysis of multiple-input-multiple-output system over rayleigh and rician fading channel yazen saifuldeen mahmood*, ghassan amanuel qasmarrogy department of communication and computer engineering, cihan university-erbil, kurdistan region, iraq abstract this paper aims to analyze the channel capacity in terms of spectral efficiency of a multiple-input-multiple-output (mimo) system when channel state information (csi) is known using water-filling algorithm and unknown at the transmitter side which it has been shown that the knowledge of the csi at the transmitter enhancing the performance, the random rayleigh and rician channel models are assumed. ergodic capacity and outage probability are the most channel capacity definitions which are investigated in this study. matlab code is devised to simulate the capacity of mimo system for different numbers of antenna nodes versus different signal-to-noise ratio (snr) values. in addition, the outage capacity probabilities for vary transmission rate and snr are discussed. keywords: channel state information, ergodic capacity, multiple-input-multiple-output, outage capacity, rayleigh, rician, water filling introduction recently, the evolving technologies demand for multiple-input-multiple-output (mimo) system is explosively growing; mimo system offers greater capacity than limited capacity of conventional single-input single-output (siso). mimo system can able to improve channel capacity, range, and reliability without requiring any additional bandwidth or transmit power. the large transmission rates associated with mimo channels are due to the fact that a rich scattering environment provides independent transmission paths from each transmit antenna to each receive antenna, i.e., multipath fading channel is produced. although fading is caused a performance poverty in wireless communication, mimo channels use the fading to increase the capacity.[1,2] mimo channel capacity is related to the shannon theoretic sense. the shannon capacity is the maximum mutual information of a single user time-invariant channel corresponds to the maximum data rate that can be transmitted over the channel with arbitrarily small error probability. in the timevarying channel, the channel capacity has multiple definitions, depending on what is known about the instantaneous channel state information (csi) at both transmitter and receiver, and whether or not capacity is measured based on averaging the rate overall channel states or maintaining a fixed rate for most channel states. when csi is known perfectly both at transmitter and receiver, the transmitter can adapt its transmission strategy relative to the channel using many power control algorithms for wireless networks.[3,4] therefore, channel capacity is characterized by the ergodic or outage. ergodic capacity can be defined as expected value of channel capacity which is suitable for fast varying channels. the outage probability is the maximum rate that can be maintained in all channel states with some probability of outage. when only the receiver has perfect knowledge of the csi, then the transmitter must maintain a fixed rate transmission strategy based on knowledge of the channel statistics only, which can include the full channel distribution just its mean and variance are known. in this case, ergodic capacity defines the rate that can be achieved through this fixed rate strategy based on receiver averaging overall channel states.[5] alternatively, the transmitter can send at a rate that cannot be supported by all channel states, in these poor channel states that the receiver declares an outage and the transmitted data are lost. the remainder of this paper is organized as follows. the next section presents the multipath fading channels (rayleigh and rician). capacity of mimo system will be studied in section 3. in section 4, ergodic capacity and outage capacity notions are explained. section 5, the simulation results with discussions will detail. finally, major conclusion of the paper is discussed. corresponding author: yazen saifuldeen mahmood, cihan university-erbil, kurdistan region, iraq. e-mail: yazen.mahmood@cihanuniversity.edu.iq.test-google-a.com received: apr 9, 2019 accepted: apr 23, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp70-74 copyright © 2019 yazen saifuldeen mahmood, ghassan amanuel qasmarrogy. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ mahmood and qasmarrogy: capacity analysis of mimo system over rayleigh and rician fading channel 71 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 70-74 fading channel models rayleigh channel the rayleigh fading channel is composed from the effects of multipath embrace constructive and destructive interference, and phase shifting of the signal. the rayleigh distribution is commonly used to describe the statistical time-varying nature of the received envelope of the flat fading signal, or the envelop of an individual multipath component, where there is no line of sight (los) path means no direct path between transmitter and receiver. the envelope of the sum of two quadrature gaussian noise signals obeys a rayleigh distribution.[6] the rayleigh distribution has a probability density function (pdf) given by:               2 22 2             0 0              0 r r e rp r r σ σ (1) where, σ2 is the time-average power of the received signal before envelope detection. rician channel when there is los, direct path is normally the strongest component goes into deeper fade compared to the multipath components. this kind of signal is approximated by rician distribution. rayleigh fading with strong los content is said to have a rician distribution or to be rician fading.[7] the rician distribution has a probability density function (pdf) given by:   2 2 2 ( ) 2 02 2 0, 0 0 0 r a r ar e i a r p r r                  σ σ σ (2) where, i 0 is the modified bessel function of the first kind and zero order. the parameter a denotes the peak amplitude of dominant signal. capacity of mimo system csi not available at the transmitter mimo system provides a significant capacity gain over a traditional siso channel, given that the underlying channel is rich of scatters with independent spatial fading. mimo systems offered increasing in channel capacity based on the diversity at both the transmitter and the receiver and also on the number of antennas at both sides. when the transmit power is equally distributed among the n transmit antennas in case of csi is perfectly known at the receiver but unknown at the transmitter, mimo capacity with n tx and m rx antennas can be given in terms of bit per second per hertz by, * 2log det /mimo mc i hh bps hz n            (3) where, ρ is the signal to noise ratio (snr), i m is the identity matrix, (*) means transpose-conjugate, and h is the m × n channel transmission matrix. this equation can be reduced as follows:[8] 21 log 1 /   =  = +  ∑ m mimo ii c bps hz n (4) where, m = min (m, n) λ i are the eigenvalues of h h*, known that the square root of λ i is the diagonal matrix, d. the “diagonalization” process has been achieved by performing the singular value decomposition of matrix h which put it in the form. h=u d v* (5) with d being a diagonal matrix, and u and v unitary orthonormal matrices. csi available at the transmitter (waterfilling algorithm) water-filling algorithm is an optimum solution related to a channel capacity improvement. this algorithm makes csi known at the transmitter side which can be derived by maximizing the mimo channel capacity under the rule that more power is allocated to the channel that is in good condition and less or none at all to the bad channels. algorithm steps:[9] 1. take the inverse of the channel gains 2. water filling has non-uniform step structure due to the inverse of the channel gain 3. at first, take the sum of the total power p t and the inverse of the channel gain. it gives the complete area in the water filling and inverse power gain 1 1n t i i p h  (6) 4. decide the initial water level by the formula given below by taking the average power allocated (average water level) 1 1n t i i p h channels    (7) 5. the power values of each subchannel are calculated by subtracting the inverse channel gain of each channel 1 1 1 n t i i i p h power allocated channels h       (8) 6. in case, the power allocated value becomes negative stop the iteration process. the capacity of a mimo system is algebraic sum of the capacities of all channels and is given by the formula below.   n 2i 1capacity log 1 power allocated * h  (9) mahmood and qasmarrogy: capacity analysis of mimo system over rayleigh and rician fading channel 72 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 70-74 ergodic capacity and outage capacity ergodic capacity mimo channels are random in nature. therefore, h is a random matrix, which means that its channel capacity is also randomly time varying. by taking the ensemble average of the information rate over the distribution of the elements of the channel matrix h, we obtain the ergodic capacity of a mimo channel. hence, the mimo channel capacity can be modeled as follows:[10] c = e{c(h)} (10) the ergodic channel capacity without using csi at the transmitter side, from (6), is given as follows: m 2 ii 1c e log 1 n               (11) similarly, the ergodic channel capacity for using csi at the transmitter side is given as follows: opt m i 2 ii 1c e log 1 n              (12) where, opt i is the power coefficient that corresponds to the amount of power assigned to the ith channel. outage capacity outage capacity is used for slowly varying channels where the instantaneous snr is assumed to be constant for a large number of transmitted symbols. unlike the ergodic capacity definition, schemes designed to achieve outage capacity allow for channel errors. hence, in deep fades, these schemes allow the data to be lost and a higher data rate can be thereby maintained than schemes achieving shannon capacity, where the data need to be received without an error overall fading states.[11] the parameter p out indicates the probability that the system can be in outage, i.e., the probability that the system cannot successfully decode the transmitted symbols. corresponding to this outage probability, there is a minimum received snr, snr min given by p out = p(snr<snr min ), for received snrs below snr min , the received symbols cannot be successfully decoded with probability 1, and the system declares an outage. in other words, the system is said to be in outage if the decoding error probability cannot be made arbitrarily small with the transmission rate of r bps/hz. the definition of the outage probability as follows: p out (r) = p(c(h)<r) (13) results and discussion mimo channel capacity and outage probability over rayleigh and rician fading channel are simulated using matlab m-file for different antenna configurations, different snr, and with and without csi at the receiver. the transmission channel matrix h is chosen randomly from the probability distribution of the considered fading channels. assuming rayleigh distribution of unity σ and rician distribution of unity σ and unity peak amplitude (a) except in figures 1 and 2, where σ has values 1 and 2. ergodic capacity the ergodic capacity average of 10,000 random generated matrix h in case of csi unknown and known at the transmitter side, i.e. without and with using water filling of the system (in terms of b/s/hz) is calculated for rayleigh and rician channels over a wide range of snr. the capacity of siso system is compared in figures 1 and 2 with 2 × 2 and 4 × 4 mimo over rayleigh channel and rician channel, respectively, for a two different values of σ. it is noticeable that the capacity is increased as snr increases and the capacity is increased for increases in number of antennas in both transmitter and receiver sides. in addition, the capacity increases as σ value increases. figure 3 shows that the water-filling algorithm has improved the transmission capacity for both channels of 4 × 4 mimo, especially for low snr. this figure depicts that the rician of unity σ and unity peak amplitude (a) has a higher capacity compared with rayleigh of unity σ. moreover, the improvement is achieved over rayleigh rather than rician channel. figure 1: single-input single-output and multiple-input-multipleoutput capacity over rayleigh channel figure 2: single-input single-output and multiple-input-multipleoutput capacity over rician channel mahmood and qasmarrogy: capacity analysis of mimo system over rayleigh and rician fading channel 73 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 70-74 outage capacity outage capacity is useful to estimate when csi is known at the receiver but not at the transmitter. therefore, the capacity is a random variable, and no matter how small the rate threshold would be, there is always a non-zero probability that the channel is incapable of supporting arbitrarily low error rates. in this section, the outage capacity probability versus transmission rate is simulated for 4 × 4 mimo system. the outage capacity probabilities are shown in figure 4 for a mimo channel for various value of snr (5, 10, 15, and 20). the figure shows the variance of the instantaneous capacity increased as the snr increased for the same system. the outage capacity probabilities are shown in figure 5 for rayleigh and rician channel with and without applying water-filling algorithm, and it has been clear that rician is better compared with rayleigh channel due to the los is existing between the antennas in the formal channel which makes a higher receiving signal level. the figure depicts the outage capacity probabilities mitigated much more when applying water filling on the rayleigh channel. conclusion in this paper, analysis and comparison of the mimo was presented when csi is known/not known at the transmitter over fading channel models by calculating the ergodic capacity and outage capacity probabilities. the results show that the capacity was enhanced when csi known at the transmitter by applying water-filling algorithm on both assumed channel models. this study concludes that water-filling algorithm gives more ergodic and outage capacity improvement over the rayleigh channel than the rician channel considering their simulation parameters although the performance of rician channel is obviously better, in general, than rayleigh channel. references 1. t. hashem and m. i. islam. “performance analysis of mimo link under fading channels”. in: 2014 17th international conference on computer and information technology”. ieee, pp. 498-503, 2014. 2. m. narayana and g. bhavana. “performance analysis of mimo system under fading channels (rayleigh and rician) using svd pca and fsvd”. journal of engineering technology, vol. 5, pp. 116-126, 2016. 3. y. liu, l. liu and c. xu. “spectrum underlay-based water-filling algorithm in cognitive radio networks”. in: 2011 international conference on electric information and control engineering, ieee, pp. 2614-2617, 2011. 4. s. yan, p. ren and f. lv. “power allocation algorithms for ofdmbased cognitive radio systems”. in: 2010 6th international conference on wireless communications networking and mobile computing. ieee, pp. 1-4, 2010. 5. q. sun, s. han, i. chin-lin and z. pan. “on the ergodic capacity of mimo noma systems”. ieee wireless communications letters, vol. 4, no. 4, pp. 405-408, 2015. 6. p. bhatnagar, j. singh and m. tiwari. “performance of mimoofdm system for rayleigh fading channel”. international journal of science and advanced technology, vol. 1, no. 3, pp. 37-40, 2011. 7. c. xiao, y. r. zheng and n. c. beaulieu. “novel sum-of-sinusoids simulation models for rayleigh and rician fading channels”. ieee transactions on wireless communications, vol. 5, no. 12, pp. 3667-3679, 2006. 8. n. kumar. “enhance the capacity of mimo wireless communication channel using svd and optimal power allocation algorithm”. international journal of electronics and telecommunications, vol. 65, no. 1, pp. 71-78, 2019. 9. h. deshmukh and h. goud. “capacity analysis of mimo ofdm system using water filling algorithm”. international journal of figure 3: the capacity with and without using water-filling algorithm over rayleigh and rician channel figure 4: the outage probability versus transmission rate with different signal-to-noise ratio threshold figure 5: outage probability versus transmission rate with signal-tonoise ratio threshold 10 db in rayleigh and rician channel mahmood and qasmarrogy: capacity analysis of mimo system over rayleigh and rician fading channel 74 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 70-74 advanced research in computer engineering and technology, vol. 1, no. 8, pp. 329-333, 2012. 10. a. paulraj, r. nabar and d. gore. “introduction to spacetime wireless communications”. cambridge university press, cambridge, 2003. 11. h. luo, d. xu and j. bao. “outage capacity analysis of mimo system with survival probability”. ieee communications letters, vol. 22, no. 6, pp. 1132-1135, 2018. . 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 research article a face recognition system based on principal component analysis-wavelet and support vector machines laith r. fleah1*, shaimaa a. al-aubi2 1department of computer science, cihan university-erbil, iraq, 2department of computer science department, college of science, salahaddin university, erbil, iraq abstract face recognition can represent a key requirement in various types of applications such as human-computer interface, monitoring systems, as well as personal identification. in this paper, design and implement of face recognition system are introduced. in this system, a combination of principal component analysis (pca) and wavelet feature extraction algorithms with support vector machine (svm) and k-nearest neighborhood classifier is used. pca and wavelet transform methods are used to extract features from face image using and identify the image of the face using svms classifier as well as the neural network are used as a classifier to compare its results with the proposed system. for a more comprehensive comparison, two face image databases (yale and orl) are used to test the performance of the system. finally, the experimental results show the efficiency and reliability of face recognition system, and the results demonstrate accuracy on two databases indicated that the results enhancement 5% using the svm classifier with polynomial kernel function compared to use feedforward neural network classifier. keywords: face recognition, feedforward backpropagation, neural network and k-nearest neighborhood, support vector machines introduction facial recognition is considered as one of the biometric systems and the importance of this field as a topic of research has increased in recent years. face recognition system uses biometric information which takes from human, but the use of this system is more easily than in another biometric system such as fingerprints, iris, and signature, especially with the uncooperative people. face recognition systems are usually used in security systems to monitor people and to discover any security breach, for example, face recognition system uses in the airport for monitoring and arrests the wanted. therefore, face recognition system is used in many security applications such as avoiding crime, monitoring through video, and also for verification of the identity of persons. principal component analysis (pca) is one of the linear transformation techniques that are used to obtain features from data or to decompress the information. it is also known as one of the famous techniques that are used to take global structures from the high-dimensional information set as well as it is applied to decompose dimensionality of the data and take unique features from the faces picture. this technique can be used also to distinguish patterns in data and express the data so as to highlight their differences and similarities between them.[1] wavelet transforms are a very effective method and specifically in image compression and many other types of data. as many coefficients of wavelet transform method become zero or very small. for this reason, wavelet transforms can consider as a very helpful tool for image compression. the primary benefit of wavelet transforms compared than other decomposition methods is that the basic functions that related with a wavelet decomposition typically have short and long support. the short support can effectively act on sharp transitions as the edges of the image; however, long support is operative in representing slow variations of the image.[2] support vector machine (svm) is one of important learning algorithms that are applied in many of applications and it is currently represented one of the very used methods in many application fields. the svm theories were introduced at first in the sixties and seventies from the previous century by the scientists vapnik and chervonenkis, but the svms method is still not implementations practical until the early.[3] cihan university-erbil scientific journal (cuesj) corresponding author: laith r. fleah, department of computer science, cihan university-erbil, iraq. e-mail: laith.flaih@cihanuniversity.edu.iq received: apr 18, 2019 accepted: apr 25, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp14-20 copyright © 2019 laith r. fleah, shaimaa a. al-aubi. this is an open-access article distributed under the creative commons attribution license. fleah and al-aubi: frs-pca and svm 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 it has been applied in many of applications in many various fields such as medical diagnosis, text or image classification and also categorization, spam categorization, detection and recognition of an object, face detection, verification and recognition, bioinformatics, signal processing, prediction, information, and image retrieval.[4] k-nearest neighbor [knn] is one of the most important algorithms used in face recognition system. this algorithm is easy to understand, but in another hand, it is useful in practice. furthermore, it is used in several of applications such as vision, proteins, computational geometry, and graphs. it also considered as one of the best 10 algorithms in data mining.[5,6] knn is considered as one of the non-parametric learning algorithms. this means that it does not produce any supposition on the implicit data distribution. this is very helpful in the real-world application, many of the practical data do not conform the classical theoretical supposition made such as linearly separable and gaussian mixtures.[5,6] the overall structure of the paper is organized as follows: section ii: introduced the structure of the system, this is included pre-processing methods, feature extraction methods, and classification methods. in section iii, the experimental results and its analysis are introduced while section iv concludes the paper. face system structure face recognition system that introduced in this paper consists of the following steps: pre-processing, feature extraction, classification, and in the final steps is identification. the stages of the system are shown in figure 1. pre-processing pre-processing stage is the first process that done in the proposed system. it is applied to improve the image and also to improve the recognition rate through remove the effect of the light on and also to correct the gradient of illumination in the image. in the following, the pre-processing techniques that applied at this stage to get the good results: normalization size of the image: two types of database are used in this thesis and each database has different size of an image; therefore, this technique is used to make all images have the same size. this is because the next stages have an arithmetic operation such as multiplication and division of array. for all these reasons, the size normalization is very significant. contrast stretching: it is also known as histogram stretching. it is considered as one of the techniques that used to enhance an image. the goal of this technique has improved the contrast in an image through stretch the range of intensity values. in other words, this is applied by raising the dynamic range of the gray levels in the image. for example, in an 8-bit system, the 256 gray levels can be shown only but if the number of gray levels in the captured image prevalence through a lesser range, the images can be improved by extended this number to a wider range. features extraction the second stage of this system is feature extraction. it is used to obtain the significant features from the picture of the face and then applied these features to the next stage (identification and classification stages). without this stage, the function of recognition and classification becomes very hard and very difficult because the picture consists of many features and most of these features cannot be applied to implement classification. in this paper, each of pca and wavelet transform techniques is used to obtain the significant features that are important to identify and classify the face pictures. after preprocessing stage is implemented, the face picture i (x, y) that is a two-dimensional matrix (n × n) is then transformed to a vector and that length is had dimension n². thus, the picture with dimension 64 × 64 will transform to a vector of dimension 4096, but this vector cannot be applied directly to recognition directly because the face of human is very comparable from each other and another reason the 4096-dimension space is big and it is needed much time to process. therefore, wavelet and pca techniques are applied to obtain a better feature from face picture and also to enhance the capability of the recognition rate. pca algorithm at first, the training set is generated and this training set consists of a number of images (m) and each image in its to be transformed to the vector (n2). the step after generated the training set is calculated the mean of images as this is described in equation 1: figure 1: the structure of a proposed system fleah and al-aubi: frs-pca and svm 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 ψ γ= = ∑ 1 1m n m i (1) γ i in the question (1) refers to the image in the training group. then, the normalized is implemented in training set to achieve the zero mean that describes the degree of different between the mean of image and image, and this is described in equation: ϕ i = γ i –ψ i 1, 2… m (2) the covariance matrix c of the training set can be obtained using equation 2. ψ = = ∑ 1 1m n m i i t  (3) i̇f a vector u satisfies the condition in question (4), the eigenvector of the covariance matrix c is considered a nonzero vector. c u k = v k u k (4) where, v k refers to the corresponding eigenvalues. the size of matrix c with the dimension of n × n is a large size and needs much computational time and size memory to obtain results from it, to solve this problem, the covariance matrix l that has small size compare than the original covariance matrix c is calculated. equation 5 describes how the matrix c can be decomposed: c = aat (5) where, a represents zero means matrix [ϕ 1 , ϕ 2 …, ϕ i ] and the eigenvectors v i can be consideration according to the following: aat v i = µ i v i (6) now, by multiply each side of equation 6 by a, we can obtain the following: aat av i = µ i av i (7) from equation (7), we can see that represents the eigenvectors of the covariance matrix c and the matrix l with dimension (m × m) can be rewritten as shown in equation (8): l = aat (8) now, we obtained the eigenvectors v i of smaller covariance matrix l that has dimension m and this way, the linear combinations of the training data of the images to the form of eigenfaces u i can be obtained from equation below: u vi k m ik k= = ∑ 1  (8) this eigenvector that has symbol u i is also called as eigenfaces because when transfer these eigenvectors form a vector of length (n) to a two-dimensional matrix with dimension (n × n) and show it. it is shown as the human face. wavelet transform it is the first step of this stage and it is used wavelet transform to remove the noise from the image with saving the quality of an image. the second benefit is to obtain some of a feature from the image and also to decompose the size of the image and by this way, thus reduced the time of the process. the output of the wavelet transform on an image, it consists of four sub-bands. these sub-bands are the following: low-low (ll), high-low (hl), low-high (lh), and high-high. the ll sub-band gives the most important features of the image and hl and lh sub-bands give the horizontal edges and vertical edges, and finally, the high sub-band gives the diagonal edges. the results that obtained after applied wavelet transform are shown in figure 2. when this technique is applied for 1 time to the image with dimensions (112 × 92), the dimensions of the four sub-bands are 56 × 46. we can conclude from the previous paragraph after used one stage of the wavelet transform, the size of the image is decreased to the half. in this work, two levels of the wavelet transform are used. classification stage a classification is the third stage of this recognition system. it is used after each of the previous stages is done. it is a very significant stage. the goals of this stage are classified the interimages based on the information that obtained through the training stage. there are a number of classification methods that can be used to do this job. in this recognition system, svms that are considered as one of the famous classification methods are used to do this function. svms are one of the most important learning algorithms which are applied in many of applications. in this work, we also aim to make a comparison between different kinds of the svms kernel to analyze its results and determine any kernels of them can give a good result with high accuracy rate and also we make a comparison with a feedforward backpropagation neural network (ffbpnn) that used as a classifier to ensure from the power of our methods. svm classifier svm is considering as learning algorithm method depended on statistical learning algorithms. it is used to figure 2: the result after used one level of the wavelet transform fleah and al-aubi: frs-pca and svm 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 classify the feature of the data by finding a maximum marginal hyperplan that separating between two classes. it also can be applied for linear and non-linear classification using different types of kernel functions. basic svm formulation svms are used to make classification among two classes to give a decision surface. this decision surface is represented a maximum distance that can depend on it to separate closest points in the classification’s class. the closest points are known as support vectors. figure 3 describes the hyperplane of support vector separating. in this part of the chapter, we will consider svms as maximum margin classifiers. to facilitate the condition that supposed, we will consider that input data can separate linearly. under this supposition, we can separate any two classes by obtained the linear hyperplane between them. figure 3 describes the good and bad separation between two classes. let us summed that, a binary classification with xi as the input data, where i = 1, 2, 3..,, l. with identical labels for the two class, now let assumed that the function of decision knows as shown in the question below: f (x) = sign (w x + b) (9) where, symbol represented the inner product and symbol b represent the bias of the function and point x take position directly on the hyperplane and it achieves the condition. w x + b = 0 (10) from calculating the left side of equation (10), we can obtain the label y i of a data, where sign refers to the label of class. the point’s x i on both sides of the hyperplane must achieve the following conditions: x i w + b < 10 (11) x i w + b > 10 (12) equations (11) and (12) can be rewritten as follows: y i (x i + b ≥ 10) (13) using the formulation of lagrangian, the svms prediction can obtain using the following equation: f x y x xi i si m ( ) ,�= <=∑ 0 (14) where, m is represented the number of support vectors, x i is a support vector, and αi is represented the corresponding lagrange multiplier, and finally, a sign of f (x) is classified every test vector. we can say that the output of classification is correct if equation (14) holds for all input points. the number of possible combinations of both weights and bias are big and sometimes not optimal. in general, in the binary classification, there is one ideal separating hyperplane is offer. on the ideal hyperplane, the margin among two data is maximized. as shown in figure 4, the margin among two sets of data is represented by d+ and d− which represented the distances between them. figure 5 describes the two separating lines, one of them is ideal and another is random.[7] identification the final stage of the system is identification. the aim of this stage is to find the nearest image for the new image figure 4: the support vector and the separating hyperplane figure 5: samples of yale database figure 3: the support vector and the separating hyperplane fleah and al-aubi: frs-pca and svm 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 that classified before that using svm or ann. knn is used to implementing that knn is not like svm or ann. it does not need to the training phase, and therefore, all the data are needed for the testing phase. the data that need for testing consist of a set of vectors and class label associated with each vector only. knn for classification in the section, we will see how can be used knn as a classifier. to do that, two data sets will create. one of them is training set and another set is testing set. the goal of this algorithm is to obtain the class label of the new point. the algorithm has various conducts depended on the value of k parameter. the first hypothesis: the value of k parameter is equal to one (k=1) to explain the first hypothesis, let consider that x is the point to be labeled and to obtain the point that closest to it. let consider the closest point y. in this way, the nn rule asks to assign the label of y to x. this appears very simplistic, but this way is correct only when the number of points is not very large. however, in other situations, when the number of data points is very big. the label of x and y may be same. to simplest the explain – let assume that the potentially biased coin. then, maybe the head will appear in next call. the same argument can be applied in this situation. now, lets us apply another case here – suppose that every point is represented in a d-dimensional plane and the number of points is very huge. this led to conclude that the density of the plane on each point of data is high. in other description, in each subspace, a large number of points is offered and consider x point is in the subspace and is many of neighbors, but in this time, consider y be the nn to the point x. we can say that, if x point and y point are very close together, then the following can propose, the probability of point x and point x related to the one class is very big, then using decision theory, the following is obtained, both points related to the one class. bellhumer et al.[8] provided a very good argument about the rule of nn. one of the important results is to obtain a very small error bound. this bound is described in question: * * (2 *) 1 c p p p p c ≤ ≤   (15) where the bayes error rate, the symbol c is represented the number of classes and the symbol p is represented the error rate of nn. the result is given the following conclusion if the number of points very huge, then the error rate of nn becomes less than twice time from the bayes error rate and this is very good results obtained from algorithm as knn. the second hypothesis: the value of k parameter is equal to two (k=2) in general, we aim to obtain the knn and then make a majority voting. as we mentioned previously, the value of k is odd when there are two numbers of classes. let’s assumed that k is equal 5 and there are three instances of c1 and two instances of c2. in this situation, knn method will label the new point as c1. this is similar when there is multiclass want to label. one of the knn techniques is not given 1 to every neighbor and another common technique is give weight to each point according to the distance that calculated. in another way, the values of each weight of point are inversely proportional to the distance of the point that will classify. from the sentence above, the neighboring points have higher values compared than the ultimate points. experimental results a number of experiments on the two different databases are used. the first experiment is applied on the yale database[8] and the second experiment is applied on the orl database.[9] both these database are very famous databases and are used by many researchers. in every database, two experiments are done. the first experiment is compared between different svms kernels to show any kernel give best results and the second experiment is between svm classifier and neural network classifier to calculate the performance of the classifier that used the system. yale experiment yale database is used in this experiment. this database is created by yale university and it is provided free on the internet and anyone can download it. this database contains from 15 subjects and each subject has 11 images take under various conditions. from that mention above, this means that the database consists of 165. this database is applied because it consists of a number of images captured at various conditions; this is included lighting condition or other things; therefore, it can be applied to test the recognition rate of the proposed system and it is also very widely used by many of researchers. in our experiment, the database is divided into two groups; the first group is for train and the second group is for the test. the training group is applied to learn the system and the training group consists of five images of the first 12 classes. the testing group is applied to calculate recognition rate of the proposed system and it consists of the other five images of the same 12 classes as well as all images of the last three classes. this is meant that 40% of the database is applied for training and 60% of the image is applied for the test. in this experiment, the combinations of the pca-wavelet transform are applied for pre-processing and feature extraction, and comparison between various types of the svms kernel is done. figure 6 describes the results of the svms classifier using various kernel functions in yale database. in the second experiment, ffbpnn is applied as classifier instead of the svms. this is done to make a comparison between the results of svms and anns classifiers as well as to calculate the performance of the system. figure 7 describes the recognition rate results of the svms classifier using polynomial kernel function and artificial neural network. orl experiment orl database is used in the second experiment. this database is created by cambridge university computer and anyone can download it from the internet without fees. this database fleah and al-aubi: frs-pca and svm 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 consists of 40 classes with 10 images for each class. this means that database consists of 400 images. in our experiment, the database is divided into two groups; the first group is for train and the second group is for the test. the training group is applied to learn the system and the training group consists of five images of the first 32 classes. the testing group is applied to calculate recognition rate of the proposed system and it consists of the other six images of the same 32 classes as well as all images of the last eight classes. these three classes are used for test only. this is meant that 40% of the database is applied for training and 60% of the image is applied for the test. figure 8 describes the results of the svms classifier using different kernel function in orl database. in the second experiment, ffbpnn is applied as classifier instead of the svms. this is done to make a comparison between the results of svms and anns classifiers as well as to calculate the performance of the system. figure 9 describes the recognition rate results of the svms classifier using polynomial kernel function and feedforward backpropagation neural network classifier. conclusions face recognition systems are one of the computer vision applications and it is significant in many of. in this work, face recognition system is designing and implementation. in this system, combines between each of wavelet transform and pca methods are applied for obtained feature from the image after that svm is applied to classify these features. the proposed system is tested using two types of databases to guarantee from the power of the system. comparison between the results 91.4285 93.333395.2 95.2 73.333 0 10 20 30 40 50 60 70 80 90 100 accuracy in yale d atabase linear quard polynomial raduis bias func�on mul�peceptrom func�on figure 6: the results of the support vector machines classifier using different kernel functions in yale database 95.2 87.619 0 10 20 30 40 50 60 70 80 90 100 svms using ploynomial kernel func�on feed forward backproga�on neural network figure 7: the accuracy results of the support vector machines classifier using polynomial kernel function and artificial neural network 88.3391.25 96.25 91.667 65.833 0 10 20 30 40 50 60 70 80 90 100 110 linear quard polynomial raduis bias func�on figure 8: the results of the support vector machines classifier using different kernel function in orl database 96.25 91.667 0 10 20 30 40 50 60 70 80 90 100 svms using ploynomial kernel func�on feed forward backproga�on neural network figure 9: the accuracy results of the support vector machines classifier using polynomial kernel function and artificial neural network fleah and al-aubi: frs-pca and svm 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 14-20 of proposed system and ffbnn of a classifier is implemented. according to the obtained results, the following conclusions can be stated: • applying wavelet-pca feature extraction algorithms enhanced the recognition rate of the system compared to the using pca only • using svm classifier based on polynomial kernel function increases the accuracy of recognition rate of the system by 5% compared to feedforward neural network classifier, especially in cases where the features of a person’s face are modified with different hairstyles or wearing glasses. the comparison between svm and ffbnn classifiers proved that svm classifier is more accurate than ffbnn classifier in recognition. references 1. s. gupta, o. p. sahu, r. gupta and a. goel. “a bespoke approach for face-recognition using pca”. international journal of computer science and engineering, vol. 2, no. 02, pp.155-158, 2010. 2. m. david. “reversible integer to integer wavelet transforms for image coding”. thesis, university of british columbia, 2002. 3. j. s. taneja. “analysis of e. coli promoters using support vector machine”. thesis, thapar institute of engineering and technology, 2006. 4. m. e. mavroforakis. “geometric approach to statistical learning theory through support vector machines (svm) with application to medical diagnosis”. thesis, national and kapodistrian university of athens, 2008. 5. x. wu, v. kumar, j. r. quinlan and j. ghosh. “top 10 algorithms in data mining”. knowledge and information systems, vol. 14, no. 1, pp. 1-37, 2008. 6. m. zuhaer and n. al-dabagh. “face recognition using lbp, fld and svm with single training sample per person”. international journal of scientific and engineering research, vol. 5, no. 5, pp. 180, may 2014. 7. r. o. duda, p. e. hart and d. g. stork. “pattern classification”. 2nd ed. new york: john wiley and sons, inc., 2000. 8. p. n. bellhumer, j. hespanha and d. kriegman. eigenfaces vs. fisherfaces: recognition using class specific linear projection. ieee transactions on pattern analysis and machine intelligence, vol. 19, no. 7, pp. 711-720, july 1997. 9. f. samaria and a. harter. d. parameterisation of a stochastic model for human face identificatio. conference: applications of computer vision. proceedings of the second ieee workshop, 1994. . 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 32-36 research article investigating the inhibitory effect of silver nanoparticles against some species of candida and pathogenic bacteria alaa m. hasan, sura m. abdul majeed, rusol m. al-bahrani department of biology, college of science, university of baghdad, baghdad, iraq abstract silver nanoparticles synthesized from aqueous extract of mushroom pleurotus ostreatus exhibited inhibitory effect at the concentration of 12.5, 25, 50, and 100 mg/ml against some pathogenic bacteria and fungi such as candida albicans, candida guilliermondii, candida krusei, candida zeylanoides, geotrichum klebahnii, staphylococcus aureus, pseudomonas aeruginosa, and escherichia coli. the maximum inhibition zone was observed against c. zeylanoides at the concentration of 100 mg/ml was 24.5 mm, while the minimum inhibition zone was observed against geotrichum at the concentration of 25  mg/ml was 8 mm and the concentration of 12.5  mg/ml was not effective against some species. keywords: pathogenic candida, pleurotus ostreatus, silver nanoparticles introduction a new dimension of the metal microbial interaction has been explored for the synthesis of metal nanoparticles such as gold, silver, cadmium, zirconia, and silica titanium.[1] silver compounds have been used to treat burns, wounds, and infections as well as in preventing bacterial colonization of prostheses and catheters.[2] various salts of silver and their derivatives are used as antimicrobial agents. nanosized silver particles exhibit antimicrobial properties. nanoparticles of silver have been studied as a medium for antibiotic delivery, and to synthesize composites for use as disinfecting filters and coating materials.[3] the nanoparticles were important due to their emission properties. these nanoparticles are used for wide range of application.[4,5] antibacterial activity is related to compounds that locally kill bacteria or slow down their growth, without being in general toxic to surrounding tissue.[6] the high bactericidal activity is certainly due to the silver cations released from ag nanoparticles (agnps) that act as reservoirs for the ag+ bactericidal agent.[7] severe fungal infections have significantly contributed to the increasing morbidity and mortality,[8] immunocompromised patients who need intensive treatment including broad-spectrum antibiotic therapy,[9,10] and candida spp. represent one of the most common pathogens which are responsible for fungal infections often causing hospital-acquired sepsis with an associated mortality rate of up to 40%.[11] currently, the majority of yeast species are resistant the available antifungal therapy[12,13] such as on polyenes (amphotericin b), triazoles (fluconazole, itraconazole, voriconazole, and posaconazole) or echinocandins (caspofungin, micafungin, and anidulafungin) which exhibit their toxicity, adverse effects, and drug interaction.[14,15] the objective of this work is to evaluate the effect of a biosynthesized agnps product against some candida spp. and bacteria species to determine how agnps interact with the growth of microbial cells. materials and methods the mushroom pleurotus ostreatus was obtained from fruit body grown on peach tree in salah-alddin province. preparation of hot aqueous extracts of mushroom the oven dried mushroom was blended; the obtained powder was soaked in distilled water at a ratio of 1:10 (w/v) and boiled with agitation at 60 ± 2°c for 30 min. the boiled mushroom powder was then left covered for 30 min. residues were then removed by filtration through gauze and further centrifugation (10,000 rpm, 30 min, and corresponding author: sura m. abdul majeed, department of biology, college of science, university of baghdad, baghdad, iraq. e-mail: suraalsadik80@gmail.com received: apr 04, 2019 accepted: apr 24, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp32-36 copyright © 2020 alaa m. hasan, sura m. abdul majeed, rusol m. al-bahrani. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ hasan, et al.: inhibitory effect of agnps against candida spp. and bacteria 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 32-36 4°c). supernatants were then collected and filtered through whatman no. 1 filter paper. after that, freeze dried extract powders were obtained using freeze dryer and stored at 4 ± 2°c.[16] biosynthesized agnps from p. ostreatus silver nitrate (1 × 10−3) agno3 stock solution was prepared in sterile deionised triple distilled water and the subsequent dilutions were made from this stock solution. the bulk amount of (10) mg/ml of aqueous extract solution of is prepared with sterile distilled water and filtered through syringe filter (0.2) µm. based on the result of a preliminary trial, 2–7 ml of 10 mg/ml of an aqueous extract of p. ostreatus (p2) were filled with sterile distilled water to a total of 10 ml. after that, the solution is added to 5 ml of (1 × 10−3) m aqueous agno3 solution and kept at room temperature and exposed under ultraviolet (uv) (365) nm (long uv). after 24 h incubation, the light yellow color of the mixture solution turned to dark yellow indicating the formation of agnps.[17] candida isolates all candida isolates candida albicans, candida guilliermondii, candida krusei, candida zeylanoides, and geotrichum klebahnii were procured from al-yarmouk teaching hospital. mycological identification was carried out for all samples by commercial carbohydrate assimilation systems such as the api 20 c test kit.[18] all yeast isolates were inoculated in a primary isolation medium such as sabouraud dextrose agar (sda) medium for 2–3 days at 37°c. antimicrobial activity of agnps (well diffusion method) the synthesized agnp was tested for antimicrobial activity by agar well diffusion method against pathogenic microbes for all the previously mentioned bacteria and yeast species. the pure cultures of bacteria and yeast were subcultured on nutrient and sda subsequently. each strain was swapped homogeneously onto the individual plates using sterile cotton swabs. wells of 10 mm diameter were done. the concentration of agnps was poured on each well. after 24 h of incubation the diameter of inhibition zone was measured. three replicates of experiments were carried out.[19] results the formation of agnps was confirmed by the uv-visible spectrophotometry, which showed a strong peak within the range of 200–500 nm, “figure 1.” intensity of particle size distribution analysis to know the size of synthesized agnps, size distribution analysis was performed using light scattering in aqueous solution. the results showed that the size of the particles ranged from 16 to 104 nm. the average size of agnps was determined to be 49 ± 16 nm as in “figures 2 and 3.” discussion antimicrobial mechanisms of nanomaterials are not fully understood, but it is proposed that when they come into contact with cells, they provoke the production of reactive oxygen specie, cell membrane disruption, mitochondrial damage, and dna damage.[20] antifungal activity by agnps has been proved against different candida species, including c. albicans, since microbial cells were inhibited using agnps.[21] furthermore, it was reported that agnps damage the structure of the cell membrane in c. albicans producing “holes” on the surface of the cells and thus inhibiting the budding process.[22] figure 1: ultraviolet-visible absorption spectra of silver nanoparticles after bioreduction by pleurotus ostreatus mushroom aqueous extract figure 2: size of silver nanoparticles figure 3: intensity of particle size distribution of pleurotus ostreatus dried mushroom aqueous extract hasan, et al.: inhibitory effect of agnps against candida spp. and bacteria 34 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 32-36 the antifungal properties of agnps against c. albicans have been demonstrated in some other studies, although reported minimum inhibitory concentration values are different from the ones we found in this work.[21-24] such differences could be due to the nature of the particles used, the difference in size being particularly important. it is known that size and shape of metallic nanoparticles influence their chemical, optical, and thermal properties.[25] the effectiveness of agnps against bacteria is clearly demonstrated, and in fact, agnps were shown to be effective against e. coli, with cells showing formation of “pits” in the cell wall. the agnps were found to accumulate in the bacterial membrane and some of them were reported to successfully penetrate into the cells.[26] similar results were found in e. coli and vibrio cholera; it was established that agnps provoked changes mainly in the cell membrane morphology, producing a significant increase in their permeability, thus affecting the proper transport through the plasma membrane, and resulting eventually in cell death. they also reported that silver nps with small diameters penetrated into the cells.[27] in our study, ag nps were found surrounding c. albicans cells, similar to the results found in bacteria.[27-29] in this study, the aqueous extracts of mushroom (p. ostreatus) gave showed (ve−) results against all microbial species when used as a control [figures 4 and 5], while the biosynthesized agnps gave showed (ve+) result as an inhibition zones for different microbial species [figures 4 and 5, tables 1,2]. table 1: the inhibitory effect of agnps (mm) against candida and bacterial species candida spp. inhibition zone (mm) 12.5 mg/ml 25 mg/ml 50 mg/ml 100 mg/ml candida albicans --10 12 16 candida guillermondii --10 15.5 20 candida krusie --14.5 15.5 19 geotrichum ktebahnii --8 14 20 candida zeylanoides 15 18.5 21 24.5 staphylococcus aureus 10 11 12 16 pseudomonas aeroginosa --12 13 14 escherichia coli 10 12 13 15 agnps: silver nanoparticles table 2: statistical analysis of the inhibitory effect of agnps (mm) on candida and bacterial isolates microbial species control agnps (mg/ml) s 12.5 25 50 100 candida albicans <0.001d 0±0.0d 10±0.57c 12±0.58b 16±0.58a candida guillermondii <0.001d 0±0.0d 15±0.58c 15.6±0.33b 20±0.57a candida krusie <0.001d 0±0.0d 14.5±0.28c 15.6±0.16b 19±0.57a geotrichumktebahnii <0.001d 0±0.0d 8±0.57c 14±0.58b 20±0.58a candida zeylanoides <0.001e 15±0.57d 18.5±0.28c 21±0.57b 24.5±0.28a staphylococcus aureus <0.001d 10±0.58c 11±0.57bc 12±0.57b 16±0.58a pseudomonas aeroginosa <0.001c 0±0.0c 12±0.58b 13±0.58ba 14±0.57a escherichia coli <0.001d 10±0.57c 12±0.58b 13±0.57b 15±0.58a the findings were described in the table represent the average of three replicates±standard error. small letters (a, b, c, d, e, ba, and ac) indicate to comparison between means in column, similar letters are non-significantly differences between means at (p≤0.05), using (lsd test). agnps: silver nanoparticles figure 4: inhibition zone of silver nanoparticles against several isolates of bacteria; (a) staphylococcus aureus, (b) pseudomonas aeroginosa (c), escherichia coli cba hasan, et al.: inhibitory effect of agnps against candida spp. and bacteria 35 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 32-36 however, despite the clear antimicrobial properties of agnps, their potential use in the clinic should be carefully evaluated since there is a lack of basic knowledge on the potentially different antimicrobial properties of agnps which may vary depending on many factors, including the method of synthesis, size, shape, functionalizing agent, and application method and also their interaction in more complex systems such as plants, animals, and humans. references 1. a. mohammed, m. girilal, s. a. mahdy, s. s. somsundar, r. venkatesan and p. t. kalaichelvan. “vancomycin bound iogenic gold nanoparticles: a different perspective for development of anti rsa agents”. process biochemistry, vol. 46, pp. 636-641, 2011. 2. j. hardes, h. ahrens, c. gebert, a. streitbuerger, h. buerger and m erren. “lack of toxicological side-effects in silver-coated megaprostheses in humans”. biomaterials, vol. 28, pp. 2869-2875, 2007. 3. j. r. morones, j. l. elechiguerra, a. camacho, k. holt, j. b. kouri and m. j. yacaman. “the bactericidal effect of silver nanoparticles”. nanotechnology, vol. 16, pp. 2346-2353, 2005. 4. b. y. geng, l. d. zhang, g. z. wang, t. y. xie and g. w. zhang. “synthesis and photoluminescence properties of znmns nanobelts”. applied physics letters, vol. 84, pp. 2157-2159, 2004. 5. k. dunn and v. edwards-jones. “the role of acticoat with nanocrystalline silver in the management of burns”. burns, vol. 30, pp. 1-9, 2004. 6. f. von nussbaum “antibacterial natural products in medicinal chemistry exodus or revival”. angewandte chemie international edition, vol. 45, pp. 5072-5129, 2006. 7. s. sarkar, a. d. jana, s. k. samanta and g. mostafa. “facile synthesis of silver nanoparticles with highly efficient anti-microbial property”. polyhedron, vol. 26, pp. 4419-4426, 2007. 8. m. a. pfaller and d. j. diekema. “epidemiology of invasive candidiasis: a persistent public health problem”. clinical microbiology reviews, vol. 20, pp. 133-163, 2007. 9. g. s. martin, d. m. mannino, s. eaton and m. moss. “the epidemiology of sepsis in the united states from 1979 through 2000”. new england journal of medicine, vol. 348, pp. 1546-1554, 2003. 10. p. g. pappas, j. h. rex, j. lee, r. j. hamill, r. a. larsen and w. powderly. “a prospective observational study of candidemia: epidemiology, therapy, and influences on mortality in hospitalized adult and pediatric patients”. clinical infectious diseases, vol. 37, pp. 634-643, 2003. 11. t. f. patterson. focus on candidemia. in: “treatment and prevention of fungal infections”. new york: applied clinical education, pp. 7-8, 2007. 12. t. c. white, s. holleman, f. dy, l. f. mirels and d. a. stevens. “resistance mechanisms inclinical isolates of candida albicans”. antimicrobial agents and chemotherapy, vol. 46, pp. 1704-1713, 2002. 13. s. perea, j. l. lopez-ribot, w. r. kirkpatrick, r. k. mcatee, r. a. santillan, m. martinez. “prevalence of molecular mechanisms of resistance to azole antifungalagents in candida albicans strains displaying high-level fluconazole resistance isolated from human immunodeficiency virus-infected patients’’. antimicrobial agents and chemotherapy, vol. 45, pp. 2676-2684, 2001. 14. m. d. levin, j. g. den hollander, b. van der holt, b. j. rijnders, m. van vliet, p. sonneveld and r. h. n. van schaik. “hepatotoxicity of oral and intravenous voriconazole inrelation to cytochrome p450 polymorphisms”. journal of antimicrobial chemotherapy, vol. 60, pp. 1104-1107, 2007. 15. l. j. worth, c. c. blyth, d. l. booth, d. c. m. kong, d. marriott, m cassumbhoy, j. ray, m. a. slavin and j. r. wilkes. “optimizing antifungal drug dosing and monitoring to avoid toxicity andimprove outcomes in patients with haematological disorders”. internal medicine journal, vol. 38, pp. 521-537, 2008. 16. h. sher, m. al-yemeni and k. khan. “cultivation of the oyster mushroom (pleurotus ostreatus (jacq.) p. kumm.) in two different agroecological zones of pakistan”. african journal of biotechnology. vol. 10, no. 2, pp. 183-188, 2011. 17. s. gurunathan, j. raman, s. n. abd malek, p. a. john and s. vikineswary. “green synthesis of silver nanoparticles using ganoderma neo-japonicum imazeki: a potential cytotoxic agent against breast cancer cells”. international journal of nanomedicine, vol. 8, pp. 4399-4413, 2013. 18. s. berardinelli and d. j. opheim. “new germ tube induction medium for the identification of candida albicans”. journal of clinical microbiology, vol. 22, pp. 861-862, 1985. 19. s. sujatha, s. tamilselvi, k. subha and a. panneerselvam. “studies on biosynthesis of silver nanoparticles using mushroom and its antibacterial activities,” international journal of current microbiology and applied sciences, vol. 2, no. 12, pp. 605-614, 2013. 20. a. j. huh and y. j. kwon. “nanoantibiotics: a new paradigm for treating infectious diseases using nanomaterials in the antibiotics resistant era”. journal of controlled release, vol. 156, pp. 128-145, 2011. 21. a. panáček, m. kolář, r. večeřová, r. prucek, j. soukupová, v. krystof, p. hamal, r. zboril and l. kvítek. “antifungal activity of silver nanoparticles against candida spp”. biomaterials, vol. 30, pp. 6333-6340, 2010. 22. k. j. kim, w. s. sung, b. k. suh, s. k. moon and j. s. choi. “antifungal activity and mode of action of silver nano-particles on candida albicans”. biometals, vol. 22, pp. 235-242, 2009. 23. k. j. kim, w. s. sung, s. k. moon, j. s. choi, j. g. kim and d. g. lee. “antifungal effect of silver nanoparticles on dermatophytes”. journal of microbiology and biotechnology, vol. 18, pp. 1482-1484, 2008. 24. m. stevanović, s. škapin, i. braćko, m. milenković, j. petković, m. filipičd and d. p. uskokovića. “poly (lactide-co-glycolide)/silver figure 5: inhibition zone of silver nanoparticles (agnps) (n) against several isolates of candida; (a) candida guillermondii; (b), candida albicans; (c), candida zeylanoides; (d), candida spp. in concentration of 100 mg/ml of ag nps the way of measuring of diameter of inhibition zone. the aqueous solution of pleurotu sostreatus (c) dc ba hasan, et al.: inhibitory effect of agnps against candida spp. and bacteria 36 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 32-36 nanoparticles: synthesis, characterization, antimicrobial activity, cytotoxicity assessment and ros-inducing potential”. polymer, vol. 53, pp. 2818-2828, 2012. 25. m. a. el-sayed. “some interesting properties of metals confined in time and nanometer space of different shapes”. accounts of chemical research, vol. 34, pp. 257-264, 2001. 26. i. sondi and b. salopek-sondi. “silver nanoparticles as antimicrobial agent: a case study on e. coli as a model for gram-negative bacteria”. journal of colloid and interface science, vol. 275, pp. 177-182, 2004. 27. a. t. le, t. t. le, h. h. tran, d. a. dang, q. h. tran and d. m. vu. “powerful colloidal silver nanoparticles for the prevention of gastrointestinal bacterial infections”. advances in natural sciences: nanoscience and nanotechnology, vol. 3, p. 045007, 2012. 28. a. de. sousa, d. mehta and r. w. leavitt. “bactericidal activity of combinations of silver-water dispersion™ with 19 antibiotics against seven microbial strains”. current science, vol. 91, pp. 926-929, 2006. 29. r. vazquez-muñoz, m. avalos-borja and e. castro-longoria. “ultrastructural analysis of candida albicans when exposed to silver nanoparticles”. plos one, vol. 9, no. 10, p. e108876, 2014. . 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 25-30 research article automatic identification of communication signals using zero-crossing based techniques emanuel s. h. qas marrogy department of communication engineering, al-mansour university college, baghdad, iraq abstract the identification of different types of modulation for any intercepted communication signal out of the vast hierarchy of possible modulation types is a key fundamental before advising a suitable type of demodulator, where this process is usually a manual option. this technique is extremely important for the purposes of communication intelligence. in this paper, a proposed methodology is suggested, validated, and tested (through computer simulations) for the automatic identification of the modulation type (analog and digital) of the intercepted communication signals. the methodology is based on the zero-based representation of signals and utilization of new algorithms for such identification. keywords: zero-crossing, communication signals, modulation, digital signal, analog signal, algorithms introduction the analysis of zero-crossing technique describes one of the several techniques, which make the use of the information pertaining to the locations in time of successive zero-crossing of a time waveform. such techniques have found applications for several signal processing and pattern recognition tasks. some of these tasks include speech analysis and recognition communications applications.[1] each of these applications has its specific features regarding the nature of the zero-crossing data measurements and the features extracted from such data. some applications utilize the number of zero-crossing, the other may count the time interval between successive zeros. by all means, the work of voelcker[2,3] remains the standard source for clearing the essence of zero-based representation of signals. inspired aspirants working under his directions (e.g. takayuki arai and yuichi yoshida)[4] and sekey[5] cleared out some difficult mathematical problems such as uniqueness, sufficiency, and completion, in addition to characterizing several classes of entire functions (ef) amenable to such representations. in this paper, we will try to extend the prowess of zerobased representations to cover important utilities in the field of communication intelligence, namely, the identification of unknown modulation of intercepted signals. further, a tested modulation scheme, based on manipulating the zeros of the baseband and carrier waveforms, will be illustrated. this scheme has been mathematically and experimentally verified to emulate most analog and digital modulation schemes. fundamentals of zero-based representation of signals this subject warrants a due introduction owing to the fact that these techniques have not been very current due to the special mathematics (mainly the theory of ef) and to the difficult problem of interpolating (reconstructing) a signal by virtue of its zero-crossings. in this paper, a brief illustration introduction of zero-based techniques exemplified by displays of actual signals generated and processed by the author by configuring the hewlett-packard spectrum analyzer and the waveform recorder under the control of the hp computer. the author developed the software to affect all the displayed signals illustrated in this paper. a band-limited (bl) signal s(z) is an ef[6] which can be described by the location of its zeros s z s o z zn n ( ) ( ) ( )( )= ∏ −= ∞ 1 1 (1) corresponding author: emanuel s. h. qas marrogy, department of communication engineering, al-mansour university college, baghdad, iraq. e-mail: emanuel.hanna@muc.edu.iq received: apr 04, 2019 accepted: apr 18, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp25-30 copyright © 2019 emanuel s. h. qas marrogy. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) marrogy: automatic identification of communication signals using zero-crossing based techniques 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 25-30 where z = t + j u; a complex variable whose real axis coincides with the real-time axis s (z) along the real axis coincides with s (t)), z n is the location of the nth zero. the time waveform wholly real zero bl signal s (t) corresponding to the equation (1) can be expressed[7] as s t a sin w t tnn m o( ) * ( ( ))( )= ∏ − −=1 2 2 2 (2) where a is a scale factor. equation (2) was utilized to effect the reconstruction of the bl signal shown in the first trace of figure 1. the second trace displays the hard-limited signal of the first trace. the third trace displays the real zeros of the signal, we can express these zero locations as z sgn s sgn s k n k k= −+ +∑ [ )( ) ( )] /( )1 21 1 (3) where n = the number of samples, s k is the digitized input signal kth in all our experimentation, the number of samples was chosen to be 400 (for due consideration to anti-aliasing). equation (2) was utilized to reconstruct the signal [the fourth trace of figure 1]. conversion of complex zeros into real zeros we note that equation (2) is valid for real zeros only, while real signals can contain, in general, complex zeros. the occurrence of complex zeros affect the “dimensionality[8] of the signal, meaning that the signal cannot be reconstructed by virtue of its real-zeros only without due consideration to its complex zeros, a methodology must exist to transform complex zeros into real zeros to allow signal reconstruction by merit of its real-zero locations. fortunately, several such methodologies do exist. one such method is to add a sine waveform which frequency is equal or greater than the highest frequency component present in the bl signal. the composite signal x(t) can be expressed as x(t) = s(t) + a cos2n (w + 1)t, 0<t<t (4) where w is the single-sided bandwidth. it is easy to show that the composite signal x(t) will have 2 (wt+1) zeros in the time interval between 0 and t. the original signal, s(t), can be recovered by subtracting the added sinusoid from the reconstructed signal s(t) = x(t) a cos2π (w + 1/t)t (5) the previous methodology is illustrated in figure 2. real-zero interpolation the technique of interpolating a minimum – bandwidth waveform to a set of real zero-crossing on the time axis was first described by voelcker[7,3] and dubbed rzi. the real-zero interpolator rzi suggested in the reference quoted, ingenious as it stands. it has been very difficult to implement practices, and the original set comprised more than 300 shift-register chips was very difficult to construct that limited zero-based technique applicability. one of the aims of this paper is to introduce a computer-simulation version, based on the same principle, easy to implement practices to reconstruct wholly real-zero signals from there zero-crossing. we shall base our simulation on the rzi version figure 3. consider the following representation[9] of the real zero signals s rz (t) s t re s t e s trz rz j rz( )( ) ( ( ) ( ))= ( (6) assume τ o , τ 1 , τ 2 ……τ i . is a sequence of points on “the realtime axis representing the zero-crossing of the bl real zero signal s rz (t). the signal which shall be reconstructed from the real zero locations is the real zero component s rz (t) of the original signal s(t). the original signal s(t) can be considered as composed of two components s rz (t) and s cz (t) as follows s(t) = s rz (t)*s cz (t) (7) where s rz (t) is the real zero signal, s cz (t) is the complex zero signal. hard limiting the signal s(t) destroys its envelope and produces sgn s t s t s t rz rz rz 1 1 [� ] ���� , , � ( ) ( ) ( ) � � � = + − > < 0 0 (8) figure 1: waveform regeneration. (1) input signal, (2) hard clipped, (3) real zeros, (4) reconstructed signal figure 2: reconstructing signal from zero-crossing. (1) original signal s(t), (2) composite signal x(t), (3) zeros of x(t), (4) reconstructed x(t), (5) reconstructed signal s(t) marrogy: automatic identification of communication signals using zero-crossing based techniques 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 25-30 we can write equation (5) in the following form s rz (t) = s rz (t) cosφ (s rz ) (t)) (9) where φ (s rz ) is the ‘instantaneous phase’ of s rz (t). each zero-crossing is represented by a delta function” at its location, then the following equation holds ϕ δ τ( ( �)) ( � )�s t n trz i= −∑2nr (10) since s rz (t) is an ef; its phase and modules are a hilbert transform pair,[6] thus h s t h trz i( ( ( �)) ( � )��ϕ δ τ= − (11) = 1 n t t iδ τ τ ( � ) �( � �� )��� − −∞ ∞ ∫− (12) = � ��� 1 t − applying the general phase-envelope relationships[9] follows that ln s t trz n nr , � �� ( �) � = −∑ 1 2  (13) =h (φ,s rz ) (t)) (14) thus integrating the output of the hilbert transform network, exponentiating its output and then multiplying it by sgn (s rz (t)) yields the interpolated signal. the previous description of the rzi versions of voeicker and sekey is limited practically by the difficulty of realization of a wideband hilbert transform network. computer simulation provides a suitable answer to this problem. computer simulation of the rzi figure 4 illustrates the developed flowchart describing the methodology for realizing real zero interpolation of a minimum bandwidth waveform fitting the real zero input sequence. figure 5 illustrates the simulation results. the first trace is a sinusoidal input clipped to yield the second trace. impulses are added to characterize the third trace. the fourth trace is the hilbert transform output. the reconstructed clipped signal is shown in the fifth trace. it is clear that it coincides with the clipped input signal. figure 3: a real zero interpolator start clipped input signal f(x) detect zero crossing z = sgn (k+1)* sgn (k))/2 x = h (z) y = ∫ � j = exp ( y ) f ( x) j * sgn f (x)) end figure 4: flowchart for the rzi figure 5: temporal waveforms in real zero interpolation. (1) input signal, (2) clipped input signal, (3) real zeros, (4) hilbert,[4] (5) integral,[5] (6) exp,[6] (7) sgn[2] marrogy: automatic identification of communication signals using zero-crossing based techniques 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 25-30 suitable demodulator can be automatically affected. figure 7 illustrates the conceptual proposed configuration which may affect automatic demodulation of unknown interceptions. to experimentally evaluate the concept illustrated above, the following procedure and algorithms were adopted: a. most widely known classical analog and digital modulation waveforms have been computer-simulated, they are zero-crossing processed in the described manner and the identification “feature” extracted; namely, the zero-pattern time course. figure 8 illustrates the results obtained. it is clear from the different processed modulation types (both analog and digital) that a unique zero-pattern is associated with each modulation type unless in some situation they have similar zero-pattern. a library of zero-pattern displays corresponding to each modulation type is thus constructed and stored. b. the real intercepted communication signal is first processed to obtain the relevant zero-pattern, as explained in the previous step. then, the displayed pattern is normalized for the same frequency as the reference pattern. c. the identification is effected automatically by comparing the interception zero-pattern display with figure 6: processing time vrs number of zeros of interception figure 7: block diagram of automatic demodulation figure 8: method of modulation recognition based on zero-pattern time course for practical applications, it would be of consequence to relate the processing (computational) load to the number of zeros necessary for the interpolation procedure and further to evaluate the effect of change of the sample size (n) on this processing time. practical results of such experimentations are exemplified in figure 6. the processing time is approximately linearly related to the zero count. modulation identification methodology based on zeropattern characterization segmenting an input intercepted communication signal into time windows encompassing a sufficiently large number of cycles of the lowest frequency present; corresponding to sufficient sample size, enables us to correlate the timedependence of the number of intercepted zero-crossing with the type of modulation. correctly and automatically identifying the type of modulation of unknown interceptions has been and will always be a subject of paramount importance in both civil and military communications. in this section, the aspirant wishes to present this contribution which is computer methodological processing of received signals (digitized and handed over to the computer), processing them in the manner described in the last section to arrive at the decision characterizing the type. of modulation; provided that it coincides with a pattern stored in a special library. change to in modulation, each coincides signal with a pattern ins stored in special library. by this means effective switching to a marrogy: automatic identification of communication signals using zero-crossing based techniques 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 25-30 figure 9: modulation identification figure 10: dsb signal generation. (1) modulating signal,(2) carrier signal, (3) real zeros, (4) padding zeros, (5) modulation signal figure 11: ask signal generation. (1) binary sequence, (2) carrier wave, (3) real zeros, (4) deleted zeros, (5) ask signal figure 12: psk signal generation. (1) binary sequence, (2) carrier wave, (3) real zeros, (4) deleted zeros, (5) psk signal the corresponding zero-patterns of the constructed modulation-type library in step (a). the algorithm adopted here for identification is the least square approximation. figure 9 illustrates the experimental results obtained. modulation/demodulation by zero manipulation another contribution presented here is a methodology for affecting different modulation/demodulation schemes using zero manipulation. figure 10 illustrates the methodology for generating an am (dsb) modulated waveform. the first and second traces in this figure illustrate computer-simulated base-band (modulating) and carrier waveforms, respectively. the third trace illustrates the processed zero-crossing of the carrier signal. the zeros of the baseband signal are added in the fourth trace, and the resulting zero sequence is applied to the proposed computer-simulated version of the real zero interpolator to yield the am (dsb) modulated waveform shown in the last trace. the same procedure is repeated for ask and psk. the results and procedures are shown in the subsequent figures 11 and 12. it is evident that such methodologies marrogy: automatic identification of communication signals using zero-crossing based techniques 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 25-30 alleviate the need for complex schemes traditionally employed in conventional modulation schemes. the process of demodulation can be affected in an inverse manner by deleting/or adding the zeros of the carrier waveform and real zero interpolating the resulting zero sequence. figure 13 illustrates the methodology and results obtained for demodulating ask-modulated signal. figure 13 is configured in a manner to illustrate the possibility of incorporating such methodology in the design of modems for data transmission. conclusion this paper shows the potency of zero-based techniques in a wide variety of applications. the success in introducing a computer-based version of the rzi may open the door easily implementing schemes, which could otherwise be very complicated and cumbersome. it is left to the reader to evaluate the impact of the ideas presented here for automatic demodulation of intercepted communication signals (without the need for manual switching of a modulation type switch). references 1. k. s. fu. school of electrical engineering purdue university w. lafayette, indiana 47907. in: spoken language generation and understanding: proceedings of the nato advanced study institute held. vol. 59. france: springer science and business media bonas, p. 221, 1979. 2. n. ishikawa and s. sugiura. “unified differential spatial modulation”. ieee wireless communications letters, vol. 3 no. 4, pp. 337-340, 2014. 3. h. voelcker. zero-crossing properties of angle-modulated signals. ieee transactions on communications, vol. 20, no. 3, pp. 307315, 1972. 4. h. zhang, a. d. m. palanivelu, r. u. nabar, s. wu and h. l. lou. “united states patent and trademark ofice. u.s. patent no. 8,195,112”. washington, dc: u.s. patent and trademark office, 2012. 5. a. sekey. “a computer simulation study of real-zero interpolation”. ieee transactions on audio and electroacoustic, vol. 18, no. 1, p. 43, 1970. 6. a. v. oppenheim and g. c. verghese. “introduction to communication, control and signal processing”. class notes for signals, systems, and inference. massachusetts: spring massachusetts institute of technology, 2010. 7. a. neubauer and j. niederholz. “u.s. patent and trademark office”. u.s. patent no. 7,564,926. washington, dc: u.s. patent and trademark office, 2009. 8. s. m. kay and r. sudhaker. “a zero crossing-based spectrum analyzer”. ieee transactions on acoustics speech and signal processing, vol. assp-34, no. 1, pp. 96-104, 2009. 9. t. s. tabatabaei, s. krishnan and a. anpalagan. “svm-based classification of digital modulation signals”. ieee international conference on systems, man and cybernetics, oct. 2010, pp. 277-280. figure 13: zero-based ask modem operation. (1) binary sequence, (2) carrier wave, (3) real zeros, (4) deleted zeros, (5) ask signal, (6) ask signal, (7) real zeros, (8) binary sequence tx_1~abs:at/add:tx_2~abs:at 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 1-5 research article association of total dietary fats and its subtypes with risk of breast cancer jwan i. jawzali* department of nursing-basic science, nursing college, hawler medical university, erbil, iraq abstract specific classes of dietary fatty acids may be important modifiers of breast cancer (b-ca) risk. aim of this study was identification risk of subtypes of dietary fat for b-ca. this case–control study carried out in rizgary teaching hospital in erbil city. data collected by interview questionnaire and included demographic and reproductive properties; anthropometric measurements; and medical history. dietary data collected by food frequency questionnaire. they were analyzed by program for mosby’s nutritrac nutrition analysis software, for calculation intake of dietary; fiber, total fat, and its subtypes, energy intake, acceptable macronutrient distribution range, and antioxidant nutrients. statistical analysis was performed using spss program. polyunsaturated fats decreased risk of b-ca while saturated and monounsaturated fats (cis form) increased risk among all and postmenopause obese women, respectively. risk of cancer increased significantly in high percentage of energy intake from monounsaturated fats, cooking oil, and dietary red meats. the study concluded that total polyunsaturated fatty acids (pufa) decrease risk of b-ca among obese menopause woman. increase risk of b-ca by cooking oils and animal origin diet may due to increased intake of saturated monounsaturated and specific pufa. these subtypes of dietary fats may promote hormones imbalance and inflammation. keywords: animal origin diet, breast cancer, monounsaturated fats, saturated fats, unsaturated fats introduction breast cancer (b-ca) is the most frequent cancer among women in kurdistan/iraq and increased in the past 10 years. it has been reported that 40% of cancers can be averted by reducing risk factors and by primary prevention.[1] role of dietary fat could result in initiation and development of breast tumor by stimulation production of estrogen and other endogenous hormone, regulation of immune function, and modulation of gene expression.[2] diet may modify risk of b-ca. specific classes of dietary fatty acids may be important modifiers of b-ca risk. unique chemical and biophysical properties of dietary fatty acids have impact on health and disease.[3] the inconsistent and limited evidence warrants research to assess the impact of consumption of fat subtypes on b-ca recurrence and mortality.[4] epidemiologic, animal experiments, and some case–control studies supported the hypothesis that dietary fat can increase b-ca risk, and still remains one of the most controversial hypotheses in nutritional epidemiology.[5] inconsistent results due to limited ranges of fat intake among populations, errors of dietary fat measurement, and high correlation between specific types of dietary fat and b-ca risk. the confounders such as body weight and huge energy intake and food components such as fiber and antioxidants can affect the exact association of total and specific subtypes of fat intake with b-ca risk.[2] fatty acids potentially associated with an increase in cancer risk in humans include saturated fatty acids (sfa), monounsaturated fatty acids (mufa), and trans-fatty acids, and two major classes of polyunsaturated fatty acids (pufa); omega-6 pufa, (pufa 18:2n6) and arachidonic acid (aa, 20:4n6). in contrast, specific pufa, n-3 pufa (α-linolenic acid), may have anticancer effects. studies have shown that sfas associated with an increased risk in contrast omega-3 pufa intake reduced risk of b-ca.[3] pufas are biologically active food components that elicit proand anti-inflammatory responses through several signaling pathways that regulate cell proliferation, apoptosis, and angiogenesis. there is increasing evidence that pufas play a role in cancer risk and progression.[6] corresponding author: jwan i. jawzali, department of nursing-basic science, nursing college, hawler medical university, erbil, iraq. e-mail: jwanjawzali@gmail.com received: jan 26, 2020 accepted: jun 26, 2020 published: jul 10, 2020 doi: 10.24086/cuesj.v4n2y2020.pp1-5 copyright © 2020 jwan i. jawzali. this is an open access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) jawzali: association total dietary fat with risk of breast cancer 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 1-5 epidemiologic studies provide unpredictable reports about the associations of dietary pufas and risks of cancer. the previous reviews evaluate overall dietary fat but do not take in to account the variation in the impact of fat subtypes. dietary fat risk to the b-ca may due to the complex composition of dietary fat. therefore, this study conducted to identify contribution of some subclasses of dietary fat to total energy and their risk to b-ca. the study also identify socio demographic and lifestyle factors which act as confounder and some antioxidant nutrients that modify the association of dietary fats and its subtype with risk of b-ca. patients and methods the study was case control, carried out in rizgary teaching hospital in erbil governorate, kurdistan region, iraq. the patients age ranged between 27 and 74 years and included 55 cases of women with diagnosed b-ca (after mastectomy), at different stage, attending outpatient unit of chemotherapy and 55 cancer-free women attending outpatient clinic of rizgary teaching hospital. sample size determined by sample size mode[7] using odds ratio (or) and percentage of controls exposed to total fats from previous case–control study conducted in the same hospital. women whose weighs changed after diagnoses and women who did not complete the interview questionnaire were excluded from the study. they were interviewed after their consent had been taken by questionnaire, which included sociodemographic properties; reproductive history, family history of b-ca, anthropometric measurements; body mass index and waist-to-hip ratio, and medical history. diet history collected by semi-quantitative food frequency questionnaire to estimate; type, frequency, and quantity of food using household measurements and standard portion size photographs of common foods. foods intake data were analyzed by program for mosby’s nutritrac nutrition analysis software, version iv (cd-rom), for calculation; total energy, dietary fat intake and its subtypes, cholesterol, dietary fibers, and some antioxidants per individual by, taking into account intake of margarine, butter, oils, and fats used in cooking and baking. calculating amount of omega 6 in cooking oil by righttrak nutrient calculator. acceptable macronutrient distribution range (amdr) represents the percentage of energy contributed by total fat calculated. food groups contributing to intake of fat, red meat, chicken, fish, and eggs, were used in association with risk of b-ca. statistical analysis all data were analyzed by spss version 22.0. descriptive statistics were used to study the mean and standard deviation and standard error for continuous variables. chi-square test was used for association measurements between categories, independents t-test used for measurement differences between the means. odds ratio (or), and binary logistic regression were used in measuring the risk of b-ca. p-value was considered significant when p ≤ 0.05 and was considered highly significant when p ≤ 0.01. results and discussion table 1 shows demographic reproductive properties of the studied samples. there was no significant difference between b-ca and control groups in age, residency, marital status, education levels, age of menarche, age at first pregnancy, and menopause age. table 2 shows no significant difference in anthropometric measurement between b-ca and controls. most were overweight and had normal value (0.8–0.85) of waist-to-hip ratio. in contrast to familial absence of cancer, this showed significant association and decrease in the risk (or=0.14**) and confident interval (ci) ranged to decrease (0.04–0.44) risk of b-ca and indicates to the importance of genetic factors, and increasing rate of b-ca incidence in iraq. this view the same results of the previous studies conducted in iraq by lafta et al. and safil et al.[8,9] nearly 60% of the studied women had family history of b-ca it is much higher than that reported in the study[10] where only 16% of the patients recorded a positive family history. table 3 shows intake of daily energy as kilocalorie (kcal./d), daily total fats and its subtypes as gram/day (g/d), their amdr, dietary fiber (g/d), and some antioxidant nutrients. there was a significant difference between means of saturated fats, unsaturated, and selenium as antioxidants. the results reveal the roles of subtypes of fats on the risk of b-ca. the most important one was pufa which represent mainly total form of two major classes of (pufa); omega-6 and omega 3. interaction with other subtype fats and antioxidants, and their amdr in all samples and largely among overweight and obese patients, decreased risk of b-ca. increased risk of b-ca was found among group with high intake of saturated fats more than 10% of total calories and ≥20 g/day of normal value of saturated fat intake, and among obese postmenopause women who had high percentage of energy intake from mufa. the finding supported by sieri et al.[11] who found positive association between b-ca and saturated fat and was more pronounced among postmenopausal women who never used hormone therapy. higher total fat intake (39% of total energy) than recommendations ensures limitation total fat intake between 20% and 35% of total caloric and restriction saturated fat consumption as stated by makarem et al.[4] furthermore, canadian cohort study showed that higher intakes of saturated fat as a percentage of total energy significantly increased the risk of b-ca death in postmenopausal women and not significantly in premenopausal women. every 5% increase in saturated fat intake, as percentage of total energy, was associated with approximately 65% increased risk of b-ca mortality.[4] otherwise, zhang et al.[12] suggested that intakes of total fat, saturated and monounsaturated fat, and n-3 and n-6 polyunsaturated fat were not associated with risk of b-ca. antioxidant nutrients (vitamins c, e, and selenium ) and their modulation effect appeared only with selenium that showed significant association with b-ca. this result is in line with pouchieu et al.[13] who suggests that specific sfas, mufas, and pufas were prospectively differentially associated with cancer risk. antioxidants may modulate these associations with interaction with other subtype fats and counteracting the potential effects of these fatty acids on carcinogenesis. significant high risk of mufa which occupies high percentage from total energy among obese patient and decreased risk by polyunsaturated and its amdr mainly among obese postmenopause women is related to body fatness jawzali: association total dietary fat with risk of breast cancer 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 1-5 and estrogen hormone production. this result reveals that the idea of other case–control studies has shown a positive role of mufa in the pathogenesis of b-ca and that body adiposity enhances the production of estrogen, hormonal imbalances in insulin-like growth factor-1 and insulin. increased bioavailable estrogens that are theorized to increase cell proliferation through activation of target genes, thereby promoting carcinogenesis. makarem et al., singh and nimbkar[4,14] concluded that reduction of dietary fat would result in a 2.5fold reduced risk of b-ca among postmenopausal women. table 4 shows significant differences in the mean levels of cooking oils, dietary red meats, poultry meats, and dietary egg between b-ca and controls. this may due to content of cooking oils (largely sunflower and corn oils intake that was abundant in studied samples) of omega-6. effectively sunflower oils showed approximately significant difference between b-ca and controls. highest percentage 77% of sunflower oil and 50% of corn oil composed of omega 6 (n3:6) linoleic unsaturated fatty acids.[15] this result explores pro-inflammatory effect of omega 6 that increases risk of b-ca as stated by prentice and sheppard[16] who concluded increased b-ca risk with increasing dietary ratio of pro-inflammatory (n-6 pufa) to anti-inflammatory (n-3 pufas) and increases in pro-inflammatory eicosanoids such as prostaglandin e2, promotes angiogenesis, and hinders apoptosis. in addition, red meats may contain other carcinogenic compounds formed during meat process and preparing that increase risk of b-ca as stated by khodarahmi and azadbakht[2] that carcinogens such as heterocyclic amines, n-nitroso compound, and polycyclic aromatic hydrocarbons, which are found largely in cooked red meat and soluble hormones or growth factor, could increase risk of b-ca and they may confound the exact relationship of the subtypes of fat with b-ca. significant difference between cases and controls in dietary egg intake may indicate to its association with risk of b-ca and it is in line with[17] that high egg intake may be associated with elevated risk of b-ca. also agree with cho et al.[18] who attributed association of high egg intake with risk of b-ca to estrogen hormone levels and concluded that animal fat intake increase b-ca risk by modulating estrogen levels among women with estrogen receptor (er)-positive cancers compare to women with er-negative cancers. significant difference in poultry meats intake among b-ca and controls conforms result of chandran et al.[19] who found positive association between poultry consumption and b-ca risk in caucasian women, with over 2-fold greater risk table 1: comparison of sociodemographic characteristics for cases and controls and their association with b-ca demographic data b-ca cases controls p-value of t-test mean±sd mean±sd age (years) 47.6±10.0 47.1±11.5 0.74 residency f % f % p-value chi-square urban rural 30 (54.5) 25 (45.5) 38 (69.1) 17 (30.9) 0.169 educational levels f (%) f (%) p-value chi-square illiterate primary school secondary school institute and above 38 (69.1) 8 (10.9) 6 (14.5) 3 (5.5) 30 (54.)5 13 (23.6) 4 (7.3) 8 (14.5) 0.068 marital status f (%) f (%) p-value chi-square married single 48 (87.3) 7 (12.7) 49 (89.1) 6 (10.9) 0.77 mean±sd mean±sd p-value t-test menarche age 13.4±1.5 13.5±0.68 1.4 age at first pregnancy 16.8±10.8 18.2±9.5 0.48 menopause age 43.6±7.6 43.7±9 0.97 f.: frequency, **p-value ≤0.01, *p-value ≤0.01 table 2: comparisons of anthropometric factors and familial history between cases and controls anthropometric b-ca cases controls p-value–t-test mean±sd mean±sd bmi (kg/m2) waist/hip ratio 31.7±5.5 0.84±0.06 30.2±4.6 0.83±0.07 0.13 0.89 bmi category f. (%) f. (%) ˂ 25 5 (9.1) 5 (9.1) 25-29 18 (32.7) 20 (36.4) > 30 32 (58.2) 30 (54.5) family history f. (%) f (%) p-value of chi-square negative 35 (63.6) 51 (92.7) 0.00** positive 20 (36.4) 4 (7.3) f: frequency, **p≤0.01, *p≤0.01 jawzali: association total dietary fat with risk of breast cancer 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 1-5 among premenopausal caucasian women and among women to er-negative tumors. this result may due to the method of preparation as reported by bao et al.[20] who concluded that b-ca risk increased when chicken was consumed with the skin, contains fats or some component produced during cooking. conclusions the study concluded that increased intake of saturated and monounsaturated fats that are derived from animal origin; red meats, poultry and eggs, and specific polyunsaturated fats mainly from cooking oil, may cause hormones imbalance, inflammation, and increase risk of b-ca, particularly among obese menopause woman. in addition, gaining high calories from monounsaturated fats and method of meat preparation and cooking may increase the risk of b-ca. acknowledgment we thank first study participants for their contributions and outpatient clinic staff of chemotherapy unit of rizgary teaching hospital for their support during data collection. references 1. v. g. guerrero, a. f. baez, c. g. c. gonzalez and c. g. m. gonzález. monitoring modifiable risk factors for breast cancer: an obligation for health professionals. revista panamericana de salud pública, vol. 41, p. e80, 2017. 2. m. khodarahmi and l. azadbakht. the association between different kinds of fat intake and breast cancer risk in women. international journal of preventive medicine, vol. 5, no. 1, pp. 6-15, 2014. 3. m. maclennan and d. w. ma. role of dietary fatty acids in mammary gland development and breast cancer. breast cancer research, vol. 12, p. 211, 2010. 4. n. makarem, u. chandran, e. v. bandera and n. parekh. dietary fat in breast cancer survival. annual review of nutrition, vol. 33, table 4: comparisons of some food items between cases and controls and their significant differences dietary data intake (g/d) controls b-ca cases p-value of t-test mean±se mean±se cooking oil 31.1±4.0 42.7±4.7 0.05* omega-6 (in cooking oilsunflower oil) 19.9±2.6 27.3±2.9 0.05* red meats 11.4±1.9 22.8±3.5 0.005** poultry meats 21.6±2.4 33.5±4.8 0.029* fish intake 4.2±0.9 5.8±1.5 0.36 egg intake 10.8±1.5 16.2±2.1 0.035* g/d: gram per day table 3: comparisons of energy, fats, and antioxidants intakes between cases and controls and their risk to b-ca nutrients b-ca cases controls p-value t-test or 95% ci mean±se mean±se total energy (kcal/day) 4653.3±1212.9 3019.1±641.2 0.23 total fat 137.8±49.4 85.2±13.9 0.31 % kcal from total fat 39.4±7.6 32.5±1.8 0.32 saturated fat intake f (%) f (%) p-value of chi-square categories 7–10% of calorie is normal ˂7% low normal >10% high 18 (32.7) 11 (20.0) 26 (47.3) 29 (52.7) 15 (27.3) 11 (20.0) 0.01** low intake of sf is reference 3.8**a 1.5–9.5 3.2* 1.13–9.2 mean±se mean±se p-value t-test or 95% ci saturated fat (sf) (g/d) 58±32.2 18.3±3.0 % kcal from saturated fat 10.8±1.6 7.9±0.6 monounsaturated (g/d) 53.7±13.6 40.2±4.6 0.35 % kcal from mafa 19.4± 4.7 18.5 ±1.5 1.7*o 1.04–2.9 polyunsaturated (g/d) 12.7± 1.9 7.3 ± 1.2 0.017* 0.93*a 0.87–0.98 polyunsaturated pafa 0.86**b 0.77–0.96 polyunsaturated pafa 0.62*o 0.4–0.97 % kcal from pafa 4.8±0.8 2.7±0.1 0.013* −0.7**a 03–0.8 cholesterol (mg/d) 400.7±194.4 126.9±18.1 0.66 dietary fiber (g/d) 56.2±105 56.8±119 0.97 vitamin c (mg/d) 521±139.6 797±400 0.52 vitamin e (mg/d) 10.7±2.1 10.6±3.4 0.98 selenium (mcg/d) 97.7±5.2 83.2±4.2 0.03* se is standard error, it is used for comparing between means, **p≤0.01, *p≤0.05, o: risk b-ca among obese, a: risk of b-ca among all sample, b: risk b-ca among all sample with antioxidant nutrients (se and vitamins c and e). mg/d: milligram/day, mcg/d: microgram/day jawzali: association total dietary fat with risk of breast cancer 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 1-5 pp. 319-348, 2013. 5. v. chajes, g. torres-mejia and c. biessy. ω -3 and ω-6 polyunsaturated fatty acid intakes and the risk of breast cancer in mexican women: impact of obesity status. cancer epidemiology, biomarkers and prevention, vol. 22, no. 2, pp. 319-326, 2011. 6. m. azrad, c. turgeon and w. demark-wahnefried. current evidence linking polyunsaturated fatty acids with cancer risk and progression, frontiers in oncology, vol. 3, p. 224, 2013. 7. sampsize project. sample size for a case-control study. 20032005 philippe glaziou. available from: http://www.sampsize. sourceforge.net/iface/s3.html. 8. r. k. lafta, e. q. saeed and s. a. isa. risk factors of breast cancer among women (a sample from baghdad). iraqi journal of community medicine, vol. 1, pp. 1-6, 2019. 9. a. m. al-safi1, b.a. ghalibyassin and e. a. al-tameemi. body mass index among women with proved or suspected breast cancer attending national center for early detection, medical city campus. iosr journal of dental and medical sciences, vol. 14, no. 5, pp. 53-59, 2015. 10. n. a. s. alwan. breast cancer among iraqi women: preliminary findings from a regional comparative breast cancer research project. journal of global oncology, vol. 2, no. 5, pp. 255-258, 2016. 11. s. sieri, v. krogh, p. ferrari, f. berrino, v. pala, a. c. m. thiébaut, a. tjønneland, a. olsen and k. overvad. dietary fat and breast cancer risk in the european prospective investigation into cancer and nutrition. the american journal of clinical nutrition, vol. 88, pp. 1304-1312, 2008. 12. c. x. zhang, s. c. ho, f. y. lin, y. m. chen, s. z. cheng and j. h. fu. dietary fat intake and risk of breast cancer: a casecontrol study in china. european journal of cancer prevention, vol. 20, no. 3, pp. 199-206, 2011. 13. c. pouchieu, m. deschasaux, s. hercberg, n. druesne-pecollo, p. latino-martel and m. touvier. prospective association between red meats intake and breast cancer risk modulation by an antioxidant supplementation in the su.vi. max randomized controlled trial. int j epidemiol, vol. 43, no. 5, pp. 1583-1592, 2014. 14. v. singh. and n. nimbkar. in: s. k. gupta (ed.), saff flower in breeding oilseed crops for sustainable production. jammu (j and k), india: sher-e-kashmir university of agricultural sciences and technology chatha, pp. 149-167, 2016. 15. h. j. murff, x. o. shu, h. li, g. yang, x. wu, h. cai, w. wen, y. t. gao and w. zheng. dietary polyunsaturated fatty acids and breast cancer risk in chinese women: a prospective cohort study. international journal of cancer, vol. 128, pp. 1434-1441, 2011. 16. r. l. prentice and l. sheppard. dietary fat and cancer: consistency of epidemiologic data and disease prevention that may follow from a practical reduction in fat consumption. cancer causes and control, vol. 1. pp. 81-97, 1990. 17. n. keum, d. h. lee, n. marchand, h. liu, h. oh, d. aune, d. c. greenwood and e. l. giovannucci. egg intake and cancers of the breast, ovary and prostate: a dose response meta-analysis of prospective observational studies. british journal of nutrition, vol. 114, no. 7, pp. 1099-1107, 2015. 18. e. cho, d. spiegelman, d. j. hunter, w. y. chen, m. j. stampfer, g. a. colditz and w. c. willett. premenopausal fat intake and risk of breast cancer. journal of the national cancer institute, vol. 95, pp. 1079-1085, 2003. 19. u. chandran, g. zirpoli, g. ciupak, s. e. mccann, z. gong, k. pawlish, y. lin, k. demissie, c. b. ambrosone and e. v. bandera. racial disparities in red meat and poultry intake and breast cancer risk. cancer causes control, vol. 24, pp. 2217-2229, 2013. 20. p. p. bao, x. o. shu, y. zheng, h. cai, z. x. ruan, k. gu, y. su, y. t. gao, w. zheng and w. lu. fruit, vegetable, and animal food intake and breast cancer risk by hormone receptor status. nutrition and cancer, vol. 64, pp. 806-819, 2012. . 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 research article evaluation of construction delay of public projects in erbil governorate noori sadeq ali, khalil ismail wali abstract delay is one of the greatest challenges facing the implementation of construction projects. the completion project time is considered one of the measure indicators for assessing the success of any project. delays usually have negative impacts of financial and social to all parties involved in the projects. the aim of the study is to find the most important causes of delay by evaluating the common causes of delays in terms of frequency, severity, and important indices in implementation of public construction projects in erbil governorate related to owners, contractors, consultants, and external factors. the data gathered through conducting a questionnaire for perception various parties involved in implementation of public construction projects. for these purpose, 104 forms of questionnaire were distributed to various parts to identify the delay causes in accordance with their opinions. it was concluded that the most significant factors causing the delay in construction of public project in erbil governorate related to the owner were “variation orders by owner during the construction” which came in the first rank, while the cause of delay related to the consultant that came in the first rank was “unclear and inadequate details in drawings,” whereas the factor related to the contractor which came in the first rank was “selecting incompetent contractor.” keywords: construction projects, delay, erbil governorate, evaluation, public project introduction the construction industry is one of the main sectors that provide important ingredients for the development of an economy. the construction is the tool through which a society achieves its goals of urban and rural development. however, it is becoming more complex due to the sophistication of construction process itself and the large number of parties involved in the construction process, i.e., clients, users, designers, regulators, contractors, suppliers, subcontractors, and consultants. modern construction projects were characterized by new standards, advanced technologies, multiparty participation, and frequent owner-desired changes. coupled with a state, there are inherent uncertainties and complexities in the physical, financial, and economic environment in which most projects are performed. such conditions have made completing projects on schedule and on budget a difficult task to accomplish, often leading to claims on cost compensation and time extensions.[1] construction time often serves as a benchmark for assessing the performance of a project and the efficiency of the project organization. the time required to complete construction of projects is often more than the specified time in contract. these “overruns” or time extensions happen due to many reasons, such as designer changes or errors, economic conditions, resource availability, and performance of projects. usually, majority of project delay occurs during construction phase, where unforeseen factors (environmental concerns and restrictions, ground conditions, etc.) are always involved. construction delays lead to increase in overall project cost, hence for the completing projects on time is beneficial to all parties involved in projects. therefore, it is essential to identify the actual causes of delay to minimize and avoid the delays and their corresponding expenses.[2] project success can be defined as meeting goals and objectives as prescribed in the project plan. a successful project means that the project has accomplished its technical performance, maintained its schedule, and remained within budgetary costs.[3] research objectives the aim of this study is to determine the most effective factors affecting the delay in public construction projects in erbil governorate projects. the main objectives of the study are as follows: corresponding author: noori sadeq ali, department of civil engineering, college of engineering, cihan university, erbil, iraq. phone: +964-750 453 75 45. e-mail: noori.ali@cihanuniversity.edu.iq doi: 10.24086/cuesj.v3n1y2019.pp18-26 copyright © 2019 noori sadeq ali, khalil ismail wali. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) received: sep 23, 2018 accepted: nov 06, 2018 published: may 13, 2019 department of civil engineering, college of engineering, cihan university-erbil, kurdistan region, iraq https://creativecommons.org/licenses/by-nc-nd/4.0/ ali and wali: evaluation of construction delay of public projects 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 1. identifying the main causes of delay in construction projects. 2. evaluating the top 10 significant causes of delay in construction projects. 3. identifying the means of minimizing delay in public construction projects. case study the research focuses on construction industry of kurdistan region in erbil governorate. the surveyed construction projects include public projects of nearly most of public sectors such as local administration, municipality, road works, irrigation works, water supply, sanitary, high education, health, and religion affairs (awqaf). the specified projects comprise schools, residential and non-residential offices, and hospitals. this study will focus on projects where the main client is the government (public construction projects). literature review types of delay delay in construction projects can be classified in various ways related to the parties involved in causing the delay as follows. excusable delay with compensation this type of delay is caused by the clients’ actions or inactions. once contractors encounter this sort of delay, they are entitled to time extension moreover as financial compensation as a result of the delays. an example of this would be the late release of drawings from the clients’ architect.[4] excusable delay without compensation this type is caused by third parties or incidents beyond the control of the client and the contractor. examples typically include acts of god, unusual weather, strikes, fires, and acts of government in its sovereign capacity.[5] in this case, the contractor is normally entitled to a time extension but no compensation for delay damages. non-excusable delay caused solely by the contractor or his suppliers, the contractor is generally not entitled to relief and must either make up the lost time through acceleration or compensate the owner.[4] many studies and researches conducted on the subject of delay in the completion time of construction projects; therefore, this part of literature review classified the studies carried out locally, regionally, and neighbor countries as follows: local studies ewadh et al.[6] studied the most effective causes of delays in execution and delivering of construction projects in specified time in karbala governorate. within 78 probable causes of delay, the most occurrence causes were delay in testing materials, awarding contracts to the lowest cost bidder, contractor’s financial difficulties, sudden rise in prices of materials, and incorrect of estimation of contract duration. muhammad[7] studied the causes of delay from the point of view of owners, engineers, and contractors. there are many causes of delay in construction projects in erbil governorate including inadequate contractor experience, waiting time for approval of tests and inspection, poor estimate of project duration by the contractor, poor technical staff qualification, financial difficulties of contractor, problems with subcontractors, unclear and inadequate details in drawings, slow payment of completed works, poor site management, and poor communication and coordination by the contractor with others. jahager[8] studied the causes of construction delay in baghdad city. he specified the most important causes of delay in construction project through a field survey of a questionnaire contained 58 causes of delay according to group of owners, contractors, and consultants. the study showed that the most important cause of delay is mistakes and discrepancies in design documents, ineffective planning and scheduling of project by contractor, poor site management and supervision by contractor, poor qualification of the contractor’s technical staff, unclear and inadequate details in drawings, inadequate designteam experience, insufficient data collection and survey before design, difficulties in financing project by contractor, obsolete or unsuitable construction methods, and unqualified workforce. bekr[9] studied the most important causes of delay in public projects in iraq through an intensive literature review and a survey which included a questionnaire used to assess the perceptions of the main participants involved in the construction of the projects. the 65 causes compiled through the literature review and the pilot study were classified into four groups: client related, contractor related, consultant related, and external factors. he concluded that the most effective delay factors affecting the time overrun in the public projects in iraq were security measures, government change of regulations and bureaucracy, official and non-official holidays, low performance of lowest bidder contractors in government tendering system, design and changes by owner, design changes by consultants, delay in progress payments by the owner, problems with local community, owner’s lack of experience in construction, and economic local and global conditions. regional studies several studies in neighbor countries and abroad have been reviewed. in jordan, sweis et al.[10] evaluated the most common causes of delays in projects using a survey distributed to consultant engineers, contractors, and owners, and interviews with senior professionals in the field. most correspondents agreed that financial difficulties faced by the contractor and too many change orders by the owner are the leading causes of construction delay. severe weather conditions and changes in government regulations and laws ranked among the least important causes. samarah and bekr[11] studied the effects of delay on project’s performance. the study was accomplished through an intensive literature review and a field study used to find the perception of major parties participating in the implementation of the construction projects in jordan. the survey which was carried out through a questionnaire covered (146) participants representing clients, contractors, and consultants. the study presented ranking of the significant factors for each group and then the significant factors of the whole survey. top ten factors causing delays for public sector projects in jordan were (1) inadequate management and supervision by the contractor, (2) client’s ali and wali: evaluation of construction delay of public projects 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 changes of the design, (3) inadequate planning and control by the contractor, (4) using lowest bid that lead to low performance, (5) changes in the extent of the project, (6) errors in design and contract documents, (7) progress payments are not made in time by the client, (8) rework due to mistakes during construction, (9) changes in the original design, and (10) low level productivity. a research survey on time performance of different types of construction projects in saudi arabia was conducted to determine the causes of delay and their importance according to each of the project participants, i.e., the owner, consultant, and contractor. 73 causes of delay were identified during this research. 76% of the contractors and 56% of the consultants indicated that average of time overrun is between 10% and 30% of the original duration. the most common cause of delay identified by all the three parties is the “change order.” furthermore, the study concluded that 70% of projects experienced time overrun and found that 45 of 76 (about 60%) projects were delayed.[12] a study of time delays and cost increases associated with construction of private residential projects in the state of kuwait was conducted. a survey response of 450 respondents was selected in 27 districts in metropolitan kuwait showed that time-delays and cost increases are the main causes of delays. the three main causes of time delays included changing orders, owners’ financial constraints, and owners’ lack of experience in the construction industry.[13] causes of delays in construction projects in turkey were studied by kazaz et al.[14] by applying a questionnaire to 71 companies working in turkey. the study showed that, of 34 factors affecting project duration, design and material change was found to be the most important predominant factor, delay of payments, and cash flow problems. abd el-razek et al.[15] studied the causes of delay in building construction projects in egypt. the study revealed that the most important causes were financing by contractor during construction, delays in contractor’s payment by owner, design changes by owner or his agent during construction, partial payments during construction, and non-utilization of professional construction/contractual management. khoshgoftar et al.[16] studied the causes of delay in iranian projects. the study showed that the main causes of delay are financial and payments of completed works, improper planning, site management, contract management, and lack of communication between parties. research methodology the data collection for the purpose of this study was conducted by the preparation of questionnaire including a list of 37 proposed causes of delay in construction projects. a handdelivered questionnaire method was used to obtain the opinion of respondents. a questionnaire was divided into three main parts. the first part was a clarification of the idea and the purpose of the survey as well as the definition of the interested area of the study. in the second part, the profile of the respondent was requested such as graduate year, number of years working in construction industry, number of projects supervised and/or executed, and the role kinds (site engineer, project manager, or any other duties) during working in construction. the third part concerned the delay causes in construction project in the form of clear statement. delay factors are classified into four categories relating to main participants: owner, consultants, contractors, and other externals. for each factor, two measures were used, frequency and severity. for each measure, the respondents have four options in the form of digits ranking from 1 to 4 as shown in table 1. the questionnaires submitted to respondents working in public project in erbil governorate who directly participated in construction of projects consisting of project managers, site engineer, and supervisors. data analysis the author used various factors related to project delay. the data was analyzed by using frequency and severity indices. frequency index the following equation was applied to analyze the causes of delay based on frequency occurrence identified by respondents. frequency index fi xi xi % � � ( )=         × ∑ ∑4 100 (1) where, xi is constant expressing weight (scale) given to ith response: 1, 2, 3, 4 fi is variable expressing frequency of ith response i is response category index of 1, 2, 3, 4. severity index the following equation was used to analyze the severity index in the manner of frequency index:        ∑ ∑ si severity index (%)=  xi 4  xi ×100 where, si is variable expressing severity of ith response. table 1: frequency and severity measurements of delay factors delay causes statements frequency severity delay scale xi 1 2 3 4 1 2 3 4 scale rarely happens sometimes happens often happens always happens little moderate great extreme factors f1 f2 f3 f4 s1 s2 s3 s4 ali and wali: evaluation of construction delay of public projects 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 important index the important index (i.i) of each cause was calculated using the following equation: research findings and discussions general characteristics of respondents’ profile the respondents include experienced engineers executed and supervised the public projects in governorate institution of most services; local administration, works and housing, municipality, roads, irrigations, water supply, sewage, electricity, religions affairs (awqaf), and health. the respondent’s year of experience ranges from 1 year to >25 years as shown in figure 1. the number of executed and supervised projects by respondents ranging between 1 and >50 projects is illustrated in figure 2. the percentage distribution of types of project ranges from 2% to 20% as shown in figure 3. analyzing the ranks of delay causes table 2 shows the causes of the delay which are related to owner, consultant, contractor, and external in terms of frequencies, severities. frequency occurrence of top 10 delay causes the frequency occurrences of top 10 highest delay causes are listed in table 3, showing that the most causes related to the contractor are “selecting incompetent contractors,” “not selecting competent subcontractors,” “contractor’s financial difficulties,” “ineffective planning and scheduling of project by contractor,” “the required skilled figure 1: respondent’s year of experience figure 2: number of projects executed and supervised by the respondents ali and wali: evaluation of construction delay of public projects 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 labor is not available,” “differing site conditions,” and “the required equipment and tools are not available.” the highest causes related to the owner are “variation orders by owner during construction,” and “delays in contractor’s payment by owner.” the “weather conditions” also listed within the highest 10 delay causes related to the external conditions. severity occurrence of top 10 delay causes the most severe causes of delay are due to contractor relating to his experience and capacity, subcontractors, and financial difficulties, while the causes related to owner are only variation order and delaying payments. only weather conditions are related to external conditions. the overall causes sorting from high to lower severity are “selecting incompetent contractors,” “variation order(s) by owner during construction,” contractor’s figure 3: types of projects executed by the respondents delay causes statements frequency of occurrence severity of occurrence importance index index rank index rank index rank causes related to the owner variation order (s) by owner during construction 71.53 2 62.85 2 44.95 4 late in revising and approving design documents 53.13 15 52.11 19 27.68 17 delay in approving shop drawing and sample materials 55.56 11 52.78 17 29.32 12 poor communication and coordination 45.14 26 45.71 30 20.63 27 slowness in decision-making process 53.29 14 53.96 13 28.76 15 conflicts between joint-ownership of the project 44.01 27 46.83 27 20.61 28 delays in contractor’s payment by owner 64.2 7 62.67 7 40.26 7 design changes by owner or his agent during construction 51.07 20 53.99 14 27.57 18 partial payment during construction 47.83 23 43.75 31 20.92 26 owner interference 51.39 19 48.26 24 24.80 21 quality assurance – control 55.00 13 52.86 16 29.07 13 bureaucracy 43.01 30 46.07 29 19.82 30 causes related to the consultant change in design by engineer or consultant 52.90 16 51.09 21 27.02 19 conflict between contract documents 39.55 35 39.34 36 15.56 36 inflexibility (rigidity) of consultant 42.25 32 43.08 33 18.20 32 poor communication and coordination by consultant with other parties 39.39 36 42.80 34 16.86 35 inadequate experience of designer 43.38 28 48.41 23 21.04 25 mistakes and discrepancies in design documents 49.15 21 50.00 22 24.58 22 design complexity 40.30 34 43.56 32 17.55 34 unclear and inadequate details in drawings 51.49 18 54.23 12 27.92 16 lack of consultant’s knowledge of available materials and equipment 45.52 25 47.76 25 21.74 24 causes related to the contractor differing site conditions 56.52 9 54.04 15 30.55 11 selecting incompetent contractors 73.57 1 71.74 1 52.78 1 not selecting competent subcontractors 68.28 3 66.67 5 45.52 2 table 2: the important index and ranking of delay causes (contd...) ali and wali: evaluation of construction delay of public projects 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 financial difficulties,” “whether condition,” “not selecting competent subcontractors,” “ineffective planning and scheduling of project by contractor,” “delays in contractor’s payment by owner,” “poor management of project site,” “the required skilled labors are not available,” and “rework due to errors during construction” as shown in table 4. important index of top 10 delay causes the important indices of top 10 delay causes are listed in table 5. the most causes of delay are related to contractor. the first three high ranked causes included “selecting incompetent contractors” with important index of 52.78% and 45.5% important index for both “not select competent subcontractors” and “contractor’s financial difficulties.” the most managers of projects are suffering from incompetent contractors and subcontractors with problem of financial and technical difficulties. the other causes are related to the owner comprising “variation order(s) by owner during construction” and “delays in contractor’s payment by owner” which are the most common cause in other neighbor countries with important indices of 45% and 40.3%, respectively. the latter cause is the “weather conditions” related to external condition with important indices of 45%. the last remained causes of delay are related to contractors such as the required labor skill, poor management, and the unavailability of equipment and tools. the most of these causes of delay and their important indices are approximately similar to those mentioned by bekr, 2015.[9] and and a study by (assaf and al-hejji, 2006)[12] as mentioned previously in literature review. ranking of delay sources the causes of delay sources as previously mentioned are related owner, contractor, consultant, and external conditions. the ranking of the frequency of occurrence, severity, and importance index is as follows. ranking of delay causes related to the owner the frequency, severity, and importance index of delay related to the owner are listed in table 6. the highest frequency is delay causes statements frequency of occurrence severity of occurrence importance index index rank index rank index rank poor management of project site 55.36 12 59.00 8 32.66 9 poor management of project contract 47.43 24 51.45 20 24.40 23 conflicts in work schedule of the subcontractors 48.91 22 52.61 18 25.73 20 contractor’s financial difficulties 66.79 4 68.12 3 45.49 3 rework due to errors during construction 51.84 17 56.15 10 29.11 14 site accidents 43.18 29 46.43 28 20.05 29 the required skilled labor is not available 58.21 8 57.97 9 33.74 8 the required equipment and tools are not available 56.44 10 54.55 11 30.79 10 ineffective planning and scheduling of project by contractor 64.45 6 64.02 6 41.26 6 causes related to external conditions new government regulations 42.65 31 42.42 35 18.09 33 strikes 31.25 37 37.86 37 11.83 37 political influence 40.67 33 47.39 26 19.27 31 weather conditions 65.94 5 67.75 4 44.68 5 table 2: (continued) table 3: frequency of top 10 highest delay causes causes of delay frequency (%) rank source selecting incompetents contractors 73.6 1 contractor variation order (s) by owner during construction 71.5 2 owner not selecting competent subcontractors 68.3 3 contractor contractor’s financial difficulties 66.8 4 contractor weather conditions 65.9 5 external conditions ineffective planning and scheduling of project by contractor 64.5 6 contractor delays in contractor’s payment by owner 64.2 7 owner the required skilled labors are not available 58.2 8 contractor differing site conditions 56.5 9 contractor the required equipment and tools are not available 56.4 10 contactor ali and wali: evaluation of construction delay of public projects 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 due to “variation orders” and “delay in contractor’s payment by owner” at 71.53% and 64.24%, respectively. the following causes are due to “delay in approving shop drawing and sample materials,” “quality assurance – control,” “slowness in decision-making process,” “late in revising and approving design documents,” “owner interference,” and “design changes by owner or his agent during construction” with frequency ranging between 51.07% and 55.56%. the remaining ownerrelated causes of delay included “partial payment during construction,” “poor communication and coordination,” “conflicts between joint-ownership of the project,” and “bureaucracy” ranging between 43.01% and 47.83%. the ranking of importance index of causes of delay is in the order as of their frequency of occurrences. regarding the important indices of delay cause, the highest two causes are “variation order(s)” and “delays in contractor’s payment” with important indices of 44.95% and 40.26%, respectively. the other causes of delay were between 20% and 29.32%. ranking of delay causes related to the consultants the ranking of delay causes related to consultants listed in table 7 shows that the highest ranked delay cause is “unclear and inadequate details in drawings,” “mistakes and discrepancies in design documents,” “lack of consultant’s knowledge of available materials and equipment,” and “inadequate experience of designer” at important indices of 27.92%, 24.58%, 21.74%, and 21.02% respectively. other causes are less effective at important indices <20% which are “inflexibility (rigidity) of consultant,” “design complexity,” “conflict between contract documents,” and “poor communication and coordination by consultant with other parties.” ranking of delay causes related to the contractor the ranking delay causes related to contractor are shown in table 8, indicating that owner, consultant and external conditions ranked high in causing the delay. therefore, selecting incompetent contractors (52.78%) have significant effects on delay of projects. the latter group of causes of delay consisting of subcontractors selecting, financial difficulties, and ineffective planning and scheduling have relatively effects on the delay of project with important indices between 40% and 50%. the other group of causes lowers the effects on delay. the important indices ranged between more than 30% to <34% which deals with labor experience, site conditions, and site management having their effects on the delay of project. the last set of causes of delay was less than 30%. they are reworking due to errors in construction, conflicts between subcontractor’s scheduling, poor management of contract, and site accidents. ranking delay related to external conditions the only four causes of delay related to external conditions rather than participants’ owner, consultant, and contractor are shown in table 9. the effective cause in this group of delay related to external causes is “weather conditions” which is not excusable as per iraqi conditions of contract of civil engineering works (1987) with importance index of 44.68%. conclusion the study focused on identifying and ranking in terms of frequency, severity, and importance of the factors of delay in construction of executed public projects. the study revealed that the top 10 delays from highest to lowest are selecting incompetents contractors, incompetent subcontractors, contractor’s financial difficulties, variation order(s) by owner, weather conditions, ineffective planning and scheduling of project by contractor, delays in contractor’s payment, the unavailability of skilled labor, poor management, finally, the unavailability of equipment and tools. table 4: severity occurrence of top 10 delay causes causes of delay severity (%) rank source selecting incompetents’ contractors 71.7 1 contractor variation order (s) by owner during construction 68.2 2 owner contractor’s financial difficulties 68.1 3 contractor whether condition 67.8 4 external condition not selecting competent subcontractors 66.7 5 contractor ineffective planning and scheduling of project by contractor 64.0 6 contractor delays in contractor’s payment by owner 62.7 7 owner poor management of project site 59 8 contractor the required skilled labors are not available 58 9 contractor rework due to errors during construction 56.2 10 contractor table 5: the important index of top 10 delay causes causes of delay important index (%) rank source selecting incompetents contractors 52.78 1 contractor not selecting competent subcontractors 45.5 2 contractor contractor’s financial difficulties 45.5 3 contractor variation order (s) by owner during construction 45 4 owner weather conditions 44.7 5 external conditions ineffective planning and scheduling of project by contractor 41.3 6 contractors delays in contractor’s payment by owner 40.3 7 owner the required skilled labors are not available 33.7 8 contractor poor management of project site 32.7 9 contactor the required equipment and tools are not available 30.8 10 contractor ali and wali: evaluation of construction delay of public projects 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 table 6: ranking of frequency of delay causes related to the owner causes of delay frequency of occurrence severity of occurrence importance index index rank index rank index rank variation order (s) by owner during construction 71.53 1 62.85 1 44.95 1 delays in contractor’s payment by owner 64.24 2 62.67 2 40.26 2 delay in approving shop drawing and sample materials 55.56 3 52.78 6 29.32 3 quality assurance – control 55 4 52.86 5 29.07 4 slowness in decision-making process 53.29 5 53.96 4 28.76 5 late in revising and approving design documents 53.13 6 52.11 7 27.68 6 owner interference 51.39 7 48.26 8 24.8 8 design changes by owner or his agent during construction. 51.07 8 53.99 3 27.57 7 partial payment during construction 47.83 9 43.75 12 20.92 9 poor communication and coordination 45.14 10 45.71 11 20.63 10 conflicts between joint-ownership of the project 44.01 11 46.83 9 20.61 11 bureaucracy 43.01 12 46.07 10 19.82 12 table 7: ranking of delay related to the consultant causes of delay frequency of occurrence severity of occurrence importance index index rank index rank index rank unclear and inadequate details in drawings 51.49 1 54.23 1 27.92 1 mistakes and discrepancies in design documents 49.15 2 50.00 2 24.58 2 lack of consultant’s knowledge of available materials and equipment 45.52 3 47.76 4 21.74 3 inadequate experience of designer 43.389 4 48.41 3 21.04 4 inflexibility (rigidity) of consultant 42.25 5 43.08 6 18.20 5 design complexity 40.3 6 43.56 5 17.55 6 conflict between contract documents 39.55 7 39.34 8 15.56 8 poor communication and co-ordination by consultant with other parties 39.39 8 42.80 7 16.86 7 table 8: ranking of delay related to the contractor causes of delay frequency of occurrence severity of occurrence importance index index rank index rank index rank selecting incompetents contractors 73.57 1 71.74 1 52.78 1 not selecting competent subcontractors 68.28 2 66.67 3 45.52 2 contractor’s financial difficulties 66.79 3 68.12 2 45.49 3 ineffective planning and scheduling of project by contractor 64.45 4 64.02 4 41.26 4 the required skilled labors are not available 58.21 5 57.97 6 33.74 5 differing site conditions 56.52 6 54.04 9 30.55 8 the required equipment and tools are not available 56.44 7 54.55 8 30.79 7 poor management of project site 55.36 8 59.00 5 32.66 6 rework due to errors during construction 51.84 9 56.15 7 29.11 9 conflicts in work schedule of the subcontractors 48.91 10 52.61 10 25.73 10 poor management of project contract 47.43 11 51.45 11 24.40 11 site accidents 43.18 12 46.43 12 20.05 12 of these 10 factors, 7 of them are related to contractor. the only two factors “variation order(s) by owner during construction,” and “delays in contractor’s payment by owner.” the weather conditions are related to external conditions. the ranking of delay factors related to owner listed from high importance index to lower (44.95–24.8%) is “variation order(s) by owner during construction,” “delays in contractor’s payment by owner,” “delay in approving shop drawing and ali and wali: evaluation of construction delay of public projects 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 18-26 sample materials,” “quality assurance – control,” “slowness in decision-making process,” “late in revising and approving design documents,” “design changes by owner or his agent during construction,” and “owner interference.” consultant-related factors based on importance index from high importance index to lower (27.92–21.04%) are “unclear and inadequate details in drawings,” “mistakes and discrepancies in design documents,” “lack of consultant’s knowledge of available materials and equipment,” and “inadequate experience of designer.” the contractor related factors of delay causes which are the most effective factors from high to lower importance index (52.78–32.66%) are “selecting incompetents contractors,” “not selecting competent subcontractors,” “contractor’s financial difficulties,” “ineffective planning and scheduling of project by contractor,” “the required labor skill are not available,” “differing site conditions,” “the required equipment and tools are not available,” and “poor management of project site.” the last source of delay related to external conditions was weather conditions 44.68%. for avoiding and minimizing the delays in construction of public projects, it is recommended to impose regulations more strict in selecting contractors and subcontractors, time payments and financial controlling, planning and scheduling and revising them, site management, more accurate initial cost estimate, and frequent progress meetings of various parties involved in construction of public projects. references 1. s. ramya and r. divya. “causes, effects and minimization of delays in construction projects”. in: national conference on research advances in communication, electrical science and structures. ncraccess, 2015. 2. s. al hammadi and m. s. nawab. “study of delay factors in construction projects”. international advanced research journal in science and technology, vol. 3, no. 4, 2016. 3. y. frimpong, j. oluwoye and l. crawford. “causes of delay and cost overruns in constructionof groundwater projects in a developing countries; ghana as a case study”. international journal of project management, vol. 21, no. 1, pp. 321-326, 2003. 4. k. a. mohammed and a. d. isah. “causes of delay in nigeria construction industry”. interdisciplinary journal of construction research in business, vol. 4, no. 2, pp. 785-794, 2012. 5. t. ghada, m. badawy and o. el-nawawy. “a model for evaluation of delays in construction projects”. international journal of innovative research in science engineering and technology, vol. 5, no. 3, pp. 688-693, mar 2016. 6. a. ewadh and k. g. al-hendawi. “causes of delay in iraq construction projects”. journal of babylon university, vol. 14, no. 4, pp. 483-494, 2007. 7. a. j. muhammad. “causes of delays in construction projects in erbil governorate”. zanco, journal of pure and applied sciences, vol. 3, no. 24, pp. 81-94, 2011. 8. q. k. jahager. “important causes in construction projects in baghdad city”. australian journal of basic and applied sciences, vol. 7, no. 4, pp. 14-23, 2013. 9. g. a. bekr. “causes of delay in public construction projects in iraq”. jordan journal of civil engineerin, vol. 9, no. 2, pp. 149162, 2015. 10. g. sweis, r. sweis, a. a. hammad and a. shboul. “delays in construction projects: the case of jordan”. international journal of project management, vol. 26, no. 1, pp. 665-674, 2008. 11. a. samarah and g. a. bekr. “causes and effects of delay in public construction projects in jordan”. american journal of engineering research, vol. 5, no. 5, pp. 87-94, 2016. 12. s. a. assaf and a. hajji. “cause of delay in large construction projects”. international journal of project management, vol. 24, no. 4, pp. 349-357, 2006. 13. p. a. koushki, k. al-rashid and n. kartamn. “delays and cost increases in the construction of private residential projects in kuwait”. construction management and economics, vol. 23, no. 3. pp. 285-294, 2005. 14. a. kazaz, s. ulubeyli and n. a. tuncbilekli. “causes of delays in construction projects in turkey”. journal of civil engineering and management, vol. 18, no. 3, pp. 426-435, 2012. 15. m. e. abd el-razek, h. a. bassioni and a. m. mobarak. “causes of delay in building construction projects in egypt”. journal of construction engineering and management, vol. 134, no. 11, pp. 831-841, nov 2008. 16. m. khoshgoftar, a. h. a. baker and o. osman. “causes of delays in iranian projects”. the international journal of construction management, vol. 10, no. 2, pp. 53-59, 2010. table 9: ranking of delay related to external conditions causes of delay frequency of occurrence severity of occurrence importance index index rank index rank index rank weather conditions 65.94 1 67.75 1 44.68 1 new government regulations 42.65 2 42.42 3 18.09 3 political influence 40.67 3 47.39 2 19.27 2 strikes 31.25 4 37.86 4 11.83 4 . 52 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 52-56 research article significance of hepatitis b virus diagnosis by real-time polymerase chain reaction over serological markers in hepatitis b virus patients amer a. khaleell*, salah t. jalall, saeed g. hussain2 1department of medical microbiology, college of health sciences-hawler medical university, erbil, iraq, 2department of medical microbiology, college of medicine-hawler medical university, erbil, iraq abstract hepatitis b virus (hbv) is the leading cause of viral hepatitis, as currently over 2 billion people have hbv infection worldwide. nucleic acid assay and quantitative hepatitis b surface antigen (hbsag) have been developed for diagnostic and therapeutic monitoring of patients with hbv infection. these tests might also show correlation between hbv dna and hbs serostatus. the study aimed to find and analyze the frequency and impact of hbsag seropositivity among patients revealed hbv dna negative level through quantitative estimation of both seromarkers. real-time polymerase chain reaction (rt-pcr) and elecsys assays were used for quantitative estimation of hbv dna and hbs antigen, respectively. a total of 256 blood samples were used from patients referred for either diagnostic purpose and/or hbv viral load monitoring after antiviral therapy. blood profile analysis showed 12.26% hbs antigen seropositivity among patients revealed negative for nucleic acid assay for hbv dna. positive hbs antigen titers ranged from 1000–50,000 coi, with seronegative anti-hbs antibody test for all samples tested positive for hbs antigen. this study delineated that negative or undetectable quantitation of hbv dna level does not exclude hbv infection; as the level might fluctuate in different phases of hbv replication. this gives an impression and raising a question about significance of replacing test for hbsag with quantitation of hbv dna pcr assay. thus, the study refers to a special hbv profile outside the classical pattern. keywords: component, hepatitis b surface antigen, hepatitis b virus, hepatitis, quantitative hepatitis b virus dna introduction hepatitis b virus (hbv) is the leading cause of viral hepatitis, as currently over 2 billion people have hbv infection worldwide. approximately 600,000 deaths occur every year as a result of the acute and chronic consequences of hbv infection.[1] chronic hbv affects nearly 350 million patients worldwide and may further progress to cirrhosis and/or hepatocellular carcinoma (hcc) in 15–40% of cases.[2] hbv infection is a dynamic process characterized by replicative and non-replicative phases based on virus-host interaction, which are present in some form of all infected patients.[3] to measure hbv dna (viral load), a laboratory measures how many hbv dna units are found in 1 ml of blood. the result is written in international units per ml (iu/ml). high levels of hbv dna indicate a high rate of hbv replication; however, low or undetectable levels–<2000 iu/ml indicate an “inactive” infection.[4] during hbv infection, markers can be detected in sera samples using immunoassays. for example, patient sera that contain anti-hbs indicates immunity from past infections or vaccinations, and a patient with anti-hbe antibodies in their sera is considered as spontaneous resolution of infection or therapy-induced improvement.[5,6] one of the serological methods using for the diagnosis of hbv is enzyme immune assays. the hepatitis b surface antigen (hbsag) and hbv dna levels were considered to be a risk factor of hepatocarcinogenesis in untreated hb patients.[7] the levels of quantified hbsag are considered as a marker with which to evaluate the host immunological control of infection and hbv replication.[8] low hbsag levels in some hbv patients are considered to indicate a high likelihood of hbv clearance and lower hepatitis activity.[7,9] the aim of this study was to estimate hbv-dna positivity states among serologically screened patients and to compare the sensitivity of molecular and serological methods for the diagnosis of hbv infections. cihan university-erbil scientific journal (cuesj) corresponding author: amer a. khaleel, college of health sciences-hawler medical university, erbil, iraq. e-mail: ameralikhaleel@yahoo.com received: mar 31, 2019 accepted: apr 17, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp52-56 copyright © 2020 amer amer a. khaleel, salah t. jalal, saeed g. hussain. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ khaleel, et al.: significance of hbv diagnosis by rt-pcr 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 52-56 materials and methods for this cross-sectional study, a total of 256 blood samples were collected from hbv patients referred to erbil central laboratory, from june 2017 to september 2017. both sexes were included (154 males and 102 females), with age range between 20 and 60 years. real-time polymerase chain reaction (rt-pcr) as a molecular and elecsys assay as a serological method were used for the diagnosis and comparing their sensitivity purposes. for rt-pcr technique, 5 ml of venous blood were collected using edta tubes and centrifuged at 1000 rpm for 5 min and patients’ plasma was separated. viral dna was extracted from a 200 µl aliquot of serum using a qiagen mini blood kit (qiagen, hilden, germany) according to the manufacture instructions. viral loads of hbv dna were estimated by rt-pcr (hbv rg pcr artus, germany). the amplification was performed in a 50 µl reaction mixture containing 30 µl of mixture of hbv rg master mix (buffer, dntp, primer, probe, and enzymes) and hbv rg inhibition control mixed with 20 µl of dna template to each reaction. the primers hbv-taq i (forward primer): caa cct cca atc act cac caac and hbv-taq 2 (reverse primer): ata tga taa aac gcc gca gac ac were used from the sequences of the conserved regions of the hbv surface gene used previously by weinberger et al.[10] for performing rt-pcr, rotorgene 3000 (a machine used routinely in erbil central laboratory for detection of variety of dnas and rnas) was used following manufacture’s instructions. the rt-pcr cycling parameters consisted of denaturation at 95°c for 15 s, 55°c for 30 s, and 72°c for 15 s. the results were considered positive if a signal was detected in cycling a fam, whereas no signal detection indicated as negative results. the quantitation of hbv dna was performed for all blood samples and the results were considered positive and significant if viral load was more than 1×105 viral copies/ ml. titers <50 iu were considered as negative for hbv dna. the screened results were categorized according to positive hbv dna titers into inactive, gray zone, and active carriers. quantitation of hbsag was measured using the elecsys hbsag quantitative assay. detection of hbsag by elecsys utilizes a sandwich principle and this assay has two stages: first, a complex is formed with two monoclonal hbsagspecific antibodies, one of which is biotinylated and the other labeled with a ruthenium complex. this complex joins to the solid phase through the interaction of biotin and streptavidin after the attachment of streptavidin-coated microparticles.[11] the mixture is subsequently aspirated into a measuring cell, where application of a voltage induces chemiluminescent emission, which is measured by a photomultiplier. all serum samples were tested at a dilution of 1:400. the elecsys hbsag quantitative assay is calibrated to give results in iu/ml.[11] statistical analysis statistical analysis was done using spss 23.0. hbv titers expressed as means ± standard error. comparisons between active and inactive carriers use t-tests. comparisons between multiple groups were performed by anova. p ≤ 0.05 was considered as statistically significant. results and discussion of the 256 cases, 154 (60%) were male and 102 (40%) were female, with age range between 20 and 60 years. rt-pcr results showed that 163 of 256 samples were hbv dna negative, from them, 20 samples showed positive results for hbsag quantitation [table 1]. the asian pacific association for the study of the liver, european association for the study of the liver, and the guidelines of the american association for the study of liver diseases refer that the disappearance of hbsag is the ideal and cornerstone during natural course of hbv infection.[12-14] from the clinical standpoint of view, the progression from chronic hepatitis b to cirrhosis is increasingly signified through the hbv viral load.[8] meanwhile, quantitative hbsag level was regarded as marker for clearance, hepatitis activity, and immunological host response to control hbv replication and the likelihood of inactive virus carrier.[9,7,15] however, little is known about correlation between hbv replication and low levels of hbsag, as reported in some patients with chronic hbv infection.[16-18] the results of the present study showed 12.26% hbs seropositivity (range 1000–5000 coi) among patients seronegative for hbv dna [table 2], with highly significant difference (p ≤ 0.001) between mean of hbv dna seropositive table 1: baseline patient’s characteristics variables all patients (n=256) gender male 154 (60%) female 102 (40%) hbv dna iu/ml 256 <50 (negative) 163 (63.67%) 101–500 22 (24.44%) 501–1000 10 (11.11%) 1001–2000 10 (11.11%) 2001–20,000 20 (22.22%) >20,000 31 (34.44%) qhbsag tested 163/256 (63.67%) <10 coi 143 (55.85%) 1000–2000 coi 3 (15%) 2001–3000 coi 4 (20%) 3001–4000 coi 8 (40%) 4001–5000 coi 5 (25%) anti-hbsab tested 20 positive 0.00 negative 20 (100%) hbs positive, hbv dna negative 20/163 qhbsag: quantitative hepatitis b surface antigen, hbv: hepatitis b virus table 2: frequencies of hbv dna seropositivity among patients negative for hbv dna assay number screened hbv dna pcr +ve hbv dna pcr−ve (n=163) hbs +ve hbs −ve n=256 93 (36.32%) 20 (12.26%) 143 (87.73%) hbv: hepatitis b virus, pcr: polymerase chain reaction khaleel, et al.: significance of hbv diagnosis by rt-pcr 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 52-56 and negative titers [table 3]. regarding to the gender positivity for pcr test, in total of 154 male patients 65 (42%) were positive and of 102 female patients only 28 (27%) were positive [table 4]. the seromarkers profile among patients negative for hbv dna revealed anti-hbs antibody seronegative among the total 20 patients positive for hbsag [table 5]. however, results of anti-hbs titers among patients negative for hbv dna showed 20 samples were positive showing different titers [table 6]. during the immune-tolerant phase, hbsag level is the highest one, and the level decreases progressively from immune tolerance to inactive phase that reflects a low replicative phase of immune control.[19] the detection of hbv infection is vital for diagnosis our group published a paper that delineated the impact of pcr dna assay for the diagnosis of hbv infection.[20] the data of this study are agree with other studies[21] for the serologic profile of hbsag seropositivity among patients screened negative for hbv dna. they reported 6% of their samples gave hbsag positive among donations negative for hbv dna level, others[8] referred that in nine patients with hcc; hbv dna was undetectable but the level of hbsag was ≥2000 coi. others reported 31.4% hbsag positive among patients negative for hbv dna.[22] however, data from another study showed that 100 of 140 patients were seronegative for hbv dna.[23] another study[24] reported that 69% of confirmed hbsag-positive donations had undetectable hbv dna levels. however, others[25,26] found that none of the hbsag-positive donors with very low or negative for hbv dna were anti-hbs antibody positive.[24,27] among explanation for such unusual hbv serologic profile, the patients may be chronically infected with hbv particularly; they are in the non-replicative phase as part of the natural course of infection.[22] the question might be raised here is about such serologic profile of hbsag positive among patients negative for hbv dna, are these patients infectious and transmissible for community,[21] referred for the need of larger sample volume (5–10 ml) necessary for extraction and concentration to determine the real hbv dna status among samples negative for dna. in this study, pearson’s correlation analysis revealed no significant correlation between serum hbv dna and hbsag levels (r = 0.331; p > 0.3) [figure 1]. different studies reported discrepant results regarding correlation between hbv dna and hbsag levels, our data were disagree with other studies who reported positive correlation.[28-34] meanwhile, our results agree with others for no correlation between hbv dna and hbsag levels.[35,36] such correlation is only weak in patients negative for hbsag.[19,30,37,38] this is because the level of hbv dna fluctuates from undetectable to >2,000,000 iu/ml.[12] this is why the level of hbsag does not fall in the same proportion or level when correlated with hbv dna level as a marker for table 3: mean titers of hbv dna among patients screened number screened hbv dna pcr +ve (n=93) hbv dna pcr −ve (n=163) p-value n=256 18,582,661.24±5,934,488.97 31.12±4.45 0.000 hbv: hepatitis b virus, pcr: polymerase chain reaction figure 1: pearson correlation between quantitative hepatitis b virus dna level and hepatitis b surface antigen. r = 0.229, p = 0.331 table 5: hbsag and anti-hbs seromarkers profile among patients negative for hbv dna serological profile hbs anti-hbs (n=20) >10 coi +ve <0.9 coi −ve <10 iu/ml −ve >10 iu/ml +ve hbv dna pcr negative (n=163) 20 (12.27%) 143 (87.73%) 20 (100%) 0.00 hbv: hepatitis b virus, pcr: polymerase chain reaction, hbsag: hepatitis b surface antigen table 4: frequencies of hbv dna positivity according to the gender gender positive pcr negative pcr total p-value male 65 (42.20%) 89 (57.79%) 154 ≤0.001 female 28 (27.45%) 74 (72.54%) 102 total 93 (36.3232%) 163 (63.67%) 256 hbv: hepatitis b virus, pcr: polymerase chain reaction table 6: distribution of hbsag and anti-hbs titers among patients negative for hbv dna hbs titers coi anti-hbsab iu/ml non-reactive <0.9 coi 1000–2000 2001–3000 3001–4000 4001–>5000 non-reactive <10 reactive >10 0.00 3 (15%) 4 (20%) 8 (40%) 5 (25%) 20 (100%) 0.00 hbv: hepatitis b virus, pcr: polymerase chain reaction, hbsag: hepatitis b surface antigen khaleel, et al.: significance of hbv diagnosis by rt-pcr 55 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 52-56 disease progression.[4] martinot-peignoux et al.[39] referred that hbv dna diminished with time from seroclearance; however, covalently closed circular dna as integrated hbv dna remained detectable in liver biopsies. it is thus critical for clinicians to be aware of the significance of hbv dna serial measurement and lifelong follow-up to establish and confirm that inactive carrier condition is maintained.[4] among major guidelines, if hbv dna is undetectable, either prophylaxis with lamivudine or strict follow-up every 1–3 months for alt and hbv dna level is indicated.[13] conclusion serology will undoubtedly continue to be widely used in the detection of hbv infection; however, hbv dna assays can serve as an important tool, especially in detecting low levels of hbv dna and in patients with the past hbv infection. data from the present study also concluded that negative or undetectable quantitation of hbv dna level does not exclude hbv infection; as the level might fluctuate in different phases of hbv replication. this gives an impression and raising a question about significance of replacing test for hbsag with quantitation of hbv dna pcr assay. thus, the study refers to a special hbv profile outside the classical pattern. references 1. world health organization. “world health organization hepatitis b factsheet”. geneva: world health organization, 2017. 2. s. baig, a. a. siddiqui, w. ahmed, h. qureshi and a. arif. “the association of complex liver disorders with hbv genotypes prevalent in pakistan”. virology journal, vol. 4, no. 1, p. 128, 2007. 3. g. fattovich. “natural history of hepatitis b”. journal of hepatology, vol. 39, pp. 50-58, 2003. 4. i. pita, a. m. horta-vale, h. cardoso and g. macedo. “hepatitis b inactive carriers: an overlooked population?” ge portuguese journal of gastroenterology, vol. 21, no. 6, pp. 241-249, 2014. 5. l. m. villar, h. m. cruz, j. r. barbosa, c. s. bezerra, m. m. portilho and l. de paula scalioni. “update on hepatitis b and c virus diagnosis”. world journal of virology, vol. 4, no. 4, p. 323, 2015. 6. i. j. radii and a. m. saud. “comparative assessment between serological and molecular diagnosis for patients groups with hepatitis b virus”. international journal of current microbiology and applied science, vol. 6, no. 3, pp. 742-748, 2017. 7. t. c. tseng, c. j. liu, h. c. yang, t. h. su, c. c. wang, c. l. chen and j. h. kao. “high levels of hepatitis b surface antigen increase risk of hepatocellular carcinoma in patients with low hbv load”. gastroenterology, vol. 142, no. 5, pp. 1140-1149, 2012. 8. m. kawanaka, k. nishino, j. nakamura, t. oka, n. urata, d. goto and g. yamada. “quantitative levels of hepatitis b virus dna and surface antigen and the risk of hepatocellular carcinoma in patients with hepatitis b receiving long-term nucleos (t) ide analogue therapy”. liver cancer, vol. 3, no. 1, pp. 41-52, 2014. 9. t. c. tseng, c. j. liu, t. h. su, c. c. wang, c. l. chen, p. j. chen and j. h. kao. “serum hepatitis b surface antigen levels predict surface antigen loss in hepatitis b e antigen seroconverts”. gastroenterology, vol. 141, no. 2, pp. 517-525, 2011. 10. k. m. weinberger, e. wiedenmann, s. böhm and w. jilg. “sensitive and accurate quantitation of hepatitis b virus dna using a kinetic fluorescence detection system (taqman pcr)”. journal of virological methods, vol. 85, no. 1-2, pp. 75-82, 2000. 11. h. j. lee, s. y. kim, s. m. lee, j. heo, h. h. kim, c. l. chang and h. c. son. “elecsys hepatitis b surface antigen quantitative assay: performance evaluation and correlation with hepatitis b virus dna during 96 weeks of follow-up in chronic hepatitis b patients”. annals of laboratory medicine, vol. 32, no. 6, pp. 420-425, 2012. 12. a. s. lok and b. j. mcmahon. “chronic hepatitis b: update 2009”. hepatology, vol. 50, no. 3, pp. 661-662, 2009. 13. european association for the study of the liver. “easl clinical practice guidelines: management of chronic hepatitis b virus infection”. journal of hepatology, vol. 57, no. 1, pp. 167-185, 2012. 14. y. f. liaw, j. h. kao, t. piratvisuth, h. l. y. chan, r. n. chien, c. j. liu and d. amarapurkar. “asian-pacific consensus statement on the management of chronic hepatitis b: a 2012 update”. hepatology international, vol. 6, no. 3, pp. 531-561, 2012. 15. h. lik-yuen chan, g. lai-hung wong, c. h. tse, h. y. chan and v. wai-sun wong. “viral determinants of hepatitis b surface antigen seroclearance in hepatitis b e antigen-negative chronic hepatitis b patients”. journal of infectious diseases, vol. 204, no. 3, pp. 408-414, 2011. 16. j. cheng, c. sun, y. chen, y. dai, x. u. zhiliang, g. sun and l. i. xiaojun. “molecular analysis on chronic hepatitis b patients with low-level hbsag”. chinese journal of laboratory medicine, vol. 32, no. 10, pp. 1128-1132, 2009. 17. j. cheng, c. sun, y. chen, z. xu, g. wang, g. sun and x. li. “analysis on clinical features and immunity in chronic hepatitis b virus infected patients with low-level hbsag”. african journal of microbiology research, vol. 4, no. 7, pp. 547-550, 2010. 18. a. jeffery-smith, j. hubb, a. oliver and c. y. tong. “an apparent low level of hepatitis b surface antigen (hbsag) in the presence of significant viral replication”. journal of clinical virology: the official publication of the pan american society for clinical virology, vol. 77, p. 111, 2016. 19. t. nguyen, a. j. thompson, s. bowden, c. croagh, s. bell, p. v desmond and s. a. locarnini. “hepatitis b surface antigen levels during the natural history of chronic hepatitis b: a perspective on asia”. journal of hepatology, vol. 52, no. 4, pp. 508-513, 2010. 20. s. t. balaky, s. g. hussain, a. a. khaleel, f. t. sabeer and a. h. mawlood. “molecular diagnosis in differentiating active and inactive forms of hepatitis b virus carriers”. journal of university of babylon for pure and applied sciences, vol. 26, no. 10, pp. 41-45, 2018. 21. m. c. kuhns, s. h. kleinman, a. l. mcnamara, b. rawal, s. glynn and m. p. busch. “lack of correlation between hbsag and hbv dna levels in blood donors who test positive for hbsag and anti-hbc: implications for future hbv screening policy”. transfusion, vol. 44, no. 9, pp. 1332-1339, 2004. 22. a. s. k. al-suraifi, a. d. j. al-rubaie, s. m. g. al-mayahie and n. m. m. al-abedy. “unusual hbv mixed genotype infections among hepatitis type b iraqi patients in wasit province/iraq”. international journal of biomedical engineering and clinical science, vol. 2, no. 1, pp. 1-7, 2016. 23. i. a. a. khaled, o. m. mahmoud, a. f. saleh and e. a. baioumi. “prevalence of hbv genotypes in egypt among hepatitis patients”. journal of american science, vol. 6, no. 11, pp. 185-190, 2010. 24. w. k. roth, m. weber, d. petersen, c. drosten, s. buhr, w. sireis and e. seifried. “nat for hbv and anti-hbc testing increase blood safety”. transfusion, vol. 42, no. 7, pp. 869-875, 2002. 25. s. sato, w. ohhashi, h. ihara, s. sakaya, t. kato and h. ikeda. “comparison of the sensitivity of nat using pooled donor samples for hbv and that of a serologic hbsag assay”. transfusion, vol. 41, no. 9, pp. 1107-1113, 2001. 26. h. hennig, i. puchta, j. luhm, p. schlenke, s. goerg and h. kirchner. “frequency and load of hepatitis b virus dna in firsttime blood donors with antibodies to hepatitis b core antigen”. blood, vol. 100, no. 7, pp. 2637-2641, 2002. 27. m. c. kuhns, a. l. mcnamara and r. perrillo. “intermittent pcr positivity for hepatitis b viral dna in surface antigen negative anti-core positive individuals”. transfusion, vol. 39, no. 10, pp. 95s-95s, 1999. 28. y. j. kim, h. c. cho, m. s. choi, j. h. lee, k. c. koh, b. c. yoo and s. w. paik. “the change of the quantitative hbsag level during khaleel, et al.: significance of hbv diagnosis by rt-pcr 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 52-56 the natural course of chronic hepatitis b”. liver international, vol. 31, no. 6, pp. 817-823, 2011. 29. j. jaroszewicz, b. c. serrano, k. wursthorn, k. deterding, j. schlue, r. raupach and m. cornberg. “hepatitis b surface antigen (hbsag) levels in the natural history of hepatitis b virus (hbv)-infection: a european perspective”. journal of hepatology, vol. 52, no. 4, pp. 514-522, 2010. 30. a. alghamdi, n. aref, m. el-hazmi, w. al-hamoudi, k. alswat, a. helmy and a. a. abdo. “correlation between hepatitis b surface antigen titers and hbv dna levels”. saudi journal of gastroenterology: official journal of the saudi gastroenterology association, vol. 19, no. 6, p. 252, 2013. 31. p. p. primadharsini and i. wibawa. “correlation between quantitative hbsag and hbv-dna in chronic hepatitis b infection”. indonesian journal of gastroenterology, hepatology, and digestive endoscopy, vol. 14, no. 1, pp. 9-12, 2013. 32. m. z. hong, w. q. huang, f. min, j. c. xu, z. lin, k. n. fang and j. s. pan. “enhanced hbsag synthesis correlates with increased severity of fibrosis in chronic hepatitis b patients”. plos one, vol. 9, no. 1, p. e87344, 2014. 33. o. günal, s. barut, i. etikan, f. duygu, u. tuncel and m. sünbül. “relation between serum quantitative hbsag, alt and hbv dna levels in hbeag negative chronic hbv infection”. turkish journal of gastroenterology, vol. 25, no. 1, pp. 142-146, 2014. 34. g. li, j. wang, y. bao, l. zheng, k. ge, x. zhou and g. chen. “clinical significance of serum hbsag levels, hbsag/hbv dna ratio, and association with liver inflammation activity in hbeag-positive chronic hepatitis b”. zhonghua gan zang bing za zhi=zhonghua ganzangbing zazhi=chinese journal of hepatology, vol. 23, no. 1, pp. 40-45, 2015. 35. a. ganji, a. esmaeilzadeh, k. ghafarzadegan, h. helalat, h. rafatpanah and a. mokhtarifar. “correlation between hbsag quantitative assay results and hbv dna levels in chronic hbv”. hepatitis monthly, vol. 11, no. 5, p. 342, 2011. 36. r. m. mukherjee, j. arava, p. b. reddy, p. n. rao, r. gupta and d. n. reddy. “role of hepatitis b virus surface antigen quantification in e antigen negative chronic hepatitis b infection”. global journal of science frontier research, vol. 13, no. 1, pp. 1-5, 2013. 37. h. park, j. m. lee, j. h. seo, h. s. kim, s. h. ahn, d. y. kim and j. y. park. “predictive value of hbsag quantification for determining the clinical course of genotype c hbeag-negative carriers”. liver international, vol. 32, no. 5, pp. 796-802, 2012. 38. a. j. thompson, t. nguyen, d. iser, a. ayres, k. jackson, m. littlejohn and g. k. lau. “serum hepatitis b surface antigen and hepatitis b e antigen titers: disease phase influences correlation with viral load and intrahepatic hepatitis b virus markers”. hepatology, vol. 51, no. 6, pp. 1933-1944, 2010. 39. m. martinot-peignoux, m. lapalus, c. laouénan, o. lada, a. c. f. netto-cardoso, n. boyer and p. marcellin. “prediction of disease reactivation in asymptomatic hepatitis b e antigennegative chronic hepatitis b patients using baseline serum measurements of hbsag and hbv-dna”. journal of clinical virology, vol. 58, no. 2, pp. 401-407, 2013. tx_1:abs~at/tx_2:abs~at 45 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 45-51 research article prevalence and characterization of hepatitis b and hepatitis c infection among blood donors in erbil goran n. saleh, chato a. taher* department of basic science, microbiology unit, college of medicine, hawler medical university, erbil, kurdistan region, iraq abstract blood transmitting infectious disease still remains a considerable global health problem. hepatitis b virus (hbv) and hepatitis c virus (hcv) are two of the most commonly transmitted infectious agents. this prospective cross-sectional study was conducted between december, 2017, and february, 2018, at the directorate of blood bank in erbil province, northern iraq. during that period, a total of 6173 blood donors donated blood; all blood donors were asked a series of questions through a structured questionnaire designed for such purpose. these patients were serologically examined for hbv and hcv. positive blood samples were further analyzed serologically and confirmed by real-time polymerase chain reaction (rt-pcr). among 6173 blood donors who were investigated for hbv, 7 (0.11%) and 98 (1.6%) were positive for hepatitis b surface antigen (hbs-ag) and hepatitis b core antibody (hbc-ab), respectively, whereas during screening for hcv, 4 (0.06%) were positive for hcv-ab. coinfection (dual infection (hbv and hcv) was positive in 1 patient (0.01%). among 98 reactive samples, 75.5% were positive for hbs antibody (hbs-ab), the remaining 24 samples (24.5%) were regarded as occult hepatitis b infection (obi), since they were positive for hbc-ab, whereas negative both for hbs-ag and hbs-ab. the diagnosis of obi could be confirmed by rt-pcr in 8 samples, 33% of samples. the overall incidence of hbv and hcv among examined blood donors was 0.5 %, and 0.06%, respectively. amidst that incidence, 0.39 % were diagnosed as obi. to prevent viral transmission through blood transfusion is needed to combine a different and sensitive method for hbv detection as well as evolve tests that have high sensitivity and specificity for serological markers. moreover, a molecular tool that is sensitive enough to detect very low copies of viral dna must also be developed. keywords: blood donors, hepatitis b surface antigen, hepatitis b surface antibody, hepatitis b, hepatitis c introduction infections with hepatitis b virus (hbv) and hepatitis c viruses (hcv) are major global health problems worldwide: it is predictable that about 350 million persons are chronically infected with hbv, and nearly 200 million people are infected with hcv worldwide. both hepatitis b and hepatitis c are among common infectious liver diseases worldwide.[1] viral hepatitis is a systemic disease primarily affecting the liver, which induces acute inflammation of the liver, resulting in a clinical illness characterized by fever, gastrointestinal symptoms such as nausea, vomiting, and jaundice. regardless of the virus type, identical histopathologic lesions are observed in the liver during acute disease.[2] both hbv and hcv infections are known for their parenteral transmission; therefore, blood transfusion is a potential route of transmission.[3,4] the risk of infection with transfusion-transmitted viruses has been reduced remarkably since the introduction of serological screening; on the other hand, a zero-risk blood supply remains the ideal aim.[5] the risk of transfusion-transmitted hbv infection has been reduced by screening all blood donations for hepatitis b surface antigen (hbs-ag) since 1970, screening for hbs-ag is still regarded as an important tool for blood screening by the world health organization (who).[6] it was accepted that the disappearance of hbs-ag indicates the clearance of hbv. meanwhile, many reports pointed to the occurrence of post-transfusion hepatitis b despite the screening for hbs-ag, necessitating the use of other markers for viral screening. for this, screening for hbc-ab was recommended by the german advisory committee blood in march 2005, especially for areas with low hbv prevalence.[7,8] recently in the major blood bank of erbil, hbc-ab screening has also been performed for all blood donors. the donated blood with negative hbs-ag but positive hbc-ab will be corresponding author: chato a. taher, department of basic science, microbiology unit, college of medicine, hawler medical university, erbil, kurdistan region, iraq. e-mail: chato.taher@gmail.com received: apr 16, 2019 accepted: apr 27, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp45-51 copyright © 2020 goran n. saleh, chato a. taher. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ saleh and taher: prevalence of hepatitis b and hepatitis c infection in erbil 46 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 45-51 discarded, these groups of blood donors are either infected but have been cured of the diseases, or else the donor is still infected with hbv that can be further confirmed and need to be further analyzed both serologically and by molecular detection using polymerase chain reaction (pcr).[9] the blood donors with hbs-ag negative serology, hbc-abs positive or negative and the presence of viral dna by pcr are known as occult hbv infected occult hepatitis b infection (obi). obi is a risky group playing an important role in hepatitis virus transmission by blood transfusion.[10] regarding the hbv prevalence in the world could be divided into three areas where the prevalence of chronic hbv infection is high (>8%), intermediate (between 2 and 8%), and low (<2%).[11] infection with hepatitis b and c is regarded as a major cause of liver cirrhosis and hepatocellular carcinoma. the who estimates that 57% of cases of liver cirrhosis and 78% of primary liver carcinomas result from a hepatitis b or c virus.[12] approximately 70%–80% of hcv-infected patients fail to clear the virus and develop chronic hcv infection, which is a risk factor for liver diseases such as fibrosis, cirrhosis, and hepatocellular carcinoma.[13,14] several studies have recorded the prevalence of hbv and hcv infection in blood donors of different iraqi cities.[15-17] according to the studies that have been conducted in the past years investigating the seroprevalence of hepatitis b and hepatitis c, the prevalence rate in our region is low which is <2%.[18,19] lack of documentary data in erbil governments and immigration of a significant number of refugees from a neighboring country to kurdistan region, as well as foreign tourists and the traveling of kurdish peoples to other countries, necessitate updated research on seroprevalence of infectious agents (hbv and hcv) among blood donors. nonetheless, the hbv vaccine was added to the expanded program of vaccination in early 2000.[20] there was an absence of data on the success of the vaccination program; therefore, we thought of evaluating the antibody for these viruses in the blood which offer an idea of the level of protection against these infectious agents. this study was conducted to find out the incidence of hbv and hcv among blood donors in erbil community. besides this, we further characterized the hbv infected individuals along with the screening for hbs-ab, evaluating the level of protection against hbv infection among our community. materials and methods study population and sampling this is a prospective and cross-sectional study investigating hbv and hcv among blood donors who donated blood in the directorate of blood bank-erbil, iraq. all volunteers who visited the blood bank to donate blood from december 2017 to february 2018 were included in the study: blood donors were selected after an interview through a structured questionnaire designed for such purpose. blood donors with low or high hemoglobin concentration, high body temperature, having any disease, weight <50 kg, cupping in past 6 months, operation in the last year, drinking alcohol, uncontrolled blood pressure, history of taking any drugs, fever, and having tattoos on their body were excluded in our study. in total, 6173 participants donated blood during the specific time and were screened for hbv and hcv. among those blood donors, 109 were reactive for hbv or hcv. fifty healthy individuals were also included in this study as a control group. each blood sample was collected in a 5 ml of a sterile tube, and then the whole blood was centrifuged, plasma was taken and stored at minus 20 freezer. the study was approved by the ethics committee at the hawler medical university, for dealing with the samples and participant information and a written informed consent was obtained from each blood donor for their participation in the study. detection of hbv serological markers elisa screening test for hbs ag and anti-hbc monolisa™ hbs ag ultra (bio-rad, france) assay is a one-step enzyme immunoassay based on the principle of the “sandwich” type using monoclonal antibodies and polyclonal antibodies selected for their capability to bind themselves to the numerous subtypes of hbs ag.[21] serologically, the samples were rechecked by elecsys hbs ag ii by device cobas e411(roche, germany). the elecsys hbsag ii assay uses monoclonal and polyclonal anti-hbs antibodies (mouse and sheep) for the hbsag determination. monolisa™ anti-hbc plus (bio-rad, france) is an enzyme immunoassay (indirect elisa type) for the simultaneous finding of total antibodies to hbv core.[22] chemiluminescent immunoassay (clia) screening test for anti-hbs clia way was used for detecting antibody of hbs by the liaison xl (diasorin, italy). amid numerous enzyme assays that give light-emitting reactions, one of the most successful tests is the improved clia regarding a horseradish peroxidase labeled antibody or antigen and a mixture of the chemiluminescent substrate, hydrogen peroxide, and enhancers.[23] detection of hcv serological markers monolisa™ anti-hcv plus version 2 or 3 (bio-rad, france) is an indirect qualitative enzyme immunoassay for the detection of infection by the hcv based on the detection of anti-hcv antibodies in serum or human plasma.[24] detection of hbv dna (pcr) the extraction of hbv dna is done by a kit ez1 virus mini kit (v2.0 qiagen, germany) using the ez1 advanced xl device (qiagen, germany).[25] the artus hbv rg pcr kit is used in the automated system for the detection of hbv dna using the pcr on rotor-gene q (qiagen, germany).[26] this is conducted for 24 samples that are negatives for anti-hbs and positives for antihbc igg, and six samples are used as a negative control. statistical analysis microsoft office excel and statistical package for the social sciences (spss) are used for data entry and analysis. results among 6173 participants, 109 were reactive for hbv and hcv. fifty negative samples were also included in this study as a control group for further analysis. among the 159 participants saleh and taher: prevalence of hepatitis b and hepatitis c infection in erbil 47 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 45-51 who were included in this study as a test and control group, the mean and standard deviation in age for the control group was 35.5 ± 8.6 and that for the test group was 40.9 ± 12. the ages of blood donors both for test and control group were grouped into six groups [tables 1 and 2]. the minimum age is 19 years, and the maximum age is 73 years for the test group and 22 and 57 years for the control group. prevalence of hepatitis b and hepatitis c serologically, based on detection of hbs-ag, hbc-ab, and hcv-ab, we decided whether the patient has hbv or hcv infection. these tests were carried out for all the participants (6173). among all examined participants, only 7 patients were reactive for hbs-ag (0.11%), whereas 98 donors were reactive for hbc-ab (1.58%). regarding hcv infection only 4 patients were positive for antihcv ab (0.065%), as described in figures 1 and 2. immune blood donors for hbv infection among the sero-reactive blood donors for hbv, we further characterize them through detection of antibody against hbs-ag (hbs-ab). should individuals of hbc-ab positive blood have hbs-ab in their blood, this indicates that these participants are cured of hbv infection and means that they have immunity to hbv infection. among the 98 sero-reactive blood donors, 74 blood donors were positive for hbs-ab (75.5 %) indicating that they are cured of natural infection. we also examined 50 healthy volunteers that are free from hbv and hcv infection, among them only 4 blood donors were positive for hbs-ab, reflecting the successful vaccination and protection against hbv infection [table 3]. diagnosing occult hbv infection as we defined obi before, blood donors with hbs-ag negative serology, hbc-abs positive or negative, and the presence of viral dna by pcr are known as occult hbv infection (obi). blood donors with only positive hbc-ab in the absence of hbs-ag and hbs-ab are regarded as obi.[10] in our study, we further characterize the positive blood donors. among the samples that are included in our study, 98 samples were positive for hbc-ab, from which 24 blood donors (24.5%) were negative for hbs-ag and hbs-ab (hbc-ab +ve, hbs-ag –ve, and hbs-ab –ve), indicating that these blood donors were defined as having occult hepatitis virus infection [table 3]. these 24 samples were further investigated for the detection of hbv dna [figure 3]. hcv incidence and dual infection in this study, as we mentioned above, besides screening for hbv infection, screening for hcv was also done for these participants. among 6173 participants, only four blood donors were positive for hcv-ab 0.06%. among hcv reactive donors, one individual was positive both for hbv infection and hcv infection, which means that he has a dual infection [table 4]. sero-status in relation to socio-demographic characteristics among the hbv and hcv positive blood donors, the highest frequency of 38 (35%) was found among those aged between 26 and 35 years (young age groups). blood donors with blood group o were found to have the highest frequency for table 1: demographic distribution of the hbv and/or bcv positive samples (positive blood donors) demographic distribution frequency (n=%) test group age groups (year) under 25 4 (3.7) 2–35 38 (35) 36–45 33 (30) 46–55 22 (20) 56–65 7 (6.4) above 66 5 (4.6) blood groups o 39 (36) a 36 (33) b 25 (23) ab 9 (8) occupations private sector employee 73 (67) public sector employee 30 (27.5) unemployed 6 (5.5) student total 109 (100) table 2: demographic distribution of the samples control group (negative blood donors) demographic distribution frequency (n=%) control group age groups (year) under 25 6 (12) 26–35 25 (50) 36–45 10 (20) 46–55 8 (16) 56–65 1 (2) above 66 blood groups o 17 (34) a 18 (36) b 10 (20) ab 5 (10) occupations private sector employee 34 (68) public sector employee 13 (26) unemployed 1 (2) student 2 (4) total 50 (100) saleh and taher: prevalence of hepatitis b and hepatitis c infection in erbil 48 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 45-51 its immunoreactivity among test group 39 (36%) [table 1], whereas those with blood group a are among those with the highest frequency among control group 18 (36%) [table 2]. most of the reactive individuals for hbv positivity were among the private sector employees [table 1]. none of these parameters were statistically significant. molecular detection of the hbv by realtime quantitative pcr (rt qpcr) for further characterization of serum samples from hbs-ag negative, hbs-ab negative, and hbc-ab positive blood donors were processed for hbv-dna detection using molecular amplification using the rt pcr. this test was done using artus® hbv rg* pcr kit 24, v1 (qiagen, germany). after extracting the nucleic acid of the hbv by ez1® virus mini kit v2.0 (qiagen, germany), we ran the purified dna of hbv by rotor-gene q (qiagen, germany). twenty-four samples were selected (antihbc igg positive and negative for anti-hbs), and six control samples (negative control) were also included in the study. out of these samples, 8 of the samples (33%) were positive, showing the dna in their blood, confirming the diagnosis of occult hepatitis b [figure 3]. discussion hepatitis b and c viruses are responsible for causing about 80% of liver cancer cases worldwide, and it could be the major cause of morbidity and mortality.[27] regarding hepatitis prevalence, the world is divided into three areas; low prevalence area, which means <2%, an intermediate area 2–8% and high prevalence area more than 8%.[28,29] most countries on the earth are still viewed as intermediate to high endemicity for hbv infection.[28,29] hcv follows the same division regarding prevalence such as low prevalence (<1.5%), moderate prevalence (1.5%–3.5%), and high prevalence area, which is more than 3.5%.[30] the current study characterizes and defines the frequency of hbv and hcv (6173) blood donors. for hbv detection, two serological markers were used (hbs-ag and hbc-ab), among those, 98 donors were reactive for hbc-ab (1.58%), whereas only 7 people were positive for hbs-ag (0.11%). for screening of hcv, hcv-ab was used and only 4 blood donors proved positive for anti-hcv-ab (0.065%), as described in figures 1 and 2. the prevalence of reactive blood donors in the current study for hbv is 1.7% (105 out of 6173) from which 1.5% is reactive against hbc-ab and 0.11% is reactive against hbs-ag, whereas for hcv is 0.06%. they are in agreement on a national level with these results by alhilfi et al., who screened 36620 blood donors in missan governorate in iraq, figure 1: serological and molecular characterization of hepatitis b virus and hepatitis c virus infection among blood donors in the major blood bank of erbil city. hbs ab – hepatitis b surface antibodies; hbc ab – hepatitis b core antibodies: pcr – a polymerase chain reaction figure 2: incidence of both hepatitis b virus and hepatitis c virus among blood donors saleh and taher: prevalence of hepatitis b and hepatitis c infection in erbil 49 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 45-51 they found 0.12% positivity for hbs-ag and 0.09% positivity for hcv-ab.[31] similar results were also found by al-rubaye et al., who screened 69915 blood donors in basra in 2013, from which they found 125 reactive individuals for hbs-ag (0.2%), and 1475 reactive individuals for hbc-ab (2.1%) and hcv-ab positivity in 0.1%.[17] on an international level, similar results to ours were also found in a neighboring country by boustani et al., which was done in ilam city in iran between 2009 and 2013: they screened 72,527 blood donors, they found hbs-ag in 102 (0.14%), and hcv-ab in 27 (0.037%).[32] moreover, another study conducted in southwestern iran, by sajjadi et al., they screened 180,304 blood donors and found hbv in 0.13% and hcv in 0.06%.[33] on the other hand, our data were much lower than that of other related studies performed in other centers. nationally, hussein et al. performed a study in duhok at 2004 found hbs-ag in 62/7900 (0.78%), and hcv-ab in 16/7900 (0.2%).[15] in addition, internationally, our result was lower than those found in india in a study conducted by gulia, they found hbv in 2.54%.[34] in libya, a study carried out by qowaider et al., they examined 78,987 blood donors in different regions in the northeast in libya, and they found hbv in 172 blood donors (0.21%), and hcv in 197 blood donors (0.24%).[35] furthermore, in turkey, hope et al., examined 53,985 blood donors, among them 2.1% were positive for hbv and 0.3% for hcv.[36] besides the blood screening, we further characterized the reactive blood donors (positive samples) among 105 positive samples for hbc ab (1.7%), 98 samples were hbc-ab positive but negative for hbs-ag (hbs-ag −ve/hbc-ab +ve), further characterization was done for these group by investigating for hbs-ab, among these 74 (75.5%) blood donors were hbs-ab positive (hbc-ab +ve, hbs-ag –ve, and hbs ab +ve) which indicated that they are cured of natural infection [figure 1 and table 3] whereas the remaining 24 blood donors (24.5 %) were hbc-ab positive but negative both for hbs-ag and hbs-ab. individuals with negative hbs-ag and hbs-ab but positive hbc-ab are defined as occult hbv infection (obi), this is because hbc-ab is regarded as a pathognomonic marker of hbv infection, particularly when hbs-ag and hbs-ab are negative. we performed hbv-dna detection assay solely on samples of blood donors who tested positive for hbc-ab but negative for both hbs-ag and hbs-ab (24 samples), including six control samples as well. among 24 samples we could confirm the presence of hcv dna only in 8 (33%) samples, using rt pcr, indicating the overall incidence of obi by dna detection to be about (0.13 %; 8 out of 6173) [figures 1 and 3]. based on the reaction graph, most of the positive sample has very low copy numbers since the reaction was arrived at between the 35 and 40 cycles [figure 3]. meanwhile, the remaining 16 (66 %) samples were hbc-ab positive but failed to be confirmed by hbv-dna table 3: distribution of hbs-ab among hbc-ab positive individuals and control group serology (hbc igg) serology (hbs antibody) p-value positive (n=) negative (n=) total (n=) positive 74 24 98 <0.05 negative 6 55 61 total 80 79 159 hbs ab: hepatitis b surface antibodies figure 3: quantitation data for cycling a. green for hepatitis b virus by real-time quantitative polymerase chain reaction table 4: the incidence and percentage of hcv reactive donors and dual infection serology (hbs antigen) serology (hcv-ab) positive negative total n= % n=% n=% positive 1 (0.6) 7 (4.4) 8 (5.0) negative 3 (1.9) 148 (93.1) 151 (95.0) total 4 (2.5) 155 (97.5) 159 (100.0) hcv-ab: hepatitis b surface antibodies saleh and taher: prevalence of hepatitis b and hepatitis c infection in erbil 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 45-51 detection. in most of the areas, these groups of blood donors are still regarded as obi, and their blood will be discarded to prevent virus transmission.[37] internationally, many studies have been published throughout the world showing the frequency of obi within hbs-ag-negative donors, the results are varied: for example in the united kingdom it was 0.56%;[38] in the united state of america it was 0.8%;[39] in iran it was 2.1%;[40] and in brazil it was 3.1%.[41] on a national level, a similar study performed in erbil city of iraq shows obi in 0.09% which is close to our finding (0.13%).[42] discarding the infected blood on the basis of hbc-ab detection, especially in areas where the prevalence of hbc-ab is high (more than 10%), will negatively affect the blood supply. in the present study, we found that the incidence of hbc-ab in the erbil city was low (1.56%, 98 out of 6173) [figure 1]. further detection of hbs-ab saved 78 of our samples (75.5%) since they were positive for hbs-ab. on the other hand, dna confirms the obi only in 8 samples (33%). in high prevalence area based on hbc-ab detection such as korea, greece, and ghana, they discarded only blood that is positive for hbvdna, consequently saving more blood units.[43,44] meanwhile, in many places, especially in low prevalence countries, blood donor samples were still considered as obi based on hbc-ab positivity, and all such donated blood was discarded to prevent transfusion-transmitted viral infection. despite the above, the failure of detecting hbv-dna might be due to a mutation in viral structure, and the false serological result might be due to the imperfection of the assays used for such purpose.[37] many researchers argued in favor of the implementation of hbc-ab or hbv dna alone or combination for the screening of donated blood.[45,46] in a large study screening, 6.5 million participants using three markers (hbs-ag, hbc-ab, and pcr for hbv dna), prioritizing the use of hbc-ab along with the dna nucleic acid testing was confirmed for the detection of a low level of hbv dna positive donors.[46] conclusion the overall incidences of hbv and hcv among examined blood donors were 0.5% and 0.06%, respectively. among that, 0.39% were diagnosed as obi. the highest avoidance of hbv blood transmission can be attained by combining a different, more sensitive, method for hbv detection with the evolving of tests that have high sensitivity and specificity for hbs ag and hbc ab detection. further, developing a molecular tool that is sensitive enough to detect a very low copy of viral dna would conserve many pints of blood. acknowledgments i would like to express my very great appreciation to the directorate of blood bank and those working in the central laboratory, as well as to all those who have helped to make possible my undertaking of this research. conflicts of interest there are no conflicts of interests to be declared. references 1. b. liu, j. li, y. han, y. liu, l. kong, y. cao and z. j. h. huang. “dynamic analysis of lymphocyte subsets of peripheral blood in patients with acute self-limited hepatitis b”. health, vol. 2, pp. 736-741, 2010. 2. j. a. hobden and k. c. carroll. “jawetz, melnick and 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“transfusion transmissible infections: maximizing donor surveillance”. cureus, vol. 10, pp. e3787-e3787, 2018. 46. s. l. stramer, s. zou, e. p. notari, g. a. foster, d. e. krysztof, f. musavi and r. y. dodd. “blood donation screening for hepatitis b virus markers in the era of nucleic acid testing: are all tests of value”? transfusion, vol. 52, pp. 440-446, 2012. tx_1~abs:at/add:tx_2~abs:at 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 review article analysis the structures of some symmetric cipher algorithms suitable for the security of internet of things devices khalid f. jasim*, reem j. ismail, abdullah a. nahi al-rabeeah, soma solaimanzadeh department of computer science, cihan university-erbil, kurdistan region, iraq abstract in the past years, the internet of things (iot) used in different applications and very wide range of fields (e.g.  cloud services, smart environments, logistics, social and personal domains, and health-care services). the iot included a variety of components and devices such as radio frequency identification devices, wireless sensors, actuators, and wireless networks. furthermore, the iot with smart devices adopted in various companies, organizations, and public services systems. for instance, some devices such as notebooks and smartphones have been used to perform different management activities and duties. these smart devices relied on data exchange and data storage resources in clouds computing services. in this context, the saved data and exchanged data required protection against hacking operations, transferred with more secure communications channels, and safe storage environment in the clouds and local storage systems. in this paper, we proposed some encryption algorithms to tackle the issue of data confidentiality in the iot applications. this research provided analysis and investigation of these encryption algorithms in light of components of the designs, versions of these algorithms, encryption keys, block size, round functions, and the techniques used in the designs. keywords: internet of things applications, block ciphers, stream ciphers, encryption algorithms, data security introduction in the previous years, the extensive growth of internet of things (iot) applications reflected, positively, on the different activities of user’s life. the basic concepts of iot were presented in 1999, in which there was massive progress in some technologies such as radio frequency identification (rfid devices), wireless sensor networks, and wireless devices. furthermore, the iot was intended to establish connections based on various objects (e.g. rfid devices, sensors, actuators, and readers).[1] the iot presented important applications in different fields (e.g. smart environments, logistics, social and personal domains, and health-care services).[2] the structure of iot based on cloud services was presented in zhou et al.,[3] and the requirements of user’s privacy and data confidentiality have been covered. in this context, the security requirements focused on location privacy, identity privacy, cloud security, layer attack, and node attack. the iot medical applications were used in the design of smart medication dispenser devices, in which can help the patients to receive their medications as specified in the treatment schedule. these smart dispenser devices designed based on iot applications, smartphone devices, and cloud computing services for data storage.[4] furthermore, some iot used in the design of e-health database systems. in these e-systems, the patient’s status can be checked and monitored more efficiently. the researchers in yang et al.[5] presented analysis study of privacy issues in these systems. then, the researchers proposed a system relied on big data and iot applications, in which this system offered data confidentiality of patients’ records and protected communications. the system relied on authentication key technique and algorithm for secure data transfer through the medical network. on the other hand, in iot technologies, huge number of smart devices are accessed various services through the internet and wireless networks and different types of sensitive information are exchanged through these networks. thus, some security issues appeared related to data confidentiality and privacy of users and smart devices.[6-8] to overcome and mitigate the security problems, some symmetric ciphers have been proposed that provide data confidentiality and privacy of users and smart devices.[9] corresponding author: khalid f. jasim, department of computer science, cihan universityerbil, kurdistan region, iraq. e-mail: khalid.jassim@cihanuniversity.edu.iq received: june 13, 2021 accepted: july 18, 2021 published: september 01, 2021 doi: 10.24086/cuesj.v5n2y2021. pp 13-19 copyright © 2021 khalid f. jasim, reem j. ismail, abdullah a. nahi al-rabeeah, soma solaimanzadeh. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ jasim, et al.: analysis cipher algorithms for the security of internet of things devices 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 in recent years, a variety of devices (e.g., sensors, smart devices, and rfid systems) have been adopted by organizations, companies, and public services. these smart devices are interconnected through iot systems. there was data transfers and data storage through these systems and networks; therefore, there was a demand to protect these data. the cryptographic algorithms (e.g., lightweight cryptographic [lwc] algorithms, lwc) can provide data security in iot systems and applications. the researchers in alahdal and deshmukh[10] conducted a survey study on lwc encryption algorithms that can support data confidentiality in iot applications. this study focused on some lwc encryptions algorithms (e.g. hash functions and block ciphers algorithms), and analyzed the features of these algorithms (e.g., energy consumption of devices, required area in designs, throughput of algorithms, and latency). furthermore, the researchers in thakor et al.[11] proposed the lwc encryption algorithms to protect and secure the communications between the smart devices which are used in iot applications. the researchers concentrated on comparing the lwc encryption algorithms in light of cost of implementations, performance of these algorithms in software and hardware implementations, and various analysis attacks as well. moreover, many users rely on some devices such as notebooks and smartphones to perform their duties, and these devices are connected via cloud computing services, in which there are data transfers among these devices, clouds, and iot applications. in such case, the exchanged data require protection and must be transferred in secure and safe communications. the lwc block ciphers algorithms have been proposed to solve the problems of data security in iot networks. for instance, some block ciphers algorithms (e.g., advanced encryption standard [aes], present, lblock, and skipjack) have been proposed as security encryption algorithms, and the features of these algorithms were investigated such as performance of algorithms, consumption of energy, time of execution during encryption/decryption operations, required memory space of these algorithms, and throughputs of these algorithms.[12] furthermore, a variety of stream ciphers and block ciphers encryption algorithms were proposed to provide data protection, secure data transmissions, and data integrity in wireless mobile communications and smartphones device as well (e.g., aes, a5, snow, and zuc).[13,14] the sections of our research paper will be presented as follows. the iot backgrounds, concepts, devices, services, algorithms, and features are presented in section 1. section 2 focused on analyzing aes cipher algorithm (e.g., secret keys, versions of aes, rounds, and math transformations). section 3 investigated some components of rc6 cipher algorithm (e.g., encryption keys, block size, the 4 registers, array s[], and encryption steps). section 4 described various elements of twofish cipher algorithm (e.g., encryption keys, function f, s-boxes, rounds, mds matrix, and encryption process). section 5 analyzed the components of speck cipher algorithm (e.g. block size, round function, logic operations [arx], parameters, and versions of speck cipher). section 6 investigated the components of lightweight encryption algorithm (lea) encryption algorithm (e.g., versions of lea, block size, encryption keys, round function with logic and rotation operations [ror and rol], and encryption process). section 7 concentrated on various components of chacha20 encryption algorithm (e.g., secret keys, state matrix, quarter round function [qrf], and process of key stream). section 8 provides analysis and discussion of iot applications and the proposed encryption algorithms that can offer data security in iot systems and applications. finally, the conclusions of our work are presented in section 9. analysis of aes cipher algorithm daemen and rijmen proposed the design of aes as encryption cipher algorithm for various types of data security applications. the design of aes relies on substitution-permutation network, linear layers, nonlinear layers, and add round key operation. this cipher system depends on three secret keys (128 bits, 192 bits, and 256 bits). furthermore, the design of aes includes three types (e.g., aes-128 with 10 rounds, aes-192 with 12 rounds, and aes-256 with 14 rounds).[15] the aes used to encrypt input block of size (128 bits), the input block arranged as 16 bytes (b0, b1, b2,…, b15), and these 16 bytes saved in state array (4 rows × 4 columns). the design of aes relies on round function; the round function includes four operations. addroundkey operation, which is responsible for adding the bytes of round key to the bytes of state array. subbytes operation, which is used to apply substitution box (s-box) on each bytes in state array. shiftrows, which shifts the bytes of row(i) in state array according to defined numbers. mixcolumns operation, in this operation each column of state array is multiplied by defined and constant matrix (mds matrix).[16] analysis of rc6 cipher algorithm the design of rc6 cipher algorithm identified by version (rc6w/r/b), in which w represented the number of bits in word w (w=32 bits), r represented the number of rounds in this cipher, and b defines the number of bytes adopted for encryption secret key. the input block size in rc6 cipher determined by 128 bits. this algorithm relied on different lengths of secret encryption key (e.g., key size equal to 128 bits, 192 bits, and 256 bits).[17,18] this algorithm is block cipher, and relies on feistel cipher structure, in which the encryption depends on input of plaintext (with 2w bits and k key), process the input by sequence of substitution and permutation operations through r rounds, then produce the block of ciphertext. rc6 includes four registers (a, b, c, and d, length of each register equal to 32 bits), these registers used to save input of plaintext/cipher text blocks during encryption and decryption processes.[19] furthermore, the array s[ ] (s[0], s[1], s[2],…, s[2r + 3], r=number of rounds) is used to save the rounds keys and these rounds keys are adopted in encryption/decryption operations. the (32 bits) words of array s[ ] are extracted from secret key bytes (128 bits, 192 bits, and 256 bits) according to key expansion algorithm.[20] in rc6, encryption process is shown in figure 1:[20] analysis of twofish cipher algorithm the twofish algorithm classified as symmetric crypto algorithm. the design adopted block ciphers technique, the sender and receiver must possess the same software of this algorithm, and jasim, et al.: analysis cipher algorithms for the security of internet of things devices 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 figure 1: encryption steps in rc6 cipher algorithm same secret encryption keys to exchange cipher text messages. the twofish cipher can operates with three types of secret encryption keys (e.g., 128 bits, 192 bits, and 256 bits), and one type of input block size which is equal to (128 bits). the inner structure of this algorithm includes 16 rounds of feistel cipher networks and function f. the function f consists of (4) s-boxes (each with 8x8 bits, and key dependent mode) and maximum distance matrix (mds with 4x4 constant values). furthermore, this function adopts bitwise rotation operations and math transformation with (pseudo-hadamard).[21,22] in the encryption process [figure 2], the input plaintext (128 bits) is partitioned into 4 words (each word with 32 bits). these 4 words are xored with 4 key words (k#0, k#1, k#2, and k#3). the 16 rounds of this algorithm are performed. in each round of the 16 rounds, the 2 words in the left side are employed as input to function (g), in which function (g) relies on 4 s-boxes (s-box#0, s-box#1, s-box#2, and s-box#3) and mds matrix. each of the 4 s-boxes process input of (8 bits) and produce encoded output of (8 bits). the 4 produced outputs are multiplied by the mds matrix. as shown in figure 2, the outputs of mds matrix are processed to be combined by using math transformation with (pseudo-hadamard). the results are added with values of 2 key words (k#2r+8 and k#2r+9, where r = round number). then, these results processed by 2 xor operations. the aforementioned processing steps are performed in each of 16 rounds and then produce the 128 bits output of cipher text.[23] analysis of speck cipher algorithm the speck cipher algorithm designed based on block ciphers techniques. this cipher algorithm constructed for light figure 3: structure of round function in speck cipher weight smart devices with small hardware requirements. the components of this light block cipher relied on basic logic operations (i.e., addition, rotation, and xor logic operations, or arx operations). in this context, these logic operations can be constructed efficiently in light weight and small hardware devices (e.g., fpga, means the field programmable gates arrays). the structure of speck cipher organized based on feistel cipher structure, employed different types of input/ output block sizes (e.g., 32 bits, 48 bits, 64 bits, 96 bits, and 128 bits), and possessed high performance when implemented using software on (64 bits) processors. the round function in speck cipher [figure 3] adopted three basic logic operations (arx: addition, rotation, and xor logic operations).[24] the round function includes left half of the input (li, i=round no. i) and right half of the input (ri, i=round no. i), ki which represents the (n bits) of round key specified for round (i), rotations operations defined by (>>> α, shift to right by α bits) and (<<< β, shift to left by β bits), modulo addition operation (), and the xor operation (). the outputs figure 2: structure of twofish cipher algorithm jasim, et al.: analysis cipher algorithms for the security of internet of things devices 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 figure 4: structure of round function in lightweight encryption algorithm cipher table 1: versions of speck cipher with different parameters block size of (input/output) in bits size of secret key in bits value of α value of β total no. of rounds 32 64 7 2 22 48 72 8 3 22 48 96 8 3 23 64 96 8 3 26 64 128 8 3 27 96 96 8 3 28 96 144 8 3 29 128 128 8 3 32 128 192 8 3 33 128 256 8 3 34 of this round function identified by the left half output (li+1) and right half output (ri+1), in which can be computed as shown in the following formulas: [25] li+1 = ((li >>> α) r)) ki, and ri+1 = (ri <<< β) li+1 (1) the speck cipher can operates with different versions, therefore the parameters (α and β) can be defined with different values (e.g. α = 7 or α = 8, and β = 2 or β = 3). various versions of speck cipher are depicted in table 1. for instance, speck 32/64 represents the version that includes block size (32 bits), secret key (64 bits), parameters (α=7 and β=2), and number of rounds (=22) speck 128/256, this version adopts block size (128 bits), secret key (256 bits), parameters (α=8 and β=3), and number of rounds (=34) analysis of lea cipher algorithm the lea encryption algorithm designed based on the concepts of block ciphers, offered high speed of data encryption, and implemented in software applications. this algorithm possessed suitable security level, in which can withstand against various types of cryptanalysis attacks (e.g. linear analysis attack, and differential analysis attack), and also has been proved as secure and efficient encryption algorithm.[26] the lea algorithm includes three versions (lea-128, lea-192, and lea-256). the first version (lea-128) relies on block size (128 bits), key size (128 bits), round number (24), and word size (32 bits). the second version (lea192) employs block size (128 bits), key size (192 bits), round number (28), and word size (32 bits). the third version (lea-256) adopts block size (128 bits), key size (256 bits), round number (32), and word size (32 bits). the design of lea constructed with round functions. the round function [figure 4] consists of (6) xors (xor logic operation), (3) additions (modular addition operation, with mod value 232), and (3) rotations (rotations with circular type operation, and identified by ror3 rotation, ror5 rotation, and rol9 rotation operations).[27] the lea encryption algorithm[28] [figure 5] was proposed to perform data confidentiality in different iot applications figure 5: lightweight encryption algorithm with encryption process (e.g. mobile based environments, cloud computing services, and embedded devices). the three versions of lea encryption algorithm (lea-128, lea-192, and lea-256) supported data confidentiality through data encryptions. for instance, encryption process starts with extracting 128 bits of plaintext, represents these 128 bits by 4 words (xi[0], xi[1], xi[2], xi[3], and each word=32 bits). in round number (i), the word xi[0] transferred into word xi+1[3] in next round (i+1). the word xi[1] xored with round key rki[1], perform modular addition on this word, apply rotation operation (rol9), then the result word transferred into word xi+1[0] in next round (i+1). the word xi[2] xored with round key rki[3], apply modular addition on this word, apply rotation operation (ror5), then the result word transferred into word xi+1[1] in next round (i+1). the word xi[3] xored with round key rki[5], perform modular addition on this word, apply rotation operation (ror3), then the result word transferred into word xi+1[2] in next round (i+1). for example, in first version (lea-128) the number of rounds equal (24), therefore the aforementioned encryption process is repeated on the jasim, et al.: analysis cipher algorithms for the security of internet of things devices 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 plaintext words (x[0], x[1], x[2], and x[3]), then after round no. 24 the 128 bits of encrypted data are produced. analysis of chacha20 cipher algorithm the stream ciphers techniques relied on composing elements of key stream based on predefined and dynamic state with initial values. in the encryption process, the elements of key stream mixed with the elements of plaintext then the output of this process represent the elements of cipher text (e.g. encrypted data). some encryption algorithms, in which relied on stream ciphers techniques, have been proposed to perform the data confidentiality in iot applications.[29] in this context, the chacha20 encryption algorithm designed based on stream ciphers techniques, and supported data confidentiality in various data security applications (e.g. iot devices, google chrome in smart devices, security of protocols in openssl, and openssh).[30] the design of this algorithm depends on a number of arx logical operations (i.e. addition operation, rotation operation, and xor logical operation). in this design, the arx logical operations constructed on (32 bits) words and it was found as efficient encryption algorithm in hardware and software implementations. for instance, each round in this algorithm constructed with 16 (xor logic operations), 16 (modulo additions operations with mod 232), and 16 (rotations operations).[31] the chacha20 cipher algorithm depends on secret key k with size (256 bits), k defined as (k=k0, k1, k2,…, k7), and it is organized to process words of (32 bits) in each time. the algorithm includes a state which is composed as matrix (4x4 matrix); this state matrix consists of 16 values and each value represented by (32 bits word). the 16 elements of state matrix defined as 4 rows (x0,x1,x2,x3; x4,x5,x6,x7; x8,x9,x10,x11; x12,x13,x14,x15). in the encryption process, the state matrix is loaded with predefined fixed values (e.g. x0=0x61707865, x1=0x3320646e, x2=0x79622d32, and x3=0x6b206574), the other elements are loaded by elements from secret key k (x4=k0, x5=k1, x6=k2, x7=k3 …, x11=k7), 32-bit counter (x12=c0), and 3 (32-bit) nonce-values (x13=n0, x14=n1, x15=n2).[32] the qrf, used in chacha20 cipher algorithm, process input of 4 words (a, b, c, d, each with 32 bits), perform sequence of operations then the result includes updated 4 words (a, b, c, d, each with 32 bits) as shown in the following algorithm 1 qrf [figure 6]. the process of key stream is shown in algorithm 2 [figure 7]. the process starts with initialize the state matrix x with values from secret key k, counter value c0, nonce-values (n0, n1, n2), and constant values as shown in state matrix [figure 8]. the 16 elements of state matrix x are processed for 20 rounds by using qrf function, and then produce the z key stream. the z key stream is used in encryption and decryption operations of input data.[33] discussion iot the iot applications designed to create connections based on different objects such as rfid devices, sensors, actuators, and figure 8: elements of (4x4) state matrix figure 6: quarter round function in chacha20 figure 7: process of key stream in chacha20 readers. these applications covered vital areas such as smart environments, logistics, social and personal domains, healthcare services, and relied on cloud services. the iot systems with smart devices have been adopted in many organizations and public services. in these systems, the exchanged data required data protection and secure data transmissions through communication systems. therefore, various encryption algorithms have been proposed that supported data confidentiality in iot systems and applications. jasim, et al.: analysis cipher algorithms for the security of internet of things devices 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 aes algorithm this encryption algorithm supported data security in different iot, internet, and cloud services applications. this algorithm adopted block ciphers concepts, three types of secret keys (e.g., 128 bits, 192 bits, and 256 bits), block size (128 bits), three types of rounds (10, 12, and 14), and relied on substitutions and permutations networks (spn technique) [table 2]. each round in the structure of this algorithm included four types of operations (i.e. add-roundkey, sub-bytes, shift-rows, and mix-columns). these operations enhanced the security of the aes algorithm. rc6 this algorithm relied on block ciphers, three types of secret keys (128 bits, 192 bits, and 256 bits), block size (128 bits), 20 rounds, and the design constructed with feistel cipher structure [table 2]. the algorithm employed 4 registers (a, b, c, and d, and each register loaded with 32 bits), state array s[ ] used for saving the values of round keys. twofish the algorithm designed with block ciphers techniques, 3 secret keys (128 bits, 192 bits, and 256 bits), and block size equal to (128 bits). the structure of this algorithm involved 16 rounds, each round contains function f. function f constructed with 4 s-boxes (s-box#0, s-box#1, s-box#2, and s-box#3), and mds matrix with fixed values. speck this algorithm classified as light block cipher and suitable for light weight smart devices. the algorithm used three types of logic operations (addition, rotation, and xor logic operations [arx])). this algorithm designed with feistel cipher structure, and included various types of block size (32 bits 128 bits), different secret keys (64 bits – 256 bits), and different number of rounds (22–34). each round employed 2 rotation operation, 2 xor operation, and 1 addition operation. lea this algorithm designed with three versions (lea-128, lea192, and lea-256). the lea-128 algorithm used secret key (128 bits), block size (128 bits), and 24 rounds. the lea-192 version adopted secret key (192 bits), block size (128 bits), and 28 rounds. the lea-256 version included secret key (256 bits), block size (128 bits), and 32 rounds. the round function in this algorithm designed with (6) xors, (3) additions, and (3) rotations (ror3 rotation, ror5 rotation, and rol9 rotation operations). chacha20 this algorithm designed based on stream ciphers techniques. the algorithm included secret key (256 bits), block size (512 bits), and 20 rounds. each round employed (16) xor, (16) additions mod (232), and (16) rotations operations (arx operations). furthermore, the algorithm used state matrix x[4, 4] with 16 elements processed in qrf function during data encryption/decryption operations. conclusion in previous years, the iot applications deployed in various fields (e.g., cloud services, healthcare systems, environment of smart cities, smart logistic applications, transportations systems, and agriculture area with smart tools). the iot devices used in these fields experienced data security issues. some encryption algorithms proposed to overcome the problems of data confidentiality during the data transmissions and data storage environments such as cloud computing services. this research focused on analyzing the proposed encryption algorithms that can be used for data protections. in this paper, different features and components were investigated such as versions of algorithms, secret keys, block size, number of rounds, types of ciphers, logic operations, math transformations, and the structures of these algorithms as well. references 1. j. gubbi, r. buyya, s. marusic and m. palaniswami. internet of things (iot): a vision, architectural elements, and future directions. future generation computer systems, vol. 29, no. 7, pp. 1645-1660, 2013. 2. l. atzori, a. iera and g. morabito. the internet of things: a survey. computer networks, vol. 54, no. 15, pp. 2787-2805, 2010. 3. j. zhou, z. cao, x. dong and a. v. vasilakos. security and privacy for cloud-based iot: challenges. ieee communications magazine, vol. 55, no. 1, pp. 26-33, 2017. 4. m. h. shukur. design a mobile medication dispenser based on iot technology. in: 3rd international conference on communication engineering and computer science (cic-cocos’19), 2019. 5. y. yang, x. zheng, w. guo, x. liu and v. chang. privacy-preserving fusion of iot and big data for e-health. future generation computer systems, vol. 86, pp. 1437-1455, 2018. table 2: analysis of the structures of encryption algorithms cipher algorithm type of cipher size of secret key (bits) block size (bits) total no. of rounds structure of cipher aes block cipher 128/192/256 128 10/12/14 spn rc6 block cipher 128/192/256 128 20 spn twofish block cipher 128/192/256 128 16 spn speck block cipher 32-128 64-256 22-34 arx lightweight encryption algorithm block cipher 128/192/256 128 24/28/32 arx chacha20 stream cipher 256 512 20 arx jasim, et al.: analysis cipher algorithms for the security of internet of things devices 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 13-19 6. s. singh and n. singh. internet of things (iot): security challenges, business opportunities and reference architecture for e-commerce. in: 2015 international conference on green computing and internet of things (icgciot), ieee, pp. 15771581, 2015. 7. m. g. samaila, m. neto, d. a. b. fernandes, m. m. freire and p. r. m. inácio. challenges of securing internet of things devices: a survey. security and privacy, vol. 1, no. 2. e20, 2018. 8. q. jing, a. v. vasilakos, j. wan, j. lu and d. qiu. security of the internet of things: perspectives and challenges. wireless networks, vol. 20, no. 8, pp. 2481-2501, 2014. 9. h. shin, h. k. lee, h. y. cha, s. w. heo and h. kim. iot security issues and light weight block cipher. in: 2019 international conference on artificial intelligence in information and communication (icaiic), ieee, pp. 381-384, 2019. 10. a. alahdal and n. k. deshmukh. a systematic technical survey of lightweight cryptography on iot environment. international journal of scientific and technology research, vol. 9, no. 3, p. 16, 2020. 11. v. thakor, m. a. razzaque and m. r. a. khandaker. lightweight cryptography algorithms for resource-constrained iot devices: a review, comparison and research opportunities. ieee access, vol. 9, pp. 77-93, 2021. 12. p. panahi, c. bayılmış, u. çavuşoğlu and s. kaçar. performance evaluation of lightweight encryption algorithms for iot-based applications. arabian journal for science and engineering, vol. 46, no. 4, pp. 4015-4037, 2021. 13. k. f. jasim and i. f. al-shaikhli. mobile technology generations and cryptographic algorithms: analysis study. in: 2015 4th international conference on advanced computer science applications and technologies (acsat), ieee, pp. 50-55, 2015. 14. k. f. jasim and i. f. al-shaikhli. analysis of confidentiality algorithms in different mobile 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choi, h. kwon, h. kim, j. park and h. seo. grover on korean block ciphers. applied sciences, vol. 10, no. 18, p. 6407, 2020. 25. l. sleem and r. couturier. speck-r: an ultra light-weight cryptographic scheme for internet of things. multimedia tools and applications, vol. 80, pp. 17067-17102, 2020. 26. iso. iso/iec 29192-2: 2019: information security lightweight cryptography part 2: block ciphers. geneva, switzerland: international organization for standardization, 2019. 27. j. song and s. c. seo. efficient parallel implementation of ctr mode of arx-based block ciphers on armv8 microcontrollers. applied sciences, vol. 11, no. 6, p. 2548, 2021. 28. y. b. kim, h. kwon, s. w. an, h. seo and s. c. seo. efficient implementation of arx-based block ciphers on 8-bit avr microcontrollers. mathematics, vol. 8, no. 10, p. 1837, 2020. 29. c. manifavas, g. hatzivasilis, k. fysarakis and y. papaefstathiou. a survey of lightweight stream ciphers for embedded systems. security and communication 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[last accessed on 2021 may 27]. 31. m. coutinho and t. c. s. neto. improved linear approximations to arx ciphers and attacks against chacha. iacr cryptol, p. 224, 2021. 32. p. mclaren, w. j. buchanan, g. russell and z. tan. deriving chacha20 key streams from targeted memory analysis. journal of information security and applications, vol. 48, p. 102372, 2019. 33. s. m. s. reza, a. ayob, m. m. arifeen, n. amin, m. h. m. saad and a. hussain. a lightweight security scheme for advanced metering infrastructures in smart grid. bulletin of electrical engineering and informatics, vol. 9, no. 2, pp. 777-784, 2020. . 70 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 research article bromodomain inhibitor jq1 as a candidate therapeutic agent in malignant pleural mesothelioma cinaria t. albadri* department of clinical medicine, trinity centre for health sciences, trinity college dublin university, college of medicine, st. james’s hospital, dublin, ireland abstract malignant pleural mesothelioma (mpm) is a rare tumor that develops from the mesothelial linings of the pleural, pericardial, and peritoneal cavities. the actual risk factor for developing the disease is exposure to asbestos in workplace. bromodomain and extraterminal (bet) domain proteins are epigenetic signaling agents that associate with acetylated histones and expedite the transcription of target genes. this study investigates whether the small molecule bet protein inhibitor jq1 specifically may be an effective therapy for mpm. reverse transcriptase polymerase chain reaction methods reveal an inclusive change in gene expression implying that jq1 is a potential inhibitor which targets the bet proteins. our results report that jq1 has tumor-suppressive effects as it significantly ceased cellular activity in mpm cell lines. we predict that jq1 may be the promising therapy for pleural mesothelioma cancers. keywords: bromodomain and extraterminal domain, jq1, malignant pleural mesothelioma introduction malignant pleural mesothelioma (mpm) is an infrequent type of cancer that affects the pleura (the external lining of the lungs and the internal lining of the chest cavity). mpm includes three main types determined by their appearance under the microscope. mpm includes three main types determined by their appearance under the microscope as: epithelial mpm which is the most popular form, sarcomatoid mpm which is meant to be the most aggressive form and biphasic mpm which mixes both epithelial and sarcomatoid mpms. pleural mesothelioma is commonly caused by the long-term exposure to asbestos fibers that induce pathogenic changes and eventually lead to cell injury, fibrosis, and possibly cancer. it has been also proposed that aside from asbestos, there are other factors which have a role in the tumor pathogenesis, such as sv40 virus infection and genetic susceptibility.[1] mpm incidence has been increasing over the world and is anticipated to grow even higher in the following 20 years due to asbestos huge spread exposure throughout the past 10 years or so.[2] it is rated that about 50–80% of pleural malignant mesothelioma in men and 20–30% in women arose in individuals who were exposed to asbestos.[3] diagnosis of this disease is pretty much difficult even for some specialists because the symptoms do not appear only after a longer period of exposure to asbestos. so far, the only approved treatment for mpm is the combined therapy of pemetrexed and cisplatin, although currently, there have been many clinical trials determining the efficacy of other drugs. bromodomain and extraterminal (bet) domain proteins play an essential role as epigenetic reader molecules that are associated with acetylated histones and aid in the transcription of target genes. bromodomain is, therefore, an acetyl-lysinebinding motif of about 110 amino acids which are found in various transcription regulators and chromatin-modifying enzymes that are encoded by specific genes.[4] research has shown that only members of the bet family (brd2, brd3, brd4, and brdt) may be prospective targets of cancer. these four bet proteins make up a subset of the larger family that consists of 46 proteins which contain 61 brds that have been determined in humans.[5] bet inhibitors are getting to become of huge interest as a potential target due to the prospect that there might be biomarkers which will recognize the patients that may possibly benefit and respond to therapy. those small molecule inhibitors conquer the acetyl-binding pockets of the bromodomains, leading in their detach from active chromatin and repression of downstream transcriptional targets.[6] corresponding author: cinaria t. albadri, st. james’s hospital, trinity centre for health sciences, trinity college dublin university, ireland. e-mail: cinariaalbadri@yahoo.com received: mar 31, 2019 accepted: apr 15, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp70-76 copyright © 2020 cinaria t. albadri. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ albadri: bromodomain inhibitor jq1 71 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 jq1 has started its journey in the research world as a novel drug in inhibiting the development of some specific cancers. jq1 successfully blocks one of the well-known cancer-causing genes of the bet proteins which are brd4, thereby preventing the early growth of some blood and lung cancers such as leukemia and multiple myeloma. jq1 is a novel thienotriazolo-1,4-diazepine having an attached bulky t-butyl ester functional group at carbon 6. this chemical compound is a vigorous, high affinity, selective bet inhibitor. jq1 detaches bet proteins from chromatin by binding to the acetyl-lysine recognition pocket of bet. jq1 has been used as a chemical probe to investigate the role of bet in the transcriptional regulation of oncogenesis.[7-9] bet inhibitors are a new promising treatment for mpm. this study will focus on the effect of the bet inhibitor (jq1) in mpm. we hypothesized that bromodomains of the bet family are expressed/overexpressed in mpm and hence investigate the effect of the compound jq1 as an inhibitor of those bet. this appears to be the first study of its kind, as far as i know that reveals the role of inhibiting bet domain proteins with jq1 in mesothelioma cancers. materials and methods cell lines in this project, two mpm cell lines were treated with the small molecule bet protein inhibitor and were assessed for antiproliferative activity as a response to that drug. the bet inhibitor used in this study was jq1. the two cell lines used were ren and h226. a brief description of the cell lines as follows: • ren is an epithelial mesothelioma cell line (fast growth rate) • nci-h226 is an epithelial squamous cell carcinoma; mesothelioma cell line (slow growth rate). cell culture cell lines were typically grown and preserved at a fixed temperature of 37°c and a humidified environment of 5% co 2 in a cell incubator. these cell lines were cultured in an appropriate cell culture media rpmi-1640 (lonza, uk) which was supplemented with 10% (v/v) fetal bovine serum, 2 mm l-glutamine, and 500 u/ml penicillin-streptomycin. cell lines were regularly passaged in t75 cm2 flasks (nunc flasks obtained from fisher scientific, ireland) as following: all cell culture procedures were done in laminar flow hoods under sterile conditions. cell media discarded and cells were washed with 5 ml phosphate buffer solution. cells were then trypsinized for 5 min at 37°c, and once detached from the flask, media were added to make up a final volume of 10 ml. cells were pipetted many times and each 1 ml transfer to two new t75 flasks labeled with the cell line, passage number, and date. further, 9 ml of media was added to each of these two flasks and cells were incubated at 37°c. in this way, cells were split at a ratio of 1:10. cell subculture for proliferation assays before drug treatment, cells were visualized under the microscope (nikon corp.) and if found to be of about 70–80% confluent, they were trypsinized with 1.5 ml trypsin-edta and 8.5 ml of media to make it up to a final volume of 10 ml. the cells were then counted using a bright-line hemocytometer (sigma, usa) and seeded at 3×103 cells/well in 96-well plate. this was done in triplicate for both cell lines (ren and h226). the plated cells were kept in the incubator at 37°c. treatment of cells once plated, cells were left to recover for 48 h. subsequently, cells were treated with jq1 bet domain inhibitor at the following concentrations (0.25 µm, 0.5 µm, 0.75 µm, 1 µm, 15 µm, 20 µm, and 25 µm) for 48 h. 4 µl of dimethyl sulfoxide (dmso) was added to the control as jq1 was dissolved in dmso vehicle. a concentrated stock (18.6 mm) of jq1 dissolved in dmso was made and further four stock dilutions (1:25, 1:10, 1:5, and 1:2) were set up from that main concentrated stock. all plates were then incubated for 48 h at 37°c. 5-bromo-2-deoxyuridine (brdu) proliferation assay a brdu assay was performed to measure the proliferative activity of the treated cells. briefly, after cell treatment with jq1 for 48 h, all media were removed from the triplicate 96-well plate, and subsequently, 10 µl brdu labeling solution was added to each well. plates were incubated at 37°c for 4 h followed by the removal of the labeling medium from the wells. 200 µl fixdenat was added to each well and plates were incubated at 25°c for 30 min followed by the removal of fixdenat solution thoroughly. 100 µl anti-brdu pod solution was added to each well and plates were incubated again at 25°c for 90 min. the antibody was then removed and wells were rinsed 3 times with 200 µl/well washing solution. finally, washing solution was removed and 100 µl/well substrate solution added to each well. plates were left for 2–3 min and thence immediately 25 µl/well of 1 m h 2 so 4 was added and measurement of plates was carried out within 5 min. the activity of proliferative cells was measured by enzyme-linked immunosorbent assay at a wavelength of 450 nm using a standard plate reader. reverse transcriptase polymerase chain reaction (rt-pcr) cell lines and tissue samples fresh-frozen mesothelial and benign pleural samples were collected following debulking surgery at glenfield hospital, leicester, and were stored in the leicestershire mesothelioma tissue bank. anonymized mpm specimens from 22 patients were transferred to st. james’ hospital, dublin. approval for the storage and use of these samples have been obtained from the st. james’ hospital/the adelaide and meath hospital research ethics committee. a panel of mesothelioma cell lines has been assembled and utilized for this project. rna isolation total rna was isolated using tri reagent® (mrc gene) protocol. samples were mixed and incubated for 5 min at room temperature followed by centrifugation at 13,500 rpm for 5 min. the rna pellet was left to air dry after the removal of the supernatant and finally resuspended in 30–50 µl of molecular grade water and stored at −80°c until further use. albadri: bromodomain inhibitor jq1 72 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 rna quantification nanodrop 1000 spectrophotometer (thermo scientific) was used to measure the purity of the isolated rna. 1 µl of each rna sample was loaded separately onto the nanodrop and analyzed. the quantity of rna per sample was measured in ng/µl. a 260/280 value of 2.0 indicates the purity of an rna sample. complementary dna (cdna) synthesis cdna was synthesized using revertaid rt according to the manufacturer’s instructions (fermentas). samples were mixed gently, incubated for 90 min at 42°c followed by 10 min at 70°c heating to terminate the reaction. the resultant cdna was then stored at −20°c until required. rt-pcr the cdna which was synthesized previously was used to detect gene expression. for pcr, the following were mixed in a new tube to set up the pcr reaction: 10 µl (2× gotaq® green [promega]), 2 µl (5 µm of each forward and reverse primers), 6 µl (molecular grade water), and 1 µl (cdna). 1 µl molecular grade water was added in the place of cdna for the negative control. pcr samples were loaded on a thermocycler. the pcr cycling conditions involved initial denaturation at 95°c for 5 min, followed by 30–35 cycles of denaturation at 95°c, annealing at the temperature 58°c, extension at 72°c, and a final elongation step at 72°c. gel electrophoresis agarose gel (2%) was used for loading all pcr products. agarose (2%) was dissolved in tris-acetate-edta (tae) in the microwave for 3 min. 4 µl of ethidium bromide was added per 25 ml of agarose solution after being cooled and then poured into a gel tray. the gel was then allowed to set at room temperature for 10–15 min. 4 µl of 100 bp dna ladder (fermentas) was loaded onto the gel in parallel with the samples. 8 µl of each pcr product and 2 µl of the corresponding 18s housekeeping gene products were loaded as well. the samples were electrophoresed using an electrophoresis unit (sigma) and 1× tae was used as a running buffer. the gels were run at 80 volts for 30 min. the resultant bands were visualized and photographed under ultraviolet light using a biospectrum imaging system (ultra violet products, fusion fx, uk). statistical analysis statistical analysis was carried out using two-tailed student’s t-test and one-way anova. data were graphed as mean ± standard error of mean using graphpad prism software. densitometry analysis was performed using tina software. results were considered significant when the p < 0.05. results screen of panel of mesothelioma cell lines for the expression of bet domain proteins a panel of 34 mesothelioma cell lines was screened for the expression of brd2 and brd4 (long and short transcript) variants. all these cell lines are mpms, except for lp9, met5a, np1 and np2, and p31 and p31cisr. beta-actin was used as a positive control which is regularly expressed in all cell lines and a negative control was included also. brd4 long transcript variant shows equal expression in all cell lines, but brd4 short transcript shows a slightly lower expression in some of the cell lines [figure 1]. brd2 also shows an equal expression in all cell lines [figure 2]. screen of primary mpm patient cells for the expression of bet domain proteins a panel of 34 primary mesothelioma cells obtained from patients was examined for the expression of brd2 and brd4 (long and short transcript) variants. these patient samples were divided into benign and three histological subtypes of mpm; epithelial, biphasic, and sarcomatoid. densitometry analysis was performed to identify the differences in expression of the brds between benign and tumor tissue following normalization against the 18s rrna expression for each sample, and a negative control was used in each pcr. brd2 showed expression across samples [figure 3a] and was significantly less expressed in the tumor samples compared to benign [figure 3b and c]. brd4 long transcript variant appeared to be expressed across the three histological subtype samples and less expressed in benign [figure 4a] and following statistical analysis, brd4 long transcript expression was significantly upregulated in figure 1: expression of brd4 long and short transcript variants in a panel of 34 mesothelioma cell lines. beta-actin is a positive control. brd4 long transcript is expressed uniformly in all cell lines and the expression of brd4 short transcript is relatively less in some of the cell lines albadri: bromodomain inhibitor jq1 73 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 the tumor samples compared to benign [figure 4b]. brd4 long transcript expression levels were significantly higher in epithelial and biphasic subtypes compared to benign, while not significant in sarcomatoid [figure 4c]. on contrary, the expression of brd4 short transcript variant appears to be almost entirely absent in the benign samples [figure 5a] and brd4 short transcript expression was significantly upregulated in the tumor samples compared to benign [figure 5b]. while not significant, a trend toward upregulation of brd4 short transcript was observed in all three histological subtypes of mpm compared to benign [figure 5c]. antiproliferative activity the brdu assay detects incorporated brdu into the cellular dna of actively proliferating cells. this assay was performed to measure changes in proliferating cells after 48 h following treatment with jq1 drug. ren-treated cells appear to show a significant decrease in proliferation activity with 5, 10, 15, 20, and 25 µm drug concentrations, while nci-h226 treated cells appear to show significant decline in proliferation with 0.25, 0.5, 0.75, 1, 15, and 25 µm drug concentrations [figure 6]. the assay was accomplished in triplicate and the statistical analyses expressed as mean of treatments using one-way anova and dunnett’s multiple comparison test. discussion mpm is a rare and aggressive form of cancer with limited treatment chances that have inconsiderable impact on survival improvement. asbestos has been known as a non-mutagenic carcinogen that induces mesothelioma and in such case, cancer may be developed as an outcome of somatic epigenome dysregulation.[10] and so, aberrant epigenetic events have been spotted in mpm. epigenome regulators include dna methylation, histone modifications, chromatin remodeling, and regulation of non-coding rnas. it is not clear why precisely epigenetic modifications are a notable feature of pleural mesothelioma, figure 2: expression of brd2 in a panel of 34 mesothelioma cell lines. beta-actin is a positive control. brd2 is uniformly expressed in all cell lines figure 3: brd2 expression levels. (a) reverse transcriptase polymerase chain reaction for brd2 levels in a panel of primary patient malignant pleural mesothelioma samples of different histological subtypes against 18s rrna housekeeping gene. (b) brd2 expression level in malignant patients is significantly less compared to benign (***p < 0.001, two-tailed t-test). (c) levels of brd2 expression in all three histological subtypes is significantly different compared to benign (*p < 0.05, **p < 0.01, ***p < 0.001, one-way anova using dunnett’s multiple comparisons test). error bars represent mean ± standard error of mean a b c albadri: bromodomain inhibitor jq1 74 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 but histone modifications have emerged as remarkable modifications in the carcinogenesis of mpm.[11] aberrant regulation of histone modification plays a role in oncogenesis by affecting gene activity. the acetylation of histones is mediated by lysine acetyltransferases. lysine acetylation is mainly recognized by bromodomains and tandem phd domains.[12] bet proteins are readers of acetyl-lysine signs. in some cancer types, bromodomain-containing proteins which identify histone acetylation signs are mistargeted or overexpressed.[13] this study focuses precisely on the role of two bet proteins, brd2 and brd4, in mpm and the effect of jq1 compound as an inhibitor for brds. the expression of these bet proteins in mpm has been evaluated in a panel of mesothelioma cell lines and in primary mpm patient cells. a panel of cell lines [figures 1 and 2] was screened to evaluate whether there are any prominent patterns of expression which need to be identified before screening of primary mpm patient cells. slight changes in expression were noted. brd4 short transcript variant is expressed weakly in some of the cell lines. among the analyzed cell lines are a cisplatin resistance cell line. cisplatin is a chemotherapeutic agent used in cancer treatment. a specific feature of mpm is that it is a cisplatin resistance tumor.[14] although there were no remarkable variations in the expression level of any of the evaluated brds, additional mpm chemoresistant cell lines need to be included so that it would be a conclusive analysis. figure 4: brd4 long transcript expression levels. (a) reverse transcriptase polymerase chain reaction for brd4l levels in a panel of primary patient malignant pleural mesothelioma samples of different histological subtypes against 18s rrna housekeeping gene. (b) brd4l expression level in tumor patients is significantly different compared to benign (***p < 0.001, two-tailed t-test). (c) levels of brd4l expression epithelial and biphasic subtypes are significantly different compared to benign, while not significantly different in sarcomatoid subtype (**p < 0.01, ***p < 0.001, one-way anova using dunnett’s multiple comparisons test). error bars represent mean ± standard error of mean a b c figure 5: brd4 short transcript expression levels. (a) reverse transcriptase polymerase chain reaction for brd4s levels in a panel of primary patient malignant pleural mesothelioma samples of different histological subtypes against 18s rrna housekeeping gene. (b) brd4s expression level in tumor patients is significantly different compared to benign (*p < 0.05, two-tailed t-test). (c) levels of brd4s expression in all three histological subtypes show no significance compared to benign (one-way anova using dunnett’s multiple comparisons test). error bars represent mean ± standard error of mean a b c albadri: bromodomain inhibitor jq1 75 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 subsequently, a panel of primary mpm patient cells including benign, epithelioid, sarcomatoid, and biphasic histological subtypes was screened [figures 3-5]. expression of the three evaluated brds varies in malignant compared to benign cells. expression of brd2 was significantly different in malignant cells compared to benign cells. in addition, all three histological subtypes showed significantly lower levels of brd2 expression compared to benign even though brd2 was expressed in benign. on contrary, brd4 long transcript expression levels are significantly higher in malignant cells compared to benign. in consistent with the results of brd4 long transcript expression levels, the expression of brd4 short transcript confirmed significantly elevated levels in malignant cells compared to benign, and this indicates that brd4 long and short transcripts are overexpressed in malignant cell. our findings have proven that brd2 is downregulated in mpm tumors, while brd4 long and short transcripts are significantly upregulated in mpm tumors. moreover, since brd4 is the main target for jq1, consequently, this makes targeting mpm with jq1 and excellent strategy. to further support our findings, the antiproliferative activity of jq1 was evaluated in vitro using two specific mpm cell lines (ren and nci-h226). these two cell lines were chosen according to their common use in human tumor xenografts, as they perform well when xenograft in mouse models and so, the evaluation of jq1 drug can be smoothly carried out in such in vivo models. jq1 is a bet inhibitor that has been selected for this study. scientists have been competing to develop novel bet protein inhibitors. a recently developed dihydroquinazoline-2-one inhibitor has been found to target bet.[15] brdu assay was the first move in analyzing the proliferative activity of jq1 compound. this assay detects brdu incorporated into the newly synthesized dna of actively proliferating cells. brdu assay was chosen to measure the proliferating activity of cells. ren and nci-h226 cells were treated with a range of jq1 concentrations [figure 6]. response of nci-h226 cells to jq1 was clearly observed, with a significant decrease in cell proliferation starting from 0.25 µm up to 25 µm concentrations compared to untreated cells. while ren cells showed a significant decrease in proliferative activity with concentrations from 5 µm up to 25 µm. dmso was used as a control to ensure that any change in proliferative activity was caused by jq1 and not dmso. dmso induced no change in the cellular activity of both cell lines and so, noted results are accurate. jq1 inhibits the proliferative activity of ren and nci-h226 cells with concentrations of 5 µm and 0.25 µm, respectively. jq1 has shown to have antiproliferative effects on mesothelioma cell lines and efficiently abrogates their clonogenic growth. exposure of sensitive cell lines with jq1 results in decreased proliferative cell activity and induction of apoptosis. in addition to confirming the effect of jq1, the brdu also provided a guideline for treatment concentrations to be used in a further study. conclusion this study has shown that bet domain proteins brd2 and brd4 are significantly altered in the tumors of patients suffering from mpm. critically levels of brd4 are significantly elevated in the tumors. jq1 is a small molecule inhibitor of brd4 and brd2. this study has shown that jq1 may be a potentially effective therapy for mpm in vitro. references 1. b. kroczynska, r. cutrone, m. bocchetta, h. yang, a. g. elmishad, p. vacek, m. ramos-nino, b. t. mossman, h. i. pass and m. carbone. “crocidolite asbestos and sv40 are co-carcinogens in human mesothelial cells and in causing mesothelioma in hamsters”. proc natl acad sci usa vol. 103, pp. 14128-14133, 2006. 2. b. w. s. robinson, a. w. musk and a. lake. “malignant mesothelioma”. lancet, vol. 366, pp. 397-408, 2005. 3. m. carbone, b. h. ly, r. f. dodson, i. pagano, p. t. morris, u. a. dogan, a. f. gazdar, h. i. pass and h. yang. “malignant mesothelioma: facts, myths, and hypotheses”. journal of cellular physiology, vol. 227, pp. 44-58, 2012. 4. s. y. wu, a. y. lee, h. t. lai, h. zhang and c. m. chiang. “phospho switch triggers brd4 chromatin binding and activator recruitment for gene-specific targeting”. molecular cell, vol. 49, pp. 843-857, 2013. 5. p. filippakopoulos, s. picaud, m. mangos, t. keates, j. lambert, d. barsyte-lovejoy, i. felletar, r. volkmer, s. muller, t. pawson, a. gingras, c. arrowsmith and s. knapp. “histone recognition and large-scale structural analysis of the human bromodomain family”. cell, vol. 149, pp. 214-231, 2012. 6. p. filippakopoulos, j. qi, s. picaud, y. shen, w. b. smith, o. fedorov, e. m. morse, t. keates, t. t. hickman, i. felletar, m. philpott, s. munro, m. r. mckeown, y. wang, a. l. christie, n. west, m. j. cameron, b. schwartz, t. d. heightman, n. la thangue, c. a. french, o. west, a. l. kung, s. knapp and j. e. bradner. “selective inhibition of bet bromodomains”. nature, vol. 468, pp. 1067-1073, 2010. 7. m. a. dawson, r. k. prinjha, a. dittman, g. giotopoulos, m. bantscheff, w. i. chan, s. c. robson, c. chung, c. hopf, m. m. savitski, c. huthmacher, e. gudgin, d. lugo, s. beinke, t. d. chapman, e. j. roberts, p. e. soden, k. r. auger, o. mirguet, k. doehner, r. delwel, a. k. burnett, p. jeffrey, g. drewes, k. lee, b. j. huntly and t. kouzarides t. “inhibition of bet recruitment to chromatin as an effective treatment for mll-fusion leukaemia”. nature, vol. 478, pp. 529-533, 2011. 8. j. e. delmore, g. c. issa, m. e. lemieux, p. b. rahl, j. shi, h. m. jacobs, e. kastritis, t. gilpatrick, r. m. paranal, j. qi, m. chesi, a. schinzel, m. r. mckeown, t. p. heffernan, c. r. vakoc, l. bergsagel, i. m. ghobrial, p. g. richardson, r. a. young, w. c. hahn, k. c. anderson, a. l. kung, j. e. bradner and c. s. mitsiades. “bet bromodomain inhibition as a therapeutic strategy to target c-myc”. cell, vol. 146, pp. 904-917, 2011. 9. j. a. mertz, a. r. conery, b. m. bryant, p. sandy, s. balasubramanian, d. a. mele, l. bergeron and r. j. sims. figure 6: 5-bromo-2-deoxyuridine proliferation assay following 48 h treatment with various concentrations of jq1. (a) ren cells show significant decline in cell proliferation at 5, 10, 15, 20, and 25 µm drug concentrations (p < 0.01, one-way anova). (b) nci-h226 cells show significant decline in cell proliferation at 0.25, 0.5, 0.75, 1, 15, and 25 µm drug concentrations (p < 0.001, one-way anova) a b albadri: bromodomain inhibitor jq1 76 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 70-76 “targeting myc dependence in cancer by inhibiting bet bromodomains”. proc natl acad sci usa, vol. 108, pp. 1666916674, 2011. 10. b. c. christensen, e. a. houseman, j. j. godleski, c. j. marsit, j. l. longaker, c. r. roelofs, m. r. karagas, m. r. wrensch, r. yeh, h. h. nelson, j. l. wiemels, s. zheng, j. k. wiencke, r. bueno, d. j. sugarbaker and k. t. kelsey. “epigenetic profiles distinguish pleural mesothelioma from normal pleura and predict lung asbestos burden and clinical outcome”. cancer res, vol. 69, pp. 227-234, 2009. 11. p. k. paik and l. m. krug. “histone deacetylase inhibitors in malignant pleural mesothelioma preclinical rationale and clinical trials”. j thorac oncol, vol. 5, pp. 275-279, 2010. 12. m. yun, j. wu, j. l. workman and b. li. “readers of histone modifications”. cell res, vol. 21, pp. 564-578, 2011. 13. n. sachini. “the role of lysine acetylation, histone acetyltransferases and bromodomain containing proteins in cancer development”. master thesis, umc utrecht, 2013. 14. v. janson. “cisplatin-resistance and cell death in malignant pleural mesothelioma cells”. doctoral thesis, umeå university, 2008. 15. s. picaud, d. d. costa, a. thanasopoulou, p. filippakopoulos, p. v. fish, m. philpott, o. fedorov, p. brennan, m. e. bunnage, d. r. owen, j. e. bradner, p. taniere, b. o’sullivan, s. muller, j. schwaller, t. stankovic and s. knapp. “pfi-1, a highly selective protein interaction inhibitor, targeting bet bromodomains”. cancer res, vol. 73, pp. 3336-3346, 2013. . 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 53-57 research article long time stability investigation for structural and magnetic properties of ti-doped barium ferrite mohammed abdul malek al saadi* department of computer and communication engineering, cihan university-erbil, kurdistan region, iraq abstract barium hexaferrite (bhf) (bafe12o19) and its substituted derivatives have been considered as the most potential magnetic candidates with considerable chemical stability and physiochemical characteristics. bhf with x ferrite ions substituted by titanium (ti-doped bthf) (batixfe12-xo19) (x=1 and x=3) was prepared from ferric oxide (fe2o3), barium oxide (bao), and titanium oxide (tio2) of purity >98%. the materials were mixed with deionized water and then dried at 1100°c and 1200°c overnight. for the formation of bafe12o19 phase, the mixture was annealed at a rate of 10°c/min in static air atmosphere until reaching 1200°c and then maintained for 10 h. structural properties of these samples were measured using x-ray diffraction (xrd) and scanning electron microscopy, while magnetic properties were measured using vibrating sample magnetometer (vsm) device. magnetic and structural characteristics are investigated after preserving ti-doped bhf samples at room temperature and ambient conditions for 12 years. the samples are characterized using the same previous techniques to find out the possible effect of long period storage on their properties. the results showed that the storage process has little effect on these properties where the granular size increased due to increased oxidation. xrd tests also showed the absence of ti at low ratios due to increased oxidation of ferrite. vsm results showed increased magnetic properties after storage due to increased iron oxide. keywords: material, electromagnetic, microwave, ferrite introduction the ferrites are homogeneous ceramic materials dark gray or black color which is made up of a blend of different types of oxides with iron oxide and have different crystalline structures.[1] it is also considered as ionic compounds and its magnetic properties due to the magnetic ions they contain. the high permeability due to the distribution of the wall of the field controlled by the sintering density and by increasing the size of particleboard.[2] barium hexaferrite (bhf) and its substituted derivatives have been considered as candidates with the most potential due to their chemical stability and suitable magnetic characteristics.[3] ferrites are ideally suited for making device such as inductor cores, circulators, memory devices, and also for various microwave application. a reduction in size and weight of power supplies can be achieved using switched-mode or resonant concepts. for the voltage conversion, several circuit designs are in use with ferrites as transformer core materials. transformer ferrites must show low-energy losses at high induction levels at higher and higher frequencies; this requires the development of new ferrite materials with constantly improving loss characteristics.[4] although the saturation magnetization of ferrites is less than that of ferromagnetic alloys, they have advantages such as applicability at higher frequency, lower price, and greater electrical resistance. ferrites are classified into two groups, namely, magnetically soft and hard. furthermore, depending on their crystal symmetry, they may be cubic, hexagonal, or orthorhombic.[5] ti, a tetravalent element, is well known candidate to cosubstitute fe3+ ions. the ionic radius of ti4+ ion is 0.605 å, which is smaller than fe3+ ion (0.645 å). furthermore, the electronegativity of ti (1.54) is small compared with fe (1.83). hence, ti4+ ions may easily substitute fe3+ ions in the structure with minimal impact on the intrinsic structure of the ferrite.[6] the main applications of these materials intend to reduce the human exposure to microwaves by means of absorbing coatings.[2,7] ti-doped bhf powder is an efficient absorber of electromagnetic waves in the microwave spectrum. to corresponding author: mohammed abdul malek al saadi, cihan university-erbil, kurdistan region, iraq. e-mail: mohammed.alsaadi@cihanuniversity.edu.iq received: apr 20, 2019 accepted: apr 23, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp53-57 copyright © 2019 mohammed abdul malek al saadi. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ al saadi: long time stability investigation for structural and magnetic properties of (ti-doped barium ferrite) 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 53-57 optimize the ti-doped bhf microwave absorption properties increase in the saturation magnetization (m s ) and a decrease in the coercivity (jh c ) are required.[8,9] these properties depend on the localization of the ti4+ in bhf structure. according to literature,[10,11] the ti4+ ions preferentially occupy the octahedral 4f 2 sites of the bhf structure when the ti-doped bhf is synthesized using the ceramic route, especially at low doping rates.[12] bhf in powder form is one of widely implicated magnetic ceramic materials by both researchers and manufacturers in many applications such as permanent magnets, microwave absorber devices, and recording media. bhf is attractive due to its high stability, excellent high-frequency response, narrow switching field distribution and its temperature coefficient of coercivity in various applications, fairly large magnetocrystalline isotropy, high curie temperature and relatively large magnetization, corrosion resistivity as well as chemical stability.[1-13] in this paper, bhf substituted by ti4+ (chose cosubstitution non-magnetic ti4+ ions for fe3+ at different levels to obtain a wide range of coercivity and high magnetic permeability) using conventional ceramic techniques is investigated by differential thermal analysis, x-ray diffraction (xrd), scanning electron microscopy (sem), and magnetic measurements by vibrating sample magnetometer (vsm). the approach to saturation law was used to determine zero-field saturation magnetization m s and anisotropy field h a . experimental ti-doped bhf (bati x fe 12-x o 19 ) is prepared by conventional ceramic techniques (the conventional ceramic methods, i.e., high-energy ball milling was employed to obtain high quality of bhf). the materials used were ferric oxide (fe 2 o 3 ), barium oxide (bao), and titanium oxide (tio 2 ) of purity greater than 98%. ratios were calculated accurately by weighing of materials involved in the preparation of ferrite samples using an electronic balance its sensitive up to 0.0001 g. after grinding, the mixed oxides individually to obtain homogeneous size distribution and to facilitate the process of granule homogeneity between these compounds during the mixing. the powdered oxides were mixed with deionized water and then placed in the furnace. a time-temperature program was started in the furnace after placing the mixture sample. heating rate was 10°c/min until it reaches the reaction temperature (tc) (1100 or 1200°c) which remained for 10 h then cooling started before collecting the samples. the crystallite phases present in the different samples were identified by xrd utilizing bruker axis d8 diffract meter using cu-ka (ƛ=1.5406) radiation operating at 40 kv and 30 ma at a rate of 2°/min. the diffraction data were recorded for 2θ values between 20° and 80°. the particles morphology was observed using the sem to study the surfaces micrographic appearance, to find out the effect of substitution on the structural properties, and to investigate the particle size measurements distribution of ti-doped bhf samples. the magnetic properties of the ferrites were measured at room temperature using a (vsm; 9600-1 ldj, usa) in maximum applied field of 15 g. from the obtained hysteresis loops, the saturation magnetization (m s ), remanence magnetization (m r ), and the coercive field (hc) were determined. the same samples were marked and preserved on laboratory shelve in ambient conditions for 12 years and then the same above-mentioned characterization routines were employed for comparison and revealing the effect of long period storage. results and discussion structure and morphology of samples xrd before 12 years figure 1 shows xrd patterns of ti-doped bhf sintering at 1100°c–1200°c for 10 h. when comparing the xrd schemes of x=0, x=1, and x=3, we find that there is a significant correlation in the results. in addition to the appearance of fe 3 o 4 phase at x=3, indicating that the increased titanium ratio generated an increase in ion exchange iron; hence, the fe 3 o 4 phase will appear. it is also clear that increasing the amount of substitution of x gives a lower relative intensity and width peaks indicating a decrease in particle size.[8] xrd after 12 years figure 2 shows the results of the same samples under the same conditions after 12 years. new and very well phases can be seen. sem before 12 years figure 3 shows the images of the surface of ti-doped bhf. image (a) shows that the interaction of this sample when x=1 at t=1100°c for 10 h did not complete and the large pores figure 1: x-ray diffraction patterns of ti-doped barium hexaferrite (a) x=0, (b) x=1, and (c) x=3 at t=1200°c for 10 h before 12 years a b c al saadi: long time stability investigation for structural and magnetic properties of (ti-doped barium ferrite) 55 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 53-57 have been shown in the surface, while the image (b) shows that the interaction will be integrated and homogeneous when t is increased to t=1200°c for 10 h. the interaction of the ferrite and particle size will be better than in the lower sintering, therefore, less porosity and better homogeneity of the sample, while image (c) shows the surface of ti-doped bhf when x=3 at 1200°c for 10 h. it is expected that increasing of replacing of iron ions by ti ions will decrease the diameter of the crystal; hence, the particle size and porosity will decrease, this is shown by the xrd (1) from the width and low height of the peaks. the pictures above and the table below have shown that grain size decreased with increasing of x and it increased with increasing of tc. the decreasing of x will be better than increasing it. table 1 shows the grain size of the samples. sem after 12 years figure 4 and table 2 show that the grain size decreased with increasing of x while it increased with increasing of tc for the same reasons mentioned above, these granular sizes correspond to the results 12 years ago. however, we note from the results that granular size increases overtime due to increase of oxidation in samples. magnetic properties of prepared samples the hysteresis loop (b-h loop) is an important property of any magnetic material and it is a measure of the lost energy per volume during a magnet cycle. the density of the magnetic saturation is dependent on the composition of the ferrite structure and it increases when the tc increases, and its effective values reduce due to the porosity of samples.[14] nanoparticles with a coercive field strength above 5 koe derived bhf are convenient for permanent magnets, but the magnetocrystalline anisotropy is too large for recording media applications. therefore, the partially substituted fe3+ ions of the hexaferrite phase, for example, with metals co 2 +, mn2+, ni2+, and ti4+, etc., were attempted to reduce anisotropy constant. before12 years figure 5 shows magnetic hysteresis loops of the ti-doped bhf samples. saturation magnetization (m s ) and coercivity figure 2: x-ray diffraction patterns of ti-doped barium hexaferrite after 12 years figure 3: scanning electron microscopy images and grain size of ti-doped barium hexaferrite (a) x=1 at 1100°c, (b) x=1 at 1200°c, (c) x=3 at 1200°c table 1: grain size of samples before 12 years number of samples t (°c) x grain size (µm) a 1100 1 0.21 b 1200 1 0.38 c 1200 3 0.28 table 2: particle size of samples after 12 years number of samples t (oc) x grain size (µm) a 1100 1 0.57 b 1200 1 0.76 c 1100 3 0.25 d 1200 3 0.59 a b c al saadi: long time stability investigation for structural and magnetic properties of (ti-doped barium ferrite) 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 53-57 (hc) extracted from the loops are given in table 3 as functions of the x and tc values. it is seen that both m s and hc decreased with increasing x. hc decreased more rapidly at lower x values, whereas m s decreased more at higher x values. figure 5 shows the hysteresis loop of ti-doped bhf at 1100°c and 1200°c, respectively, for 10 h. figure 5 shows that mr and ms increased with increasing of tc and they decreased with increasing of x,[4] while 11 hc decreased with increasing of x and tc; therefore, the samples will change from hard to soft material when the concentration and temperature increase. these parameters of all samples are shown in table 3. after 12 years it is known that magnetic property of ferrite is influenced by the chemical composition and ion distribution in the crystallographic sites. in the basic structure of m-type barium ferrite, fe3+ ions occupy five different interstitial sites: one tetrahedral site 4f1 (↓), one bipyramidal site 2b (↑), and three octahedral sites12k (↑), 2a (↑), and 4f2 (↓). in addition to the reduction of the magnetocrystalline anisotropy due figure 5: x-ray diffraction of ti-doped bhf: (a) x=1 at 1100°c figure 4: scanning electron microscopy images and grain size of ti-doped barium hexaferrite (a) x=1 at 1100°c after 12 years table 3: the magnetic parameters of ti-doped barium hexaferrite before 12 years x t (°c) hc (koe) ms (emu/g) mr (emu/g) type of material 0 1100 4.5 1.35 0.42 hard 1 = 5.96 1.3 0.76 hard 3 = 0.06 1.1 0.37 soft 0 1200 6.9 16.3 8.94 hard 1 = 2.7 3.85 1.58 hard 3 = 0.04 2.95 0.56 soft table 4: the magnetic parameters of ti-doped barium hexaferrite after 12 years x t (°c) hc (koe) ms (emu/g) mr (emu/g) type of material 1 1100 1790 g 30 14.26 hard 3 1100 168 g 20.7 3.25 soft 1 1200 909.1 g 32.73 10.17 hard 3 1200 145.5 17.87 2.45 soft a b c d a b c d al saadi: long time stability investigation for structural and magnetic properties of (ti-doped barium ferrite) 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 53-57 to the substitution of iron by titanium, a decrease in the hc has resulted. this is in agreement with the magnetic results, figures 5 and 6. the magnetic hysteresis curves show that the original hard magnetic behavior is shifted to soft magnetic by means of high doping content. in effect, it is clearly observed that ti-doped bhf samples exhibit a decrease of hc and ms. it is clear from the patterns below that samples were not significantly affected by the time factor, where they retained their properties. the hard sample remained hard and soft remained soft. furthermore, we will see that hc and mr decreased with increasing of x and tc, while ms will decrease when x increases, but it will increase at x=1 when tc increases while it will decrease at x=3 when tc increases. the ratio of ti is affected by the increasing of oxidation of ferrite due to the storage factor; hence, the samples with the low substitution ratio of x are close to the pure bhf. figure 6 and table 4 show b-h loop and the magnetic parameters of our samples after 12 years. conclusions ti-doped bhf (bati x fe 12-x o 19 ) prepared by conventional ceramic techniques. in addition to the appearance of fe 3 o 4 phase at x=3, indicating that the increased titanium ratio generated an increase in ion exchange iron; hence, the fe 3 o 4 phase will appear. it is also clear that increasing the amount of substitution of x gives a lower relative intensity and width peaks indicating a decrease in particle size. the results show that the deviation in the value of d is very low, not exceeding 0.007, due to convergence of fixed network values between fe3+ (0.63 a°) and ti4+ (0.68a) ions. the density of the magnetic saturation is dependent on the composition of the ferrite structure and it increases when the temperature increases, and its effective values reduce due to the porosity of samples. the magnetic parameters (mr, mrs, and hc) are increasing with increasing temperature. bhf has been substituted by ti4+ with formula (baiti x fe 12-x o 19 ), at different x and temperature. the forms of the b-h loop shown the material changes from the solid to the soft material at high concentrations as well as to all temperatures. references 1. c. kittle. “introduction to solid state physics”. 5th ed. new york: john wiley and sons, 1976. 2. z. yang, c. s. wang, x. h. li and h. x. zeng. “(zn.ni.ti) substituted ba-ferrite particles with improved temperature coefficient of coersivity”. materials science and engineering b, vol. 90, pp. 142-145, 2002. 3. g. mendoza, k. k. johal, h. mancha, j. l. escalante and m. m. cisneros. “magnetic properties of zn-sn substituted ba-ferrite powders prepared by ball milling”. matrials research bulletin, vol. 36, pp. 2597-2603, agu 2001. 4. a. j. b. fuller. “ferrites of microwave frequencies”. london, united kingdom: peter peregrinus, 1987, pp. 14-16. 5. m. b. amin and j. r. jams. “tech. for utilization of hexagonal in radar absorbers”. part 1. radio and electron engineering, vol. 51, no. 5, pp. 209-218, 1981. 6. j. li, h. zhang, y. liu and h. yang. “co-ti co-substitution of m-type hexagonal barium ferrite”. materials research express, vol. 2, no. 4, p. 046104, 2015. 7. pamela quiroz, bernd halbedel, angel bustamante, juan c. gonzález. “effect of titanium ion substitution in the barium hexaferrite studied by mössbauer spectroscopy and x-ray diffraction”. hyperfine interact, vol. 202, p. 106, aug 2011. 8. b. halbedel, d. hülsenberg, belau, st., schadewald, u.: cfi/ber. dkg vol. 82, no. 13, 182-188, 2005. 9. nibruchnnay, umberto “microwave ferrite and application” electro. mater., chap. 17, pp. 451,459,460, 1994. 10. rohde and schwarz. news 199/09, pp. 70-72, 2009. 11. p. a. mariño-castellanos, j. anglada-rivera, a. cruz-fuentes and r. lora-serrano. “magnetic and microstructural properties of the ti4+-doped barium hexaferrite”. journal of magnetism and magnetic materials, vol. 280, pp. 214-220, 2004. 12. v. a. m. brabers, a. a. e. stevens, j. h. j. dalderop and z. simga. “ultrasonic attenuation in a batife11o19 single crystal”. journal of magnetism and magnetic materials, vol. 196-197, pp. 312-314, may 1999. ىلع زينغنملاو مويناتيتلا ريثأت» ريتسجام ةلاسر فيان ديعس .13 «ةينيسلا.ةمزحلا تايدمل ةقيقدلا تاجوملل ةصام داومك مويرابلا تيارف .2001 ، رابنالا ةعماج ، مولعلا ةيلك 14. j. matutes, s. diaz, m. mirabal and s. a palomaress. “synthesis by coperecipitation and study of ba-hexaferrite powders”. scripta materialia, vol. 42, pp. 295-299, aug 2000. figure 6: x-ray diffraction of ti-doped barium hexaferrite: (a) x=1 at 1100°c, (b) x=1 at 1200°c, (c) x=3 at 1100°c, (d) x=3 at 1200°c a b c d tx_1~abs:at/add:tx_2~abs:at 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 9-12 research article likelihood approach for bayesian logistic weighted model: missing completely at random case dler h. kadir* department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq abstract increasing the response rate and minimizing non-response rates represent the primary challenges facing researchers performing longitudinal and cohort research, especially can be seen in the area of pediatric medicine. when there are missing data, complete case analysis makes findings bias. inverse probability weighting (ipw) is one of the many available approaches for reducing bias using complete case analysis. here, a complete case is weighted by probability inverse of complete cases. the data were collected from the neonatal intensive care unit at erbil maternity hospital from 2012 to 2017. in total, 570 babies (288 male and 282 females) were born very preterm. the aim of this paper is to use ipw on the bayesian logistic model developmental outcome. the mental development index approach was used for assessing the cognitive development of those born very preterm. almost half of the information for the babies was missing, meaning that we do not know whether they have cognitive development issues. we obtained greater precision in results and standard deviation of parameter estimates which are less in the posterior weighted model in comparison with frequent analysis. further, research is needed using methods such as bootstrapping, sandwich, resampling, and jackknife methods for dealing with missing data. keywords: likelihood, logistic weighting, missing data, preterm infants introduction increasing the response rate and minimizing non-response rates represent the primary challenges facing researchers performing longitudinal and cohort research. this can especially be seen in the area of pediatric medicine, whereby birth cohorts are often utilized for epidemiological research and randomized clinical trials of parental interventions. the contrition of participants lowers the strength of the research, as well as leading to bias in the results.[1] a non-response usually has more medical and socioeconomic risks and can have systematic variations regarding interest disorders, causing a biased estimate of an adverse outcome.[2] it is also frequently the case that data are missing in social research. they present ambiguities in statistical analyses of a different type to that of the usual imprecisions of samples, which become lower with increases in sample sizes. therefore, greater assumptions are required to permit inferences to be reached. over the past 10 years, there has been much theoretical research into ways of analyzing missing data sets. missing data can be defined as a value that is not recorded for a variable in the observation of interest. almost all branches of scientific research face this issue and will occasionally have to deal with missing data. frequently, the missing data appear as incomplete data on a subject. usually, the following analysis uses only a subject with a complete case measurement (complete case analysis). this proves expensive not only with regard to reductions in sample sizes, as an unclear variance estimate, lower statistical power, and the parameters which are thought to be possibly biased until a complete case analysis represents a random sampling of the focus population. to ensure that bias is considered, it is important to understand the mechanisms and the patterns of the missing data relevant to the research. the missing data mechanisms, as stated by rubin, indicate the link between the missing values and the observed data.[3] it is obvious that bias cannot only occur when there are a systematic dropout and non-response, it can additionally stem from a different sampling probability resulting from the study design. a non-participants rate of, perhaps, 10% might not produce a stronger bias unless the non-response more powerfully relates to the parameters of interest.[4] corresponding author: dler h. kadir, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq. e-mail: dler.kadir@su.edu.krd doi: 10.24086/cuesj.v4n2y2020.pp9-12 copyright © 2020 dler h. kadir. this is an open-access article distributed under the creative commons attribution license cihan university-erbil scientific journal (cuesj) received: jul 7, 2020 accepted: jul 25, 2020 published: aug 13, 2020 mailto:dler.kadir%40su.edu.krd?subject= kadir: likelihood approach for bayesian logistic weighted model 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 9-12 inverse probability weighting (ipw) an ipw method will directly model the missingness instead of modeling missing data observations.[5] ipw is occasionally termed “inverse propensity weighting.” when a probability score is projected for all subjects, the interest covariate observed values are weighted through the inverse of the relevant probability scores. response probability is able to be modeled with logistical regression models and uses the inverse of the probability scores as one of the factors of adjustment. the ipw adjustment permits a greater number of variables to be employed for predicting non-responses. the more appropriate variable set needs to be used to discover the model, which is the best fit for predicting non-response. this results in a “smooth” adjustment factor distribution, with no need for choosing an arbitrary cut-point.[6] yet, the ipw may possess an extreme value, causing an adjustment factor which might possess a highly covariate weight, and thus, a high covariate weight-adjusted estimates. this issue can be resolved by trimming the adjustment factor or trimming nonresponse adjusted weights. however, such remedies might increase bias possibilities. weighted posterior distributions from a bayesian perspective, we can gather the posterior distribution π(|x) through the combination of two types of information concerning the random variable . one source is given by the observed data which are summarized by the likelihood function, and the other information source is the previous information regarding its distribution π(). weighted posterior distributions can be defined through the replacement of the likelihood function by its ipw counterpart, as discussed in the previous section. the weighted posterior distribution is as follows: � � � � �ipw ipwx l x| |� � � � �� � �� (1) next, we propose ipws evaluated as in the above equation of the form ˆ ˆipw ( ) pw( ; , )θ=i i ipw nx x f . it is apparent that from the part of lipw (x|), we can obtain first-order property of the actual function likelihood under the model assumption; therefore, this has validity for bayesian estimates in a standard manner. one benefit of weighting is that it uses other pseudolikelihood functions, leading to posterior distributions which belong to the same family of those obtained through the use of the genuine likelihood function. therefore, the weighted posterior distributions vary from the genuine posterior distributions for the estimated values. it can seem that there is a conflict between the method and a proper bayesian perspective. this is because the weighted likelihood function is not immediately driven by a probabilistic model; rather, it is driven by adaptive weights. the data still tell a story; however, some values are not consistent with the required models, and we are unable to simply delete outliers, yet it still contributes to the posterior estimate.[7] preterm data for using logistic regression the data were collected from the neonatal intensive care unit at erbil maternity hospital from 2012 to 2017. in total, 570 babies (288 males and 282 females) were born very preterm. we have considered the infants born before 28 weeks. the mental development index approach was used for assessing the cognitive development of those born very preterm. almost half of the information for the babies was missing, meaning that we do not know whether they have cognitive development issues. now let i1 p 0  =   i if the data was collelected with probability r if the there is no response (2) • the procedure, therefore, involves: fitting a binary logistic regression, responding to the research under observation (1 if observed, 0 if not) with the more appropriate variable set as an explanatory variable. • obtaining the fitted probability for all infants, p i , i ∈(1,…,n) • calculation of the ipw for all infants ipw i = 1/p i ) and uses ipw to fit weighted bayesian logistic regression models using winbugs software. through the assumption that the posterior weighting model is correct, we can obtain consistent parameter estimates to know the effect of the outcome model. yet, the major issue in weighted data analysis is that the weight is not representative of the actual subject number; however, only an expected number might be applicable if the statistical weight features every detail regarding the sampling probability. identical samples appear from simple random sampling (whereby every individual of the same sizes is able to be sampled with an equal probability).[4] an additional issue with using ipw is where a missingness predictor distribution in full cases varies from incomplete cases. the ipw will then greatly vary since complete case analyses, where the missingness predictor observation is nearer the center of the observation distributions in the incomplete case which might obtain a larger weight. this will, therefore, cause a larger standard error.[8] when the weight can account for the missing data, parameters must be predicted. the complete case data variance estimator makes the assumption that the weight is known and ignores any uncertainty in estimations about them.[9] seaman et al. recommend the use of sandwich estimators in accounting for uncertainties in the weights. in reality, the true asymptotic uncertainty is frequently more when a true weight is utilized than when they are estimated. thus, ignoring uncertainties in a fixed weight might cause a standard error.[8] here, the weights value was altered in all iterations using markov chain monte carlo. the weight calculated from the variable weights sampled from posterior distributions instead of being obtained from a fixed value. therefore, in all iterations of the outcome models, various weights values were gained. ignoring any uncertainty between variable and fixed weights was examined to determine if there were any issues. consequently, uncertainties were included in the weight value using variable weights. in addition, weights were standardized (and multiplied by the number of observations/total number of the complete population). this results from the total of the weights being equal to the sums of the sample sizes. if the weights are not standardized, the sum of the weight is then equal to the entire kadir: likelihood approach for bayesian logistic weighted model 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 9-12 population instead of the total amount of observations, meaning that uncertainty (i.e., standard deviation and standard error) in the model without standardization will be underestimated. thus, in this project, the weight is standardized. in these analyses, complete cases are weighted by the inverse probability of there being complete cases. two logistic regression models were operated at the same time for both outcome and response. a covariate of mother birth age, sex, gestational age, and birth weight z-scores was used in the response model. let, r i denotes the outcomes (response (infants where the developmental questionnaire was responded to)/ non-response [infants where the developmental questionnaire was not responded to]). the outcome was modeled with the assumption of the bernoulli distribution. r i ~ bernoulli(q i ) (3) where, 0logit 1   = +  −  i i q a ax q when statistical models for weight have been recognized, it can then be used in developmental delay model analyses to be run alongside the dataset. nonetheless, weighting is vital and, thus, the amount of information is required to account for the weight relying on particular parameters being considered. inference is normally changed by multiplying the contributions of every infant to a statistic by its statistical weight. for modeling outcome and weight, each individual’s outcome variable value was multiplied by the individual’s weight. ipw for each of the infants is calculated using 1/probability of responses; the ipw was then standardized. therefore, the likelihood functions created in winbugs and is thus: * (1 )= −y ipw y* ipwil p p (4) p is the probability of babies surviving with developmentally delay issues, y is the outcome variable, and ipw is the ipw. five hundred seventy infants featured in the response model. yet, only 235 babies were used in the outcome model analysis (those babies known to be alive). we have used ipws as adjustment factors for babies about which we do not have cognitive developmental delay information. we put weights on the likelihood function using winbugs software. we repeated the analysis using frequentist logistic regression using variable weights. we obtained greater precision in results and standard deviation of parameter estimates as it is shown in table 1, which are less in the posterior weighted model in comparison with frequent analysis. discussion we have developed a likelihood-based approach to place weights which were calculated from response models into outcome models using logistic regression. it is noticed that unweighted models produce biased results. we have realized that a weighted model would provide more precise results in terms of odds ratio and uncertainty. ipw is one of the many available approaches for reducing bias using complete case analysis. here, a complete case is weighted by probability inverse of complete cases. even though weighting is often used in designing and analyzing surveys, using it in analyses of missing data not as well recognized, as the ipw fact parameter estimates can be inefficient in regard to probability-based analysis.[10] the resulting estimate is frequently sensitive to the exact type of models for the response probabilities.[11] in reality, the greatest concern of the data analysis is the efficacy of the ipw approach regarding the likelihood approach. although some methods have been suggested for obtaining more efficient and robust estimates, this approach has not yet been effectively developed to handle more than one situation. alternative methods could have been used in this analysis, for example, doubly robust ipw and multiple imputation. in addition, one disadvantage of this paper is the assumption that the weight is known; we have ignored uncertainties in the weight. to calculate ipw estimator standard error, methods like robust standard error created by weighted models could have been used. other methods such as bootstrapping, sandwich, resampling, and jackknife methods are also possibilities, although these methods require intensive computations.[4,12] references 1. d. wolke, b. sohne, b. ohrt and k. riegel. follow-up of preterm children: important to document dropouts. lancet, vol. 345, no. 8947, p. 447, 1995. 2. s. johnson, s. e. seaton, b. n. manktelow, l. k. smith, d. field, e. s. draper, n. marlow and e. m. boyle. telephone interviews and online questionnaires can be used to improve neurodevelopmental follow-up rates. bmc research notes, vol. 7, p. 219, 2014. 3. d. b. rubin. inference and missing data. biometrika, vol. 63, no. 3, pp. 581-592, 1976. 4. m. hofler, h. pfister, r. lieb and h. u. wittchen. the use of weights to account for non-response and drop-out. social psychiatry and psychiatric epidemiology, vol. 40, no. 4, pp. 291299, 2005. 5. l. lazzeroni, n. schenker and j. taylor. robustness of multipleimputation techniques to model misspecification. united states: proceedings of the survey research methods section, american statistical association, pp. 260-265, 1990. 6. b. l. carlson and s. williams. a comparison of two methods to adjust weights for non-response: propensity modeling and weighting class adjustments. united states: proceedings of the annual meeting of the american statistical association, 2001. 7. c. agostinelli and l. greco. weighted likelihood in bayesian inference. new york: proceedings of the 46th scientific meeting table 1: posterior odds ratio and the standard deviation gained from the binary model using variable weights estimates of parameters odds ratio sd 95% credible interval constant 0.54 0.11 0.34, 0.79 gestational age (centered)a 0.79 0.06 0.67, 0.94 mother’s age 0.987 0.02 0.91, 1.02 birth weight z-score 1.05 0.04 0.97, 1.14 female 0.52 0.18 0.27, 0.98 agestational age centered around 28 weeks kadir: likelihood approach for bayesian logistic weighted model 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 9-12 of the italian statistical society, 2012. 8. s. r. seaman and i. r. white. review of inverse probability weighting for dealing with missing data. statistical methods in medical research, vol. 22, no. 3, pp. 278-295, 2013. 9. s. r. seaman, i. r. white, a. j. copas and l. li. combining multiple imputation and inverse‐probability weighting. biometrics, vol. 68, no. 1, pp. 129-137, 2012. 10. d. clayton, d. spiegelhalter, g. dunn and a. pickles. analysis of longitudinal binary data from multiphase sampling. journal of the royal statistical society, vol. 60, no. 1, pp. 71-87, 1998. 11. r. j. little and d. b. rubin. statistical analysis with missing data. chichester: wiley, p. 5, 1987. 12. l. h. curtis, b. g. hammill, e. l. eisenstein, j. m. kramer and k. j. anstrom. using inverse probability-weighted estimators in comparative effectiveness analyses with observational databases. medical care, vol. 45, no. 10, pp. s103-s107, 2007. . 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 research article cooperating newton’s method with series solution method for solving system of linear mixed volterra-fredholm integral equation of the second kind talaat i. hasan* department of mathematics, college of basic education, salahaddin university, erbil, iraq abstract in this paper, for the 1st time, we use newton’s method with series solution method (ssm) for solving system of linear mixed volterrafredholm integral equations of the second kind (slmvfie-2). in this work, we use a new technique for studying the numerical solutions for slmvfie-2 which is newton’s method with ssm, first solving this system using ssm and after that cooperation newton’s method with ssm, suggesting an algorithm for the technique. the new results are achieved and some new theorems have proved for convergence of the method, several numerical examples are tested for illustrating the usefulness of the technique; the numerical results are obtained and compared with the exact solution, computing the least square error, and running times which are criterion of discussion. for programming the technique, we use general matlab program. keywords: newton’s method, series solution method, system of linear mixed volterra-fredholm integral equations of the second kind, taylor series introduction integral equations have been one of the significant and principal instruments in various areas of sciences such as applied mathematics physics, biology, and engineering. on the other hand, it has many applications in different areas of science, involving potential theory, electricity, and quantum mechanics.[1,2] in the past 20 years ago, many kinds of problems in integral equation making from various phenomena in applied mathematical physics. the mixed volterra-fredholm integral equations can be constructed form ordinary differential equations with changed argument,[3-5] which arising from the theory of parabolic boundary value problems. in addition, this type of problems appears in various physical and biological problems.[6,7] recently, many kinds of integral equations constructed and reformulated; the approximating methods take an important role for finding the numerical solution for these classes of problems in integral equations. it has many advantages witnessed by the increasing frequency of the integral equations in literature and in many areas, because some problems have their mathematical representation appear directly.[8,9] then, for the reasons, many scientists applied different techniques for finding numerical solution of integral equations, maleknejad, in 2006, solved system of volterra-fredholm integral equations using computational method; chen, in 2013, studied the approximate solution for mixed linear volterra-fredholm integral equation. wazwaz, in 2011, solved volterra integral equation of the second kind using series solution method (ssm). young, in 2015, used newton’s raphson for finding the approximate solution of non-linear equations. hassan, in 2016, found numerical solution of linear system volterra-fredholm integral equation. saleh, in 2017, studied system of volterra-fredholm integral equation of the second type, for extending this work, the system of linear mixed volterra-fredholm integral equations of the second kind (slmvfie-2) is studying and finding the numerical result of it by cooperating the nm with ssm and using power functions u x xi m i m( ) ,= for m=0, 1, 2, 3,…q. some definitions and theorems in this section, discussing some definitions and theorem, which relate to the study. cihan university-erbil scientific journal (cuesj) corresponding author: talaat i. hasan, department of mathematics, college of basic education, salahaddin university, erbil, iraq. e-mail: talhat.hassan@su.edu.krd received: nov 13, 2018 accepted: dec 24, 2018 published: may 13, 2019 doi: 10.24086/cuesj.v3n1y2019.pp27-33 copyright © 2019 talaat i. hasan. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ hasan: cooperating newton’s method with series solution method 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 definition (1) we assume slmvfie-2 as follows: u x f x k x t u t dtdxi i j n ij a x ij j a b ( ) ( ) ( , ) ( )= + = ∑ ∫ ∫ 1  (1) for i=1, 2, 3,…r. where f i (x) and k ij (x,t) are analytic functions on domain h= {(x,t), a≤t<x≤b} such that u i (x) is the unknown functions. definition (2)[4,5] a real value function u(x) is called analytic if it has derivatives of all order such that the taylor series at a point b in its domain. u x u b k x b k k k ( ) ( ) ! ( ) ,= − = ∞ ∑ 0 converges to u(x) in the neighborhood of b. definition (3)[10] let f(x)=0 be the non-linear function and x 0 be initial solution to the exact solution and then the newton’s method is defined as the form x x f x f xn n n n = − ′− − − 1 1 1 ( ) ( ) , n=1, 2,…. which generating the sequence {x n }. using ssm for solving slmvfie-2 suppose that the numerical solution of slmvfie-2 be analytic function and can be written of the form u x u xi im i m m q ( ) ( )= = ∑ 0 for i=1, 2, 3,…r (2) substituting it in equation (1), we get α λ αip m q i m i ij j n ij im m q i mu x t f x k x t u t dtdx = = = ∑ ∑ ∑= + 0 1 0 ( ) ( ( )) ( , ) ( ) aa b a x i i i i i i iq i q i u x u x u x u x t f x ∫∫ + + + + = α α α α0 0 1 1 2 2( ) ( ) ( ) ... ( ) ( ( ))) ( , )[ ( ) ( ) ( ) .. + + + + ∫∫∑ = λ α α α ij ij i i a b a x j n i i i i k x t u t u t u t 0 0 1 1 1 2 2 .. ( )]+ αiq i qu t dtdx (3) from equation (3), in the right-hand side calculating the integrals for variables t and x. getting the system of (r×(q+1) equations. collecting and equating the terms of the same powers of u xi m( ) in both sides. using gauss elimination method, solving the resulting in equation (3) to obtain the values of a im such that the determinate of algebraic system does not equal to zero. by putting the values of a im in equation (2), we get the approximation solution of equation (1). the calculation more terms we use will enhance the level of accuracy of the approximate solutions. in this work, we use power functions u x xi m i m( ) = for m=0, 1, 2, 3,…q. new theorem (1) let v: b(a)→b(a) be a continuous linear integral operator on the banach space b(a) f i (x) be analytic function on [a,b] and k ij (x,t) be continuous on h={(x,t), a≤t<x≤b} where |k ij (x,t) u 1 (t)−k ij (x,t)u 2 (t)−|≤d i |u 1− u 2 | such that 0<d i <1, then vn is contractive mapping. proof: suppose that [ ] 0 0 ( ( )) ( ) ( , ) ( ) , 0, , p x i i ij i i iv u x f x k x t u t dtdx x p p 1.= + <∫∫ to prove that vn is contraction mapping for any u 1 (s), u 2 (s)  b (a) for sufficient large n. v u x v u x f x k x t u t dtdx f x k i ij xp i ij ( ( )) ( ( )) ( ) ( , ) ( ) ( ( ) 1 2 0 1 0 − = + + ∫∫  00 2 0 0 1 2 0 xp ij xp x t u t dtdx k x t u t u t dtdx ∫∫ ∫∫= − ( , ) ( ) ) ( , )( ( ) ( ) v u x v u x k x t u t k x t u t dtdx d ij ij xp i ( ( )) ( ( )) ( , ) ( ) ( , ) ( )1 2 1 200 − = − ≤ ∫∫ [[ ] p u ui 2 1 22 − by suppose |k ij (x,t)u 1 (t)−k ij (x,t)u 2 (t)|≤d i |u 1 −u 2 |, then we get v u x v u x d u u dtdx d u u dtdx d p i xp i xp i ( ( )) ( ( )) [ 1 2 1 2 00 1 2 00 − ≤ − = − = ∫∫ ∫∫ ii u u 2 1 22 ] − now, v u x v u x v f x k x t v u t dtdx v i ij xp 2 1 2 2 0 1 0 ( ( )) ( ( )) ( ( )) ( , ) ( ( )) [ − = + ∫∫  (( ( )) ( , ) ( ( )) ] f x k x t v u t dtdx i ij xp − ∫∫ 00 2 = −∫∫ k x t v u t v u t dtdxij xp 0 1 2 0 ( , )( ( ( )) ( ( ))) v u x v u x d d p u u dtdx d p u i i x i p i i 2 1 2 2 0 2 1 2 0 2 2 1 2 2 ( ( )) ( ( )) [ ] [ ] − ≤ − ≤ − ∫∫ uu dtdx d p u u xp i i 2 00 2 4 1 24∫∫ ≤ −[ ] using mathematical induction, we obtain v u x v u x d p u u n n i n i n n ( ( )) ( ( )) [ ] .1 2 2 1 22 − ≤ − since by suppose 0<d i <1 and p i <1, then we obtain 0<d i p i <1 and 0 12< <d pi n i n therefore 0 2 1 2 < <d p i n i n n [ ] then, by definition of contraction vn is contraction on. hasan: cooperating newton’s method with series solution method 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 new theorem (2) suppose that the hypotheses of theorem (1) are holds; then, system of mixed volterra-fredholm integral equation of the second kind has a unique solution. proof: suppose that u 1 (x) and u 2 (x) be any two solution of slvfie-2. u x f x k x t u t dtdxi ij xp 1 0 1 0 ( ) ( ) ( , ) ( )= + ∫∫ and u x f x k x t u t dtdxi ij xp 2 0 2 0 ( ) ( ) ( , ) ( )= + ∫∫ using theorem (1), we get v u x v u x d p u u n n i n i n n ( ( )) ( ( )) [ ]1 2 2 1 22 − ≤ − since lim [ ] , n i n i n n d p →∞ = 2 2 0 then we obtain vn(u 1 (x))=vn(u 2 (x)), therefore u 1 (x)=u 2 (x) hence, slvfie-2 has a unique solution. algorithm for ssm • step 1: let the numerical solution of equation (1) as the form u x u xi im i m m q ( ) ( )= = ∑ 0 , for i=1, 2,…,r. • step 2: by substituting equation (2) in equation (1) and calculating the integrals of variables. • step 3: from both sides collect and equate the terms of the same power for m=1, 2, 3,…,q. • step 4: by solving the system of algebraic equations with unknown coefficients a im . • step 5: substitute the values of a im , in equation (2) for getting the numerical solution of slmvfie-2. numerical examples and results in this section, illustrating the technique from numerical examples. example (1): solve lsmvfie-2 u x e x e xt u t u t dtdxx x 1 2 1 1 2 0 1 0 2 1( ) ( ) ( )( ( ) ( ))= − + − + −∫∫ u x xe x e x x u t u t dtdxx x 2 3 1 3 2 1 2 0 1 0 2 2 3 8 3 ( ) ( )( ( ) ( ))= − + − + −∫∫ using ssm, where the exact solutions u 1 (x)=−2ex and u 2 (x)=−2xex? figure 2: (a) graphs of the exact u 1 (x)=−2sin(x) and the approximate solutions. (b) graphs of the exact u 2 (x)=−2cos(x) and the approximate solutions. (source: created by researcher) aq1 ba figure 1: (a) graphs of the exact u 1 (x)=−2ex and the approximate solutions. (b) graphs of the exact u 2 (x)=−2xex and the approximate solutions. (source: created by researcher) aq1 b a hasan: cooperating newton’s method with series solution method 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 solution: let u x u xm m m q 1 1 1 0 ( ) ( )= = ∑ then, using ssm, obtaining the values of coefficients a 10 =−2, a 11 =−2, a 12 =−1, a 13 =−0.3333, a 14 =−0.083333 and a 15 =−0.03333, for u x u xm m m q 2 2 2 0 ( ) ( )= = ∑ then, using ssm, obtaining the values of coefficients a 20 =0, a 21 =−2, a 22 =−2, a 23 =−1, a 24 =−0.33333 and a 25 =−0.08333 example (2): solve lsmvfie-2 �� ( ) ( ) sin( ) ( )(sin( ) cos( )) u x x x x x t t u dtdx x 1 2 2 0 0 2 2 2= − − + − ∫ ∫  �� ( ) ( ) cos( ) ( )(sin( ) cos( )) u x x x x t t t u dtdx x 2 10 0 2 2= − − + − ∫ ∫  using ssm, where the exact solutions u 1 (x)=−2sin(x) and u 2 (x)=−2cos(x)? table 1: comparison between the exact and numerical results x m exact solution of u 1 (x)=−2ex approximate value of u 1 (x) absolute error m+1 m+1 i i ie = u (x) u (x) exact solution of u 2 (x)=−2xex approximate value of u 2 (x) absolute error m+1 m+1 i i ie = u (x) u (x) 0.2 1 3 5 7 9 11 13 15 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.40000000 −2.44266666 −2.44280533 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 4.2805×10−2 10.3884×10−4 1.8298×10−7 1.2986×10−9 1.1653×10−12 6.7659×10−14 5.3362×10−16 3.6521×10−18 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.45344832 −0.48898815 −0.48856177 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 3.5611×10−2 4.2735×10−4 6.7527×10−7 2.5998×10−9 5.8432×10−11 3.4562×10−13 4.6783×10−15 2.3351×10−17 table 2: comparison between the exact and numerical results x m exact solution of u 1 (x)=−2sin (x) approximate value of u 1 (x) absolute error m+1 m+1 i i ie = u (x) u (x) exact solution of u 2 (x)=−2cos (x) approximate value of u 2 (x) absolute error m+1 m+1 i i ie = u (x) u (x) 8 π 1 3 5 7 9 11 13 15 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.78539816 −0.76211785 −0.76536743 −0.76536686 −0.76536686 −0.76536686 −0.76536686 −0.76536686 2.0031×10−2 1.5507×10−4 5.7028×10−6 2.6223×10−8 1.7146×10−10 2.2083×10−12 5.0679×10−14 3.6582×10−16 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −2.00000000 −1.84578731 −1.84776922 −1.84775903 −1.84775906 −1.84775906 −1.84775906 −1.84775906 1.5224×10−1 1.9716×10−3 1.0159×10−5 2.8005×10−7 4.8013×10−9 5.6301×10−11 3.9014×10−13 4.3382×10−15 table 3: comparison between the exact and numerical results x q exact solution of u 1 (x)=−2ex approximate value of u 1 (x) absolute error p p 1 1e = u (x) u (x) exact solution of u 2 (x)=−2xex approximate valueof u 2 (x) absolute error p p 2 2e = y (x) y (x) 0.2 1 3 5 7 9 11 13 15 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.44280551 −2.8698700 −2.4779392 −2.4424024 −2.44280875 −2.44280551 −2.44280551 −2.44280551 −2.44280551 4.2702×10−1 3.5134×10−2 1.4041×10−4 3.2468×10−6 5.8432×10−8 3.4562×10−9 4.6783×10−11 2.3351×10−13 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.48856110 −0.79987234 −0.4964455 −0.48553062 −0.48858445 −0.48856110 −0.48856110 −0.48856110 −0.48856110 3.1131×10−1 1.2214×10−2 3.0304×10−3 2.3342×10−5 6.3489×10−7 5.7741×10−9 4.5560×10−10 3.2981×10−12 hasan: cooperating newton’s method with series solution method 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 solution: let u x u xm m m q 1 1 1 0 ( ) ( )= = ∑ then using ssm, obtaining the values of coefficients a 10 =0, a 11 =−2, a 12 =0, a 13 =0.3333, a 14 =0, a 15 =−0.01666, and a 16 =0, for u x u xm m m q 2 2 2 0 ( ) ( )= = ∑ then using ssm, obtaining the values of coefficients a 20 =−2, a 21 =0, a 22 =1, a 23 =0, a 24 =−0.08333, a 25 =0, and a 26 =0.002777. newton’s method cooperate with ssm for solving slmvfie-2 since the numerical solution u x u xi im i m m q ( ) ( )= = ∑ 0 is analytic function, then using taylor series expansion at a point x 0 it can be written of the form u x u b k x b u b u b x b u b x b i i k k k i i i( ) ( ) ! ( ) ( ) ( )( ) ( ) ! ( = − = + − + − = ∞ ∑ 0 0 1 1 2 2 )) ( ) ! ( ) ...2 3 3 3 + − + u b x bi which is similar to u x u b k x b u b u b x b u b x b i i k k k i i i( ) ( ) ! ( ) ( ) ( )( ) ( ) ! ( = − = + ′ − + ′′ − = ∞ ∑ 0 1 2 )) ( ) ! ( ) ...2 3 3 + ′′′ − + u b x bi table 4: comparison between the exact and numerical results x q exact solution of u 1 (x)=−2sin (x) approximate value of u 1 (x) absolute error p p 1 1e = u (x) u (x) exact solution of u 2 (x)=−2cos (x) approximate value of u 2 (x) absolute error p p 2 2e = y (x) y (x) 8 π 1 3 5 7 9 11 13 15 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.76366864 −0.65484390 −0.78998921 −0.76887972 −0.76539865 −0.76536922 −0.76536686 −0.76536686 −0.76536686 1.1122×10−1 2.6323×10−2 5.2214×10−3 3.2213×10−5 1.1426×10−6 5.7892×10−8 3.4753×10−9 5.2253×10−11 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.84775906 −1.52331434 −1.84992540 −1.84790077 −1.84779887 −1.84775968 −1.84775906 −1.84775906 −1.84775906 3.2444×10−1 4.2234×10−2 2.5971×10−4 4.1812×10−5 6.2342×10−7 4.6638×10−8 5.6754×10−10 3.8932×10−11 table 5: comparison between the results which depending on lse and rt at appoint number of terms criterion of ssm for u 1 (x)=−2ex criterion of ssm for u 2 (x)=−2xex x q lse rt lse rt 0.2 3 2.5480×10−4 0:0:1.852 3.2361×10−4 0:0:1.852 5 3.6531×10−8 0:0:2.224 4.3452×10−8 0:0:2.224 7 2.8874×10−10 0:0:3.036 2.6674×10−10 0:0:3.036 9 6.4456×10−13 0:0:4.368 9.8678×10−14 0:0:4.368 lse: least square error, rt: running times, ssm: series solution method if we truncating it in the terms of order two, we get u x u b u b x bi i i( ) ( ) ( )( )= + ′ − and put h=x−b such that u i (x)=0, then we get h u b u b i i = − ′ ( ) ( ) then, we obtain this iteration formula: u x u x u x u xi m i i m i m + = − ′ 1( ) ( ) ( ) ( ( )) (4) for m=0, 1, 2, 3,… where m is the number of iteration. cooperating newton’s method with ssm for solving slmvfie-2 the main aim of this section is reformulating and cooperating nm with ssm for getting the approximate solutions of the problem. these proses are accelerating the convergence to exact solution for the problem. first solving slmvfie-2 using ssm. then, cooperating nm with ssm to obtain the new sequence u xi m m ( ){ } = ∞ 1 of approximate solutions for the problem using the relationship in equation (4). u x u x u x u x mi m i i m i m + = − ′ =1 0 1 2 3( ) ( ) ( ) ( ( )) , , , ,...for nm on ssm algorithm input: a, b, x, m, e for i=1 to q for m=0 to max hasan: cooperating newton’s method with series solution method 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 step 1: using ssm to obtain the numerical solutions u x u xi im i m m q ( ) ( )= = ∑ 0 end for end for for i=1 to q for m=0 to max step 2: getting the new sequence using the relationship in equation (4). step 3: compute the absolute error e u x u xi m i m i + += −1 1( ) ( ). if e ei m+ <1 go to output end if end for end for output: the numerical results of the technique u xi m+1( ) and ei m+1 . table 7: comparison between the results for cooperating mn with ssm which depending on lse and rt at appoint number of terms criterion of ssm for u 1 (x)=−2ex criterion of ssm for u 2 (x)=−2xex x q lse rt lse rt 0.2 3 3.5573×10−6 0:0:1.852 5.3428×10−6 0:0:1.852 5 4.2582×10−10 0:0:2.224 3.6680×10−12 0:0:2.224 7 2.4006×10−12 0:0:3.036 6.2314×10−13 0:0:3.036 9 5.8062×10−15 0:0:4.368 7.7451×10−16 0:0:4.368 lse: least square error, rt: running times, ssm: series solution method tables 8: comparison between the results for cooperating mn with ssm which depending on lse and rt at a point number of terms criterion of ssm for u 1 (x)=−2sin (x) criterion of ssm for u 2 (x)=−2cos (x) x q lse of ssm rt lse rt 8 π 3 4.4116×10 −5 0:0:1.7734 3.5411×10−4 0:0:1.7734 5 3.2984×10−8 0:0:2.1453 4.6905×10−7 0:0:2.1453 7 5.5205×10−12 0:0:2.9784 4.8834×10−11 0:0:2.9784 9 6.8752×10−16 0:0:3.5621 5.9672×10−15 0:0:3.5621 lse: least square error, rt: running times, ssm: series solution method table 6: comparison between the results which depending on lse and rt at a point number of terms criterion of ssm for u 1 (x)=−2sin (x) criterion of ssm for u 2 (x)=−2cos (x) x q lse of ssm rt lse rt 8 π 3 6.3489×10 −3 0:0:1.7734 2.0036×10−2 0:0:1.7734 5 5.7823×10−6 0:0:2.1453 3.4437×10−6 0:0:2.1453 7 3.5539×10−9 0:0:2.9784 5.7284×10−9 0:0:2.9784 9 5.6127×10−11 0:0:3.5621 4.3582×10−12 0:0:3.5621 lse: least square error, rt: running times, ssm: series solution method numerical examples and results in this section, illustrating the technique from numerical examples. test example: solving example (1) using new technique. solution: by cooperating nm with ssm, we get the following results in table 1. test example: solving example (2) using new technique. solution: by cooperating nm with ssm, we get the following results in table 2. conclusion in this paper, the approximate solutions of slmvfie-2 by cooperating newton’s method with ssm for solving slmvfie-2 are discussed. several examples were solved for illustrating the technique and numerical results were achieved. after testing the numerical technique and its algorithm on numerical examples, the results are written in tables 1-4 for ssm and cooperating newton’s method with ssm. when the number of terms m increasing, the absolute errors are decreasing and the convergence is satisfactory. in addition, the least square error and running times are written in tables 5-8 which are criterion hasan: cooperating newton’s method with series solution method 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 27-33 of discussion for the technique which depending on the number of terms m. then, we conclude that the cooperating newton’s method with ssm is faster and better than ssm for getting the approximate solutions. therefore, this technique is successful for getting the numerical solutions for this class of problems. references 1. z. chen. “an approximate solution for a mixed linear volterra fredholm integral equation”. applied mathematics letters, vol. 25, pp. 1131-1134, 2013. 2. m. paripour and m. kamyar. “numerical solution of nonlinear volterra-fredholm integral equations by using new basis functions”. communications in numerical analysis, vol. 1, no. 17, pp. 1-12, 2013. 3. h. ibrahim, f. attah and t. gyegwe. “on the solution of volterrafredholm and mixed volterra-fredholm integral equations using the new iterative method”. applied mathematics, vol. 6, no. 1, pp. 1-5, 2016. 4. a. d. polynine and a. v. manzhivove. “handbook of integral equations”. 2nd ed. united states of america: crc press, 2008, p. 1238. 5. a. m. wazwaz. “linear and non-linear integral equations methods and applications”. berlin heidelberg: springer-verlag, 2011. 6. b. g. pachpatte. “on mixed volterra fredholm type integral equations”. journal pure applied mathematics, vol. 17, no. 4, pp. 488-496, 1986. 7. s. saleh, t. i. hasan and n.a. sulaiman. “fixed point and its improvement for the system of volterra-fredholm integral equations of the second kind”. journal of matematika, vol. 33, no. 2, pp. 191-206, 2017. 8. k. maleknejad and m. r. yami. “a computational method for system of volterra fredholm integral equations”. applied mathematics and computation, vol. 183, pp. 589-595, 2006. 9. m. rahmani. “integral equations and their application”. great britain: athenaeum press ltd., 2007. 10. t. young and j. m. martin. “introduction to numerical method and matlab for engineers”. athens, oh: ohio university, free software foundation, 2015. author queries??? aq1:kindly cite figures 1 and 2 in the text part . 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 research article a new scheme for removing duplicate files from smart mobile devices: images as a case study ammar asaad, ali adil yassin alamri* department of computer, education college for pure sciences, university of basrah, 61004 basrah, iraq abstract the continuous development of the information technology and mobile communication world and the potentials available in the smart devices make these devices widely used in daily life. the mobile applications with the internet are distinguished simple, essay to use in any time/anywhere, communication between relatives and friends in different places in the world. the social application networks make these devices received several of the duplicate files daily which lead to many drawbacks such inefficient use of storage, low performance of cpu, ram, and increasing consumption battery. in this paper, we present a good scheme to remove from the duplicate files, and we focus on image files as a common case in social apps. our work overcomes on the above-mentioned issues and focuses to use hash function and huffman code to build unique code for each image. our experiments improve the performance from 1046770, 1995808 ns to 950000, and 1981154 ns in galaxy and huawei, respectively. in the storage side, the proposed scheme saves storage space from 1.9 gb, 1.24 gb to 2 gb, and 1.54 gb, respectively. keywords: duplicating images, hash code, mobile device, performance, storage management introduction one of the key things that can help us to build a scheme to get rid of this issue is the features extracted from images. it is difficult to observe and understand these features by a human{shyu, 1998 #2343}. these features divide into two types global and local.[1] image matching is the method used either global features which describe the whole image and can describe a whole image by only one vector, or local features, which focus on important details in the image and it is more powerful. to apply both methods, we need to deal with the content of the images and extract information from them; this means that both methods consume time to treat with images and need to have hardware that has ability and efficiency to get these features. this leads us to the fact that the features extraction in the mobile device will consume a lot of time, random memory, and cup time thus increasing the consumption of the battery; as a result, effect on daily use for the user.[2] hence, the process of finding the best and most efficient methods of retrieval and matching files is the most important research topics in the era of mobile applied technology.[3] matching is one of the essential tasks that are used to remove duplicated files. mostly, it is selected the appropriate features to reduce the computational time, and it denotes to find the same file/s (image) save(s) in a database that is/are same to an input file. matching is one of the indispensable processes in several applications.[4] in the past decade, mobile technology and internet have become a hotspot in scientific research and daily life.[5] social media consider a common element between internet and mobile devices; it plays the main role to keep in touch with our colleagues/relatives and discovering new persons.[6] in the past few years, social network was not used in mobile devices in widely manner, but with the rapid development in the internet and mobile technologies, the social media become one of the important factors in human life.[7] at present, there are several social networks that work in mobile devices and personal computers such as facebook, whatsapp, twitter, viber, imp, and others called mobile apps.[5] the users of these apps exchange many types of files such as photos, audios, and videos. ultimately, these files are stored in duplicity way led to lose of storage space of the mobile device. in this paper, we focus on the removal of duplicate images issued from social media apps to mobile device based on proposed image matching scheme that suits with mobile environment and deals of the whole image. the main objectives of our purpose scheme are as follows: corresponding author: dr. ali adil yassin alamri, department of computer science, education college for pure science, university of basrah, basra, 6100, iraq. e-mail: en.uobasrah.edu.iq/ali.yassin@uobasrah.edu.iq received: apr 08, 2019 accepted: apr 14, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp5-13 copyright © 2019 ammar asaad, ali adil yassin alamri. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 1. our work can eliminate this repetition of images saved in mobile’s storage from different social media apps with high performance 2. our scheme is offered cost of consumption energy of battery and less used for both ram and cpu 3. we use a hash function algorithm (message-digest algorithm 5 [md5]) and huffman code algorithm to build unique code for each image which is used to delete all reputation images 4. our proposed scheme is applied on real-word data using two types of mobile devices (huawei cun-u29, galaxy a5 [2017] sm a520f) and we achieve good results to increase storage space and improve the processing time. organization of the paper, section 2 shows the relevant literature on the topic of image matching. section 3 refers to the primitive tools of the proposed scheme while section 4 explains the proposed scheme. section 4 focuses on experimental results and discussions. the paper is concluded in section 5. related works this section briefly discusses duplication files in a mobile device, refers the several studies of deduplication schemes as follows. attractively, there are many fields to use deduplication such as paging files, image redundancies, and memory sectors.[8,9] this field has been applied primarily on data backup services such as microsoft office and google drive.[10] in addition, some researches in the field of removing duplicate files focus on multiple deduplication schemes.[10] at present, with rapid development in cloud computing and mobile device, there is a platform to support low latency network access and extra storage called mobile cloud computing (mcc).[10] besides, mcc enables to support mobile in extra storage based on payas-you-go within cloud computing principle, but this technique leads to pay cost by the customer,[9,11] in the same time, the duplicate files are stayed in the storage of the cloud. storer et al.[12] suggested a good scheme eliminated redundancies files depending on its type. however, the method of deleting duplicate files is selected manually. it works well with known files and suffers to deal with new files that do not exist in the index file of their work.[10] haustein et al.,[13] the patent is about selecting a deduplication method depending on the file type and deduplication rate, where the redundant deletion is done by the server. they used the ratio of duplication files and then the implemented scheme operates based on this ratio to deduplication files from the server. their works had many drawbacks by working in server-side and they proposed in a theoretical manner without executed in real-world data. widodo et al.[10] presented a scheme to reduce energy consumption and the amount of data by detection duplicate files. this scheme suffers from low of duplicate detection performance and deduplication throughput for a few files in the beginning. although their works focused on using cloud storage to duplicate files, the duplications files are stilled in devices’ storage. there are several researchers focused on detection of duplicate data using hash functions such as md5 and sha-256.[14,15] these schemes cannot face the collisions growing the size of the signature increases as well as the processing time is increased with the size of signatures.[16] in this paper, we present a scheme that has been applied as mobile apps to remove the duplicate files imported to mobile by social media applications such as facebook, whatsapp, viper or mobiles camera. the image files are one of most commonly used in social networks world, therefore, we focus on the image files as a case study in our experimental results relied on two types of mobile devices “samsung and hawawi.” our work focus to use hash function and hffuman code to build unique code for each image and overcomes the drawbacks of collisions grows as the size of the signature increases. however, the scheme generates a good search index file used to insert an image or ignore in the repeating case. moreover, our scheme is offered cost of consumption energy of battery and less used for both ram and cpu of the mobile device. in addition, our work can manage the storage of mobile and cloud storage (based on some of google’s application such as google drive and google photos) by removing duplicate images. primary tools in this section, we demonstrate the main tools used in our work to achieve the vital goals of the research. huffman code huffman coding considers one of the most important algorithms proposed and published by david a. huffman.[17] it is based on the frequency of the amount of a data item. huffman algorithm works using the tree data structure to generate an optimal binary tree called huffman tree to acquire encoding form.[17,18] the technical working of this algorithm uses a lower number of bits to encode the input data that exist more frequently.[19] practically, huffman considers an easy-touse algorithm because of its simpler mathematical computation to obtain the several parameters.[20] huffman coding algorithm consists of two phases: building binary tree and generating code. the result of the first phase is constructed through the occurrence frequency of each item. to achieve the code of the huffman algorithm, each branch has a set (0 or 1) based on its direction left (0) or right (1).[19,21] figure 1 explains the mechanism work of huffman code algorithm. figure 1: explain the huffman code algorithm asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 md5 (hash) md5 is one of the most algorithms worked to compress data files such as images, videos, and audios. the output of this algorithm is a fixed value (128-bit) when the algorithm is implemented once or many times on the same data.[22,23] this algorithm was developed by the rivest[24] and considered simple to compute the md5 value. the md5 processes 512-bit blocks and breaks into 32-bit words. it includes 64 rounds. the md5 steps as follows:[8] the first process is to add the bits to the block to equal the value of (448 mod 512), start with 1, followed by 0’s, the second process is adding the length of the message to the value (448 mod 512), the third step set the md5 register values (a, b, c, and d), each one 32 bits (hexadecimal number), and the fourth step is the heart of the algorithm; its four pressure functions each of these pressure functions have logical operations [figure 2]. proposed protocol our proposed work consists of two phases: creating index file and removing daily images. the first phase focuses on detecting and creating an index file (if see table1 explain the notification of symbols) without duplicate images, while the second one is implemented to remove images on a daily basis. our work focuses on a vital part of smartphone when receives daily hundreds of photo files, video clips, audio, and others. there are many of these files repeated in mobile’s storage. the proposed scheme plays the main role in deleting duplicated photos that caused to lose a large space of storage. each a new photo checked inside if (index file ), it removes from storage when it saved in if previously. otherwise, a new photo inserts to the if. as a result, the mobile device works more efficiently and performance as well as reduce energy consumption and disposal of redundancy of photos in the memory. the limited of hardware components of the mobile device makes the proposed scheme dealing with photos in light and high-performance manner. therefore, we used md5 hash function and huffman code to build unique code for each photo. figure 3 explains the main aims of our work. figure 2: algorithm mechanism and steps table 1: notification of symbols symbol description md mobile device such as smartphone and tablet… ms mobile’s storage (external, internal) img i image that is saved inside md s set of images (img i ) p i img i ’s path p set of img’s path size() function to get size of each img if index file has path of img and its details code 1 number created by convert hash to assci code and compute code by multiply assci by position code 2 number obtained by huffman coding and convert to decimal number h md5 hash function h i hash code for img i uc i final code resulted by xor code 1 and code 2 delduplicate() function to delete all duplicate img and save one only search() function to detect duplicate img dfds() function to delete img from device storage deleterowif() function to delete a row from if save() save details in index file (uc i , p i , size (p i ),) getimg() get img i based on its p i getimages() get all p i asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 creating index file phase in the era of information technology, all smartphones have several folders related to photos which received from different mobile applications such as whatsapp and facebook. therefore, there are many duplicated images saved in mobile storage. these duplicated images have several drawbacks such slow mobile completion, the difficulty of communication, loss of flexibility to handle more than one application and slow the mobile response of the received orders from another device as well as power consumption due to slow performance. however, this phase is used once to create an index file and removing all duplicate images from mobile storage. the main steps of this phase are as follows: a. generate set of all mobile’s images s = {img 1 , img 2 ,… img n } and their paths p = {p 1 ,p 2 ,…p n } based on extension of images file such as jpeg, bmp. b. create unique code for each image (img i ∈ s) based on hash (eq.1 and eq2.) and huffman code functions as follows. h i = h(img i ) (1)    n 1 i i j=1 code = ascii(h (j)) * position h j (2) • apply huffman code function • code 2 = huffman(hi), • unique code = code 1 xor code 2 (3) • figure 4 shows a code generation technique. c. create index file (if) that each record consists of uc i , p i , size(img i ). algorithm 1 demonstrates the mechanism of generating if to all mobile device images, [figure 4]. algorithm 1 of creating index file input: {pi} 1 ≤i 1; where n is number of p output: index file compute h 1 based on equation (1) compute uc 1 based on equation (2,3) save (uc 1 , p 1 , size(p 1 )) for i=2 to n do table 2: some of the rules have been implemented on images entered into md rules details 1 this rule describes receiving a picture through more than one program, and this image is in if. all copies are deleted 2 this rule describes receiving a picture through more than one program, and this image is not found in if. a single copy is saved and the rest of the copies are deleted 3 this rule describes receiving a picture through a single program and this image is not found in if. this version is saved 4 this rule describes receiving a picture through a single program, and this image is found in if. this version is deleted 5 this rule explains the reception of the same image more than once through a single program, and this image does not exist in if. hence, one copy is saved and the rest is deleted 6 this rule explains the reception of the same image more than once through a single program, and this image is in if. all copies are deleted figure 4: demonstrates the process of generating unique code figure 3: (a) receiving daily images, (b) applying our work, and (c) the results of work a b c asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 compute hi as in equation (1) compute uci as in equation (2, 3) if (search (uci) =true) dfds (pi) else save (uci, pi, size(pi)) end for end algorithm 1 of creating index file. removing daily images phase all smartphones receive many messages on a daily these messages may be pictures or audio clips or video files or other and thus affect the storage space of the device as well as the performance of the device. our scheme helps us get rid of the repetition for the images and thus let the machine work better. the removing of the repeated images depends on the code of image which has been computed in the first phase [figure 5]. the main steps of this phase are as follows: a. generate set paths p’ = {p’ 1 , p’ 2 ,…p’ n } of mobile’s image that incoming to device after creating an index file. these images have been imported to the device in daily. b. create set of unique code (uc’ = {uc’ 1 , uc’ 2 ,…uc’ n }) for each a new image based on the same procedure in step 2 of create index file phase. c. check uc’ i ∈ if(uc); if the result is false then extract a new record (p’ i , uc’ i ) of img’i added to if; otherwise, remove img i from mds. tables 2 and 3 and figure 3 denote to the table 3: some scenarios for receiving images and how to process them img whatsapp viper messenger download telegram if add to if non add and removed from mds rule apply √ √ × √ √ √ × √ rule 1 √ √ × √ √ × √ × rule 2 √ × × × × × √ × rule 3 × √ × × × √ × √ rule 4 √ × × × × × √ × rule 5 × √ × × × √ × × rule 6 asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 mechanism of processing (adding/removing to if) images which incoming to a mobile device (md). algorithm 2 of adding new images input: {pi} 1≤i≤n; where n is number of p, output: add new images to index file for i=1 to n do compute h i as in equation (1) compute uc i as in equation (2, 3) if (search (uc i ) =true) dfds (p i ) else save (uc i , p i , size(p i )) end for end algorithm 2 of adding new image experimental results in this part, we evaluate the proposed scheme and use the images of the mobile device to guarantee the reproducibility of practical results. the experiments are implemented using android studio 3.3.1 running on windows 10 64–bit operating system with an intel core i5-2450m cpu at 2.50ghz 250ghz, 8 gb ram, and 2.4 ghz cpu, and mobile huawei cun-u29 ram 956.52 mb internal storage 4.15 gb with external storage 8 gb, battery 2200 mah, android version 5.1 (lollipop_mr1), and another mobile galaxy a5 (2017) sm-a520f, android version 8.0 (oreo), battery 3000 mah, processor arm64-v8a 8 core, and total ram 2815 mb. our results have passed in many steps as follows. processing time calculation of index file creation we used 2600 images to create index file (if) in huawei cun-u29 mobile device. after that, we divided images into n groups (we obtained 13 groups based on the total number of table 5: add and remove received images day huawei cun-u29 galaxy a5 (2017) sm-a520f received add remove received add remove 1 37 7 30 63 13 50 2 27 7 20 18 8 10 3 18 8 10 12 8 4 4 19 15 4 20 12 8 5 12 8 4 50 30 20 figure 5: explaining the main steps of the proposed scheme table 4: processing time of index file creation group huawei cun-u29 galaxy a5 (2017) sm-a520f size of group (kb) time of processing (nanosecond) size of group (kb) time of processing (nanosecond) 1-200 585080 32421400159 281353 10395266647 201-400 191979 11017601159 134797 5059068621 401-600 19271 1432455537 318393 11132803762 601-800 22118 1641043162 178462 6512452069 801-1000 18069 1359571455 132321 4641873378 1001-1200 16480 1335055147 12960 758972849 1201-1400 88118 5374266081 6760 401045846 1401-1600 202894 11694264538 1601-1800 34670 2326661993 1801-2000 5332 578131154 2001-2200 4897 540429308 2201-2400 69854 3991726847 2401-2600 81739 4908648533 asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 figure 8: the mobile apps of our proposed scheme images and each group consists of 200 images). in the other side, we implemented our work to another mobile device named galaxy a5 (2017) sm-a520f contained 1310 images to construct index file (if). table 4 and figure show the performance of the calculation of index file creation in both devices. daily phase apply we implemented our proposed scheme in real-word data saved on a mobile device that has properties as refereed in the previous section. figure 6 shows the main operation in this phase. the proposed scheme has been implemented as mobile apps figure 8 in several days to apply the main operations such as detecting and removing redundancy figure 7: processing time of index file creation (a) galaxy a5 (2017) sm-a520f device for 1310 images (b) huawei cun-u29 device for 2600 images aq1 a b figure 6: (a) it shows the process of receiving the image, generating the code, checking if and making the decision, add of image in if, (b) it shows the process of receiving the image from social media apps, generating the code, checking if, and making the decision to delete image a b asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 figure 11: it shows the time spent searching for one image in the middle and one at the end. before and after deleting duplicate files. in the two devices aq1 images, adding new images. table 5 and figure 8 describe the central operations for 5 days. furthermore, we test the mobile storage based on our proposed work and notice good results as figure 9 displayed the storage of mobile devices (galaxy a5 [2017] sm-a520f, huawei cun-u29) before and after implementing our proposed scheme. moreover, we calculate the time of retrieved image on devices as in figure 10. figure 10: (a) before and after apply for our work at galaxy a5 (2017) sm-a520f and (b) before and after apply for our work at huawei cun-u29 a b figure 9: add and remove images received (a) huawei cun-u29 device for 5 days received images (b) galaxy a5 (2017) sm-a520f device for 5 days received images a b asaad and alamri: a new scheme for removing duplicate files from smart mobile devices 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 5-13 conclusions one of the most important challenges for smartphone users is to keep the device working as high as possible. low storage space is the most important issue. we solved this issue by the used hash function (md5), huffman code to generating unique code for each image and save this code in image in the index file and use it to rid of a duplicate from device’s storage. removing images have the same code and maintain one copy. the proposed scheme get a good result in remove this duplicate images in fast. we applied it at two devices (galaxy a5 [2017] sm-a520f, huawei cun-u29). as a final result, the performance of device’s cpu is better that makes the device work in the best way. as future work, in relation to the image matching schemes, the user may add something to the image, and as a result, new image obtained, but it still the same image with simple change. in future work, we will try to solve this issue. references 1. c. r. shyu, c. brodley, a. kak, a. kosaka, a. aisen and l. broderick. “local versus global features for content-based image retrieval”. in proceedings. ieee workshop on contentbased access of image and video libraries (cat. no. 98ex173), 1998, pp. 30-34. 2. d. a. lisin, m. a. mattar, m. b. blaschko, e. g. learned-miller and m. c. benfield. “combining local and global image features for object class recognition”. in 2005 ieee computer society conference on computer vision and pattern recognition (cvpr’05)-workshops, 2005, pp. 47-47. 3. m. agarwal, a. singhal and b. lall. “3d local ternary co-occurrence patterns for natural, texture, face and bio medical image retrieval”. neurocomputing, vol. 313, pp. 333-345, 2018. 4. s. s. hwang and s. h. hong. “pre-extraction of features and environment variable-based database filtering for fast image matching on mobile”. mobile and wireless technologies, vol. 2016, pp. 223-230, 2016. 5. j. machajdik and a. hanbury. “affective image classification using features inspired by psychology and art theory”. in proceedings of the 18th acm international conference on multimedia, 2010, pp. 83-92. 6. n. lee, c. kim, w. choi, m. pyeon and y. kim. “development of indoor localization system using a mobile data acquisition platform and bow image matching”. ksce journal of civil engineering, vol. 21, pp. 418-430, 2017. 7. l. wang and h. wang. “improving feature matching strategies for efficient image retrieval”. signal processing: image communication, vol. 53, pp. 86-94, 2017. 8. k. miller, f. franz, m. rittinghaus, m. hillenbrand and f. bellosa. “{xlh}: more effective memory deduplication scanners through cross-layer hints”. in presented as part of the 2013 {usenix} annual technical conference ({usenix}{atc} 13), 2013, pp. 279-290. 9. t. liu, f. chen, y. ma and y. xie. “an energy-efficient task scheduling for mobile devices based on cloud assistant”. future generation computer systems, vol. 61, pp. 1-12, 2016. 10. r. n. widodo, h. lim and m. atiquzzaman. “sdm: smart deduplication for mobile cloud storage”. future generation computer systems, vol. 70, pp. 64-73, 2017. 11. e. ahmed, a. gani, m. sookhak, s. h. ab hamid and f. xia. “application optimization in mobile cloud computing: motivation, taxonomies, and open challenges”. journal of network and computer applications, vol. 52, pp. 52-68, 2015. 12. m. w. storer, k. greenan, d. d. long and e. l. miller. “secure data deduplication”. in proceedings of the 4th acm international workshop on storage security and survivability, 2008, pp. 1-10. 13. n. haustein, c. a. klein, u. troppens and d. j. winarski. “method of and system for adaptive selection of a deduplication chunking technique”. google patents, 2009. 14. b. zhu, k. li and r. h. patterson. “avoiding the disk bottleneck in the data domain deduplication file system”. in fast usenix conference, 2008, pp. 1-14. 15. d. meister and a. brinkmann. “multi-level comparison of data deduplication in a backup scenario”. in proceedings of systor 2009: the israeli experimental systems conference, 2009, p. 8. 16. y. won, r. kim, j. ban, j. hur, s. oh and j. lee. “prun: eliminating information redundancy for large scale data backup system”. in 2008 international conference on computational sciences and its applications, 2008, pp. 139-144. 17. m. sharma. “compression using huffman coding”. ijcsns international journal of computer science and network security, vol. 10, pp. 133-141, 2010. 18. r. li, y. zhao, q. xu and x. qi. “research of subnetting based on huffman coding”. in international conference on cloud computing and security, 2018, pp. 606-616. 19. o. yue-long, l.n. zhang and n. yu. “researching on md5’s characteristics based on software reversing”. the journal of china universities of posts and telecommunications, vol. 17, pp. 127130, 2010. 20. i. c. lin and l. c. yang. “a noise generation scheme based on huffman coding for preserving privacy”. in international conference on security with intelligent computing and big-data services, 2017, pp. 149-160. 21. r. patel, v. kumar, v. tyagi and v. asthana. “a fast and improved image compression technique using huffman coding”. in 2016 international conference on wireless communications, signal processing and networking (wispnet), 2016, pp. 2283-2286. 22. s. h. kim, j. jeong and j. lee. “selective memory deduplication for cost efficiency in mobile smart devices”. ieee transactions on consumer electronics, vol. 60, pp. 276-284, 2014. 23. y. tian, k. zhang, p. wang, y. zhang and j. yang. add “salt” md5 algorithm’s fpga implementation. procedia computer science, vol. 131, pp. 255-260, 2018. 24. d. pamula and a. ziebinski. “hardware implementation of the md5 algorithm”. 9th ifac workshop on programmable devices and embedded systems, roznov pod radhostem, czech republic, feb. 2009. author queries??? aq1:kindly cite figure 7 and 11 in the text part tx_1~abs:at/add:tx_2~abs:at 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 32-37 research article a comparison between new modification of aadaptive nadaraya-watson kernel and classical aadaptive nadaraya-watson kernel methods in nonparametric regression: a simulation study hazhar t. a. blbas1, wasfi t. kahwachi2 1department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region, iraq, 2research center, tishk international university-erbil, kurdistan region, iraq abstract nonparametric kernel estimators are mostly used in a variety of statistical research fields. nadaraya-watson kernel (nwk) estimator is one of the most important nonparametric kernel estimators that is often used in regression models with a fixed bandwidth. in this article, we consider the four new proposed adaptive nwk regression estimators (interquartile range [iqr], standard deviation [sd], mean absolute devotion, and median absolute deviation) rather than (fixed bandwidth, adaptive geometric, adaptive mean, adaptive range, and adaptive median). the outcomes in both simulation and actual data in leukemia cancer showed that the four new aadaptive nwk estimators (iqr, sd, mean absolute devotion, and median absolute deviation) are more effective than the kernel estimations with fixed bandwidth in previous studies based mean square error criterion. keywords: nonparametric regression, kernel regression, new aadaptive nadaraya-watson estimators, leukemia cancer, acute myeloid leukemia introduction nonparametric regression models aim to precisely determine the relationship between explanatory and response variables in various statistical situations; however, they are substantially less successful than parametric approaches when fitting a normal distribution. on the opposite side, nonparametric models are highly efficient in populations that do not fit normal distribution.[10] they may be used to a wide range of data types such as ordinal, nominal, ratio, and interval data.[17] suppose the regression model for a given data points ( , ) ( ) x y ri i i n � �1 is y f x ii i i� � � � � �� ; , , , ,1 2 3 (1`) where ℇ i is an observation error with zero mean variance σ2 and m is unknown regression function. smoothing is a significant aspect in the nonparametric regression process and is a technique for expressing the dependent variable’s pattern.[11] there have been four main factors for doing a nonparametric regression approaches for fitting data depending on.[17] first, to know the relationship between predictor and response variable. second, to enable predictions of future findings without using a fixed parametric model. third, by studying the results of individual locations, it offers a method for detecting spurious findings. finally, working to solve the absent values by replacing or merging neighboring values of the independent variables x.[18] kernel regression assume that x 1 , x 2 ,…, xn from a random variable describe a sample of size (n) with density f(x).[1] bring the kernel density function of f(x) at point x: ( ) 1 1ˆ = −  =     ∑ n i h i x x f x k nh h (2) kernel regression (nadaraya-watson estimator) was established by nadaraya in 1965 and watson in 1964 which is corresponding author: hazhar t. a. blbas,department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region, iraq. e-mail: hazharstat@gmail.com received: september 4, 2021 accepted: october 1, 2021 published: october 30, 2021 doi: 10.24086/cuesj.v5n2y2021.pp32-37 copyright © 2021 hazhar t. a. blbas, wasfi t. kahwachi. this is an openaccess article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) blbas and kahwachi: new modification of anwk in nonparametric regression 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 32-37 one of the most frequently used technique in nonparametric.[5,6,12] both nadaraya and watson indicated the general estimator of ( )ˆ f x in nonparametric regression related to smoothing bandwidth (h) and kernel (k) in bellow formula. ( ) 1 1 ˆ = = −     = −      ∑ ∑ n i ii n i i x x k y hf x x x k h (3) ( ) 1 ˆ = = ∑ n i i i f x w y (4) u x x h i� �� � � � � � (5) the smoothing or bandwidth parameter h is used to control the smoothness of the approximate graph, and the kernel weights as identified by w x x h k x x h i i n i i n i � �� � � � � � �� � � � � � � � � � 1 1 (6) the most important factor to consider in nonparametric regression is choosing bandwidth and kernel function. on the other hand, the selection of bandwidth is much more essential than the choice of kernel function on estimation.[8] the smoothed function can be expressed by scanning each data point with a weighted kernel function and then evaluating the input at each point. the kernel function is penalized based on its range from the centered location, and the degree of this penalty is defined by the bandwidth.[8] a narrow (small) bandwidth of h results in a wiggly curve and a wealth of noise in estimation, whereas a vast (big) bandwidth of h results in a flat curve and over-smoothed curves in estimation. one of the assumptions of kernel density function is a symmetric that is often applied with a standard normal density.[13] the kernel’s functions of k can be used to one of several frequently used functions, namely epanechnikov, triangle, quartic, gaussian, uniform, and tricube (triweight).[3] gaussian is a common and practical kernel density function[2] which is used in this paper as shown below. k u e uu� � � � �� �� ��1 2 2 2 � / ; , (7) ( ) 2 1 2 1 1 1 22ˆ 1 1 22 π π = = − −    = − −     ∑ ∑ n i ii h n i i x x exp y hf x x x exp h (8) if you already have over one predictive variable, lengthy distributions, or multi modal distributions, fixed nadaraya watson (fnw) is not always the best option.[9] in this situation, we can use the variable nadaraya-watson kernel (vnwk) estimator with a variable bandwidth i(x i ) as seen below: ( ) 1 1 ( ) ( ) 1 ( ( ) ˆ ) = =  −    =  −     ∑ ∑ n i i i i i vnw n i i i i x x y k h x h x f x x x k h x h x (9) [14] showed a formula to compute h(x i ) in 1982, h x h f xi i � � � ( ) (10) while f(x i ) is determined by the kernel function estimator which is a probability density function of x i. a new algorithm developed for the abramson design estimator by silverman in 1986, which he called an aadaptive nwk (anwk) function estimator. for a fixed h, the previous kernel function estimator was used as a first stage,[4] which is represented by ( )ˆ if x and he established local h factor θi as: ˆ ( ) α θ −   =      i i g f x g (11) while g represents the geometric mean of ˆ )( if x with g≠0 and a illustrates the sensitivity parameter between 0 and 1 (0≤α≤1).[14] selected the value of α=0.5 because its gives an accurate predictive results. moreover then, silverman provided an adaptive h as seen below: h x hi gi� � �� (12) in 2010,[15] it is introduced a change to the anw approach that they have used arithmetic mean x of ( )ˆ if x instead of using g for figuring out the h factor in nwk estimator. ˆ ( ) α θ −   =      i i x f x x (13) in 2014,[7] it modified the anw approach that they have used range r of ( )ˆ if x instead of using g or x for figuring out the h factor in nwk estimator. ˆ ( ) α θ −   =      i i r f x r (14) in 2019,[16] it proposed another change for the anw approach that he used median instead of using geometric, mean, or range for calculating the h factor in nwk estimator. ( )ˆ α θ −   =      i i me f x me (15) new proposed nwk function estimator in this study, a new changes for the adaptive nadarayawatson approach was proposed, which used four different statistical techniques such as interquartile range (iqr), standard deviation (sd), mean absolute deviation (mad), and median absolute deviation (mead) of ( )ˆ if x instead of using geometric mean, arithmetic mean, range, or median for figuring out the h factor in nwk estimator. blbas and kahwachi: new modification of anwk in nonparametric regression 34 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 32-37 ˆ ˆ ˆ ˆ ( ) ( ) ( ) ( ) α α α α θ − − − −                          =                   i i i i p i f x iqr f x sd f x mad f x mead (16) where iqr is an abbreviation for iqr, sd represents sd, mad is an acronym for mad, and mead stands for median absolute deviation. mean square error (mse) in this paper, mse is used as an estimation criterion to find out the difference between (classical) traditional and newly proposed nwk estimators. as mentioned below, the best estimator will be the one with lowest mse value. 2 1 1 ( )ˆ = = −∑ n i i i mse y y n (17) materials and methods leukemia cancer data are used to undertake the performance of all proposed methods such as anw irq, anw sd, anw mad, and anw mead in real application which collected from january 2015 to december 2020 at nanakali hospital for blood in erbil city of iraq. furthermore, the cd45 outcome as an explanatory variable and platelet (plt) as a response variable in aml type of leukemia cancer from 30 patients in table 1 is used to compare between proposed methods and classical methods. since simple regression cannot be met due to assumptions such as linearity and autocorrelation, nonparametric regression is used for both classical and proposed approaches based on mse. table 2 compares the findings of classical methods to a new proposed modification of the anw kernel estimator for iqr, sd, mad, and mead based on improving the prediction accuracy of the anw kernel estimator. the proposed mead approach has the smallest mse in various bandwidths and sample sizes, followed by mad, sd, and iqr, respectively. we achieved the same results as in the simulation analysis, namely that all new proposed methods have smaller mse than all classical methods. simulation study a simulation analysis was carried out to compare the efficiency between traditional nadaraya-watson and new proposed methods of anw estimators using the r (4.0.2) language software. an explanatory variable and response variable with adding noise to an exponential wave are simulated in this nonlinear regression function to identify that proposed methods outperforms classical models. y x x ni i i i� � � � �exp ~ ( , )3 0 1� (18) where xi was chosen at random from a uniform distribution on the interval [–2, 2] and generated different samples of size (25, 50, 75 and 150) with different fixed h (0.5, 0.75, 1, and 1.5). in this paper, comparison between classical methods such as fnw, vnw geometric, anw mean, anw median, anw range with new proposed methods such as anw irq, anw sd, anw mad, and anw mead kernel estimations were computed by gaussian kernel function with 20000000 repetitions in each estimation. figure 1 represents the results between classical and new proposed methods at h (5, 15, and 30) for real data. on the table 1: aml type of leukemia cancer from 2015 to 2020 cd45 plt cd45 plt cd45 plt cd45 plt cd45 plt 70 17 80 54 62 13 80 89 74 61 70 51 88 31 60 15 90 6 77 16 80 7 58 16 32 62 88 137 80 155 94 79 54 40 42 71 60 2 55 24 40 44 70 95 54 19 35 58.9 70 31 85 68 74 46 86 5 43 33 80 229 table 2: mse values between classical methods and proposed methods of nwk estimators in real data (n=30) h fixed nw variable nw g anw m anw me anw r anw iqr anw sd anw mad anw mead 5 1890.7 1751.1 1768.2 1781 1787.9 1609.8 1609.7 1574.3 1572.4 10 2071 2010.9 2017.1 2015.6 1964.9 1909.1 1827.6 1803.2 1802.8 15 2214.5 2176.2 2179.1 2179.2 2046.3 1983.4 1906 1889.1 1889 20 2297.3 2277.5 2278.5 2279.3 2111.5 2021.5 1945.7 1928.5 1927.2 25 2340.4 2331.2 2331.6 2332.1 2152.3 2029.6 1966.8 1947.9 1945.8 30 2365.4 2360.9 2361.1 2361.4 2174.6 2033.7 1978 1958 1955.5 blbas and kahwachi: new modification of anwk in nonparametric regression 35 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 32-37 other hand, figures 2 and 3 illustrate the simulation results for both classical and proposed methods in nonparametric regression functions with sample size of 75 and at h (0.5, 07.75, 1, and 1.5), respectively. table 3 compares simulation results between all classical (traditional) methods and new proposed update of adaptive nw kernel such as iqr, sd, mad, and mead aimed at figure 1: left hand classical methods and right hand proposed methods at h=5, 15, and 30 for real data. (a) cd45 versus plt at h=5, (b) cd45 versus plt at h=5, (c) cd45 versus plt at h=15, (d) cd45 versus plt at h=15 (e) cd45 versus plt at h=30 fcd45 versus plt at h=30 improving the prediction accuracy of anwk estimator. according to mse criteria, the new proposed methods outperform classical methods of using various sample sizes and initial bandwidth values. while the new proposed iqr is better than other classical approaches, it is less effective than other proposed methods such as sd, mad, and mead, respectively. on the other hand, dc b f a e blbas and kahwachi: new modification of anwk in nonparametric regression 36 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 32-37 the proposed method using mead has less mse than mad, in turn mad is less than sd, and sd is less than iqr in different bandwidth and sample size. conclussion 1the new proposed method estimator for median absolute deviation (mead) was more reliable than any of the other classical methods for both simulation and actual data depending on mse criteria because it is able to reduce the effect of outliers on the fitting kernel model. 2new proposed method estimator for mad was more reliable than any of the other classical methods for both simulation and specific data because the absolute mean difference in mad gives lower value of mse. figure 2: left hand classical methods and right hand new proposed methods at h=0.5 and 0.75 with ample size 75, (a)-x versus y at h=0.5 and n = 7, ( b)-x versus y at h=0.5 and n = 75, (c)-x versus y at h=0.75 and n = 75, (d)-x versus y at h=0.75 and n = 75 figure 3: left hand classical methods and right hand new proposed methods at h=1 and 1.5 with sample size 75, (a)-x versus y at h=1 and n = 75 (b)-x versus y at h=1 and n = 75, (c)-x versus y at h=1.5 and n = 75 (d)-x versus y at h=1.5 and n = 75 table 3: mse values between classical methods and new proposed methods of nwk estimators in simulation data h n fixed nw variable nw g anw m anw me anw r anw iqr anw sd anw mad anw mead 0.5 25 1.1997 1.2769 1.3439 1.3116 1.7164 1.0449 1.0124 0.9336 0.9317 50 1.5189 1.5314 1.6769 1.4909 2.2494 1.5075 1.5106 1.4193 1.3552 75 1.3763 1.5314 1.6357 1.4530 2.1951 1.3629 1.4166 1.3108 1.2386 150 1.3899 1.5314 1.6141 1.3753 2.3476 1.1274 1.3872 1.2582 1.1485 0.75 25 1.7391 1.5314 1.8532 1.8887 2.1838 1.2308 1.5290 1.3999 1.3796 50 1.9968 1.5314 2.1020 1.9050 2.5395 1.9694 1.7755 1.6786 1.6309 75 1.9562 1.5314 2.1081 1.9022 2.5044 1.8816 1.6998 1.5897 1.5259 150 1.9938 1.5314 2.1887 1.8970 2.7280 1.6904 1.6997 1.5504 1.4423 1 25 2.1124 1.5314 2.1651 2.1877 2.3063 1.6398 1.6593 1.5147 1.5114 50 2.4127 1.5314 2.4600 2.3296 2.7635 2.2906 1.9823 1.8860 1.8617 75 2.4083 1.5314 2.4799 2.3478 2.7244 2.2515 1.9349 1.8337 1.7994 150 2.5004 1.5314 2.6277 2.4347 2.9452 2.1524 1.9254 1.7770 1.7158 1.5 25 2.4158 1.5314 2.4393 2.4446 2.4028 2.0489 1.8624 1.7373 1.7350 50 3.0111 1.5314 3.0178 2.9693 3.0401 2.5985 2.2131 2.1163 2.1072 75 2.9941 1.5314 3.0051 2.9563 2.9868 2.5805 2.1996 2.1043 2.0916 150 3.1563 1.5314 3.2067 3.1444 3.1738 2.5569 2.1742 2.0404 2.0173 dc ba dc ba blbas and kahwachi: new modification of anwk in nonparametric regression 37 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 32-37 3new proposed method estimator for sd was more reliable than any of the other classical methods for both simulation and current data using mse criteria. 4new proposed method estimator for iqr was more reliable than any of the other classical methods for both simulation and real data based on mse criteria. 5median absolute deviation (mead) has less mse than mad, in turn mad is less than sd, and sd is less than iqr in different bandwidth and sample size in either simulation or real data. 6in both simulation and real data, both proposed and classical methods are improved by reducing the initial bandwidth values and sample size. reerences 1. m. hollander, d. a. wolfe and e. chicken. nonparametric statistical methods. john wiley & sons, inc., hoboken, new jersey, 2014. 2. w. härdle. applied nonparametric regression. cambridge university press, cambridge, england, new york, 1997. 3. m. memmedli and m. yildiz. comparison study on smoothing parameter and sample size in nonparametric fuzzy local polynomial regression models. in: 2012 iv international conference “problems of cybernetics and informatics” (pci), 2012. 4. w. hardle. applied nonparametric regression. biometrics, vol. 50, no. 2, p. 592, 1994. 5. a. christmann and i. steinwart. consistency and robustness of kernel-based regression in convex risk minimization. bernoulli, vol. 13, no. 3, pp. 799-819, 2007. 6. e. a. nadaraya. on estimating regression. theory of probability and its applications, vol. 9, no. 1, pp. 141-142, 1964. 7. g. s. watson. smooth regression analysis, sankhya the indian journal of statistics series a, vol. 26, no. 4, pp. 359-372, 1964. 8. m. p. wand and m. c. jones, kernel smoothing. springer, boston, ma, 1995. 9. h. takeda, s. farsiu and p. milanfar. kernel regression for image processing and reconstruction. ieee transactions on image processing, vol. 16, no. 2, pp. 349-366, 2007. 10. w. l. martínez and a. r. martínez. computational statistics handbook with matlab. chapman & hall, london, 2008. 11. d. conn and g. li. an oracle property of the nadaraya-watson kernel estimator for high–dimensional nonparametric regression. scandinavian journal of statistics, vol. 46, no. 3, pp. 735-764, 2018. 12. b. w. silverman. density estimation for statistics and data analysis estimation density. kluwer academic publishers, london, 1986. 13. m. hanif, s. shahzadi, u. shahzad and n. koyuncu. on the adaptive nadaraya-watson kernel estimator for the discontinuity in the presence of jump size. süleyman demirel üniversitesi fen bilimleri enstitüsü dergisi, vol. 22, no. 2, p. 511, 2018. 14. i. s. abramson. on bandwidth variation in kernel estimates-a square root law. the annals of statistics, vol. 10, no. 4, pp. 12171223, 1982. 15. d. li and r. li. local composite quantile regression smoothing for harris recurrent markov processes. journal of econometrics, vol. 194, no. 1, pp. 44-56, 2016. 16. s. demir and ö. toktamiş. on the adaptive nadaraya-watson kernel regression estimators. hacettepe journal of mathematics and statistics, vol. 39, no. 3, pp. 429-437, 2010. 17. h. a. khulood and i. al turk lutfiah. modification of the adaptive nadaraya-watson kernel regression estimator. scientific research and essays, vol. 9, no. 22, pp. 966-971, 2014. 18. t. h. ali. modification of the adaptive nadaraya-watson kernel method for nonparametric regression (simulation study). in: communications in statistics-simulation and computation. taylor & francis group, united kingdom, pp. 1-13, 2019. tx_1~abs:at/add:tx_2~abs:at 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 review article the comparative study of biaxial bending analysis of steel sections using aisc and eurocode approaches mohammed m. saleh1*, dlshad k. ahmed2, ali r. yousef 3 1department of civil engineering, engineering collage, universita della calabria, cosenza, italy, 2department of civil engineering, cihan university-erbil, kurdistan region, iraq, 3department of civil engineering, engineering collage, salahaddin university, erbil, iraq abstract this study investigates the capacity of the steel section using both aisc and eurocode approaches. three types of steel sections were subjected to biaxial bending by applying loads to both main axes and examined by both approaches. the concept of fisher was also adopted as an approach. this concept proposed that only the compression flange could withstand lateral loading and the torsional influence could be ignored. the findings suggested that the eurocode approach is more conservative in the design of steel sections subject to biaxial bending as it takes into account the level at which the load is applied, the type of the section whether rolled or welded and its height-to-width ratio (lateral buckling effect). the aisc approach considers the shear center of the section as the level at which the loads are applied. the conservatism of the results was more pronounced when the section is close to h-section. fisher`s concept of structural design of biaxial bending of structural steel is more conservative than both aisc and eurocode approaches of analysis. keywords: aisc specification, biaxial bending analysis, eurocode, fisher`s concept, steel sections introduction due to many factors, such as good mechanical properties, quick and easy construction and economy, steel structures are commonly used for construction. overhead crane runway girders are examples of biaxial bending in which bending moments applied to both major and minor axis. when the loads applied through the shear center, twisting would not develop[2] as shown in figure 1. for this case, aisc 360 commentary,[1] biaxial bending equation given by the aisc 360 eq. (h1-1b) by supposing of axial load that equal to one. the interaction equation then reduces to m m m m ry cy rz cz � � 1 (1) where m ry = applied bending moment (y-axis), m cy = nominal moment capacity (y-axis), m rz = applied bending moment (z-axis), m cz = nominal moment capacity (z-axis). for the lrfd method, mc = φ b m n , while for asd method, mc = m n /ω b , φ b = 0.9 = factor for resistance in flexure, ω b = 1.67 = safety factor for flexure. regarding to the bending about the strong axis, the eurocode method focused on several factors that could affect the flexure strength of the beams. the elastic critical moment is valuable for the study of lateral-torsional beam buckles. the maximum bending moment value provided by the beam is described as this quantity, far from imperfections of some kind. 2 2 1e wcr t z t e i m g i e i l l g i π π  = +    (2) where i t is the torsion constant, iz is the second moment of area in (weak axis), i w the warping constant, l is the length between laterally braced cross-sections of the beam, and e and g are the longitudinal modulus and the shear modulus of elasticity, respectively. because of the different loading types and bending moment diagrams, broad equations can be found in practical implementations and are suitable for a range of conditions, the most widely used in the formulation adopted by the commission,[3] acceptable to members subject to bending moment on a strong axis with a mono-symmetric cross-section corresponding author: mohammed m. saleh, department of civil engineering, engineering collage, universita della calabria, cosenza, italy. e-mail: mohammed@civil-eng.it received: oct 16, 2020 accepted: dec 20, 2020 published: dec 30, 2020 doi: 10.24086/cuesj.v4n2y2020.25-32 copyright © 2020 mohammed m. saleh, dlshad k. ahmed, ali r. yousef. this is an open access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 on a weak axis. for the calculation of the elastic critical moment under criteria not covered by boissonnade et al.[3] ( ) ( ) 0.52 z 222 2 g 2 gz z w t cr 1 2 2 3 j 3 jw z zz k l c z c ze i k i g i m c c z c zk i e ik l       − −     π    = + + −          π             (3) the shape of the bending moment diagram gives the coefficient of c 1 , c 2 , and c 3 , support conditions give the coefficients k z and k w which are called the effective length factor, z g = (z a z s ), where z a and z s , comparable to the centroid of the cross-section, are the locations of the point of applied load and the shear center; these amounts are positive if the compressed portion is placed and negative if the tension portion is placed. while aisc requirements lead us to calculate the capacity of the bending moment without taking into account the level of load application,[1] even the steel section is rolled or welded.[4] the lateral load is also added to the compression flange of the member, as shown in figure 2. in this case, the conceptual solution proposed by fisher should be used,[5] this concept assumed that only the compression flange can survive lateral loading and the torsional impact can be ignored. in the case of i-shapes, the plastic section modulus of a z-axis flange is provided by z z t z= 2 (4) where z z is plastic section modulus about the z-axis. the moment capacity of one flange about the y-axis is nt y tm f z= × (5) linear additions of the moment terms in[1] as shown in figure 3 typically lead to results which are too conservative.[6] based on the definition on biaxial bending, we will extend this to one case of study using both aisc and eurocodes to investigate and demonstrate the deference between them regarding the level at which the load is applied, the type of the section whether rolled or welded and its height-to-width ratio by evaluating the results using robot structural analysis software. figure 1: pure biaxial bending[2] figure 2: lateral load applied on the upper flange methodology to demonstrate the exact difference between aisc and eurocode analysis methods regarding biaxial bending, the case of study struggles with the loads added to the upper flange as shown in figure 1, and fisher’s concept[5] will be regarded. the case study includes a simply supported beam with a span of 10.0 m, measured center-to-center of support, with the load being applied at midspan, as shown in figure 4, the beam under consideration is subjected to vertical load and the lateral load with the value 25 kn and 5 kn, respectively. the beam has yielding strength (f y = 355 mpa), modulus of elasticity (e = 21x103 mpa) and detailed properties are shown in table 1. for the common base of comparison, the load factors in both approaches will be based on that of aisc standards.[7] 1.2 dl + 1.6 ll (6) the applied factored loads (moments) are shown in table 2. the dead load includes the self-weight of the member as uniformly distributed which is calculated by the software. aisc methods of analysis for aisc methods of analysis in the robot structural analysis software, modification factor should be defined for moment gradient c b in the member definition section which is equal to 1.32 for single point load act on simply supported beams laterally braced at the supports only. the lateral buckling length coefficient that represents the lateral bracing is equal to 1.0 (no lateral bracing for the beam). the results of the analysis are shown in tables 3-6. classification of section for local buckling � f b t � � 9 58. width-to-thickness ratio for a flange. � fp e fy � �0 38 9 24. . limiting slenderness for compact flange. � � f fp � the flange is non-compact. saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 table 1: cross-section properties cross-section properties symbul values unit symbol description a x 139.2 cm2 cross-section area a y 55.2 cm2 shear area – y-axis a z 86.88 cm2 shear area – z-axis i y 104264.9 cm4 second moment of area about the y-axis i z 2443.48 cm4 second moment of area about the z-axis z y 3435.12 cm3 plastic section modulus about the y (major) axis s y 2880.246 cm3 elastic section modulus about the y-axis z z 342.6 cm3 plastic section modulus about the z (minor) axis s z 212.477 cm3 elastic section modulus about the z-axis d 72.4 cm cross-section height b f 23 cm cross-section width t f 1.2 cm thickness of flange t w 1.2 cm web thickness table 2: internal forces internal forces aisc vertical load lateral load applied factored loads 40 kn 8 kn applied moment y-axis z-axis 116 kn.m 20 n.m figure 3: simple linear interaction for biaxial bending[2] figure 4: the case of the study � w w h t � � 58 3. width-to-thickness ratio for a web. � wp e fy � �3 76 91 45. . limiting slenderness for compact web. � � w wp � ∴ the web is compact. parameters of lateral buckling analysis c b = 1.32 lateral-torsional buckling modification factor, l b =10 m laterally unbraced length of a member (lateraltorsional buckling), c = 1 for doubly symmetric section. r i c s ts z w y = = 5 49. cm, h0 = d – 2tf = 70 cm l r e f p z y = =1 76 1 79. . limiting laterally unbraced length for the limit state of yielding. 2 2 yts r y y 0 y 0 0.7f1.95r e jc jc l 6.76 0.7f s h s h e     = + +        = 5.3 m limiting laterally unbraced length for the limit state of inelastic lateraltorsional buckling. q lb>lr ∴ f c e l r jc s h l r cr ltb b b ts y b ts ( ) . .� � � � � � � � � � � � � � � �2 2 0 2 1 0 078 112 13 mpa (7) fcr: critical stress (lateral-torsional buckling), j: torsional constant, c: coefficient, sy: elastic section model. m f s ny ltb cr ltb y( ) ( ) .� � � 322 95 kn.m nominal lateral-torsional buckling strength. eurocode method of analysis in this method of analysis, the level where the load is applied must be defined, set the level of the applied load at z = 1 in the member definition section beside the shape of bending moment in the load type section, the results of the analysis are shown in tables 7-9. class of section flange: c/t f = 8.68 > 10 235 8 1 f y = . ∴ the flange is classified to the third class. saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 � lt y y cr w f m � � 2 14. non-dimension slenderness ratio for lateraltorsional buckling. lateral buckling curve is d from table 11. α lt =0.76 imperfection factor for lateral buckling curves coefficient for calculation of x lt. � � � � lt lt lt lt � � �� � � 1 0 16 2 2 2 . reduction factor for lateraltorsional buckling. m w f b rd lt y y m . .� � � � 1 161 39 kn.m buckling resistance moment. results the main differences between aisc and eurocode analysis results are explained in this section. concern to the nominal strength, based on the results of the analysis, significant differences between the two approaches are demonstrated in table 10. as shown in table 10, the difference between the design moment strengths of the y-axis is 129.26 kn.m, where the section capacity regarding the aisc approach is approximately 80% higher than the eurocode approach. the main reason for this would be due to the level of applied load, as this condition is not recognized by the aisc standard. aisc specifications take into consideration the level of applied load through the cross-section shear center, while this level would be under or above or just through the shear center considered by the eurocode approach, this effect is demonstrated in figure 5. the level of application of the load has a considerable effect on the elastic critical moment m cr . to show this effect more, several sections with deferent depth and width but the same thickness of flange and web as listed below were tested. section 1: total depth of beam = 724 mm, width of beam = 230 mm. section 2: total depth of beam = 574 mm, width of beam = 250 mm (similar i section). section 3: total depth of beam = 474 mm, width of beam = 400 mm (similar h section) for all sections assumed that the level of the applied load at the upper part of the beam just on the flange with actual member definition for both approaches, the outcomes for this condition are shown in figure 6. this figure indicates that section 3 which is similar to h sections would be the critical situation. to show an explanation for this huge difference between the two approaches, for both approaches the level of applied load places at the shear center of the cross-section (center of the section), and the modification factor c b sets to one. the results of the analysis are shown in figure 7. the results are getting closer so that the first reason behind this difference in the computation is recognized which the aisc approach does not consider the level of applied table 3: cross-section properties required for aisc approach symbol values unit symbol description a x 139.2 cm2 cross-section area a y 55.2 cm2 shear area y-axis a z 86.88 cm2 shear area z-axis j 66.82 cm4 torsional constant cw 3096768.81 cm6 warping constant i y 104264.9 cm4 moment of inertia of a section about the y-axis i z 2443.48 cm4 moment of inertia of a section about the z-axis z y 3435.12 cm3 plastic section modulus about the y (major) axis s y 2880.246 cm3 elastic section modulus about the y-axis z z 342.6 cm3 plastic section modulus about the z (minor) axis s z 212.477 cm3 elastic section modulus about the z-axis d 72.4 cm height of cross-section b f 23 cm width of cross-section t f 1.2 cm flange thickness t w 1.2 cm web thickness r y 27.37 cm radius of gyration y-axis r z 4.19 cm radius of gyration z-axis table 4: internal forces (aisc approach) internal forces symbul values unit symbol description section m rz −20 kn.m required flexural strength v ry −4 kn required shear strength v rz −20 kn required shear strength web: c w /t w = 57.53 < 72 235 58 58 f y = . ∴ the web is classified to the first class. lateral-torsional buckling analysis (general method [6.3.2.2]) l cr,upp = 10 m upper flange, lateral bracing, c1=1.26 / c2=0.55 / c3 =1.73 factors, i w =3096768.8 cm6 warping constant, z g = 36.2 cm from the shear center to the point of applied load. ( ) ( ) ( ) ( ) 0.52 z 2 2 z w tz 2cr 1 2 g 3 j2 w z z z 2 2 g 3 j k l k i g ie i m c c z c z 223.05k i e ik l c z c z          π  + + = − − =  π      −     kn.m critical moment for lateral-torsional buckling. saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 table 5: nominal strengths (aisc approach) nominal strengths symbul amounts unit symbol description section respect to the y axis m py 1219.47 kn.m nominal plastic bending moment [f] m ny [yd] 1219.47 kn.m nominal flexural strength in the limit state of yielding [f3.1] m ny [ltb] 322.95 kn.m nominal lateral-torsional buckling strength [f3.1] m ny 1[ltb] 244.66 kn.m nominal lateral-torsional buckling strength (c b =1.0) [f3.1] m ny [flb] 1203.48 kn.m nominal strength for local buckling of a compression flange [f3.2] m ny 322.95 kn.m nominal flexural strength [f3] v nz 1850.54 kn nominal shear strength [g2.1] respect to the z axis m pz 121.62 kn.m nominal plastic bending moment [f] m nz [yd] 120.69 kn.m nominal flexural strength in the limit state of yielding [f6] m nz [flb] 118.53 kn.m nominal strength for local buckling of a compression flange [f6.2] m nz 118.53 kn.m nominal flexural strength [f6] v ny 1175.76 kn nominal shear strength [g2.1] table 6: design strengths (aisc approach) design strengths symbol amounts unit symbol description section respect to the y axis fib*m py 1097.52 kn.m design plastic bending moment [f] fib*m ny [yd] 1097.52 kn.m design flexural strength in the limit state of yielding [f3.1] fib*m ny [ltb] 290.65 kn.m design lateral-torsional buckling strength [f3.1] fib*m ny 1[ltd] 220.19 kn.m design lateral-torsional buckling strength [f3.1] fib*m ny [flb] 1083.13 kn.m design strength for local buckling of a compression flange [f3.2] fib*m ny 290.65 kn.m design flexural strength [f3] fiv*v nz 1665.49 kn design shear strength [g2.1] respect to the z axis fib*m pz 109.46 kn.m design plastic bending moment [f] fib*m nz [yd] 108.62 kn.m design flexural strength in the limit state of yielding [f6] fib*m nz [flb] 106.68 kn.m design strength for local buckling of a compression flange [f6.2] fib*m nz 106.68 kn.m design flexural strength [f6] fiv*v ny 1058.18 kn design shear strength [g2.1] verification formulas uf (h1_1b) 0.59 m ry /(fib*m ny ) + m rz /(fib*m nz ) verified uf (g2_1) 0 v ry /(fiv*v ny ) verified uf (g2_1) 0.01 v rz /(fiv*v nz ) verified load unless when the load applied just on the top flange and for the case l b > lr then the square root of engineers[7] may conservatively takes one which does not take one in the analysis of robot structural analysis software. in addition, other factors cause. such as the type of section (rolled or welded) and the depth to width ratio(h/b), which is very effective in the case of lateral torsional buckling table 11. the imperfection factor α lt which is given in table 12 covers the effect of lateral-torsional buckling that results in the redaction factor x lt for the design moment capacity as shown in velikovic et al.[8] saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 table 7: cross-section properties required for eurocode approach symbol values unit section a x 139.2 cm2 a y 55.2 cm2 a z 84 cm2 i x 65.509 cm4 i y 104264.9 cm4 i z 2443.48 cm4 w ely 2880.246 cm3 w elz 212.477 cm3 h 72.4 cm b 23 cm t f 1.2 cm t w 1.2 cm r y 27.37 cm r z 4.19 cm a nb 1 (6.2.2.2) e ta 1 (6.2.6.(3)) table 8: internal forces internal forces symbol values unit symbol description section m y, ed 111.84 kn.m bending moment m y.ed m z, ed −18.75 kn.m bending moment m z.ed figure 6: comparison of design moments about y-axis with actual conditions figure 7: comparison of design moments about y-axis with the load at the shear center for both approaches � � � � lt lt lt lt � � �� � ���� � �� 0 5 1 0 2 2 . . (8) � � � � lt lt lt lt � � �� � 1 2 2 2 (9) m w f b rd lt y y m . � � � 1 (10) the rolled section performs better than the welded section. table 2 shows that, because of the lateral load, the difference in the bending moment between two approaches around the z-axis is about 31.25. robot structure analysis software does not follow fisher’s concept, however, in most cases, robot structure analysis software does not follow fisher’s concept, in most cases, the eurocode approach is closer to the concept of fisher. fisher’s concept is based on (2) and (3), where the nominal strength is figure 5: the influence of the level of the applied load on the capacity of the section saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 table 9: design forces and verifications design forces symbol values unit symbol description section m b, rd 161.39 kn.m buckling resistance moment (6.3.2.1) m y, pl, rd 1219.47 kn.m plastic resistance moment (6.2.5.(2)) m y, el, rd 1022.49 kn.m elastic resistance moment (6.2.5.(2)) m y, c, rd 1022.49 kn.m moment resistance (6.2.5.(2)) v y, c, rd 1131.38 kn plastic shear resistance (6.2.6.(2)) respect to the z axis z m z, pl, rd 121.62 kn.m plastic resistance moment (6.2.5.(2)) m z, el, rd 75.43 kn.m elastic resistance moment (6.2.5.(2)) m z, c, rd 75.43 kn.m moment resistance of a compressed section part (6.2.5.(2)) v z, c, rd 1721.66 kn plastic shear resistance (6.2.6.(2)) verification formulas global stability check of member ufb[m y m z ] 0.94 m y, ed /(x lt *m y, rk /gm1) + m z, ed /(m z, rk /gm1) (6.3.3.(4)) ratio rat 0.94 efficiency ratio section ok table 10: comparison between aisc and eurocode results approaches aisc eurocode design strength notation yaxis (kn.m) zaxis (kn.m) yaxis (kn.m) z-axis (kn.m) design buckling resisting moment m b , r d x x 161.39 x design lateral-torsional buckling strength f ib *m ny 290.65 x x x moment resistance of a compressed section part m z, c , r d x x x 75.43 design strength for local buckling of a compression flange f ib *m nz x 106.68 x x table 11: imperfection factor for lateral torsional buckling[8] buckling curve a 0 a b c d imperfection factor 0.13 0.21 0.34 0.49 0.76 z z cm t y= = = 2 342 6 2 171 3 3 . . (11) m z f nmm knm nt t y � � � � � �171300 355 60811500 60 81. . . (12) the design moment capacity for section 1 is ∅m nz =0.9(m nt )=0.9(60.81)=54.72 kn.m, and for sections table 12: buckling curves for lateral-torsional buckling (general method) section limits buckling curve i or h sections rolled h/b≤2 a h/b≤2 b i or h sections welded h/b≤2 c h/b≤2 d other sections ---d figure 8: comparison between the three approaches for design moment about z-axis saleh, et al.: biaxial bending analysis of steel sections using aisc and eurocode approaches 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 25-32 2 and 3 are 63.1 kn.m and 155.9 kn.m, respectively. the comparison between the three approaches is shown in figure 8 fisher’s concept is more conservative than aisc and eurocode methods of analysis. as shown, it is safer to use fisher’s concept or eurocode approach in robot structure analysis software, figuure 9 where the safety factor is defined as the ratio of the moment capacity of the section to the applied moment. conclusions based on the results of steel sections which analyzed by aisc and eurocode standards, and fisher’s concept, the following points can be drawn: 1. eurocode method of analysis results in more conservative design strength about y-axis of the section as it takes into consideration the level where the loads are applied. on contrary, the aisc method of analysis does not take this effect into consideration unless when the load applied just on the top flange and for the case l b > lr then the square root of equation (7) may conservatively takes one which does not take one in the analysis of robot structural analysis software 2. eurocode considers the lateral-torsional effect in terms of height/width ratio and the type of the section, rolled or welded, which results in the safety factor, which is not covered by aisc standards. the effect of lateral-torsional buckling and member type is much clear when the section is close to h-section 3. for the three types of sections (1, 2, and 3) considered in this study, eurocode predicts design strength with safety factors of 1.98, 1.94, and 1.41, respectively, while such safety factors were 1.59, 1.54, and 1.26 for aisc approach. this concludes the adoption of eurocode method of analysis in structural design of biaxial bending case when using robot structure analysis software 4. fisher’s concept of structural design of biaxial bending of structural steel is more conservative than both aisc and eurocode approaches of analysis. references 1. a. ansi. aisc 360-16, specification for structural steel buildings, 2016. 2. a. aghayere and j. vigil. structural steel design: pearson india, 2008. 3. n. boissonnade, r. greiner, j. p. jaspart and j. lindner. rules for member stability in en 1993-1-1: background documentation and design guidelines, 2006. 4. l. laim, j. p. c. rodrigues and l. s. da silva. experimental and numerical analysis on the structural behaviour of cold-formed steel beams. thin walled structures, vol. 72, pp. 1-13, 2013. 5. m. fisher. steel design guide 7: industrial buildings, roofs to anchor rods. chicago, il: american institute of steel construction, 2005. 6. y. l. pi and n. s. trahair. inelastic torsion of steel i-beams. journal of structural engineering, vol. 121, pp. 609-620, 1995. 7. a. engineers. minimum design loads for buildings and other structures, standard asce/sei 7-10. reston, va: american society of civil engineers, 2013. 8. m. velikovic, l. s. da silva, r. simões, f. wald, j. p. jaspart, k. weynand. eurocodes: background and applications design of steel buildings worked examples, joint research center, 2015. figure 9: the difference in the factor of safety for biaxial bending manner tx_1~abs:at/add:tx_2~abs:at 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 19-24 research article study the prevalence, characteristics of mothers, and associated risk factors of pica eating among pregnant women: a cross-sectional study yaseen m. galali1,2* 1food technology department, college of agricultural engineering sciences, salahaddin university-erbil, erbil, krg-iraq, 2department of nutrition and dietetics, cihan university-erbil, erbil, krg-iraq abstract eating disorder is a common phenomenon accompanying pregnancy including ingestion non-nutritive substances. causes of pica are uncertain. the purpose of this study, therefore, is to assess prevalence, characteristics, and risk factors of pica among kurdish pregnant women. a cross-sectional study was conducted in iraqi kurdistan and the information regarding sociodemographic information, prevalence, and characteristics, and risk factors were collected. four hundred women were registered and interviewed for this study. data showed that the prevalence of pica was 60.5% (242), and the most frequent forms of pica eaten were pagophagia, geophagia, and amylophagia by 29.7% (119), 25.3% (101), and 12.5% (50), respectively. data showed that pica is related to mother education (p = 0.011), mother occupation (p = 0.002), living place (p = 0.011), health complication during pregnancy (p = 0.018), and affects breastfeeding (p = 0.040). baby gender, mother age, iron supplementation, history of child abortion, and economic satisfaction were not significantly associated to pica eating (pe). the results also showed that psychological factors for pe was reduce nausea, ameliorate stress, and anxiety and very fewer pregnant used to control hunger. mother should check for any health complication before pregnancy and receive enough and regular check-up as well as educate themselves regarding consequences of pica. furthermore, psychological consultation is important throughout pregnancy period. keywords: eating disorder, pagophagia, pica eating, pregnancy, risk factors introduction normal eating disturbance is very common during pregnancy and majority mothers experience it. pica eating (pe) is a common eating disorder during pregnancy characterized by consistently ingesting non-nutritive substances.[1] common types of pe include consumption of clay or mud (geophagy), eating ice or ice frost (pagophagy), and consumption of uncooked starch-like row rice (amylophagy).[2] on the other hand, some people practice polypica which known as eating different non-nutritive substances.[3] data from studies have shown that the prevalence of pe is dissimilar among populations. according to the data available, the prevalence of pe in pregnant women ranges between 0.02% in denmark and 74% in kenya.[4] even in some population like malawi, pregnancy is recognized through practicing pica. etiology of pe has been ascertainly known, however, it has been attributed to several factors including health problem such as nutrients deficiency (i.e., iron), exposure to toxins (i.e., lead), and pathogens, gastrointestinal disorder,[5] suppress nausea and vomiting, as well as cultural backgrounds.[6] this complication may cause not only merely discomfort to pregnant women but can also interfere with appetite, adequate dietary intake, reduce intake nutritious foods, and essential nutrients. furthermore, non-food substance might contain an amount of nutrients which is not tolerated, cause malabsorption, and/or containing toxic substances. consequently, it can lead to serious health complications.[7] it can also influence breastfeeding and affect fetus and child health.[8] there is luck of investigation of pe in pregnant women in iraqi kurdistan in terms of prevalence, characteristics, and risk factors and comparing to other populations. therefore, the aims of the study are to study the prevalence of pe, type, and risk factors among kurdish pregnant women its relation to sociodemographic information. also, the investigation of the characteristics of the mothers and their associated risk factors of pe. corresponding author: yaseen m. galali, food technology department, college of agricultural engineering sciences, salahaddin university-erbil, erbil, krg-iraq. e-mail: yaseen.galali@su.edu.krd received: oct 20, 2020 accepted: dec 20, 2020 published: dec 30, 2020 doi: 10.24086/cuesj.v4n2y2020.19-24 copyright © 2020 yaseen m. galali. this is an open access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) galali: pica eating among pregnant women 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 19-24 methodology study design the study was carried in the period between july and august 2020 through interviewing mothers vising maternal care unit at erbil maternity hospital clinics in erbil city, capital of kurdistan region of iraq. electronic form was developed for those who were not able to interview face to face from other districts such as duhok, sulaimani, and halabja due to covid-19 pandemic movement restriction. the inclusion criteria included mothers with no previous history of lifelong health complications such as cancer, cardiovascular diseases, endocrine diseases, and agree to participate. the study was divided into four sections; the first section was on sociodemographic information (age, qualification, residency, occupation, and level of income satisfaction). the second section included obstetric-related information; pregnancy complication (preeclampsia, anemia, hypertension, gestational diabetes, vomiting or nausea, bleeding, and premature contractions), child delivery type, and parity. the third section included assessment of iron taking before and during pregnancy and child feeding type (breastfeeding, formula feeding, and mixture of breastfeeding and formula feeding). the fourth section assessed pe practice questionnaire during pregnancy including non-food items such as ice, flower or tree leaves, clay, soil, hair, cigarette ashes, row rice, coffee grounds or tea, tobacco, charcoal, and/or any other non-food items ingested during pregnancy period. the reasons for pe were also studied including suppress hunger, ameliorate stress and anxiety, and attenuate nausea and vomiting. sample size sample size was calculated using the yemane’s formulation with 50% prevalence at the range for finite targeted population.[9] n=n/(1+ne2)= 400 where, n=corrected size of the targeted sample, e=margin of error 0.05 based on the research condition. n=population size which is estimate to be around 100,000 without any health complication data collection analysis the research was registered at the department of nutrition, university of cihan, erbil. permission from hospitals and clinics administration offices was taken to interview the participants. the collected data were organized double-checked for appropriateness before submitting for analysis. the data were then transported to excel spreadsheet and statistical packages for analysis. the data were then statistically analyzed for significant differences using statistical tests including logistic regression and chi-square from version 22 of spss statistical package for life sciences (spss inc., chicago, il, usa). results data from sociodemographic information of participants showed that pregnant women aged between 20 and 29 were 193 (48.1%) and age between 30 and 39 were 118 (29.4%); birth gender showed male 210 (52%) and female (47.5%). the majority of the mothers had education; high school 135 (33.8%), university 181 (45.1%), but 43 (10.6%) were not education. more than half of the participants (61.3%) were housewife and 126 (31.5%) were employed. regarding the economic level, only 63 (15.7%) were satisfied with low level. the rest moderately 218 (54.4%) and highly 119 (29.9%) satisfied. the majority of the participants were from cities 273 (68.3%). it is interesting that the delivery types like normal vaginal delivery were 185 (46.3%) and cesarean delivery was 215 (53.7%) [table 1]. the prevalence of the pe among studied population was 60.5% (242) and it seems that overtime is reducing. this means that in the first trimester scored the highest with 72.4% and it was reduced to 26.8% and 0.8% in the second and third trimesters, respectively. regarding taking iron supplementation, the data showed that 43.7% of the participants taking iron supplement in a month before pregnancy and 39.6% never taken [figure 1]. on the other hand, taking iron supplement during pregnancy is table 1: sociodemographic characteristics of participants variables count percent birth gender male 210 52.5 female 190 47.5 mother age <19 31 7.6 20–29 193 48.1 30–39 118 29.4 >40 59 14.8 education level of mother elementary 42 10.4 high school 135 33.8 university 181 45.1 non-educated 43 10.6 mother occupational status housewife 245 61.3 employed 126 31.5 student 29 7.2 levels of income satisfaction high 119 29.9 moderate 218 54.4 low 63 15.7 residency city 273 68.3 town 50 12.5 village 77 19.2 delivery type nvd* 185 46.3 cesarean 215 53.7 *normal vaginal delivery galali: pica eating among pregnant women 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 19-24 more prevalent than before pregnancy. in the 1st month, 21.3% take it, after 4th month taking iron seemed to be 16.6% and gradually decreasing to 2.7% in the last month of pregnancy. the most common pica type seemed to be pagophagia (i.e., ice) by 29.7% followed by geophagia (clay), amylophagia (raw starch and rice), coffee phagia (coffee and tea ground), and trichophagia by 25.3%, 12.5%, 8.4%, and 7.9%, respectively [table 2]. the data also showed that there was association between pe with studied variables [table 3]. it seemed that pica is most common in mother give males (33.6%) comparing to females (113%) although not significantly difference (p = 0.351). pica also most common among women aged 20–29 by 30.1%, whereas the least practiced pica is among women over 40 (9.7%) but not significantly different (p = 0.264). mothers educational level had significant influence on pica. it was found that pe is prevalent significantly (p = 0.011) among university and high school educated (22.3% and 21.4%, respectively) comparing to elementary (6.4%) and non-educated women (7.1%). furthermore, pica practice is significantly practiced in housewife (41.7%) comparing to students (4.4%). the results also discovered that pe is significantly associated with residency (p = 0.011), health complication during pregnancy (p = 0.018), and types of infant feeding (p = 0.04) [table 3]. the regression modeling was applied at significant level p < 0.05 for forwarded variables for predicting pica. the factors included table 2: non-food items which used as pica eating by interviewed mothers non-food items n % pagophagia (ice) 119 29.7 geophagia (clay) 101 25.3 amylophagia (raw rice) 50 12.5 coffee phagia (tea or coffee ground) 33 8.4 trichophagia (hair) 32 7.9 cigarette ash 29 7.3 charcoal 25 6.4 flower or leaves 11 2.6 variables pica eating p-value*** yes no n (%) n (%) baby gender 0.351 male 134 (33.6) 76 (19) female 113 (28.2) 77 (19.2) mother age 0.264 <19 22 (5.6) 8 (2.1) 20–29 120 (30.1) 72 (18.1) 30–39 66 (16.4) 52 (13) >40 39 (9.7) 20 (5.1) mother educational level 0.011 elementary 28 (6.4) 14 (3.2) high school 93 (21.4) 43 (9.9) university 96 (22.3) 84 (19.5) no education 31 (7.1) 12 (2.8) mother occupational status 0.002 housewife 167 (41.7) 79 (19.7) employed 63 (15.7) 63 (15.7) student 18 (4.4) 11 (2.8) levels of economic satisfaction 0.137 high 74 (18.5) 45 (11.3) moderate 128 (31.9) 90 (22.5) low 45 (11.3) 18 (4.4) pregnancy occurrence 0.232 wanted 116 (28.9) 131 (20.1) unwanted 81 (32.9) 72 (18.1) parity 0.314 first 71 (17.8) 48 (12) second 79 (19.7) 56 (13.9) three and more 97 (24.3) 49 (12.3) residency urban 156 (39.1) 117 (29.2) 0.011 town 33 (8.3) 17 (4.2) rural 57 (14.4) 19 (4.9) history of abortion or stillbirth 0.239 yes 96 (24.1) 51 (12.7) no 151 (37.7) 102 (25.5) iron supplementation before pregnancy 0.567 yes 77 (19.2) 44 (10.9) no 170 (42.6) 109 (27.3) table 3: association between pregnant women characteristics and pica eating (n=400) (contd...) 1 2 3 4 5 6 7 8 9 noiron before 43.7 2.7 3.7 3.9 2.0 2.6 0.9 0.5 0.3 39.6 during 21.3 7.5 10.5 16.6 9.7 7.5 5.1 3.6 2.7 15.5 0 5 10 15 20 25 30 35 40 45 50 p ar tic ip an ts % months figure 1: iron supplement before and during pregnancy time galali: pica eating among pregnant women 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 19-24 in the regression analysis included mothers education, occupation, health complication during pregnancy, residency, and breastfeeding. only living place (residency) was found significant p = 0.012 [table 4]. regarding factors for practicing pica, 21.3% of participants declared that the main factor is for ameliorating nausea; relieve stress and controlling hunger respectively. tea or coffee ground was frequently used to ameliorate nausea, relieve stress, anxiety, and control hunger by 21.2%, 9.4%, and 10.6%, respectively. discussion this study aimed discovering the prevalence of pica and its association with factors among kurdish pregnant women. the results showed that the prevalence of pica was 60.5%. this number seemed to be higher comparing to other countries. the previous studies have reported that the prevalence of pica is different among countries, for instance, danish women 0.02%, iranian women 8.33%,[1] argentinian women 23.2%,[10] and sudanese women 404%[11] to as high as 63.7% in tanzanian women.[7] pe kurdish women are only lower than tanzania. the factors might push them to practice pica, particularly ameliorating nausea and suppress stress and anxiety. the study found that the first trimester is the highest practiced period of pica. lopez et al. (2012) found similar result noticed in argentinian pregnant women. the most common form of pe was pagophagia, geophagia among pregnant women. kariuki et al. (2016) have found that these two types of pica were the most practiced. however, they should no continue since they might affect blood and minerals availability in diet despite having some inconclusive in vitro studies that support that geophagia releases minerals of the soil but not significant.[12] the study also found that pica practice related to higher mother educational level and mother occupation, urban residency, and health complication [table 2]. the previous study by konlan et al. (2020) found similar result that pica is more prevalent in urban and educated individuals but not significantly different. this may be related to higher level of stress and anxiety that they face during pregnancy and work. therefore, they practice to reduce stress. the previous study stated that pica is more practiced in depressed pregnant women comparing to non-depressed.[13] it is interesting that cesarean delivery is increasing and pica is more common in the women undergo cesarean delivery. this can be kind of mimicry and fear of pain of normal vaginal delivery. ezzeddin et al. (2015) also discovered that cesarean delivery is increasing in iranian pregnant women and it is connect to pica practice. it is also noticed that pe is associated with feeding baby [table 3]. pe was significantly higher in women who they feed breastfeed and formula comparing to women who only breastfeed. this could be associated to the unhealthy pica practice and health complication faced during pregnant and unable to feed the baby properly. torgersen et al. (2010) found that eating disorder interferes with mother’s breastfeeding practice. despite possessing protective role of iron supplementation toward pica, but no significant (p > 0.05) association was found. previous research concluded same result.[4] data in literature have confirmed that pe is mainly due to compensate low level of iron or ameliorate stress and anxiety.[14] in our study, pica seems to have more connection to relieve psychological stress and nausea. the study also noticed that there is no signification correlation between pica and mother age, residency, baby gender, mother age, history of abortion or stillbirth, birth delivery, and pregnancy occurrence, despite having dissimilarities in each variable. the previous studies[15] found that there is no significant relationship between pica and parity and level of income.[16] regarding the factors of pica, there were significant differences between the factors and pe. the data showed that variables pica eating p-value*** yes no n (%) n (%) iron supplementation during pregnancy 0.199 yes 180 (44.9) 119 (29.9) no 68 (16.9) 33 (8.3) health complication during pregnancy 0.018 yes 194 (49) 105 (26) no 53 (13) 48 (12) delivery 0.131 nvd* 107 (27) 78 (19) cesarean 140 (35) 75 (19) infant feeding 0.04 breastfeeding 51 (14.1) 52 (13) formula feeding 86 (20.1) 44 (11.1) mixture feeding** 110 (27.5) 56 (14.1) *nvd: normal vaginal delivery. **mixture feeding (breastfeeding and formula feeding). ***p value level<0.05 considered significantly different using chi-square or fisher’s exact test table 3: (continued) table 4: logistic regression model with pica as the dependent variable p-value or 95% ci lower upper mother educational level 0.158 1.202 1.552 0.931 mother occupational status 0.121 1.293 1.788 0.935 residency 0.012* 0.704 0.927 0.534 health complication during pregnancy 0.055 1.556 2.445 0.99 infant feeding 0.156 0.748 1.118 0.5 *significant level at p<0.05. results of binary logistic regression. or: odds ratio ci: confidence interval galali: pica eating among pregnant women 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 19-24 majority of the participants practiced pica to reduce nausea and ameliorate stress and anxiety as discussed earlier, especially tea and coffee ground and rose and leaves (figure 2). the data also confirmed the fact that these items are non-food and not usually used to suppress hunger but rather reduce stress and nausea that are common in all pregnant women.[14] conclusion the present study conducted to assess pica prevalence, types, and factors of pe. pe seems to be associated with mother education and occupation, health complication, residency, and then influences infant feeding. the most common types of pe found to be pagophagia, geophagia and common risk factors were nausea, stress, and anxiety. therefore, it is important to educate women about consequences of pica before pregnancy and more attention is required to control their health complications. psychological aspects of pregnant women should be taken into consideration as well as they should be educated to control anxiety and stress beside their physiological aspects as pica can be mainly consequence of physiological and/or psychological problems. references 1. n. ezzeddin, r. zavoshy, m. noroozi, h. jahanihashemi and s. h. riseh. prevalence and risk factors for pica during pregnancy in tehran, iran. eating and weight disorders, vol. 20, no. 4, pp. 457-463, 2015. 2. k. d. konlan, j. a. abdulai, k. d. konlan, r. m. amoah and a. doat. practices of pica among pregnant women in a tertiary healthcare facility in ghana. nursing open, vol. 7, no. 3, pp. 783792, 2020. 3. d. f. moore and d. a. sears. pica, iron deficiency and the medical history. the american journal of medicine, vol. 97, no. 4, pp. 390393, 1994. 4. a. boatin, b. wylie, m. p. singh, n. singh, k. yeboah-antwi and d. hamer. prevalence of and risk factors for pica among pregnant women in chhattisgarh, india. american journal of obstetrics and gynecology, vol. 206, no. 1, p. s299, 2012. 5. s. l. young, m. j. wilson, d. miller and s. hillier. toward a comprehensive approach to the collection and analysis of pica substances, with emphasis on geophagic materials. plos one, vol. 3, no. 9, p. e3147, 2008. 6. s. l. young, s. s. khalfan, t. h. farag, j. a. kavle, s. m. ali, h. hajji, k. m. rasmussen, g. h. pelto, j. m. tielsch and r. j. stoltzfus. association of pica with anemia and gastrointestinal distress among pregnant women in zanzibar, tanzania. the american journal of tropical medicine and hygiene, vol. 83, no. 1, pp. 144-151, 2010. 7. c. n. nyaruhucha. food cravings, aversions and pica among pregnant women in dar es salaam, tanzania. tanzania journal of health research. vol. 11, no. 1, pp. 29-34, 2009. 8. l. torgersen, e. ystrom, m. haugen, h. m. meltzer, a. von holle, c. k. berg, t. reichborn-kjennerud andc. m. bulik. breastfeeding practice in mothers with eating disorders. maternal and child nutrition, vol. 6, no. 3, pp. 243-252, 2010. 9. t. yamane. statistics, an introductory analysis. 2nd ed. new york: harper and row, 1967. 10. l. b. lópez, m. marigual, n. martín, m. mallorga, e. villagrán, m. e. zadorozne, m. l. p. de portela and c. r. o. soler. characteristics of pica practice during pregnancy in a sample of argentine women. journal of obstetrics and gynaecology, vol. 32, no. 2. pp. 150-153, 2012. 11. s. ahmed, h. abdullahi and i. adam. practice of pica among pregnant women in khartoum, sudan. international journal of gynecology and obstetrics, vol. 118, no. 1, pp. 71-72, 2012. 12. p. w. abrahams, t. c. davies, a. o. solomon, a. j. trow and j. wragg. human geophagia, calabash chalk and undongo: mineral element nutritional implications. plos one, vol. 8, no. 1, p. e53304, 2013. 13. n. ezzeddin, r. zavoshy, m. noroozi, m. e. sarichloo, h. jahanihashemi and m. notroozi. the association between postpartum depression and pica during pregnancy. global journal 0 5 10 15 20 25 ice clay raw rice tea or coffee ground hair cigarette ash charcoal rose or leaves ice clay raw rice tea or coffee ground hair cigarette ash charcoal rose or leaves ameliorate nausea 3.5 3.5 1.8 21.2 2.4 5.3 0.6 10.6 relieve stress and anxiety 5.9 5.9 1.2 9.4 1.2 2.9 0.6 4.1 control hunger 1.8 1.8 0.6 10.6 1.2 1.2 0.6 2.4 figure 2: association between pica items and factors of pica eating galali: pica eating among pregnant women 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (2): 19-24 of health science, vol. 8, no. 4, pp. 120-126, 2016. 14. a. c. b. da cunha, s. f. patricio, l. p. f. akerman, p. s. maynarde and c. saunders. pica in the pregnancy and related psychological aspects. temas em psicologia, vol. 25, no. 2, pp. 631-646, 2017. 15. a. m. santos, g. r. g. benute, r. m. y. nomura, n. o. santos, m. c. s. de lucia and r. p. v. francisco. pica and eating attitudes: a study of high-risk pregnancies. maternal and child health journal, vol. 20, no. 3, pp. 577-582, 2016. 16. l. kariuki, c. lambert, r. purwestri and h. k. biesalski. trends and consequences of consumption of food and non-food items (pica) by pregnant women in western kenya. nfs journal, vol. 5, pp. 1-4, 2016. . 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 1-6 research article effect of drinking water hardness on kidney stones formation in ranya district akram o. esmail1*, bahast a. qadir2, hawnaz q. hamad2 1department of soil and water, college of agriculture, salahaddin university, erbil, iraq, 2department of biology, college of sciences, raparin university, raparin-sulaimani, iraq abstract this study was conducted in rania district, raparin university, during september 2018–march 2019, to test the relation between water hardness and kidney stone formation. the investigation depended on questionnaire form which was distributed on 100 person in raparin (rania, hajiawa, and chwarqurna) and patients whom vested the rania clinical during december 1, 2018–january 22, 2019 which were 238 patients and only 20 of them had kidney stones developing which represent 8.4% of the total kidney diseases. the results indicated to significant effect of gender at level of significant 5% on kidney stones formation, 10% of male, and 18% of female having kidney stones. the results of chi-square test indicated to highly significant effect of age on kidney stone formation at level of significant (0.001). the kidney stone formation increased from 19.23% to 75% with an increase in age class from (14–34) to (54 or more) year. the negative correlation coefficient value of (r = −0.63*) was recorded between water hardness and stone risk index due to the high magnesium content of drinking water in the studied area. keywords: ca-hardness, component, mg-hardness, stone risk index, total water hardness introduction the urolithiasis regards as a very complex disease, many factors affecting on urinary calculus or formation of kidney stones.[1] hard water means the water which contains more minerals in comparing with ordinary water. the most important ions are calcium and magnesium. hardness is expressed in terms of calcium carbonate (caco 3 ) mg/l. the degree of hardness increases with increasing the concentration of calcium and magnesium in water if the concentration of them <75 m/l is considered as a soft water, 76–150 mg/l moderately hard, and more than 150 mg/l regards as a hard water.[2] numerous studies were conducted in the kurdistan region about water quality and its suitability for drinking purpose, most of them included total hardness, magnesium hardness, and calcium hardness water which directly related to kidney stones developing or forming or regards as one of the factors related to kidney stone formation in the world depending on type of water hardness and magnesium and bicarbonate concentration in drinking water.[1,3] several factors increase the risk for kidney stones forming or developing which were summarized as inadequate water or fluid intake and dehydration, reduce in urinary volume, and increase in concentrations or levels of certain chemicals in urine such as calcium, oxalate, and uric acid to high level or decrease of magnesium to low level or too low and some medical conditions such as reflux and medullary sponge kidney urinary tract infections in additional to tubular acidosis. anything that reduces or blocks the urine flow like urinary obstruction and genetic abnormalities also increase the risk of kidney stones developing.[4] drinking water hardness in the kurdistan region studied by numerous researchers as follow: the water harness in sulaimani governorate the results indicated that the total hardness was ranged (from 110 to 280) mg caco 3 /l, it means most of the studied water hard and may have negative effect of kidney stone developing.[5] the water harness study of (chaq–chaq) kliassan stream in sulaimani city showed that the total hardness was ranged from 110 to 355 mg/l; it means most of studied water hard corresponding author: akram othman esmail, department of soil and water, college of agriculture, salahaddin university, erbil, iraq. e-mail: akram.esmail@su.edu.krd received: apr 14, 2019 accepted: apr 26, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp1-6 copyright © 2020 akram o. esmail, bahast a. qadir, hawnaz q. hamad. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ esmail, et al.: effect of drinking water hardness on kidney stones 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 1-6 to very hard and could have negative effect of kidney stone formation.[6] the water hardness of karees, springs, and streams in erbil governorate was studied by esmail,[7] the results explained that the total hardness was ranged from 151 to more than 1000 mg/l with a mean of 321 mg/l. it means most of the studied water was hard to very hard. the water hardness of kapran near erbil city was studied, the results indicated that the total hardness was ranged between 135.48 and 306.93 mg means must studied moderate hard to very hard.[8] the water hardness of springs at halgwrd mountain was ranged between 93 and 122 mg/l caco 3 , it means that the water of this location is must soft water and suitable for drinking purpose depending on.[9,10] the water hardness in erbil was ranged (138–446) mg caco 3 /l, it means most of the studied water was hard to very hard.[11] the value of total hardness of the water in bator and southwest and south erbil was ranged between 173 and 900 mg caco 3 /l, this indicated that most studied water had hard to very hard class or not suitable for drinking purpose[9] or may have negative effect kidney stone developing.[8,12] the mean of total hardness, ca-hardness, and mg-hardness was (248.82–339.30), (207.49–291.21), and (41.34–97.57) mg/l, respectively, for drinking water in halabja as mentioned by schwart et al.[13] the studied water hardness in dohuk governorate had the values ranged between 281 and 972 mg/l as caco 3 ; it means most of the studied water had hard to very hard class.[14] the water hardness of the studied water in sulaimani governorate recorded the total hardness of (200–580) mg/l as caco 3 means; it refers to that the studied water has hard to very hard class and may have negative effect on kidney stone forming.[15] the inverse relationship was recorded between drinking water hardness and kidney stones developing (urolithiasis) for 2302 patients in various geographical regions of the united state.[16] the workers found the positive correlation between drinking water hardness and kidney stones forming and developing (urolithiasis) in 1000 general hospitals in the united states of america. on the other hand, the weak correlation was recorded between kidney stone-forming and drinking water hardness as mentioned by churchill et al.[17] and shuster et al.[18] the positive correlation was recorded between total water hardness and kidney stones forming in the united kingdom, due to the water hardness values in south and east of england.[19] the relation between drinking water quality (hardness) and the stone risk index (sri) (kidney stone forming) for 24 provincial capitals of iran, was studied by basiri et al.[1] the results indicated that the water hardness was ranged between 57 and 874 ppm; the sri was ranged between 0.00 and 0.0198. the non-significant correlation was recorded between total water hardness of tap water and kidney stone formation, while the negative significant correlation was recorded between mg-hardness and kidney stone formation with the correlation coefficient value of r = −0.51*. in the study conducted in iran from 20 capital cites depending on large number of samples (21200) the negative significant correlation means increase in water hardness cased decrease in risk of kidney stone forming since 30% of total hardness is magnesium hardness which causes decrease in stone formation. since the drinking water of the kurdistan region is hard, especially the water of springs, wells, and rivers in raparin district is hard to very hard which ranged from 150.46 to 1414.80 with the mean value of 337.71 mg/l as caco 3 . it means the water hardness of water in the kurdistan region is above the recommended standard value of who.[9] for the above reasons, the aims of this investigation are to study the effect of: 1. drinking water hardness on kidney stones developing or formation. 2. amount of water intake per day, gender, age, and family history on developing kidney stones. materials and methods for scientific research about the effect of drinking water hardness on kidney stones forming, the data were collected from december 1, 2018 to january 20, 2019 in one clinic of rania city specialized in kidney diseases. the number of patients visited the mentioned clinic was 238 patients and only 20 of them had kidney stones developing or forming. the other source of data collecting was the distribution of questionnaire form on 100 persons (50 male and 50 female) inside and outside of raparin university, which included (hajyawa, chwarqwrna, and ranya), the questionnaire form was shown from table 1 which prepared as mentioned from 20. the survey was conduct on water hardness in the kurdistan region, while 52 water samples in raparin district was depended in calculating risk index of stone formation. the water sample was analyzed from raparin during 2018 by dawson,[20] the total water hardness, magnesium hardness, and calcium hardness we determined according to following models and stone risk formation for 52 samples of water were calculate using following equation as mentioned by basiri et al.[1] stone risk index = sri = [(ca mg/l)/(mg mg/l)]/ (bicarbonate (mg/l). table 1: the questionnaire form age gender male female source of drinking water spring well amount of red meat eat high medium low amount of water intake l/day 3–4 2–3 1–2 l/day family history yes no esmail, et al.: effect of drinking water hardness on kidney stones 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 1-6 total hardness = [ca mg/l*2.50+mg mg/l*4.20]. mg-hardness = (mg mg/l*4.20). ca-hardness = total hardness−mg-hardness. the statistical analysis was conduct using chi-square test depend on the spss program version (22) and regression correlation curve was drown using excel program. results as shown from table 2, the gender affected significantly on kidney stones forming at level of significance (0.05), the statistical analysis indicated that the calculated chi-square value (3.81) is more than tabulated value (3.50). the results indicated that 10% of male and 18% of female having kidney stones forming depending on total sample size (100). on the other hand, if the percentage was calculated separately for male and female, the percentage of samples having kidney stones forming was 20 and 30% for male and female, respectively. table 3 indicates to the highly significant effect of age on kidney stone formation at the level of significant (0.001) with the calculated chi-square value of 15.50. the kidney stone formation increased from 19.23% to 75% with increase in age. table 4 indicates to the effect of source of drinking water on kidney stone was not significant in spite of the percentage of kidney stone in case of a source of drinking spring water was 20%, while the percentage was 29.415 in case of drinking well water. table 5 explains that the amount of consumed meat per person had not affected significantly on kidney stones. chisquare test refers to non-significant effect since the calculated chi-square value (1.29) is less than tabulated value (3.40). table 6 explains that the amount of drinking water (l/day/person) had not affected significantly on kidney stone formation. chi-square test refers to non-significant effect since the calculated chi-square value (1.290) is less than table value. table 2: the effect of gender on kidney stone gender kidney stone percentage of patients having kidney have have not from subsample from total (n=100) male 10 40 20 10 female 18 32 30 18 total 28 72 28 calculated chi-square=3.81 tab. chi-square=3.50 p≥0.05 table 3: explains the effect of age on kidney stone age (year) have kidney stone have not kidney stone percentage of patients having kidney stone 14–34 16 67 19.23 34–54 9 4 69.23 54 or more 3 1 75.00 calculated chi-square value (x2)=15.50*** tab. chi-square=3.20 p≥0.001 table 4: indicates to the effect of source of drinking water on kidney stone source of water have kidney stone have not kidney stone percentage of patients having kidney stone spring 3 12 20.00 well 25 60 29.41 calculated chi-square=3.00 tab. chi-square=3.50 p≥0.15 table 5: indicates to the effect of eating meat on kidney stone meat consumption have kidney have not kidney stone % of patients having kidney stone high 7 25 21.88 medium 12 23 34.29 low 9 24 27.27 calculated chi-square=1.29 tab. chi-square=3.40 p≥0.53 table 6: indicates to the effect amount of drunken water (l/day) on kidney stone-forming amount of drunk water l/day have kidney stone have not kidney stone percentage of patients having kidney stone low (1–2) 16 33 32.65 medium (2–3) 6 15 28.57 high (3–4) 6 24 20.00 calculated chi-square=1.47 tab. value=3.20 p≥0.47 esmail, et al.: effect of drinking water hardness on kidney stones 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 1-6 table 7: effect of family history on kidney stone formation family history have kidney stone have not kidney stone percentage of patients having kidney stone yes 11 23 32.35 no 17 49 25.76 calculated chi-square=0.48 tab. value=3.50 p≥0.49 table 8: explains some important chemical properties of water samples in raparin district location concentration (mg/l) sri hardness (mg/caco 3 /l) mg2+ ca2+ hco 3 mg2+ ca2+ hco 3 1-spring 17.92 61.80 301.65 0.42 73.63 153.88 227.52 2-river 10.27 44.18 286.88 0.55 42.22 110.01 152.23 3-spring 56.93 267.50 3057.93 0.06 233.97 666.08 900.05 4-well 30.49 128.20 1653.10 0.09 125.32 319.22 444.54 5-well 23.64 102.80 547.48 0.29 97.16 255.97 353.13 6-well 23.02 91.86 474.46 0.31 94.60 228.73 323.33 7-spring 19.93 59.78 318.54 0.34 81.92 148.85 230.77 8-well 17.53 44.62 256.75 0.36 72.06 111.10 183.16 9-river 31.42 65.08 1164.49 0.07 129.12 162.05 291.17 10-spring 11.94 50.48 238.39 0.65 49.07 125.70 174.77 11-spring 17.11 88.30 454.08 0.42 70.33 219.87 290.20 12-spring 20.81 88.52 402.05 0.39 85.52 220.41 305.94 13-well 12.55 209.80 990.03 0.62 51.59 522.40 573.99 14-well 15.72 146.00 735.66 0.46 64.61 363.54 428.15 15-well 19.68 69.40 418.22 0.31 80.88 172.81 253.69 16-well 11.81 63.16 273.65 0.72 48.53 157.27 205.80 17-well 21.60 96.34 506.91 0.32 88.78 239.89 328.66 18-well 24.24 83.22 408.64 0.31 99.63 207.22 306.84 19-well 30.36 100.80 513.80 0.24 124.78 250.99 375.77 20-spring 28.30 89.88 433.77 0.27 116.30 223.80 340.10 21-spring 26.56 78.10 387.17 0.28 109.15 194.47 303.61 22-spring 94.99 411.40 1808.65 0.09 390.42 1024.39 1414.80 23-spring 26.74 65.44 349.90 0.26 109.88 162.95 272.83 24-spring 28.30 78.90 409.86 0.25 116.30 196.46 312.76 25-spring 26.44 69.08 356.67 0.27 108.65 172.01 280.66 26-spring 26.71 70.06 371.55 0.26 109.79 174.45 284.24 27-spring 12.22 40.26 193.68 0.62 50.21 100.25 150.46 28-spring 63.29 234.80 1091.90 0.12 260.11 584.65 844.77 29-spring 38.04 108.90 575.29 0.18 156.34 271.16 427.51 30-well 12.88 55.38 257.73 0.61 52.92 137.90 190.82 31-well 15.78 60.48 296.95 0.47 64.86 150.60 215.45 32-well 14.42 55.46 271.51 0.52 59.28 138.10 197.38 33-spring 29.57 95.16 459.33 0.26 121.52 236.95 358.47 34-spring 31.03 113.70 527.47 0.25 127.54 283.11 410.65 35-river 41.88 99.14 551.32 0.16 172.13 246.86 418.99 36-spring 20.29 64.86 315.92 0.37 83.40 161.50 244.90 37-spring 13.08 43.92 220.33 0.56 53.76 109.36 163.12 (contd...) esmail, et al.: effect of drinking water hardness on kidney stones 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 1-6 table 7 explains that the family history of the studied samples person had not affected significantly on kidney stone formation. chi-square test refers to non-significant effect since the calculated chi-square value (0.484) is less than tabulated value. figure 1 shows the correlation between sri and total hardness of 52 water resources (rivers wells and springs) in raparin district. the coefficient of determination (r2) and correlation coefficient (r) between them was 0.426 and −0.65*, respectively. the sri, coefficient of determination (r2), and correlation coefficient (r) were determined from the data recorded in table 8. discussion table 2 shows that the gender affected significantly on kidney stone formation the highest value (18%) was recorded from female, while (10%) was recorded from male this may be due to human activity of male more than female or may be due to biological difference male and female and lifestyle. table 3 refers to highly significant effect of age on kidney stone (0.001). the kidney stone information increased from 19.25% to 75% for age class of (14–34) year to age class of more than (54) year, respectively, since every 10 years the kidney stone formation is possible as mention by basiri et al.[1] and kumar et al.[4] the statistical analysis using chi-square test also indicated to significant effect of age on kidney stone formation since the calculated chi-square value (15.50) was more than tabulated value which was equal to 3.50. on the other hand, the reverse results can be obtained depending on total sample size of this study because the sample size increased with decrease in age. table 4 explains that non-significant effect of source of drinking water on kidney stone formation or there was no significant difference between water hardness of spring water and well water since both of them are hard water to very hard water.[9] the mg-hardness represents 31% of total hardness as shown from table 8, similar results were obtained by basiri et al.[1] in the main cities of iran which caused decrease in risk of stone formation. the significant negative correlation coefficient was recorded between the risk of stone formation and total hardness of 52 water samples in raparin district with the value of r = −0.65** as shown from figure 1 which decrease in the effect of total hardness of drinking water in kidney stone formation. these results agree with those recorded by basiri et al.[1] table 5 explains that consumed meat not affected significantly on kidney stone formation in spite of the figure 1: relation between total hardness of drinking water and stone risk index of kidney 38-spring 18.40 69.06 328.12 0.42 75.61 171.96 247.57 39-well 10.76 61.48 270.35 0.77 44.24 153.09 197.33 40-spring 17.15 53.76 265.11 0.43 70.48 133.86 204.34 41-well 18.06 77.78 355.94 0.44 74.23 193.67 267.90 42-well 19.45 60.42 304.45 0.37 79.95 150.45 230.39 43-well 23.62 80.14 388.39 0.32 97.06 199.55 296.61 44-well 21.78 76.40 406.50 0.32 89.52 190.24 279.75 45-well 27.19 91.88 451.03 0.27 111.76 228.78 340.54 46-well 34.02 69.10 412.85 0.18 139.82 172.06 311.88 47-spring 17.22 60.98 297.50 0.44 70.77 151.84 222.61 48-well 24.38 84.04 416.33 0.30 100.22 209.26 309.48 49-well 41.02 84.52 553.15 0.14 168.58 210.45 379.03 50-well 25.16 101.50 476.59 0.31 103.42 252.74 356.16 51-well 24.96 92.58 442.01 0.31 102.59 230.52 333.11 52-spring 26.51 118.10 613.05 0.27 108.95 294.07 403.02 min 10.27 40.26 193.68 0.06 42.22 100.25 150.46 max 94.99 411.40 3057.93 0.77 390.42 1024.39 1414.80 average 25.33 93.82 535.83 0.35 104.11 233.61 337.71 location concentration (mg/l) sri hardness (mg/caco 3 /l) mg2+ ca2+ hco 3 mg2+ ca2+ hco 3 table 8: (continued) esmail, et al.: effect of drinking water hardness on kidney stones 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 1-6 difference in the percentage of kidney stone formation which ranged between (21.88% and 34.29%). table 6 indicates to decrease in kidney stone formation from 32.65% to 20% with increase amount drunk water per day or water intake per day per person, this may be due to increase water may be prevent precipitation of crystals such as calcium, oxalate, and uric acid in the kidney then decrease stone formation. as shown from table 7, the family history caused an increase in percentage of kidney stone formation from 25.76% to 32.35% but not reached to the significant effect. its appear from the above discussion that there is more than one factor responsible for kidney stone formation for this reason, sometime the result for single factor is not significant, while the combination among the studied factors may cause the kidney stone formation. conclusion and recommendations the kidney stone formation affected significantly by gender and amount of drunken water (l/day/person). the total water hardness was not affected significantly on kidney stone formation since magnesium hardness represents 31% of total hardness which caused decrease in the risk of kidney stone formation. the drinking water in raparin district is hard to very hard, but they have not risk on kidney formation due to high magnesium and bicarbonate of the studied water resources. we highly recommend that: • it is necessary to increase sample size. • it is better to have the same number of studied samples for each age classes. • it is better to have the same number of male and female number in the suture studies. references 1. a. basiri, n. shakhssalim, a. r. khoshdel, h. pakmanesh and m. h. radfar. “drinking water composition and incidence of urinary calculus”. iranian journal of kidney disease, vol. 5, no. 1, pp. 15-20, 2011. 2. a. o. esmail. “limitation of some ground water suitability in erbil plain for different uses”. m. sc. thesis, university of salahaddin. in arabic, 1986. 3. a. o. esmail. “economical importance of water quality in kurdistan”. vol. 5. charmo a quarterly academical social and cultural magazine. university of sulaimani, college of agriculture, pp. 13-20, 2015. 4. v. kumar, a. abbas and j. aster. “robbins basic pathology”. 9th ed. usa: elsevier, p. 928, 2012. 5. s. m. al-shahwani. “a study of the primary productivity of sarchnar spring”. msc. thesis, university of sulaimani, college of science, 1980. 6. a. o. esmail. “role of ionic activity in relation between ionic strength and electrical conductivity of ground water in erbil plain”. journal of dohuk university, vol. 4, no. 1, pp. 73-77, 2001. 7. r. o. rasheed. “a limnlogical study on the some water system in erbil province”. msc. thesis, salahaddin university, college of science, department of biology, 1994. 8. m. a. k. chnaray. “hydrology and hydrochemistry of kapran basin, erbil north of iraq”, ph.d. thesis, university of baghdad, college of science, department of geology, 2003. 9. a. shahab, s. qi, m. zaheer, a. rashid, m. a. talib and u. ashraf. “hydrochemical characteristics and water quality assessment for drinking and agricultural purposes in district jacobabad, lower indus plain, pakistan”. international journal of agricultural and biological engineering, vol. 11, no. 2, pp. 115-121, 2018. 10. f. h. aziz and d. g. a. ganjo. “nutrient status and algal flora in the halgoard (hasarost) mountain area within erbil province, iraq”. journal of dohuk university, vol. 8, no. 3, pp. 103-127, 2003. 11. a. q. nabi. “immunological and bacteriological studies on some well water with in hawker city, kurdistan region, iraq”. m.sc. thesis university of salahaddin, erbil, iraq, 2005. 12. h. o. salh. “role of ion paring and activity in classification irrigation water”. m sc. thesis. salahaddin university, erbil, college of agriculture, department of soil and water, 2008. 13. b. f. schwart, j. bruce, s. leslie and m. l. stoller. “rethinking the role of urinary magnesium in calcium urolithiasis”. journal of endourology, vol. 15, pp. 235-250, 2001. 14. m. s. s. dohuki. “evaluation of some well and spring water in dohuk governorate for irrigation and drinking purpose”. msc. thesis, university of salahaddin, erbil, college of science, department of biology, 1997. 15. g. a. m. rasool. “effect of water quality on nutrient availability for corn”. msc. thesis university of sulaimani, college of agriculture, department of soil and water sciences, 2000. 16. r. sierakowski, b. finlayson and r. landes. “stone incidence as related to water hardness in different geographical region of the united state”. urological resarch, vol. 7, pp. 157-160, 1979. 17. d. churchill, d. bryant, g. fodor and m. h. gault. “drinking water hardness and urolithiasis”. annals of internal medcicine, vol. 88, pp. 513-540, 1978. 18. j. shuster, b. finlayson, r. schaeffer, r. sierakowski, j. zoltek and s. dzegede. “water hardness and urinary stone disease”. journal of urology, vol. 128, pp. 422-450, 1989. 19. d. j. barker and s. p. donnan. “reginal variations in the indices of upper urinary tract stones in england and wales”. the british medical journal, vol. 1, pp. 67-70, 1978. 20. b. m. salam. “limiting water suitablity in ranya district for different uses.” msc. thesis salahaddin uinv. college of agricultural engineering sciences, department of soil and water, 2018. tx_1~abs:at/add:tx_2~abs:at 35 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 35-38 research article assessment of minor disorders among pregnant women attending malafandi primary health-care center/erbil-city ayad a. rashid1, samir m. othman2, dara a. al-banna3, aram m. bra1 1department of community health, college of health technology, cihan university-erbil, kurdistan region, iraq, 2department of community medicine, college of medicine, hawler medical university, erbil, iraq, 3department of nursing, faculty of nursing, tishk international university. erbil, kurdistan region, iraq abstract minor disorders are the discomforts associated with the pregnancy such as nausea, vomiting, backache, heart burn constipation, varicose vein, hemorrhoids, frequency of urination, fatigue, and leg cramps. many women experience some minor disorders during pregnancy; these discomforts should be adequate as they may occur due to hormonal changes and postural changes. the aim of this study was to assess minor disorder among pregnant women. a  descriptive study was conducted in malafandi primary health-care center, from december 2020 to january 2021, 100 pregnant women were taken, and the data were collected by direct interview. the highest percentage of them with 25–34 years old graduated from high school with middle economic status. all of them had minor disorders nausea and vomiting, backache, heartburn, leg cramps, varicose veins, and hemorrhoids: fatigue, frequency of urination, constipation, and loss of appetite. there is a statistically significant association between age group with nausea and vomiting, and hemorrhoids (p ≤ 0.001 and 0.002, respectively). the findings of this study indicated that the most common minor disorder in pregnancy came from fatigue, while the lowest came from constipation. keywords: hormonal change, minor discomforts, minor disorders, pregnancy, primary health-care center introduction minor disorders are the discomforts associated with the pregnancy such as nausea, vomiting, backache, heart burn, constipation, varicose vein, hemorrhoids, frequency of urination, fatigue, and leg cramps. many women experience some minor disorder during pregnancy these discomforts should be adequate as they may occur due to hormonal changes and postural changes.[1] nausea and vomiting during pregnancy known as “morning sickness” affects approximately 80% of pregnant women, nausea, and vomiting a common minor disorder in 1st trimester of pregnancy that the cause is unknown but sometimes due to an increase of human chorionic gonadotrophin and estrogens. backache is cause by the relaxation of the pelvic joint and faulty posture. constipation causes of growing uterus create pressure vertebrectomy. veins are causes of action of progesterone on the blood vessels causing relaxation, making them more circular with sluggish blood flow. fatigue is cause by the increase release of hormones and adds of the fetus. heartburn caused by the regurgitation of acid gastric content into esophagus. frequency of urination causing extra weight and pressure of baby pressing on the bladder and pelvic floor also causes incontinence. hemorrhoids cause of pressure the gravid uterus, and a tendency to constipation.[2] understanding the common disorder of pregnancy to advise the women on strategies that help to cope with the condition and minimize the effects she experiences. the severity and occurrence of minor discomfort differ from mother to mother and other factors, including maternal age and parity.[3] the national institute for care and health excellence report in 2008 shows that minor discomforts are very common and are reported by 50–80% of pregnant women. nausea occurs in 80–85% of all pregnancies during the first-trimester. a clinical gastroenterology report in 2007 shows that heartburn occurs from 30% to 50% of pregnancies, with prevalence approaching 80% in some populations.[4] this study is a benefit to benefit for pregnant women for their health protection and awareness of that disorder which has more effect on women and babies and it is important to find out the most common minor discomfort among pregnant women in erbil city. the aim of the study corresponding author: dara a. al-banna, department of nursing, faculty of nursing, tishk international university, erbil, kurdistan region, iraq. e-mail: dara.albanna@hmu.edu.krd received: july 31, 2021 accepted: february 22, 2023 published: may 1, 2023 doi: 10.24086/cuesj.v7n1y2023.pp35-38 copyright © 2023 ayad a. rashid, samir m. othman, dara a. al-banna, aram m. bra. this is an open-access article distributed under the creative commons attribution license (ccby-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) rashid, et al.: assessment of minor disorders among pregnant women 36 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 35-38 was to assess minor disorders among pregnant women in malafandi primary health-care center/erbil-city. materials and methods a cross-sectional descriptive study was conducted in malafandi primary health-care center in erbil city from december 2020 to january 2021 among 100 pregnant women who recurred by non-probability (purposive) method and were attending for antenatal care unit or medical check-up during pregnancy in malafandi primary health-care center. data were collected using questionnaire by interview technique with pregnant women in the antenatal care unit. pregnant women were excluded from this study that has a specific chronic diseases like cardiovascular disease, psychiatric disease and having gestational diabetes mellitus, urinary tract infection, hypertension induce pregnancy, placenta previa, sever anemia, oligohydramnios, and polyhydramnios. before starting the interview, permission was taken from pregnant women and clarifying the purpose of this study. the data were analyzed by spss version 23 using descriptive statistical analysis of frequency, percentage, mean and standard deviation, and inferential statistical analysis using chi-square and fisher’s exact tests.[14] results table 1 shows that the majority within each age group among pregnant women were 25–34 years old which represented 41%, while the lowest age group was 35–44 years old which represented 19%. the highest percentages of educational level were graduated from high school, which represented 29%, while the lowest percentages of educational level were table 2: reproductive variable data of the 100 pregnant women age (years) of married f % age (years) of married 1–7 65 65 8–15 23 23 16–23 12 12 gravid of group 1 40 40 2–4 39 39 ≥5 21 21 para of group nullipara 46 46 primiparous 20 20 multiparous 28 28 grand multipara 6 6 abortion of group none 75 85 1–2 17 15 >2 8 8 weight group 43–62 31 31 63–82 48 48 >83 21 21 trimester first trimester 32 32 second trimester 45 45 third trimester 23 23 type of pregnancy planned 34 34 unplanned 66 66 chronic disease none chronic disease 92 92 hypertension 6 6 diabetes mellitus 2 2 total 100 100 graduated from an institute which represented 9%. the most percentage among occupational was housewife which given 82%, while the lowest percentage among occupational were public-employee which given 7%. then, the majority of economic statuses were middle which represented 86%. table 2 indicated that the majority of the highest percentage within age groups of married 1–7 which represented 65%, while the lowest age of married were 16–23 which represented 12%. the majority of gravid women were 1 which represented 40%, while the lowest number of gravid women gravid was ≥5 which represented 21%. the majority of para were nullipara (a woman [or female animal] that has never given birth) which represented 46%, while the lowest table 1: sociodemographic characteristics of 100 the pregnant women sociodemographical data f % age group 15–24 40 40 25–34 41 41 35–44 19 19 education illiterate 22 22 primary school 24 24 high school 29 29 institute 9 9 university 16 16 occupation housewife 82 82 self-employee 11 11 public-employee 7 7 economical status high 9 9 middle 86 86 low 5 5 total 100 100 rashid, et al.: assessment of minor disorders among pregnant women 37 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 35-38 table 4: association between age group and some of minor disorder minor pregnancy discomforts age group/years of pregnant woman p-value 15–24 25–34 35–44 f % f % f % nausea and vomiting yes 28 58.3 18 37.5 2 4.2 <0.001 vhsno 12 23.1 23 44.2 17 32.7 backache yes 33 39.8 36 43.4 14 16.9 0.397 nsno 7 41.2 5 29.4 5 29.4 heart burn yes 9 27.3 15 45.5 9 27.3 0.135 nsno 31 46.3 26 38.8 10 14.9 leg cramps yes 17 34 23 46 10 20 0.458 nsno 23 46 18 36 9 18 varicose vein yes 7 25 12 42.9 9 32.1 0.056 nsno 33 45.8 29 40.3 10 13.9 hemorrhoids yes 4 14.8 13 48.1 10 37 0.002 hsno 36 49.3 28 38.4 9 12.3 fatigue yes 38 39.6 39 40.6 19 19.8 0.613 nsno 2 50 2 50 0 0 frequency of urination yes 37 39.8 38 40.9 18 19.4 0.947 nsno 3 42.9 3 42.9 1 14.3 constipation yes 6 27.3 11 50 5 22.7 0.386 nsno 34 43.6 30 38.5 14 17.9 anemia yes 8 28.6 16 57.1 4 14.3 0.123 nsno 32 44.4 25 34.7 15 20.8 loss of appetite yes 16 36.4 22 50 6 13.6 0.223 nsno 24 42.9 19 33.9 13 23.2 of para were grand multiparous (a patient who has had ≥5 births (live or stillborn) at ≥20 weeks of gestation) which represented 6%. the majority of abortions were (none) which represented 75%, while the lowest of abortions were >2 which represented 8%. the majority of women’s weights were 63–82 which represented 48%, while the lowest of women weight <83 which represented 21%. about 45% of the women were second trimester, while 23% of the women were the third trimester. the majority type of pregnancies was unplanned which represented 66%, while 34% of the women were planned. about 92% of the women have non-chronic disease, while 2% of the women have diabetes mellitus. table 3 demonstrate that all of the study sample had minor disorders as nausea and vomiting, backache, heart burn, leg cramps, varicose vein, hemorrhoid’s, fatigue, frequency of urination, constipation, and loss of appetite. the highest percentage (96%) had fatigue and the lowest percentage (22%) had constipation. table 4 shows that was significant statistical association between age group and nausea and vomiting (p ≤ 0.001). there is a statistically significant association between age group hemorrhoids (p = 0.002). discussion the highest age groups of women were between 25 and 34 years old, this finding agrees with a descriptive study done by nazik and eryilmaz.[5] agree with the of the development of a scale for pregnancy‐related discomforts done by shinkawa et al., in 2012.[6] agree with study of associations between nausea, vomiting, fatigue and health-related quality of life of women in theearly pregnancy: the generation r study bai et al., 2016.[7] and agree with a study entitled: associations table 3: assessment of the minor disorder during pregnancy minor disorders items yes no f % f % 1. do you have nausea and vomiting? 48 48 52 52 2. do you have backache? 83 83 17 17 3. do you have heart burn? 33 33 67 67 4. do you have leg cramps? 50 50 50 50 5. do you have varicose veins? 28 28 72 72 6. do you have hemorrhoids? 27 27 73 73 7. do you have fatigue? 96 96 4 4 8. do you have a frequency of urination? 93 93 7 7 9. do you have constipation? 22 22 78 78 10. do you have anemia? 28 28 72 72 11. do you have loss of appetite? 44 44 56 56 12. are you take any medication? 49 49 51 51 13. have you ever smoked during pregnancy? 7 7 93 93 14. have you had previous experiences about minor disorder of pregnancy? 60 60 40 40 total 100 100 100 100 between nausea, vomiting, fatigue and health-related quality of life of women in early pregnancy: the generation r study done by bai et al. 2016 and along with other study done by haider et al. under the title of risk factors of urinary tract infection in pregnancy[8,9] and also agree with the study done by yikar and nazik running with the title of the “effects of prenatal education on complaints during pregnancy and on quality of life” in 2019.[10] concerning educational level the heights percentages of educational level graduated from high school, while the lowest percentages of educational level were rashid, et al.: assessment of minor disorders among pregnant women 38 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 35-38 graduated from institute disagree with research associations between nausea, vomiting, fatigue, and health-related quality of life of women in the early pregnancy: the generation r study bai et al., 2016 who showed the most representative was graduated from the institute and college[7] and in agreement with study done by tshiama et al.[12] the majority of the most percentage among occupational was housewife, while the lowest percentage among occupational were public-employee. then, the majority of economical statuses were middle which disagree with the research, which were associations between nausea, vomiting, fatigue, and health-related quality of life of women in the early pregnancy: the generation r study bai et al., 2015[7] and agrees with the study of nausea, vomiting, fatigue, and health-related quality of life of women in early pregnancy the generation r study done by bai et al., 2015. [8] the highest percentage within age groups of married were 1–7, while the lowest age of married were 16–23 this agree with study incidence of pregnancy-related discomforts and management approaches to relieve them among pregnant women, done by nazik and eryilmaz.[5] the majority of gravid was 1, while the lowest of gravid were ≥5, the majority of para were (nullipara), while the lowest para were (grand multiparaous). the majority of abortions were (none), while the lowest of abortions were >2. the majority of women’s weight was 63–82, while the lowest of women weight >83 these findings agree with the study incidence of pregnancyrelated discomforts and management approaches to relieve them among pregnant women, done by nazik and eryilmaz. [5] the present study showed that less than half of the women were the second trimester, while others of the third trimester. the majority type of pregnancy was unplanned this agree with the study incidence of pregnancy-related discomforts and management approaches to relieve them among pregnant women, done by nazik and eryilmaz. the majority of the women do not have the chronic diseases.[5] the highest percentage had fatigue and the lowest percentage had constipation. this finding agrees with descriptive study done by nazik and eryilmaz[5] and agrees with the study of nausea, vomiting, fatigue, and health-related quality of life of women in early pregnancy: the generation r study done by bai et al., 2015 and agree with the study of development of a scale for pregnancy-related discomforts done by shinkawa et al., 2012.[6] the findings of the current study agreed with the study under the title “associations between nausea, vomiting, fatigue and health-related quality of life of women in early pregnancy: the generation r study” done by bai et al. 2016[7] it is supported by the study done in 2020 entitled “pregnant mother of trimester ii and iii pregnant to change physiological”[11] and a randomized clinical trial study done by poskus et al, in 2022 under the title of “preventing hemorrhoids during pregnancy: a multicenter, randomized clinical trial”[13] the finding of the study showed no significant association between the age group and (fatigue, frequency of urination, constipation, anemia, loss of appetite, backache, heartburn, leg cramps, and varicose veins). the present study showed a significant association between age group and nausea, vomiting, and hemorrhoid’s.[8] conclusion this study concluded that the majority of the study sample indicated that the most common minor disorder in pregnancy came from fatigue while the lowest came from constipation and there is an association between age group and nausea and vomiting. there is a statistically significant association between age groups with nausea and vomiting and hemorrhoids pain. recommendation this study recommended to encouraging the pregnant women to follow-up during pregnancy to prevent any complications any complication due to minor disorders to educate them about the minor disorders of pregnancy and their home selfmanagement to help pregnant mothers to manage their minor disorders at home itself. references 1. p. delma. minor disorder among pregnant women. indian journal of scientific research, vol. 5, no. 9, p. 1551, 2014. 2. p. ian. nursing practice: knowledge and care. john wiley and sons inc., uk. malden, pp. 259-261, 2014. available from: https:// www.books.google.iq/books?isban=8180613623 [last accessed on 2016 dec 01]. 3. p. nitanjali. assessment minor disorder of pregnancy and home remedies. indian journal of scientific research, vol. 4, no, 1, pp. 674, 2005. 4. s. h. maqsood. self-management of pregnant women regarding minor discomforts primary health care centers. hawler medical university/college of nursing, erbil, p. 2, 2015. 5. e. nazik and g. eryilmaz. incidence of pregnancy‐related discomforts and management approaches to relieve them among pregnant women. journal of clinical nursing, vol. 23, no. 11-12, p. 1736-1750, 2014. 6. h. shinkawa, m. shimada, k. hirokane, m. hayase and t. inui. development of a scale for pregnancy-related discomforts. journal of obstetrics and gynecology research, vol. 38, no. 1, p. 316-323, 2012. 7. g. bai, i. korfage, e. hafkamp-de groen, v. w. v. jaddoe, e. mautner and h. raat. associations between nausea, vomiting, fatigue and health-related quality of life of women in early pregnancy: the generation r study. plos one, vol. 11, no. 11, p. e0166133, 2016. 8. g. bai, e. h. groen, h. a. moll, v. w. jaddoe, i. korfage and h. raat. nausea, vomiting, fatigue and health-related quality of life of women in early pregnancy: the generation r study. quality of life research, vol. 24, p. 21, 2016. 9. g. haider, n. zehra, a. munir and a. haider. risk factors of urinary tract infection in pregnancy. the journal of the pakistan medical association, vol. 60, no. 3, p. 213, 2010. 10. s. k. yikar and e. nazik. effects of prenatal education on complaints during pregnancy and on quality of life. patient education and counseling, vol. 102, no. 1, p. 119-125, 2019. 11. e. mail. pregnant mother of trimester ii and iii pregnant to change physiological. journal kebidanan, vol. 9, no. 2, p. 83-88, 2020. 12. c. tshiama, g. bongo, o. nsutier and m. b. babintu. the lay knowledge regarding the prevention of complications related to childbirth: perceptions of congolese pregnant women. qualitative research in medicine and healthcare, vol. 6, no. 1, p. 8740, 2022. 13. t. poskus, z. sabonyte-balsaitiene, l. jakubauskiene, m. jakubauskas, i. stundiene i, g. barkauskaite, m. smigelskaite, e. jasiunas, d. ramasauskaite, k. strupas and g. drasutiene. preventing hemorrhoids during pregnancy: a multicenter, randomized clinical trial. bmc pregnancy and childbirth, vol. 23, no. 11-12, p. 374, 2022. 14. o. polit and b. hunger. nursing research, principle and methods. 6th ed. lippincott, philadelphia, pa, pp. 354-698, 1999. tx_1~abs:at/add:tx_2~abs:at 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 28-33 research article immunohistochemical and molecular studies of p53 and kras protein and their relations to colorectal carcinoma merza h. homady1*, tanya s. salih2, mariamm m. al-jubori3, mustafa d. younus2 1department of biomedical sciences, cihan university-erbil, kurdistan region, iraq, 2department of general biology, cihan university-erbil, kurdistan region, iraq, 3college of medicine, babylon university, iraq abstract the study included 50 tissue blocks embedded in paraffin wax (16 females and 34 males), obtained from a patients group with colorectal cancer (crc), as well as 35 tissue blocks that were embedded in paraffin wax from normal colon (ulcerative colitis) as controls. a relatively few oncogenes and most prominently tumor-suppressing genes, kirsten rat sarcoma virus (kras), and p53 genes have been mutated into a significant part of crcs, and a broad collection of mutated genes has been defined in crc subsets. current findings showed very significant differences between patients and control subjects in the p53-positive rate (p < 0.001). tp53 pro/pro genotype positivity was higher in the control group than in the patient group and this was a significant difference (p < 0.001) with an odds ratio of <1. the genotype  pro/pro was considered to be protective against colorectal carcinoma preventively fractured 0.767. the positive rate of p53 arg/arg genotype in patients was more frequent and statistically significant (p < 0.01), because the odd ratio was more than one. the genotype arg/arg would be considered a colorectal carcinoma risk factor. we conclude that p53 overexpression is used as an indicator of p53 mutation (as identified by immunohistoric chemistry) and kras protein expression was negatively impaired for all the patients in the current study. keywords: kirsten rat sarcoma virus, carcinoma, stage, colorectal, histology introduction carcinogenesis and colorectal cancer (crc) development are a multistage process where colorectal epithelial cells are transported through small adenoma, large adenoma, and subsequently into adenocarcinoma.[1] the development of crc involves several steps, which occur through the development of several genetic changes, including chromosome defects, gene mutations, and the epigenetics of various genes which control and differentiate cell growth, apoptosis, and angiogenesis. crc generally develops from a benign polyp called an adenoma and the subsequent abnormal cell growth, leading to carcinoma, which can spread to other areas of the body.[2,3] tumor suppressing genes and oncogenes mutations and probably several pathways lead to lesion pathology transitions and tumor drive toward malignancy and metastasis.[3] crc is not one disease; it involves a heterogeneous complex of diseases with various genetic and epigenetic changes.[4] modifications of various genes include activating k-ras oncogene, mutating, and removing p53 antioncogene.[5] in particular, mutations of the tp53 tumor suppressor gene and kras oncogene contribute to more than 80% of cases of crc.[6] tp53 encodes the tumor suppressor protein p53, which is a cell cycle regulator; mutations in tp53 leading to crc commonly occur in exons 5-8 .[7] in mitogen active protein kinase signals, kras encodes a gtpase; crcrelated kras mutations are common in codons 12 and 13, both within exon.[8] then, the aim of this study is, therefore, to clarify the proposed role for mutations in crc of the p53 and the kras genes through the determination of frequent crc mutation of kras and p53 by estimating the rate of protein over-expression employing immunohistochemical (ihc) staining and anti-kras antibodies and anit-p53, respectively. corresponding author: merza h. homady, department of biomedical sciences, cihan university-erbil, kurdistan region, iraq. e-mail: merza.homady@ cihanuniversity.edu.iq received: may 16, 2021 accepted: june 13, 2021 published: may 20, 2021 doi: 10.24086/cuesj.v5n1y2021.28-33 copyright © 2021 merza h. homady, tanya s. salih, mariamm m. al-jubori, mustafa d. younus. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) homady, et al.: immunohistochemical and molecular studies of p53 and kras protein 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 28-33 materials and methods this study was conducted in the faculty of science, university of kufa, molecular laboratories and al-sadar teaching hospital in al-najaf province. each paraffin block was processed using hematoxylin and eosin stain for histopathological assessment, mutations in p53 (exon 4 codons 72) were detected by polymerase chain reaction (pcr), mutations detected throughout restriction fragment length polymorphism-pcr (rflp-pcr) in krason (exon 2 codon 12), protein expression had been detected in p53 and the immunohistochemical method of kras. mouse monoclonal anti-human kras, p53, and p53 were used to detect kras as primary antibodies, and p53 proteins were described by al-juborii(20l5).[9] staining results: a visual scoring system was used by a light microscope based on the number of positively stained neoplastic cells in each sample of tissue. the intensity and staining pattern was assigned to each slide. total intensity ratings were evaluated by counting the percentage of positive cells in 100 malignant cells at 40× total objective magnification. the immunostaining was calculated as a percent by the number of malignant cells immunostaining (semi-quantitative scoring). the intensity of stain was measured by counting the positive cell percentage in 100 malignant cells in 25 fields representing the most positive area with an objective magnification of 40×. the staining pattern (qualitative assessment) was either faint (staining pattern that could only be detected using higher magnification, ×40 objective) or dense (a diffuse cytoplasmic staining pattern and easily be seen by lower power objective ×4). kras tissue evaluation: three scoring scales were applied at ×40 objective,[10,11] as follows: score 0: none of the cells revealed positively for tumor marker stains 1score l: 10% of positive tumor cells (weak). 2score 2: <50% of positive cells (moderate). 3 score i3: ≥50% of positive cells (strong). p53 tissue evaluation: p53 nuclear protein expression scored according to: [12] score 0: 0–l0% positive staining of tumor cells considered negative staining; score l+: 11–25% i positive staining of tumor cells, mild or weak staining; score 2+: 26–50% positive staining of tumor cells, moderate staining; and score +3i: >50 positive staining of tumor cells, strong staining. results immunohistochemical expression of p53 and kras the present findings showed a highly significant difference in p53-positive rate between patients and control subjects (p < 0.001), table l. however, the result was limited to patients with colonic carcinoma (46%) whereas none of the control subjects exhibited a positive p53 expression (0%). the p53 ihc expression was detected as brown nuclear dots, as shown in figures l-3. the score of p53 was as follows: score l accounted for 2 (9%) of total positive cases; score 2 was seen in 18 (78%) of figure l: immunohistochemical section p53 expression in the form of brown nuclear stain score 1 (arrow). notice that <10% of the cells are stained (10×) figure 2: immunohistochemical section p53 expression in the form of brown nuclear stain score 2 (arrow). notice that around 10–50% of the cells are stained (10×) figure 3: immunohistochemical section p53 expression in the form of brown nuclear stain score 3 (arrow). notice that >50% of the cells are stained (10×) homady, et al.: immunohistochemical and molecular studies of p53 and kras protein 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 28-33 total positive cases; and score 3 was present in 3 (l3%) of total positive cases [figure 4]. the present results did not show any significant difference in the mean age of patients with positive p53 expression in comparison to patients with negative p53 expression. moreover, patients with rectosigmoid location showed the highest rate of p53 expression (53.33%). in addition, there was no significant association between the site of colonic tumor, stage or grade, and p53 expression. kras expression the immunohistochemical results were negative in all cases of colonic adenocarcinoma and there was not significant correlation between greyscale and stage of tumor (r = −0.l50, p > 0.05), [figure 5]. in addition, these findings did not show any significant differences in mean gray scale between patients with positive p53 expression and patients with negative p53 expression p53 genotype determined by pcr the rate of positivity for tp53 pro/pro genotype was more frequent in control than in patients’ groups and this difference was statistically significant (p < 0.00l), as shown in table 2. the risk was estimated by the use of odds ratio statistic which was 0.097 with a 95% confidence interval of 0.034–0.28l, this was due to odds ratio which is <l. pro/pro genotype would be considered protective against colorectal carcinoma with a preventive fraction of 0.767, as shown in figure 6. the rate of positivity for the p53 arg/arg genotype was more frequent in the patients group than in the control group and this difference was statistically significant (p = 0.019), as shown in table 3. risk was estimated by the use of adjusted odds ratio statistic which was 14.2 with a 95% confidence interval of 0.792–245.7 because odds ratio is >1. among these findings, the arg/arg genotype would be considered a risk factor for colorectal carcinoma. the etiologic fraction was not estimated because one of the cells contains zero counts, as shown in figure 7. these results did not reveal any significant association between arg/arg genotype, in patients group, and table 2: age and agnor in colorectal carcinoma patients in relation to pro/pro genotype pro/pro n mean sd p-value age negative 34 49.824 18.300 positive 16 54.875 12.366 0.323 total 50 51.440 16.674 agnor negative 34 6.097 0.817 positive 16 6.238 0.784 0.569 total 50 6.142 0.801 table 3: age and agnor in colorectal carcinoma patients in relation to arg/arg genotype arg/arg n mean sd p-value age negative 42 51.405 16.200 positive 8 51.625 20.220 0.973 total 50 51.440 16.674 agnor 42 6.088 0.737 negative positive 8 6.425 1.099 0.280 total 50 6.142 0.801 table 1: the rate of positive p53 ihc expression in control and patients groups p53 control colonic carcinoma no. % no. % positive 0 0 23 46 negative 35 l00 27 54 total 35 l00 50 l00 p<0.00l, corrected χ2=l9.803; df=i score 1 2 9% score 2 18 78% score3 3 13% figure 4: the proportions of p53 scores figure 5: the spearman correlation between gray scale intensity and stage homady, et al.: immunohistochemical and molecular studies of p53 and kras protein 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 28-33 clinicopathological parameters including age, gender, site of tumor, grade, stage, and agnor count. the rate of positivity for p53 arg/pro genotype was more frequent in patient groups than in control group and this difference was statistically significant (p < 0.001), as shown in table 4. value was estimated by the use of odds ratio statistic which was 5.236 with a 95% confidence interval of 1.852– 14.807 (odds ratio is >l). therefore, we can conclude that arg/pro genotype would be considered a risk factor for colorectal carcinoma (etiologic fraction was 0.657). evaluation of kras mutation using rflp-pcr colonic tissue obtained from all patients with colorectal carcinoma was proved to be negative for kras mutation by the use of the rflp-pcr technique. discussion role of immunohistochemical p53 expression in colorectal carcinoma the present study showed that p53 immunohistochemical expression was not correlated with grade or stage of tumor in patients with colorectal carcinoma. to explain that, one may suppose that p53 mutation is an early event in colorectal carcinoma and hence can be seen in low-grade, intermediategrade, and high-grade lesions and can be seen in all stages of disease.[13] another opinion is that p35 occurs very early in the progression of the tumor during time of conversion from adenoma to carcinoma, and hence, it will be a fixed mutation pattern in all grades and stages and eventually no association will be obtained between p53 mutation and grade of tumor and between p53 mutation and stage of the disease.[14] importantly, the absence of p53 expression in benign tumors indicated that p53 can be considered as a good marker for malignant colorectal tumors. however, and in harmony with several studies;[l3,14] this marker was not useful for the classification of the different histopathological grades and hence prognosis of the disease. the reported pattern of p53 expression in the different stages has been interpreted into different and sometimes contradicting directions. if we consider that only 50% of poorly differentiated tumors expressed p53, p53 might be correlated with poor outcome and bad prognosis. on the other hand, differences between the different histological grades were not significant abolishing the prognostic value of this marker. this might not be surprising if we know that several studies indicated that p53 nuclear staining does not always rule out the presence of mutated malfunctioned p53 protein.[15] menezes[14] studied 82 patients with colorectal carcinoma for the immunohistochemical expression of p53, bcl-2, and ki67 and concluded that there was no significant correlation between the expressions of these markers separately or in conjunction, table 4: arg/pro genotype in colorectal carcinoma patients and control group among this study there was no significant association between arg/pro genotype, in patients group, and clinicopathological parameters including age, gender, site of tumor, grade, stage, and agnor count arg/pro colonic carcinoma control p-value 95%ci no. % no. % odds ratio lower upper ef positive 26 52.00 6 17.14 negative 24 48.00 29 82.86 0.001 5.236 1.852 14.807 0.657 total 50 100.0 35 100.00 figure 6: pcr amplification of the tp53 codon 72 (electrophoresis in 2% agarose get) in 13 colorectal adenocarcinoma samples and control samples. lanes 1 and 10 markers, lanes 3, 4, 5, 6, 7, 12, and 13 positive for pro allele (177 bp). lanes 2, 8, 9, and 11 negative for pro allele. lanes 14 and 15 negative control figure 7: pcr amplification of the tp53 codon 72 (electrophoresis in 2% agarose get) in 13 colorecta1 adenocarcinoma samples and control samples. lane 1 and 10 markers, lanes 3, 5, 6, 12, and 14 positive for arg allele (141 bp). lanes 2, 4, 7, 8, and 9 negatives for arg allele. lanes 11, 13, and 15 negative control homady, et al.: immunohistochemical and molecular studies of p53 and kras protein 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 28-33 concerning the grade of tumor. asaad[16] conducted a study on the role of p53 and cyclin d1 in 41 patients with colorectal carcinoma and reported that p53 immunohistochemical expression was not correlated to grade or stage of tumor. another study conducted by malik[17] on 50 patients with colorectal carcinoma and found no association between grade of tumor and p53 mutation. it was reported that by ghavamnasiri,[18] there was no significant association between p53 protein expression and some common clinicopathologic variables such as age, gender, site of tumor, pathologic type, and stage of the disease. the lack of significance reported herein might indicate that the defensive role of p53 expression is accompanied by other pathways that define the prognosis and grade of the tumor. kras mutations, for example, were one of the targets which support that direction.[19] on the contrary, several works of literature had reported significant associations between p53 expression and grade and stage of tumor: [20,21] from the above discussion, one can conclude that the role of p53 as a prognostic factor in colorectal carcinoma is still controversial and need thorough investigation of the contrary to the proven fact of the prognostic value for p53 expression in several other solid tumors. role of agnor in colorectal carcinoma the present study showed that agnor score was significantly higher the patient group in comparison with the control one, and also, it was significantly correlated with the grade of tumor, being higher with less differentiated tumors. these findings can be attributed to the fact that malignant tumor cells are often characterized by aberrant chromatin patterns such as polyploidy and aneuploidy, and this abnormal chromatin pattern will be reflected in the form of high score nuclear organizing regions. arginine and proline genotype polymorphism in colorectal carcinoma the present study showed that pro/pro genotype was protective against colorectal carcinoma while arg/arg and arg/pro genotypes were risk factors for colorectal carcinoma, on the other hands, the present study failed to establish any significant correlation between above-mentioned genotypes and various clinicopathological characteristics of patients enrolled in the study. this may be attributed to the following suggestion: arg/arg genotype may be transcribed into less functioning or malfunctioning p53 protein and as it is well known that p53 is the guardian of the genome, the cell will be rendered liable for targeting by many mutagenic agents with the ultimate acquisition of the neoplastic phenotype. onrat[22] conducted a study on 35 patients with colorectal carcinoma and reported that individuals homozygous for the arg allele have a higher frequency than other alleles and that colon cancer may be related to arg allele frequency. dastjerdi[23] also studied tp53 gene polymorphism in iran and stated that a significant difference between cases and controls was found for the arginine/arginine genotype compared with (grouped) arginine/proline and proline/proline genotypes (odds ratio = 1.451 [1.002–2.103], p = 0.048) and that arginine/arginine genotype may be correlated with overexpression of p53 and increased risk for crc. these findings support the findings of the current study. in the present study, none of the tissue samples taken from the paraffin block of patients with colorectal carcinoma was positive for kras mutation. smith[24-26] stated the kras mutations were significantly more common in rectal than in colon tumors, indicating differences in the pathways of carcinogenesis in these tissues. p53 and kras mutations were rarely found together in the same tumor, suggesting different genetic pathways leading to tumor formation, a finding which strongly supported the finding of the present study. it was reported that the kras and p53 mutations rarely coexist in the same tumor,[27] indicating that these alterations do not represent a synergistic evolutionary pathway, again these findings are similar to the findings of the present study. conclusions from the achieved results of this study, we can conclude the following: p53 is an independent prognostic marker and be regarded as an early event in the pathogenesis of crc. the p53 and kras mutations are not found together in the same tumor and neither kras gene mutations nor its protein overexpression were detected in colorectal carcinoma. such findings necessitate the need for conducting a specific study focusing on kras genes expression and protein overexpression in colorectal carcinoma. references 1. e. r. fearon and b. vogelstein. a genetic model for colorectal tumorigenesis. cell vol. 61, no. 5, pp. 759-767, 1990. 2. a. russo, s. rizzo, g. bronte, n. silvestris, g. colucci, n. gebbia, v. bazan and f. fulfaro. the long and winding road to useful predictive factors for anti-egfr therapy in metastatic colorectal carcinoma: the kras/braf pathway. oncology, vol. 77, no. suppl 1, pp. 57-68, 2009. 3. f. coppedè, a. lopomo, r. spisni and l. migliore. genetic and epigenetic biomarkers for diagnosis, prognosis and treatment of colorectal cancer. world journal of gastroenterology, vol. 20, no. 4, p. 943, 2014. 4. b. leggett and v. whitehall. role of the serrated pathway in colorectal cancer pathogenesis. gastroenterology, vol. 138, no. 6, pp. 2088-2100, 2010. 5. w. jin, m. q. gao, z. w. lin and d. x. yang. multiple biomarkers of colorectal tumor in a differentia1 diagnosis model: a quantitative study. world journal of gastroenterology, vol. 10, no. 3, pp. 439442, 2004. 6. s. d. markowitz and m. m. bertagnolli. molecular basis of colorectal cancer. new england journal of medicine, vol. 361, no. 25, pp. 2449-2460, 2009. 7. b. iacopetta. tp53 mutation in colorectal cancer. human mutation, vol. 21, no. 3, pp. 271-276, 2003. 8. y. imamura, t. morikawa, x. liao, p. lochhead, a. kuchiba, m. yamauchi, z. r. qian, r. nishihara, j. a. meyerhardt, k. m. haigis, c. s. fuchs and s. ogino. specific mutations in kras codons 12 and 13, and patient prognosis in 1075 braf wild-type colorectal cancers. clinical cancer research, vol. 18, no. 17, pp. 4753-4763, 2012. 9. m. b. al-jubori. immunohisto chemical and molecular study of colorectal cancer patients. ph.d. thesis, college of science, kufa university, iraq, 2015. 10. p. uribe, l. andrade and s. gonzalez. lack of association between braf mutation and mapk erk activation in melanocytic nevi. homady, et al.: immunohistochemical and molecular studies of p53 and kras protein 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 28-33 journal of investigative dermatology, vol. 126, no. 1, pp. 161-166, 2006. 11. l. m. mcglynn, t. kirkegaard, j. edwards, s. tovey, d. cameron, c. twelves, j. m. s. bartlett and t. g. cooke. ras/raf-1/mapk pathway mediates response to tamoxifen but not chemotherapy in breast cancer patients. clinical cancer research, vol. 15, no. 4, 1487-1495, pp. 2009. 12. e. ioachim, m. c. michael, m. salmas, k. damala, e. tsanou, m. m. michael, v. malamou-mitsi and n. e. stavropoulos. thrombospondin-1 expression in urothelial carcinoma: prognostic significance and association with p53 alterations, tumour angiogenesis and extracellular matrix components. bmc cancer, vol. 6, no. 1, pp. 1-8, 2006. 13. p. f. rambau, m. odida and h. wabinga. p53 expression in colorectal carcinoma in relation to histopathological features in ugandan patients. african health sciences, vol. 8, no. 4, pp. 234238, pp. 2008. 14. h. l. de menezes, m. j. jucá, e. g. a. de gomes, b. l. nunes, h. o. costa and d. matos. analysis of the immunohistochemical expressions of p53, bcl-2 and ki-67 in colorectal adenocarcinoma and their correlations with the prognostic factors. arquivos de gastroenterologia, vol. 47, no. 2, pp. 141-147, 2010. 15. s. strano, s. dell’orso, s. di agostino, g. fontemaggi, a. sacchi and g. blandino. mutant p53: an oncogenic transcription factor. oncogene, vol. 26, no. 15, pp. 2212-2219, 2007. 16. n. y. asaad, m. a. kandil and n. m. mokhtar. prognostic value of cyclin d1 and p53 protein in colorectal carcinoma. therapy, vol. 42, p. 48, 2000. 17. a. malik, r. n. mishra, b. fanthome, r. rao and s. r. patrikar. role of cd34, vascular endothelial growth factor, and p53 in neoangiogenesis as correlated with stage of disease in colorectal carcinoma. medical journal armed forces india, vol. 67, no. 4, pp. 320-325, 2011. 18. n. mr. ghavam, t. m. seylanian, e. rezaei, k. ghafarzadegan and h. malekifard. expression of p53 in colorectal carcinoma: correlation with clinicopathologic features. vol. 10, no. 1, pp. 38-42, 2007. 19. a. conlin, g. smith, f. a. carey, c. r. wolf and r. j. c. steele. the prognostic significance of k-ras, p53, and apc mutations in colorectal carcinoma. gut, vol. 54, no. 9, pp. 1283-1286, 2005. 20. c. v. georgescu, a. saftoiu, c. c. georgescu, r. ciurea and t. ciurea. correlations of proliferation markers, p53 expression and histological findings in colorectal carcinoma. journal of gastrointestinal and liver diseases, vol. 16, no. 2, p. 133, 2007. 21. a. hegazy, s. a. daoud, w. s. ibrahim, k. el-atrebi, m. saker and n. abdel-wahab. role of ki-67, p53 and bcl-2 in advanced colorectal carcinoma (histopathological and immunohistochemical study). academic journal of cancer research, vol. 7, no. 3, pp. 168-172. 2014. 22. s. t. onrat, e. ellidokuz, a. küpelioğlu and e. durhan. frequency of tp53 codon72 polymorphism in cases with colon cancer. turkish journal of cancer, vol. 39, no. 1, p. 72, 2009. 23. m. n. dastjerdi. tp53 codon 72 polymorphism and p53 protein expression in colorectal cancer specimens in isfahan. acta medica iranica, vol. 49, no. 2, pp. 71-77, 2011. 24. g. smith, f. a. carey, j. beattie, m. j. v. wilkie, t. j. lightfoot, j. coxhead, r. c. garner, r. j. c. steele and c. r. wolf. mutations in apc, kirsten-ras, and p53-alternative genetic pathways to colorectal cancer. proceedings of the national academy of sciences, vol. 99, no. 14, pp. 9433-9438, 2002. 25. s. m. salama, i. a. a. ibrahim, n. shahzad, s. al-ghamdi, n. ayoub, a. s. alrashdi, m. a. abdulla, n. salehen and m. bilgen. hepatoprotectivity of panduratin a against liver damage: in vivo demonstration with a rat model of cirrhosis induced by thioacetamide. apmis, vol. 126, no. 9, pp. 710-721, 2018. 26. e. bagheri, k. saremi, f. hajiaghaalipour, f. l. faraj, h. m. ali, m. a. abdulla, s. l. khaing and n. salehen. synthesis of novel derivatives of quinazoline schiff base compound promotes epithelial wound healing. current pharmaceutical design, vol. 24, no. 13, pp. 1395-1404, 2018. 27. d. calistri, c. rengucci, i. seymour, a. lattuneddu, a. m. polifemo, f. monti, l. saragoni and d. amadori. mutation analysis of p53, k‐ras, and braf genes in colorectal cancer progression. journal of cellular physiology, vol. 204, no. 2, pp. 484-488, 2005. . 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 21-25 research article levels of apelin, endoglin, and transforming growth factor beta 1 in iraqi women with polycystic ovary syndrome noor alhuda k. ibrahim1, wasnaa j. mohammad1, sanan t. abdawahab2* 1department of basic sciences, college of nursing, baghdad university, baghdad, iraq, 2department of medical laboratory techniques, dijlah university college, baghdad, iraq abstract polycystic ovary syndrome (pcos) is one of the most common causes of infertility in women of reproductive age. the aim of the study was to determine the level of apelin, insulin resistance (ir), transforming growth factor beta 1 (tgf-β1), and endoglin in women with polycystic ovary syndrome. fifty pcos patients and 40 non-pcos infertile patients were recruited. the fasting serum levels of folliclestimulating hormone (fsh), luteinizing hormone (lh), testosterone (t), prolactin, fasting blood glucose, insulin, and apelin at the early follicular phase were measured. levels of apelin, lh, lh/fsh, t, and fasting insulin, as well as homeostatic model assessment of ir (homa-ir) in pcos patients, were significantly higher than in the control group. correlation analysis showed that apelin level was positively correlated with body mass index and homa-ir. apelin levels and tgf-β1 were significantly increased in pcos patients while show decrease levels of endoglin. keywords: apelin, endoglin, transforming growth factor beta introduction polycystic ovary syndrome (pcos) is the most common endocrine disease affecting women of reproductive age. the prevalence of pcos varies according to the diagnostic criteria used, with estimates ranging from 9% in women of reproductive age, according to nih criteria, up to 18%, with the rotterdam criteria.[1] it is primarily characterized by ovulatory dysfunction and hyperandrogenism,[2] but the clinical presentation is heterogeneous and patients may present some of various signs and symptoms. this heterogeneity seems to be modulated by multiple factors such as prenatal androgen exposure, nutritional status in the uterus, genetic factors, and ethnicity, insulin resistance (ir) of puberty and/or exaggerated adrenarche and changes in body weight.[3] environmental factors, such as obesity, appear to exacerbate the underlying genetic predisposition. concerning ethnicity, the presence of hirsutism is less frequent in asian patients (around 10%), compared to caucasian ones (70%).[4] obesity is a prevalent characteristic of pcos, ranging from 12.5% to 100%, with a pooled estimated prevalence of 49%, as shown by a recent meta-analysis.[5] the presence of obesity may exacerbate the metabolic and reproductive disorders associated with the syndrome, including ir, dyslipidemia, and metabolic syndrome.[6] a meta-analysis[7] has shown that women with pcos have higher levels of triglycerides (tg), low-density lipoprotein (ldl)-cholesterol and total cholesterol (tc), and lower high-density lipoprotein (hdl)-cholesterol levels compared with control women, regardless of body mass index (bmi). in addition, pcos women present higher risk for type 2 diabetes. pcos is also associated with a clustering of cardiovascular risk factors.[8] however, there is no definitive evidence for increased cardiovascular events, nor data showing that pcos alone leads to increased cardiovascular risk independent of associated risk factors. in fact, more rigorous cohort studies of longterm cardiovascular outcomes and clinical trials of risk factor modification are required for women with pcos. in addition, evidence suggests clinical phenotypes are related with different metabolic risks. in this sense, ir seems to be a specific feature of the classic phenotype and, to a lesser extent, of the ovulatory phenotype. non-hyperandrogenic phenotype behaves as a separate group that is metabolically similar to non-pcos women.[7] cihan university-erbil scientific journal (cuesj) corresponding author: sanan t. abdawahab, department of medical laboratory techniques, dijlah university college, baghdad, iraq. e-mail: sanan.thaer@duc.edu.iq received: apr 11, 2019 accepted: apr 22, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp21-25 copyright © 2020 noor alhuda k. ibrahim, wasnaa j. mohammad, sanan t. abdawahab. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ ibrahim, et al.: apelin, endoglin, and tgf-β1 in polycystic ovary syndrome 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 21-25 adipose tissue is considered not only a storage tissue but also aproper endocrine organ, metabolically active.[6] ir is considered the main pathogenic factor in the background of increased metabolic disturbances in women with pcos which can explain hyperandrogenism, menstrual irregularity, and other metabolic manifestations seen in this disease.[7] however, it is not a diagnostic criterion for pcos.[8] obesity alters the expression, circulating levels, and signaling mechanisms of adipose-secreted factors and these changes have been linked to the development of ir, type 2 diabetes, dyslipidemia, atherosclerosis, cancer, and other diseases.[9] apelin is an endogenous cytokine,[10] an adipokine,[11] a regulatory peptide,[12] and a neuropeptide[13] that has been linked to obesity and ir. apelin is a 36-amino acid peptide (ap-36) generated from a larger precursor the 77-amino acid proapelin. the human preproapelin gene is located on chromosome x at locus xq25-q26.1.[10] collected data from both the clinical and basic settings show that ap (1) correlates with states of ir and obesity, (2) stimulates glucose utilization, (3) decreases insulin secretion, and (4) negatively regulates catecholamine-mediated lipolysis. ap seems to be a key regulator in glucose and lipid metabolism and may be associated with ir and possibly implicated in pcos which is known to be associated with increased ir[14] and from hence the possible link to apelin levels. endoglin (seng)/cd105 is a transmembrane auxiliary receptor for transforming growth factor beta (tgf-b) it works as a non-signaling coreceptor of the tgf-b modulating its responses. there are a number of reports suggesting that soluble endoglin may be regarded as a biomarker of endothelial dysfunction, for instance, atherosclerosis, hypercholesterolemia,[15] type 2 diabetes mellitus (t2dm), and hypertension.[16] tgf-b1 is a multifunctional cytokine that is produced by a variety of cells and its level dysregulated in women with pcos, which might play a role in the pathophysiology of this syndrome. substantial evidences implicate that tgf-b1 activation is closely associated with endothelial dysfunction and hypertension.[17] there is scattered evidence for the association between tgf-b1, visceral adiposity, metabolic syndrome, nonalcoholic fatty liver disease, and t2dm.[15] methods this case–control study was conducted at the obstetrics and gynecology department in medical city in baghdad. it was conducted on ninety women during the period from march 2018 to september 2018. these were divided into two groups: group i: fifty overweight and obese women with pcos (pcos group) and group ii: forty overweight and obese women without pcos (non-pcos group). the recruited women were counseled and written informed consent was obtained from each woman before her participation in the study. diagnosis of pcos was made according to the rotterdam criteria[18] in the presence of at least two of the following: (a) oligomenorrhea and/or anovulation, (b) biochemical and/or clinical hyperandrogenism, and (c) ultrasound appearance of polycystic ovaries (multiple cysts >12 in number of 2–9 mm size),[3] bmi p 25 kg/m2. samples were obtained between days 2 and 5 of normal spontaneous or withdrawn menstrual cycle. blood allowed to clot for 10–20 min at room temperature and centrifuged at (2000–3000 rpm) for approximately 20 min to separate the serum from the cells. 2 ml serum was stored at _20_c until the time of ap-36 and hormonal profile assay. hormonal evaluation included a baseline plasma determination of luteinizing hormone (lh), follicle-stimulating hormone (fsh), dehydroepiandrosterone sulfate, tsh, e2, and free testosterone. all hormones were measured by radioimmunoassay methods using commercial kits (dsl inc., webster, texas, usa). the fasting insulin was measured by radioimmunoassay methods using commercial kits (the intra and inter-assay coefficients of variation were 1.98% and 0.84%, respectively) (immunospect corporation, owensmouth ave, canoga park, ca). fasting glucose was measured by the glucose oxidase method (yellow springs instrument, yellow springs, oh, usa). tc, tg, and hdl were assayed using fully automated clinical chemistry autoanalyzer system konelab 20i ldl was calculated according to friedewald formula: ldl = tc−(tg/5+hdl).[19] ir was calculated by homeostatic model assessment (homa) using the following formula: homa-ir = ([fg in mg/dl_0.05551]_ fi in mu/ml)/22.5. a diagnosis of ir established for homa-ir values >2.71 nmol_mu/l[20] equal (fg in mg/dl_fi in mu/ ml)/405. ir was diagnosed according to the fg to fi ratio. if a participant’s fg/fi was 4.5 or less, she would be classified as being ir and if her fg/fi was >4.5, she would be classified as being non-ir.[21] results the results obtained are shown in table 1 and table 2. the group mean±sd p-value pcos control age (year) 28.28±5.11 33.27±6018 0.00033* bmi (kg/m2) 32.16±2.28 29.07±1.40 0.00001* hirsutism score 13.79±1.2 5.33±0.75 0.00001* tgf-β1 (ng/ml) 19.34±2.73 12.29±1.19 0.00001* table 1: the mean and sd of biochemical and immune markers level in the patients group and healthy control group (contd...) ibrahim, et al.: apelin, endoglin, and tgf-β1 in polycystic ovary syndrome 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 21-25 table 2: correlation between apelin, transforming growth factor beta, and endoglin with other parameters in the pcos disease patients the group correlation coefficient (r) tgf-β1 (ng/ml) s-endoglin ng/ml apelin (pg/ml) age (year) 0.0917 −0.0948 −0.1572 bmi (kg/m2) 0.3333 0.0379 0.0353 hirsutism score −0.2653 0.0955 −0.1134 tgf-β1 (ng/ml) 0.2007 0.1903 s-endoglin ng/ml 0.2007 −0.0502 apelin (pg/ml) 0.1903 −0.0502 fasting plasma glucose (mg/dl) 0.1447 −0.0654 −0.0531 insulin (iu/ml) −0.0978 −0.0906 0.0132 homa 0.1547 −0.1586 0.1233 prolactin (pg/ml) −0.1503 0.3305 −0.223 fsh (miu/ml) −0.0841 −0.0611 −0.1269 lh (miu/ml) −0.0413 0.0798 0.2071 shbg (nmol/l) 0.2081 −0.189 0.198 dhea-s (mg/ml) −0.1289 −0.0002 −0.1991 total testosterone (ng/ml) 0.0887 −0.1985 −0.0872 free testosterone (pg/ml) 0.1543 −0.4447 0.0568 h-crp (μg/ml) 0.0484 −0.1559 −0.0966 cholesterol mg/dl −0.1114 0.0285 0.323 triglyceride (mg/dl) 0.354 0.1584 0.0839 hdl (mg/dl) −0.1063 0.1887 −0.0182 ldl (mg/dl) −0.1063 −0.1289 0.43 *(p<0.05), **(p<0.01), ns: non-significant. pcos: polycystic ovary syndrome, bmi: body mass index, tgf: transforming growth factor, homa: homeostatic model assessment, fsh: follicle-stimulating hormone, lh: luteinizing hormone, shbg: sex hormone-binding globulin, dhea: dehydroepiandrosterone, h-crp: high sensitivity c-reactive protein, hdl: high-density lipoprotein, ldl: low-density lipoprotein the group mean±sd p-value pcos control s-endoglin ng/ml 3.37±1.25 5.02±1.34 0.00001* apelin (pg/ml) 220.92±16.87 367.05±37.87 0.00001* fasting plasma glucose (mg/dl) 94.76±20.71 90.0±14.7 0.00001* insulin (iu/ml) 18.76±4.33 12.0±6.5 0.00001* homa 2.44±1.16 1.72±1.01 0.001375* prolactin (pg/ml) 13.71±1.25 13.14±1.56 0.02867* fsh (miu/ml) 7.04±1.13 6.03±1.32 0.000099* lh (miu/ml) 12.13±1.77 5.88±1.2 0.00001* shbg (nmol/l) 30.25±2.51 54.12±4.3 0.00001* dhea-s (mg/ml) 2.21±0.59 0.95±0.3 0.00001* total testosterone (ng/ml) 0.91±0.41 0.59±0.22 0.000017* free testosterone (pg/ml) 2.65±0.66 1.064±0.26 0.00001* h-crp (μg/ml) 5.75±1.34 2.3±0.7 0.00001* cholesterol mg/dl 172.6±49.9 166.0±27.3 0.254423 ns triglyceride (mg/dl) 155.86±34.69 103.0±32.2 0.00001* hdl (mg/dl) 39.14±10.09 39.92±7.15 0.347843 ns ldl (mg/dl) 103.42±36.31 101.9±24.2 0.413274 ns *(p<0.05), **(p<0.01), ns: non-significant. pcos: polycystic ovary syndrome, bmi: body mass index, tgf: transforming growth factor, homa: homeostatic model assessment, fsh: follicle-stimulating hormone, lh: luteinizing hormone, shbg: sex hormone-binding globulin, dhea: dehydroepiandrosterone, h-crp: high sensitivity c-reactive protein, hdl: high-density lipoprotein, ldl: low-density lipoprotein table 1: (continued) ibrahim, et al.: apelin, endoglin, and tgf-β1 in polycystic ovary syndrome 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 21-25 discussion pcos is one of the most common causes of infertility in women of reproductive age. ir is a main pathophysiologic feature in these patients.[2] apelin (apln) is a recently discovered adipokine involved in the regulation of various metabolic functions. its receptor, aplnr, is expressed in reproductive tissues; however, its role in human ovarian cells is unknown. pcos patients are more vulnerable to develop diabetes, cardiovascular diseases, and metabolic syndrome.[3] ir is prevalent in women with pcos independently of obesity and is critically involved in reproductive and metabolic complications of the syndrome.[10] several tests have been developed to measure ir, some very reliable but complex like the hyperinsulinemic euglycemic glucose clamp and others less precise but easier and less invasive like homa-ir.[22] new markers are needed to reach a more reliable assessment of ir. to date, several surrogate markers have been proposed in literature to facilitate and improve the determination of ir. many new proteins are strongly involved with pcos physiopathology and ir such as some adipocytokines (adiponectin, visfatin, vaspin, and apelin), copeptin, irisin, pai-1, and zonulin. many other proteins have been proposed as potential new markers of ir in pcos such as resistin, leptin, retinol-binding protein 4 (rbp4), kisspeptin, and ghrelin, but their role is still controversial.[14] adipose tissue is implicated in the secretion of several hormones such as adiponectin, resistin, leptin, visfatin, apelin, and rbp4 called adipocytokines that are involved in energy homeostasis and metabolism.[11] apelin is a peptide isolated from bovine stomachs, but it is expressed in several other organs and also in visceral and subcutaneous tissues.[4] in our present study, however, serum apelin levels are significantly higher in pcos women compared with controls. moreover, we also observed that apelin level is significantly and positively correlated with bmi and homa-ir. apelin was found to be higher in pcos patients by gören et al.[23] but without a significant correlation with homa-ir. olszanecka-glinianowicz et al.[18] reported an inverse association between apelin and glucose, insulin, and homa-ir values, supporting the role of apelin in the regulation of insulin sensitivity. apelin levels were higher in non-obese pcos patients, suggesting a compensatory mechanism for metabolic consequences of ir. lower serum concentrations of apelin were found in pcos subjects by altinkaya et al.[19] with positive correlation with bmi, insulin, homa-ir, tg, and free testosterone, speculating that apelin can be used as a marker for insulin sensitivity. conversely, sun et al.[20] observed significantly enhanced apelin concentration in pcos patients with positive association with bmi and homa-ir; treatment with drospirenone-ethinylestradiol plus metformin improved ir and apelin levels. discrepant findings among the published studies may be attributed to the differences in ethnicity, age, study design, sample size, genetic characteristics of populations, and assessment methodology. the increase of lh level probably plays an important role in the pathological mechanism of the higher androgens production in the ovaries, which can interfere with the maturation of the oocyte.[21] in the present study, the serum lh levels are significantly higher, while fsh level is lower in pcos women compared with controls. these results further confirm that high lh level and relative insufficiency of fsh are the characteristics of pcos. in line with our results, tal et al. found increased tgf-b1 and decreased seng in pcos. these data suggest that increased tgf-b1 bioavailability may contribute to the pathogenesis of pcos and its increased risk for ovarian hyperstimulation.[17] by contrast, the study conducted by irani et al. found that seng protein concentration was not different between pcos and non-pcos control group at baseline.[15] women with pcos have abnormal increase in tgf-b1 bioavailability (tgfb1/seng) due to an increase in serum tgf-b1 and decrease in seng levels.[17] conclusion polycystic ovary syndrome (pcos) is a condition that affects a woman’s hormone levels. levels of tgf β1, lh, lh/fsh, t, and, as well as homeostatic model assessment of ir (homa-ir) in pcos patients, were significantly higher than in the control group, while show decrease levels of apelin and endoglin levels. therefore the correlation analysis showed that apelin level was negatively correlated with body mass index (bmi) and homa-ir. references 1. e. karimi, a. moini, m. yaseri, n. shirzad, m. sepidarkish, m. hossein-boroujerdi and m. j hosseinzadeh-attar. “effects of synbiotic supplementation on metabolic parameters and apelin in women with polycystic ovary syndrome: a randomised doubleblind placebo-controlled trial”. the british journal of nutrition, vol. 119, no. 4, pp. 398-406, 2018. 2. g. bozdag, s. mumusoglu, d. zengin, e. karabulut and o.b. yildiz. “the prevalence and phenotypic features of polycystic ovary syndrome: a systematic review and meta-analysis”. human reproduction, vol. 31, pp. 2841-2885, 2016. 3. i. r. indran, z. huang, l.w. khin, j. k. y. chan, v. viardot-foucault and e. l. yong. “simplified 4-item criteria for polycystic ovary syndrome: a bridge too far”? clinical endocrinology (oxford), vol. 89, no. 2, 202-211, 2018. 4. x. z. zhang, y. l. pang, x. wang and y. h. li. “computational characterization and identification of human polycystic ovary syndrome genes”. scientific reports, vol. 8, no. 1, pp. 12949, 2018. 5. n. naderpoor, s. shorakae, a. joham, j. boyle, b. de courten and h. j. teede. “obesity and polycystic ovary syndrome”. minerva endocrinologica, vol. 15, no. 1, pp. 37-51, 2015. 6. j. kataoka, e. c. tassone, m. misso, a. e. joham, e. stenervictorin, h. teede and l. j. moran. “weight management interventions in women with and without pcos: a systematic review”. nutrients, vol. 9, no. 9, pp. e996, 2017. 7. m. h. dahan and g. reaven. “relationship among obesity, insulin resistance, and hyperinsulinemia in the polycystic ovary syndrome”. endocrine, vol. 64, no. 3, pp. 685-689, 2019. 8. i. lazúrová, z. lazúrová, j. figurová, s. ujházi, i. dravecká, j. mašlanková and m. mareková. “relationship between steroid hormones and metabolic profile in women with polycystic ovary syndrome”. physiological research, vol. 68, no. 3, pp. 457-465, 2019. ibrahim, et al.: apelin, endoglin, and tgf-β1 in polycystic ovary syndrome 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 21-25 9. e. koivuaho, j. laru, m. ojaniemi, k. puukka, j. kettunen, j.s. tapanainen, s. franks, m. r. järvelin, l. morin-papunen, s. sebert and t. t. piltonen. “age at adiposity rebound in childhood is associated with pcos diagnosis and obesity in adulthood-longitudinal analysis of bmi data from birth to age 46 in cases of pcos”. international journal obesity (london), vol. 43, no. 7, pp. 1370-1379, 2019. 10. i. falcão-piresi, n. gonçalves, t. henriques-coelho, d. moreiragonçalves, r. roncon-albuquerque and a. f. leite-moreira. “apelin decreases myocardial injury and improves right ventricular function in monocrotaline-induced pulmonary hypertension”. american journal of physiology heart and circulatory physiology, vol. 296, no. 6, pp. h2007-h2014, 2009. 11. b. masri, b. knibiehler and y. audigier. “apelin signalling: a promising pathway from cloning to pharmacology”. cellular signalling, vol. 17, pp. 415-426, 2005. 12. l. li, g. yang, q. li, y. tang, m. yang, h. yang and k. li. “changes and relations of circulating visfatin, apelin, and resistin levels in normal, impaired glucose tolerance, and type 2 diabetic subjects”. experimental and clinical endocrinology diabetes, vol. 114, pp. 544-548, 2006. 13. j. boucher, b. masri, d. daviaud, s. gesta, c. guigné, a. mazzucotelli, i. castan-laurell, i. tack, b. knibiehler, c. carpéné, y. audigier, j. s. saulnier-blache and p. valet. “apelin, a newly identified adipokine up-regulated by insulin and obesity”. endocrinology, vol. 146, pp. 1764-1771, 2005. 14. k. polak, a. czyzyk, t. simoncini and b. meczekalski. “new markers of insulin resistance in polycystic ovary syndrome”. journal endocrinology invest, vol. 40, no. 1, pp. 1-8, 2017. 15. m. irani, d.b. seifer, r. v. grazi, n. julka, d. bhatt, b. kalgi, s. irani, o. tal, g. lambert-messerlian and r. tal. “vitamin d supplementation decreases tgf-β1 bioavailability in pcos: a randomized placebo-controlled trial”. the journal clinical endocrinology metabolism, vol. 100, no. 11, pp. 4307-4314, 2015. 16. n. m. rashad, a. i. amin, a. e. ali and m. h. soliman. “influence of obesity on soluble endoglin and transforming growth factorb1 in women with polycystic ovary syndrome”. middle east fertility society journal, vol. 23, pp. 418-424, 2018. 17. r. tal, d. b. seifer, a. shohat-tal, r.v. grazi and h. e. malter. transforming growth factor-β1 and its receptor soluble endoglin are altered in polycystic ovary syndrome during controlled ovarian stimulation”. fertility sterility, 100, no. 2, pp. 538-543, 2013. 18. m. olszanecka-glinianowicz, p. madej, m. nylec, a. owczarek, w. szanecki, p. skałba and j. chudek. “circulating apelin level in relation to nutritional status in polycystic ovary syndrome and its association with metabolic and hormonal disturbances”. clinical endocrinology (oxford), vol. 79, no. 2, pp. 238-242, 2013. 19. s. ö. altinkaya, s. nergiz, m. küçük and h. yüksel. “apelin levels in relation with hormonal and metabolic profile in patients with polycystic ovary syndrome”. european journal obstetrics and gynecology reproductive biology, vol. 176, pp. 168-672, 2014. 20. x. sun, x. wu, y. zhou, x. yu and w. zhang. “evaluation of apelin and insulin resistance in patients with pcos and therapeutic effect of drospirenone-ethinylestradiol plus metformin. medical science monitor, vol. 21, pp. 2547-2552, 2015. 21. c. k. nath, b. barman, a. das, p. rajkhowa, p. baruah, m. baruah and a. baruah. “prolactin and thyroid stimulating hormone affecting the pattern of lh/fsh secretion in patients with polycystic ovary syndrome: a hospital-based study from north east india”. journal family medicine and primary care, vol. 8, no. 1, pp. 256-260, 2019. 22. b. wiweko, i. indra, c. susanto, m. natadisastra and a. hestiantoro. “the correlation between serum amh and homa-ir among pcos phenotypes”. bmc research notes, vol. 11, no. 1, pp. 114, 2018. 23. k. gören, n. sağsöz, v. noyan, a. yücel, o. cağlayan and m. s. bostancı. “plasma apelin levels in patients with polycystic ovary syndrome”. journal of the turkish german gynecological association, vol. 13, no. 1, pp. 27-31, 2012. . 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 research article cytotoxic effects of pistacia khinjuk seed extracts on different cell lines and its mitogenic effects on blood lymphocyte in vitro reshna k. ahmad1*, kawa f. dizaye2, asaad a. al-asady3 1department of basic science, unit of anatomy, college of medicine, hawler medical university, erbil, iraq, 2department of basic science, unit pharmacology, college of medicine, hawler medical university, erbil, iraq, 3department of anatomy and histology, college of medicine, duhok university, duhok, iraq abstract reports indicated that extract pistacia khinjuk has anti-inflammatory, antipyretic, antibacterial, and antiviral, in treating of diarrhea and throat infections and has hepatoprotective effects against acetaminophen and carbon tetrachloride. this study was undertaken to investigate the possible cytotoxic effects of methanolic and aqueous seeds extract of p. khinjuk on different tumors (rhabdomyosarcoma [rd] and murine mammary adenocarcinoma [ahmed-mohammed-nahi-2003 (amn-3)]) and normal cell lines (murine fibroblast) and its mitogenic effects on blood lymphocytes. the cytotoxic effects of p. khinjuk seed extracts were evaluated on two tumor cell lines, rd and murine mammary adenocarcinoma (amn-3) and one normal cell line, murine fibroblast (l20b). moreover, the mitogenic effects of the plant extract were studied, on human blood lymphocytes. both methanolic and aqueous seed extracts of p. khinjuk significantly induced tumor cell lines and the normal cell line proliferation, especially in highest concentrations. the results show that the extracts induced significant increases in human blood lymphocyte proliferation at 72 h. this activity of plant extracts recommends it as a good mitogenic agent in researches; in conclusion, seed extracts of p. khinjuk induced proliferation of all tested cell lines. high concentrations of both aqueous and methanolic seed extracts of p. khinjuk showed mitogenic effects. keywords: mitogenic effect, pistacia khinjuk, proliferative effect, tissue culture introduction conventionally, pistacia khinjuk used as tonic and expectorant, it has been used in cough, fever, and asthma.[1] few investigations have been reported the effect of different crude and isolated compound extracts of p. khinjuk on different diseases. some species of pistacia have been used in folk medicine as anti-inflammatory, antipyretic, antibacterial, and antiviral, in treating diarrhea and throat infection.[2] kordali et al. tested the antifungal activity of crude extracts obtained from the leaves of pistacia vera, pistacia terebinthus, and pistacia lentiscus.[3] ozcelik et al. used extracts of p. vera and studied antibacterial, antifungal, and antiviral properties of it, it showed little antibacterial activity, a noticeable antifungal activity, and significant antiviral activity.[4] ljubuncic et al. studied the effects of aqueous extracts prepared from the leaves of p. lentiscus in experimental liver disease.[5] taran et al. (2009) studied the anthelmintic effect of p. khinjuk against protoscoleces of echinococcus granulosus, the results of this study indicate in vitro anti-echinococcal activity of the essential oil of p. khinjuk.[6] derwich et al. (2010) studied antibacterial activity of the essential oil isolated from the leaf of p. lentiscus, the study was conducted to determine the phytochemistry and antibacterial activities of pistacia leaves oil against both gramnegative and gram-positive bacteria.[7] in another study, taran et al. investigated the antimicrobial activity of the leaves of p. khinjuk and they found that some major constituents of essential oil from the aerial parts of p. khinjuk extracts showed antibacterial and antifungal activities.[1] dizaye examined the effect of aqueous extract of p. khinjuk on acetaminophen and carbon tetrachloride-induced acute liver toxicity in albino rats, he concluded that an aqueous extract of p. khinjuk has hepatoprotective effects against acetaminophen and carbon tetrachloride.[8] tohidi et al. investigated the antibacterial and wound healing activity in both p. khinjuk and pistacia atlantica; cihan university-erbil scientific journal (cuesj) corresponding author: reshna k. ahmad, department of basic science, unit of anatomy, college of medicine, hawler medical university, erbil, iraq. e-mail: reshna.kamal@hmu.edu.krd received: apr 23, 2019 accepted: apr 29, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp13-20 copyright © 2020 reshna k. ahmad, kawa f. dizaye, asaad a. al-asady. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 the results suggested that antibacterial and wound healing activities of p. khinjuk are better than p. atlantica at the same concentrations.[2] according to our knowledge, no data are available regarding the cytogenetic and mitogenic activities of p. khinjuk. therefore, this study was designed to investigate the cytogenic effects of methanolic and aqueous seed extract of p. khinjuk on two tumor cell lines (rhabdomyosarcoma [rd] and murine mammary adenocarcinoma) and one normal cell line (murine fibroblast). moreover, the mitogenic effects of p. khinjuk was studded on human blood lymphocytes. materials and methods plant collection p. khinjuk were collected from erbil (shaqlawa) governorate. plants were deposited to be identified, the identification done by the department of pharmacognosy, college of pharmacy of hawler medical university, iraq. seeds of p. khinjuk were separated and dried at room temperature according to al-barazanjy. the dried seeds were grind into powder by electrical grinder (mesh no. 0.5 mm), and the powder was kept in plastic bags in a deep freezer (−20°c) until use.[9] preparation of aqueous extracts from p. khinjuk seeds aqueous extract was prepared from the grind powder according to al-barazanjy. then, crude dried extract was placed in labeled, tightly sealed plastic tubes, and stored at −20°c until used.[9] for the following experiments, 1 g of aqueous seeds extract was dissolved into 100 ml phosphate buffer saline (pbs), the suspension then filtered and sterilized using 0.2 µm sterile millipore filter and kept in a deep freeze in (−20°c) until use. preparation of methanolic extracts from p. khinjuk seeds methanolic extraction was prepared according to harborn (1984), then the extract was stored in a tightly sealed plastic test tube at −20°c until use.[10] cell lines rhabdomyosarcoma (rd) cell line this is a human cell line derived from a biopsy specimen obtained from a pelvic rhabdomyosarcoma of a 7-year-old caucasian girl.[11] passages 258–263 of rd cell line were used throughout this study and rpmi-1640 was used for maintaining the cells. ahmed-mohammed-nahi-2003 (amn-3) cell line this cell line is a murine mammary adenocarcinoma cell line derived from a spontaneous mammary adenocarcinoma of female balb/c mice.[12] the cells were maintained in rpmi1640 medium. passages128–135 of amn-3 cell line were used throughout this study. l20b cell line this cell line is a murine cell line derived from mouse l cells (fibroblasts) expressing the human poliovirus receptor.[13] passage numbers (14–18) of the l20b cell line were used in this study and it was maintained in rpmi-1640 medium containing 10% bovine calf serum. cytotoxicity assay the cytotoxicity protocol was depending on flick and gifford, 1984.[14] cells were plated in a 96-well flat-bottomed plate, after adhesion, serial dilutions of aqueous and methanolic extract separately three replicates for each concentration were placed in three columns for each type (200 µl from each extract) to the appropriate wells and incubated for 24, 48, or 72 h at 37°c, 5–10% co 2 in a humidified environment. untreated cells were used as controls. then, the supernatants were decanted off and cells in each well were washed with pbs twice from, and 50 µl of 0.01% neutral red dye was added to each well and reincubated for 2 h; at the end of incubation, excess dye was removed by washing the wells twice with 150 µl pbs, then 125 µl of extraction dye solution was added.[10] the optical density (od) of each well was read using an enzyme-linked immunosorbent assay reader at a transmitting wavelength on 492 nm. mitogenic activity of p. khinjuk seeds extracts in vitro human blood culture was prepared by collecting blood sample by vein puncture and transferred into heparinized tubes containing media (rpmi-1640); then, different extracts of p. khinjuk were added to blood culture tubes (three tubes for each) instate of polyhydroxyalkanoates (pha) and three tubes prepared with pha as control positive and other three without pha as negative control. all tubes incubate in co 2 incubator and shacked gently each 15 min at the 1st h, 30 min at the 2nd h, and ones at the 3rd and 4th h then each 12 h and incubated for 48 and 72 h; then, the cells were harvested, slides prepared and stained according to suman et al., 2006.[15] statistical analysis significance level was ascertained by one-way analysis of variance, followed by student-newman-keuls multiple tests. results were expressed as the mean ± standard error of the mean. p < 0.05 was considered statistically significant. all statistical procedures were performed with spss software version 16. results cytotoxic effects of aqueous and methanolic extract of p. khinjuk seeds on rd tumor cell line tables 1 and 2 show the effect of both aqueous and methanolic seed extracts of p. khinjuk on rd tumor cell line. the inhibitory effect was found in the proliferation of rd tumor cells when treated with 78.125, 312.5, 625, and 1250 µg/ml of seed aqueous extract for 24 h. however, an increase in the proliferation of rd cells was detected when these cells treated for 24 h with 10,000 µg/ml, in which the od was 0.4773 ± 0.0115. this induction effect was also detected at 48 h treatment with the same plant extract, in which a significant increase was observed at ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 48 h of treatment, while the highest level of proliferation was detected in concentration of 10,000 µg/ml at 48 h of exposure with od of 0.1687 ± 0.0064 when compared with its control (0.1133 ± 0.0003) [figure 1]. in contrast, a non-significant difference was observed when this type of cell line was treated with aqueous seed extract of p. khinjuk for 72 h in comparison with its control. whereas the effect of methanolic extract of the same plant on rd tumor cell line was stimulation in growth of the cells at 24 h and 48 h of exposure, particularly at the higher concentrations (5000 µg/ml and 10,000 µg/ml), and the results showed non-significant differences at 72 h of treatment. as shown in figures 2 and 3, treatment of the cells with the highest concentrations of both extracts caused remarkable changes in the confluent monolayer appearance. figure 2: rhabdomyosarcoma tumor cells treated with 10,000 µg/ml pistacia khinjuk aqueous seeds extract 48 h (×400) figure 1: confluent monolayer of untreated rhabdomyosarcoma tumor cells (×400) table 2: cytotoxic effect of methanolic seeds extract of p. khinjuk seeds on the growth of rd tumor cell line concentrations(µg/ml) exposure period 24 h 48 h 72 h control 0.1327±0.0101a 0.1470±0.0012a 0.1427±0.004ab 78.125 0.1547±0.0009b 0.1543±0.0022a 0.1704±0.0053b 156.25 0.1637±0.0015b 0.1543±0.0009a 0.1627±0.0067ab 312.5 0.147±0.0052b 0.1540±0.0017a 0.1667±0.0064ab 625 0.1613±0.0018b 0.1533±0.0035a 0.1607±0.0046ab 1250 0.1633±0.0047b 0.1550±0.0038a 0.1547±0.0015ab 2500 0.1633±0.0019b 0.1653±0.0018b 0.1370±0.0131a 5000 0.164±0.0027b 0.1650±0.0012b 0.1533±0.0026ab 10,000 0.1667±0.0009b 0.1690±0.0036b 0.1537±0.0067ab p. khinjuk: pistacia khinjuk, rd: rhabdomyosarcoma. *similar letters indicate no significant differences. *different letters indicate significant differences at p<0.05 table 1: cytotoxic effect of aqueous extract of p. khinjuk seeds on the growth of rd tumor cell line concentrations (µg/ml) exposure period 24 h 48 h 72 h control 0.3987±0.008b 0.1133±0.0003a 0.1127±0.0019a 78.125 0.3567±0.024a 0.1413±0.0032b 0.117±0.0051a 156.25 0.3520±0.0144a 0.1477±0.0067bc 0.1097±0.0082a 312.5 0.3440±0.0016a 0.1467±0.0027bc 0.1157±0.0113a 625 0.3467±0.0043a 0.1467±0.0027bc 0.1233±0.0035a 1250 0.3463±0.0018a 0.1637±0.0059bc 0.1293±0.0027a 2500 0.3997±0.0023b 0.1667±0.0023c 0.132±0.0044a 5000 0.3977±0.0009b 0.1413±0.0032c 0.1277±0.0062a 10,000 0.4773±0.0115c 0.1687±0.0064c 0.1193±0.0049a p. khinjuk: pistacia khinjuk, rd: rhabdomyosarcoma. *similar letters indicate no significant differences. *different letters indicate significant differences at p<0.05 ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 cytotoxic effects of aqueous and methanolic extract of p. khinjuk seeds on amn-3 tumor cell line the aqueous extract was more effective against the proliferation of amn-3 cells at 24 h of treatment than other treatment periods. the treatment for both 48 and 72 h showed non-significant differences in cell proliferation when compared with the control group, but the treatment for 24 h showed significant decreases in the proliferation of amn-3 cells and the effect was started from 78.125 µg/ml to highest concentration of 10,000 µg/ml, table 3. table 4 shows the effect of methanolic seeds extract of p. khinjuk on amn-3 tumor cells which exhibited a significant increase in proliferation of tested cell line in all concentrations at all periods of treatment, particularly at the highest concentrations of 5000 µg/ml and 10,000 µg/ml. figure 4 shows the effect of aqueous seeds extract on amn-3 cells at 24 h treatment, in which some cells with different changes in their shape and size were seen, the monolayer appearance is absent. figure 5 shows the shape and size of amn-3 cells similar to that of untreated cells in control [figure 6]. figure 3: rhabdomyosarcoma tumor cells treated with 10,000 µg/ml pistacia khinjuk methanolic seeds extract 48 h (×400) table 3: cytotoxic effect of aqueous extract of p. khinjuk seeds on the growth of amn-3 tumor cell line concentrations(µg/ml) exposure period 24 h 48 h 72 h control 0.3190±0.2355a 0.1030±0.0089a 0.2267±0.0034a 78.125 0.1027±0.0029b 0.1080±0.0027a 0.2097±0.0018a 156.25 0.0963±0.0015b 0.1067±0.0015a 0.2233±0.0150a 312.5 0.1067±0.0038b 0.0960±0.0025a 0.2303±0.0064a 625 0.1060±0.0051b 0.1070±0.0031a 0.2594±0.0091a 1250 0.1103±0.0060b 0.1062±0.0029a 0.2920±0.0089a 2500 0.1160±0.0035b 0.1340±0.0031a 0.2910±0.0035a 5000 0.1097±0.0019b 0.1270±0.0038a 0.2930±0.0076a 10,000 0.1137±0.0023b 0.1113±0.0033a 0.2937±0.0033a *similar letters indicate no significant differences. *different letters indicate significant differences at p<0.05. p. khinjuk: pistacia khinjuk, amn-3: ahmed-mohammed-nahi-2003 table 4: cytotoxic effect of methanolic extract p. khinjuk seeds on the growth of amn-3 tumor cell line concentrations (µg/ml) exposure period 24 h 48 h 72 h control 0.0820±0.0025a 0.1030±0.0089a 0.2267±0.0034a 78.125 0.1007±0.0029b 0.1133±0.0026ab 0.2277±0.0069a 156.25 0.0957±0.0020b 0.1210±0.0012abc 0.2307±0.0113ab 312.5 0.1023±0.0038b 0.1217±0.0020bc 0.2410±0.0164abc 625 0.1027±0.0033b 0.1207±0.0035bc 0.2443±0.0085abc 1250 0.1020±0.0015b 0.1347±0.0037c 0.2823±0.0033abc 2500 0.0997±0.0012cd 0.1540±0.0049e 0.2990±0.0006abc 5000 0.1093±0.0018d 0.1367±0.0024d 0.3040±0.0025bc 10,000 0.1103±0.0015c 0.1160±0.0065c 0.3093±0.0064c p. khinjuk: pistacia khinjuk, amn-3: ahmed-mohammed-nahi-2003. *similar letters indicate no significant differences. *different letters indicate significant differences at p<0.05 ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 cytotoxic effects of aqueous and methanolic extract of p. khinjuk seeds on l20b cell line table 5 shows the effect of p. khinjuk aqueous seeds extract on the growth of l20b cells. during the first 2 days of incubation, the aqueous extract did not stimulate the growth of l20b cells as compared with control [figure 7]. only a significant induction of cell proliferation was detected when treated with 10,000 µg/ml for 48 h of exposure. after 72 h of treatment, the aqueous extract significantly inhibited cell growth at almost all concentrations. this effect is shown in figure 8, in which the cells exhibited different changes in cell shape. some rounded, binucleated, and multinucleated cells were detected when treated with 10,000 µg/ml. viability of l20b cells after treatment with the methanolic extract is shown in table 6. treatment of cells with concentrations of 2500 µg/ml and 10,000 µg/ml for 24 h caused a significant increase in cell viability. all the other concentrations have no significant effects on cell viability. non significant differences were detected when the cells treated with78.125, 156.25, 312.5, and 625 µg/ml at 48 h, while the cells when treated with higher concentrations 1250, 2500, 5000, and 10, 000µg/ml at 48h a significant increases were table 5: cytotoxic effect of aqueous extract of p. khinjuk seeds on the growth of l20b cell line concentrations (µg/ml) exposure period 24 h 48 h 72 h control 0.0947±0.0013a 0.1±0.0061a 0.0993±0.0003d 78.125 0.0940±0.0027a 0.1±0.0015a 0.0707±0.0012a 156.25 0.0947±0.0032a 0.1003±0.0009a 0.0777±0.0007b 312.5 0.0957±0.0039a 0.1030±0.0006a 0.0797±0.0007b 625 0.960±0.0031a 0.1030±0.0006a 0.003±0.0019b 1250 0.0963±0.0017a 0.1077±0.0032a 0.0777±0.0019b 2500 0.0940±0.0015a 0.1083±0.0015a 0.0887±0.0032c 5000 0.1140±0.0135a 0.113±0.0007a 0.1030±0.0025d 10,000 0.1153±0.0019a 0.1753±0.0058b 0.0910±0.0021c *similar letters indicate no significant differences.*different letters indicate significant differences at p<0.05. p. khinjuk: pistacia khinjuk, figure 6: confluent monolayer of untreated ahmed-mohammednahi-2003 tumor cells (×400) figure 5: ahmed-mohammed-nahi-2003 tumor cells treated with 5000 µg/ml of pistacia khinjuk methanolic seeds extract at 72 h of exposure (×400) figure 4: ahmed-mohammed-nahi-2003 tumor cells treated with 10,000 µg/ml of pistacia khinjuk aqueous seeds extract at 24 h of exposure (×400) figure 7: confluent monolayer of untreated l20b cells (×400) ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 detected in cell viability. the results also showed insignificant differences at 72 h of exposure. figure 9 shows the induction effect of this extract 10,000 µg/ml for 48 h on l20b cells with different changes such as rounded and binucleated and multinucleated cells. mitogenic activity of aqueous and methanolic extracts of p. khinjuk on human blood lymphocyte aqueous and methanolic extracts of seeds of p. khinjuk 10,000 µg/ml were used to detect its effect as mitogen in blood lymphocyte cultures for both 48 and 72 h exposure; the results were compared with positive control group which was treated with pha as a mitogen and also were compared with the negative control group (group that treated with pbs). significant induction was detected in all treated group with 10,000 µg/ml p. khinjuk seeds extract for 48 h when compared with negative control group, and non significant differences was detected when the cells treated with aqueous and methanolic extracts for 72 h in all groups when compared with positive control group, but a significant induction was recorded in mitotic index (mi) of all groups when compared with the negative control group, table 7. discussion effects of aqueous and methanolic seeds extracts of p. khinjuk on rd, amn-3, and l20b cell lines when both cell lines (rd and amn-3) treated with seed extract of p. khinjuk, a significant increases in cell proliferation were detected for both aqueous and methanolic extracts, except exposure of both types of cell lines for 72h to aqueous extract, this increase might be due to the fact that extractions from some herbs can restrain cancer cells and induce its proliferation. cao et al. estimated that extractions from some herbs can restrain hepatocarcinoma, and the herbal extractions of flavonoid and arsenic trioxide cannot only figure 8: l20b cells treated with 10,000 µg/ml of pistacia khinjuk aqueous seeds extract at 72 h of exposure (×400) figure 9: l20b cells treated with 10,000 µg/ml of pistacia khinjuk methanolic seeds extract at 48 h (×400) table 6: cytotoxic effect of methanolic extract of p. khinjuk seeds on the growth of l20b cell line concentrations (µg/ml) exposure period 24 h 48 h 72 h control 0.1±0.0061a 0.1001±0.0061a 0.993±0.0003ab 78.125 0.1±0.0012a 0.1003±0.0067a 0.0913±0.0017a 156.25 0.1003±0.0019a 0.1003±0.0049a 0.0920±0.0006a 312.5 0.1127±0.0029a 0.1027±0.0024a 0.0937±0.0026a 625 0.1113±0.0009a 0.1323±0.0024a 0.1037±0.0047ab 1250 0.1270±0.0050a 0.2417±0.0197d 0.1033±0.0018ab 2500 0.1777±0.0112c 0.2430±0.0103bd 0.1120±0.0027b 5000 0.1317±0.0038a 0.2073±0.0049d 0.0910±0.0012b 10,000 0.1550±0.1617b 0.2977±0.0089e 0.1030±0.0060ab *similar letters indicate no significant differences. *different letters indicate significant differences at p<0.05. p. khinjuk: pistacia khinjuk table 7: mean±sem for mitotic index of human blood lymphocyte when p. khinjuk used as mitogenic factor exposure period positive control (pha) negative control (pbs) p. khinjuk methanolic seeds extract 10,000 µg/ml p. khinjuk aqueous seeds extract 10,000 µg/ml 48 h treatment 0.4893±0.00445a 0.2438±0.00185d 0.3564±0.02775c 0.3767±0.00845c 72 h treatment 0.4906±0.00561a 0.2862±0.29093b 0.3822±0.00740a 0.3807±0.00491a *similar letters indicate no significant differences. *different letters indicate significant differences at p<0.05. p. khinjuk: pistacia khinjuk, pha: polyhydroxyalkanoates, pbs: phosphate buffer saline, sem: standard error of the mean ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 restrain the proliferation of hepatocarcinoma cells but also induce apoptosis of hepatocarcinoma cells.[16] rui-chuan et al. reported that chinese medicine can induce cytodifferentiation of hepatocarcinoma, also rhubarb acid could restrain cell proliferation of various tumors such as mastocarcinoma, lung cancer, hepatocarcinoma, and colon carcinoma, and has synergistic effects when used in combination with mutamycin.[17] it could inhibit synthesis of dna, promote apoptosis of cancer cells, and also it has been found that the inhibitory effect of repression of rhubarb on cancer cells is closely related with cell signal transduction.[18] tohidi et al. and azadpour et al. found that the extract of p. khinjuk showed faster healing compared with control groups, faster wound contraction rate may be due to the presence of flavonoids, which is responsible for the free radical scavenging activity that is believed to be one of the most important components of wound healing that also be an evidence for the increased number of cells when treated with p. khinjuk extracts which may be caused rearrangement of the cell and induction in there proliferation by the presence of some of those phytochemical compounds such as flavonoids.[2,19] in general, oleic and linoleic acids are known for their anti-inflammatory properties. linoleic and alpha-linoleic acid provide lipids necessary for cell membrane repair and cellular respiration.[20] tavakoli and pazhouhanmerhr found that p. khinjuk fruits when analyzed contain linoleic and linoleic acid, and the presence of these two types of essential fatty acids may be caused induction in cell proliferation when used for cell membrane repair.[21] moulos et al., 2009, demonstrated that exposure of lewis lung carcinomas to mastic oil (mastic oil from p. lentiscus) caused a time-dependent alteration in the expression of 925 genes.[22] mirian et al., 2015, found that after the exposure of cell lines to methanolic extracts of three plants, the cells shows high survival when they treated with p. khinjuk extract in comparison with other two extracts, and this property may be due to the presence of essential oils.[23] the effect of those extracts on l20b normal cells was also an induction of cell proliferation at high concentrations of all types of extracts and this induction in currently applied extract, lacks selectivity, and does not spare normal (non-malignant) cells from cancer cells, it also often induces resistance of tumor cells to antineoplastic agents, generally in the form of “multidrug resistance” to structurally unrelated compounds.[24] mitogenic activity of aqueous and methanolic extracts of p. khinjuk seeds on human blood lymphocyte aqueous and methanolic extracts from both leaves and seeds of p. khinjuk were tested as a mitogen on blood lymphocyte. the results of the present study showed non-significant differences were found between the values of mi of each human lymphocyte that treated for 72 h either by aqueous or methanolic seeds extracts of p. khinjuk and positive control group (pha), the same results were obtained after the treatment with methanolic leaves extract for 48h. these results indicated that the activity of all these extracts is similar to that of pha. the increase in a number of cells when treated with p. khinjuk extracts may be caused by rearrangement of the cells and induction in their proliferation according to the presence of some phytochemical compounds such as flavonoids.[2] tavakoli and pazhouhanmerhr found that p. khinjuk fruits when analyzed contain linoleic and linoleic acid and the presence of these two types of essential fatty acids may be caused induction in cell proliferation when used for repairing of the cell membrane.[21] ghosh and gaba mentioned that p. khinjuk fruits and leaves have wound healing properties which may be due to the presence of different phytochemical compounds in these extracts.[25] conclusions methanolic and aqueous seed extracts of p. khinjuk induced proliferation of all tested cell lines. high concentrations of both aqueous and methanolic seed extracts of p. khinjuk showed mitogenic effects. references 1. m. taran, m. sharifi, e. azizi and m. khanahmadi. “antimicrobial activity of the leaves of pistacia khinjuk”. journal of medicinal plants, vol. 9, no. 6, pp. 81-85, 2010. 2. m. tohidi, m. khayami, v. nejati and h. meftahizade. “evaluation of antibacterial activity and wound healing of pistacia atlantica and pistacia khinjuk”. journal of medicinal plants research, vol. 5, no. 17, pp. 4310-4314, 2011. 3. s. kordali, a. cakir, h. zengin and m. e. duru. “antifungal activities of the leaves of three pistacia species grown in turkey”. fitoterapia, vol. 74, no. 1-2, pp. 164-167, 2003. 4. b. ozcelik, m. aslan, i. orhan and t. karaoglu. “antibacterial, antifungal, and antiviral activities of the lipophylic extracts of pistacia vera”. microbiological research, vol. 160, no. 2, pp. 159161, 2005. 5. p. ljubuncic, h. song, u. cogan, h. azaizeh and a. bomzon. “the effects of aqueous extracts prepared from the leaves of pistacia lentiscus in experimental liver disease”. journal of ethnopharmacology, vol. 100, no. 1-2, pp. 198-204, 2005b. 6. m. taran, e. azizi, a. shikhvaisi and n. asadi. “the anthelmintic effect of pistacia khinjuk against protoscoleces of echinococcus granulosus”. world journal of zoology, vol. 4, no. 4, pp. 291-295, 2009. 7. e. derwich, a. manar, z. benziane and a. boukir. “gc/ms analysis and in vitro antibacterial activity of the essential oil isolated from leaf of pistacia lentiscus growing in morocoo”. world applied sciences journal, vol. 8, no. 10, pp. 1267-1276, 2010. 8. k. f. dizaye. “hepatoprotective effects of the aqueous extract of pistacia khinjuk on acetaminophen and carbon tetrachlorideinduced acute live of toxicity in albino rats”. the asa newsletter. asa 08-3, issue no. 126, jun, 2008. 9. r. k. al-barazanjy, k. dizaye and a.a. al-asadye. “cytotoxic and cytogenetic effects of salvia officinalis on different tumor cell lines”. middle east journal of internal medicine, vol. 6, no. 4, pp. 15-25, 2013. 10. j. b. harborne. “phytochemical methods”. 2nd ed. london: chapman and hall, 1984. 11. r. m. mcallister, j. melnyk, j. z. finklestein, e. c. adams and ahmad, et al.: cytotoxic effects of pistacia khinjuk seed extracts on different cell lines blood lymphocyte 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 13-20 m. b. gardner. “cultivation in vitro of cells derived from a human rhabdomyosarcoma”. cancer, vol. 24, no. 3, pp. 520-526, 1969. 12. a. m. h. al-shamary. “the study of newcastle disease virus effect in treatment of transplanted tumors in mice”. m.sc. thesis, college of veterinary medicine, university of baghdad, iraq, 2003. 13. e. duizer, k. j. schwab, f. h. neill, r. l. atmar, m. p. g. koopmans and m. k. estes. “laboratory efforts to cultivate noroviruses”. journal of general virology, vol. 85, pp. 79-87, 2004. 14. d. a. flick and g. e. gifford. “comparison of in vitro cell cytotoxic assays for tumor necrosis factor”. journal of immunological methods, vol. 68, no. 1-2, pp. 167-175, 1984. 15. g. suman, r. naravaneni and k. jamal. “in vitro cytogenetic studies of cypermethrin on human lymphocytes”. indian journal of experimental biology, vol. 44, pp. 233-239, 2006. 16. y. cao, q. h. xia, h. meng and a. p. zhong. “antitumor and synergistic effect of chinese medicine “bushen huayu jiedu recipe” and chemotherapy on transplanted animal hepatocarcinoma”. world journal of gastroenterology, vol. 11, no. 33, pp. 52185220, 2005. 17. c. rui-chuan, s. jin-hua, o. gao-liang, c. ke-xia, l. jin-quan and x. xiao-guang. “induction of differentiation in human hepatocarcinoma cells by isoverbascoside”. planta medica, vol. 68, pp. 370-372, 2002. 18. r. h. cichewicz, y. zhang, n. p. seeram and m. g. nair. “inhibition of human tumor cell proliferation by novel anthraquinones from daylilies”. life sciences, vol. 74, pp. 1791-1799, 2004. 19. m. azadpour, m. rezaei, m. taati, m. g. dehnoo and b. ezatpour. “antioxidant, antibacterial, and wound-healing properties of methanolic extract of pistacia khinjuk”. comparative clinical pathology, vol. 24, pp. 379-385, 2014. 20. z. djerrou, z. maameri, y. hamdi-pacha, m. serakta, f. riachi, h. djaalab and a. boukeloua. “effect of virgin fatty oil of pistacia lentiscus on experimental burn wounds healing in rabbits”. african journal of traditional complementary alternative medicines, vol. 7, no. 3, pp. 258-263, 2010. 21. j. tavakoli and s. pazhouhanmehr. “fatty acid composition of oils from fruits of three pistacia species growing in iran”. chemistry of natural compounds, vol. 46, no. 4, pp. 525-526, 2010. 22. p. moulos, o. papadodima, a. chatziioannou, h. loutrari, c. roussos and f. n. kolisis. “a transcriptomic computational analysis of mastic oil-treated lewis lung carcinomas reveals molecular mechanisms targeting tumor cell growth and survival”. bmc medical genomics, vol. 2. no. 68, pp. 1-15, 2009. 23. m. mirian, m. behrooeian, m. ghanadian, n. dana and h. sadeghi-aliabadi. “cytotoxicity and antiangiogenic effects of rhus coriaria, pistacia vera and pistacia khinjuk oleoresin methanol extracts”. research in pharmaceutical science, vol. 10, no. 3, pp. 233-240, 2015. 24. m. beljanski and s. crochet. “mitogenic effect of several interleukins, neuromediators and hormones on human glioblastoma cells, and its inhibition by the selective anticancer agent pb-100”. deutsche zeitschrift für onkologie, vol. 28, pp. 14-22, 1996. 25. p. k. ghosh and a. gaba. “phyto-extracts in wound healing”. journal of pharmacy and pharmaceutical science, vol. 16, no. 5, pp. 760-820, 2013. tx_1~abs:at/add:tx_2~abs:at 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 research article the estimation of (covid-19) cases in kurdistan region using nelson aalen estimator sami a. obed1*, parzhin a. mohammed1, dler h. kadir1,2 1department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq, 2department of business administration, cihan university-erbil, kurdistan region, iraq abstract it is described how the nelson–aalen estimator may be used to control the rate of a nonparametric estimate of the cumulative hazard rate function based on right censored as well as left condensed survival data, furthermore how the nelson–aalen estimator can be utilized to estimate various amounts. this technique is mostly applied to survival data and product quality data similar to the incorporated relative mortality in a multiplicative model with outer rates and the cumulative infection rate in a straightforward epidemic model. it is shown that tallying measures produce a structure that permits to a brought together treatment of all these different conditions, and the main little and massive sample properties of the assessor are summarized. this estimator is a weighted average of the nelson-aalen reliability estimates over 2  time periods. the suggested estimator’s suitability and utility in model selection are reviewed. morover, a real-world dataset is evaluated to demonstrate the proposed estimator’s suitability and utility. this work proposes a simple and nearly unbiased estimator to fill this gap. the information was gathered from the ministry of health’s website between october 1, 2020, and february 28, 2021. the results of the nelson allen estimator demonstrated that the odds of surviving were higher during a short period of time after being exposed to the virus. as time passes, the possibilities become slimmer. the closer the estimate comes to value 1 from 0.5 upward, the greater the chances of surviving the infection. keywords: covied-19, death, estimator, nelson allen estimator, survival analysis introduction nelson-aalen estimation is useful to separate how a assumed population develops through time. this strategy is frequently applied to survival data and product feature information. this makes the virus lethal since it can be encountered by someone who is infected without showing signs. many papers/articles had been written on the novel coronavirus (covid-19) within a relatively short time after its existence.[1] this research will answer questions such as: what are the affected people’s odds of survival? what is the survival probability’s time frame? how can a person survive a viral outbreak, particularly in the impacted areas? the nelson-aalen estimator, or more generally visualizing the hazard function over time, is not a very popular approach to survival analysis. that is because in evaluation to the survival function clarification of the curves is not so simple and intuitive. however, the risk function is of great significance for more advanced methods to survival analysis, for example, the cox regression. that is why it is vital to appreciate the concept. moreover, it will try to provide some insights about it. we can say that the cumulative hazard functions (chfs).[2] the nelson-aalen estimation is a method for estimating a public’s hazard function without considering that the data originates from a specific distribution. the nelson-aalen approach produces the chf from the hazard function, which is then used to obtain the survival function. it is a non-parametric approach of generating results due to the lack of parameters necessary in this model. this research will serve as a foundation for health policy formulation, resulting in fewer untimely deaths and an overall improvement in quality of life.[2] the cumulative hazard is one of the most reliable parameters in the analysis of time-to-event data with corresponding author: sami a. obed, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq e-mail: muhsin.hmeadi@cihanuniversity.edu.iq received: july 7, 2021 accepted: september 6, 2021 published: september 30, 2021 doi: 10.24086/cuesj.v5n2y2021. pp 24-31 copyright © 2021 sami a. obed, parzhin a. mohammed, dler h. kadir, this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) obed, et al.: the estimation of (covid-19) cases in krg 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 independently right-censored and left-truncated survival times. cumulative transition intensity is another name for it. the well-known nelson–aalen estimator is frequently used to estimate it nonparametric ally. in this context time-simultaneous confidence bands are perhaps the best interpretive tool for accounting for related estimating uncertainty in this situation.[3] methodology this section studied some basic concepts of survival analysis; survival function, hazard function, and some tests and methods used to analyze survival data. coronavirus is a virus whose genome consists of a single strand of ribonucleic acid. this is to distinguish this disease from other coronavirus outbreak in the past or future. this was a result of genetic comparison of the animal and the infected person. numerous review methods and strategies were designed during the course of this study to collect data/ materials relevant to the study in order to gain a thorough understanding of the subject matter.[4] a survival analysis can be used to evaluate not just the likelihood of manufacturing equipment failure based on the number of hours it has been in operation, but also to distinguish between different operating situations. if the chance varies depending on whether the equipment is operated outdoors or indoors, for example.[4] as a result, it is important to remember that the overall quality of the sample frames and methods utilized determines the trustworthiness of the survey estimates. in general, the broader the sample frame, the greater the result’s quality and validity.[5] nelson–aalen estimator the nelson–aalen estimator is a nonparametric estimator which might be used to estimate the cumulative hazard rate function from censored survival information the nelson– aalen estimator might be applied and exemplify its use in one specific case. moreover, it is shown how checking measures give a system, which permits to a bound together treatment of every one of these different circumstances, and summarizes the main properties of the nelson–aalen estimator.[2] the nelson-aalen analysis strategy has a place with the enlightening strategies for survival analysis, for example, life table analysis and kaplan-meier investigation. the nelsonaalen approach can rapidly provide you with a bend of combined peril and gauge the hazard capacities dependent on unpredictable time intervals.[6] an “occasion” can be the failure of a non-repairable component, the death of a human being, or any occurrence for which the experimental unit remains in the “failed” state (e.g., death) from the point at which it changed on.[4] survival function estimator[5] let t 1 < t 2 <… < t k represent the observed death times in a sample of n subjects from a homogeneous population with survival function s(t). consider s(t) as a discrete function with probability mass at each t i ,; i =1;…; k. therefore it can be written that s q q q qti i j j i ( ) � �� � �� �� �� � � �� � ��1 1 1 11 2 1 (1) where q j is the probability of subject death in the interval [t j -1, t i ]conditional of being alive at t j -1 that is, q j can be written as q p tj � �( [ t j -1,t t tj j) / ≥ -1) (2) suppose that d i deaths occurs at t i and there are n i subjects at risk at t i , kme is obtained from (1) and (2) as ( ) < < − = = −∏ ∏ . ( ) / / (1 i i i i i km t i ii t t i t t n d d s n n ) (3) it is a staircase function of t with a jump at each failure time. the nae is given by h d n r t h d ntj k k j t jk j k kk j ( ) , exp� � � � �� � �� ��� ��1 1 (4) where h(t) is the chf and is also a staircase function of t. it is noted that the above definitions extend the original definitions of the nae to the censored observations.[1] censoring categories for the nelson-aalen analysis[3] there are some types of censoring of survival data: 1. left censoring: we know that an event occurred at t * t when it is reported at time t=t(i) (i) 2. right censoring: when an occasion is reported at time t=t(i), we see that it happened at time t * t(i) if it happened at all 3. interval censoring: we know that an event occurred during [t(i-1); t(i)] when it is recorded at time t=t(i) 4. exact censoring: we know that an incident happened exactly at t=t(i) because it is reported at time t=t(i) (i). independent censoring for the nelson-aalen method the nelson-aalen technique requires that the explanations be unbiased. second, the censoring should be consistent – if two random persons are enrolled in the research at time t-1 and the other is censored at time t while the other lives, each must have the same chance of surviving at time t.[7] independent censorship can be divided into four categories: • simple type i: all individuals are censored at the same time, or individuals are followed for a set period of time • progressive type i: at the same time, everybody is censored (for illustration, when the study terminates). • type ii: the research will attempt until all of the n measures have been validated • random: the period when censoring occurs is unrelated to the time it takes to survive.[8] obed, et al.: the estimation of (covid-19) cases in krg 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 the nelson-aalen method and the chf the nelson-aalen methodology allows for population comparisons based on their hazard curves. when studying chfs, the nelson-aalen estimator should be used instead of the kaplan-meier estimator. the kaplan-meier estimator should be used when evaluating cumulative survival functions. the chf is a function that calculates the probability of: h(t) = � �t ti (5) d r i i with d i being the number of observation dropping at time ti and r i the number of observation at hazard (still in the study) at time ti.[9] results and discussion descriptive statistics table 1 gives the results of nelson allen estimator from the first of october (2020) till 28th of february (2021) applied to a data set of size 61716 cases, and shows that the most of the percentage of disease in this study are on october (2020) (46.1%) and deaths (28.9%). after that on november (2020) the percent estimate to life (33.6%) and (27.9%) was death. on the other hand, the highest estimated time until the death on december (2020) and equal (37.5%). the lowest estimated time for death which is equal to (2.1%) on february (2021). application nelson aalen survival estimate formula/notations e t m t� � � � 0 1 � nelson aalen survival estimate (sna(t) �t t m � � � 0 1 estimate of the chf. m day of observation t m date/time death observed d m number of deaths at tm n m number of people available to die at tm �m m m d n � hazard rate or probability that a life fails at tm. analysis and interpretation of data a value of 0.5 in the nelson aalen survival model suggests an equal chance of life and death. this survival model was chosen to analyze the data since it is a non-parametric technique with a non-informative censoring estimator assumption. from 0.5 to 1, the closer the value is to 1, the better the chance of survival, and vice versa. nelson aalen survival analysis of the data is shown in table 2 calculation of nelson aalen survival analysis. in the kurdistan region, there were 61171 confirmed cases and 2476 deaths, according to table 3. the total reported case number is the total number of people who have been infected, died, or who have been recovered/discharged. out of the 61171 cases reported, 28234 were found to be infected in october. the remaining 28% of cases were those that had a resolution this month. in figure 1, observation/investigation of cases was approved out from october 1 to february 28; 2021. the descriptive statistics is concerned largely with summary calculations and graphical displays of results/data to derive reasonable decisions. the horizontal axis shows the time to event, and the vertical axis shows the number of cases and deaths in this plot, drops in the survival curve occur whenever the covid-19 takes effect in a patient. thus, any point on the survival curve shows the probability that a patient on a given treatment will not have experienced relief by that time. explanation: in figure 2, the survival estimate and the range of time it applies have been clearly stated for better understanding. in the same vein, the value of m which ∑ µt=1 applies has also been shown. that is if there are no lives remaining to be censored among the observed lives when the investigation ended, and the estimation of the last value of the estimate will be zero which means sure death at that time. from the analysis carried out in table 3, at the initial day of the investigation, the estimate shows the value of one (1). from the analysis, it is evident that as time passes by, the estimate figure activates to reduce, meaning that more table 1 : descriptive statistics for qualitative variables months status % october (2020) disease 46.1 deaths 28.9 november (2020) disease 33.6 deaths 27.9 december (2020) disease 11.6 deaths 37.5 january (2021) disease 4.4 deaths 3.4 february (2021) disease 4.6 deaths 2.1 october (2020) november(2020) december (2020) january (2021) deaths 717 691 929 86 cases 28234 20591 6768 2703 0 5000 10000 15000 20000 25000 30000 35000 figure 1: monthly deaths and cases growth rate obed, et al.: the estimation of (covid-19) cases in krg 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 day of observation cases death hazard rates cumulative hazard rates estimation 1 684 21 0.0307 0.0307 0.97 2 640 21 0.0328 0.06351 0.938 3 494 16 0.0324 0.0959 0.909 4 630 19 0.0302 0.12606 0.882 5 839 31 0.0369 0.16301 0.85 6 793 30 0.0378 0.20084 0.818 7 886 29 0.0327 0.23357 0.792 8 793 31 0.0391 0.27266 0.761 9 838 12 0.0143 0.28698 0.751 10 769 19 0.0247 0.31169 0.732 11 589 24 0.0407 0.35244 0.703 12 720 26 0.0361 0.38855 0.678 13 900 23 0.0256 0.41411 0.661 14 912 20 0.0219 0.43604 0.647 15 974 28 0.0287 0.46478 0.628 16 932 18 0.0193 0.4841 0.616 17 722 23 0.0319 0.51595 0.597 18 797 32 0.0402 0.5561 0.573 19 1032 26 0.0252 0.5813 0.559 20 1010 24 0.0238 0.60506 0.546 21 1078 15 0.0139 0.61897 0.538 22 1308 17 0.013 0.63197 0.532 23 1053 20 0.019 0.65096 0.522 24 860 26 0.0302 0.6812 0.506 25 828 25 0.0302 0.71139 0.491 26 1283 23 0.0179 0.72932 0.482 27 1597 24 0.015 0.74434 0.475 28 1297 27 0.0208 0.76516 0.465 29 1285 18 0.014 0.77917 0.459 30 1002 25 0.025 0.80412 0.447 31 689 24 0.0348 0.83895 0.432 32 1054 28 0.0266 0.86552 0.421 33 1137 26 0.0229 0.88839 0.411 34 1235 23 0.0186 0.90701 0.404 35 962 28 0.0291 0.93612 0.392 36 1195 23 0.0192 0.95536 0.385 37 997 33 0.0331 0.98846 0.372 38 771 21 0.0272 1.0157 0.362 39 787 21 0.0267 1.04238 0.353 40 900 24 0.0267 1.06905 0.343 41 765 27 0.0353 1.10434 0.331 42 772 22 0.0285 1.13284 0.322 43 1170 29 0.0248 1.15763 0.314 44 686 28 0.0408 1.19844 0.302 table 2: results of estimation of covid-19 cases (contd...) obed, et al.: the estimation of (covid-19) cases in krg 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 day of observation cases death hazard rates cumulative hazard rates estimation 45 573 27 0.0471 1.24556 0.288 46 571 28 0.049 1.2946 0.274 47 659 23 0.0349 1.3295 0.265 48 645 25 0.0388 1.36826 0.255 49 576 22 0.0382 1.40646 0.245 50 530 21 0.0396 1.44608 0.235 51 567 23 0.0406 1.48664 0.226 52 361 20 0.0554 1.54204 0.214 53 495 17 0.0343 1.57639 0.207 54 488 21 0.043 1.61942 0.198 55 500 21 0.042 1.66142 0.19 56 435 25 0.0575 1.71889 0.179 57 379 17 0.0449 1.76375 0.171 58 403 26 0.0645 1.82826 0.161 59 259 14 0.0541 1.88232 0.152 60 304 14 0.0461 1.92837 0.145 61 415 14 0.0337 1.9621 0.141 62 389 23 0.0591 2.02123 0.132 63 331 17 0.0514 2.07259 0.126 64 248 9 0.0363 2.10888 0.121 65 236 8 0.0339 2.14278 0.117 66 298 9 0.0302 2.17298 0.114 67 377 10 0.0265 2.19951 0.111 68 372 12 0.0323 2.23176 0.107 69 356 9 0.0253 2.25704 0.105 70 287 13 0.0453 2.30234 0.1 71 248 11 0.0444 2.3467 0.096 72 197 6 0.0305 2.37715 0.093 73 183 9 0.0492 2.42633 0.088 74 167 9 0.0539 2.48022 0.084 75 201 9 0.0448 2.525 0.08 76 222 3 0.0135 2.53851 0.079 77 238 11 0.0462 2.58473 0.075 78 215 6 0.0279 2.61264 0.073 79 215 8 0.0372 2.64985 0.071 80 156 4 0.0256 2.67549 0.069 81 118 8 0.0678 2.74329 0.064 82 165 3 0.0182 2.76147 0.063 83 278 5 0.018 2.77945 0.062 84 163 8 0.0491 2.82853 0.059 85 150 3 0.02 2.84853 0.058 86 164 3 0.0183 2.86683 0.057 87 128 4 0.0313 2.89808 0.055 88 118 9 0.0763 2.97435 0.051 89 132 2 0.0152 2.9895 0.05 table 2: (continued) (contd...) obed, et al.: the estimation of (covid-19) cases in krg 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 day of observation cases death hazard rates cumulative hazard rates estimation 90 140 3 0.0214 3.01093 0.049 91 123 3 0.0244 3.03532 0.048 92 153 1 0.0065 3.04185 0.048 93 122 4 0.0328 3.07464 0.046 94 68 2 0.0294 3.10405 0.045 95 97 1 0.0103 3.11436 0.044 96 102 6 0.0588 3.17319 0.042 97 107 6 0.0561 3.22926 0.04 98 104 3 0.0288 3.25811 0.038 99 80 4 0.05 3.30811 0.037 100 140 5 0.0357 3.34382 0.035 101 139 3 0.0216 3.3654 0.035 102 68 6 0.0882 3.45364 0.032 103 126 2 0.0159 3.46951 0.031 104 88 0 0 3.46951 0.031 105 158 1 0.0063 3.47584 0.031 106 69 4 0.058 3.53381 0.029 107 78 3 0.0385 3.57227 0.028 108 43 2 0.0465 3.61878 0.027 109 110 3 0.0273 3.64606 0.026 110 97 2 0.0206 3.66668 0.026 111 100 4 0.04 3.70668 0.025 112 68 3 0.0441 3.75079 0.023 113 77 4 0.0519 3.80274 0.022 114 85 2 0.0235 3.82627 0.022 115 49 1 0.0204 3.84668 0.021 116 58 1 0.0172 3.86392 0.021 117 94 1 0.0106 3.87456 0.021 118 67 2 0.0299 3.90441 0.02 119 70 5 0.0714 3.97584 0.019 120 73 2 0.0274 4.00324 0.018 121 68 1 0.0147 4.01794 0.018 122 48 1 0.0208 4.03877 0.018 123 50 2 0.04 4.07877 0.017 124 107 2 0.0187 4.09747 0.017 125 54 2 0.037 4.1345 0.016 126 48 3 0.0625 4.197 0.015 127 56 3 0.0536 4.25058 0.014 128 51 1 0.0196 4.27018 0.014 129 24 4 0.1667 4.43685 0.012 130 38 2 0.0526 4.48948 0.011 131 70 2 0.0286 4.51805 0.011 132 64 3 0.0469 4.56493 0.01 133 69 0 0 4.56493 0.01 134 59 0 0 4.56493 0.01 table 2: (continued) (contd...) obed, et al.: the estimation of (covid-19) cases in krg 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 day of observation cases death hazard rates cumulative hazard rates estimation 135 98 3 0.0306 4.59554 0.01 136 53 1 0.0189 4.61441 0.01 137 106 3 0.0283 4.64271 0.01 138 72 4 0.0556 4.69827 0.009 139 138 2 0.0145 4.71276 0.009 140 115 1 0.0087 4.72145 0.009 141 117 2 0.0171 4.73855 0.009 142 131 0 0 4.73855 0.009 143 77 1 0.013 4.75153 0.009 144 150 2 0.0133 4.76487 0.009 145 157 1 0.0064 4.77124 0.008 146 129 2 0.0155 4.78674 0.008 147 176 3 0.017 4.80379 0.008 148 199 6 0.0302 4.83394 0.008 149 202 0 0 4.83394 0.008 150 78 0 0 4.83394 0.008 151 237 0 0 4.83394 0.007 table 2: (continued) table 3: reported cases covid-19 monthly in kurdistan region months cases deaths october (2020) 28234 717 november (2020) 20591 691 december (2020) 6768 929 january (2021) 2703 86 february (2021) 2875 53 deaths are being noted. this indicates that there are balances of surviving the disease at the first period of the infection, provided the infected persons take adequate medical aid. invariably, as shown in table 2, the more the virus stays in the body, the lower the probability of survival or the chance of recovery from the disease. it’s worth noting that the survival rate follows a specific pattern. as the value of m increases, the estimate values drop. this is due to the fact that the number of cases and deaths included in the study follow the same trend. both the number of cases and the number of deaths recorded increased during the study period. in practice, if the results of this study are used to predict a future epidemic of this virus, an average of 2476 infected people out of a 61171infected people are predicted to die, because the survival chance for that duration is 0.97. this projection will be accurate or true if interventions targeted at reducing risk factors and ensuring adequate access to health care are not implemented properly and efficiently. when life’s quality is poor, it will be brief. figure 2: the nelson aalen estimate of s(t) ( ) 1 0 0.97 0 1 0.93 1 2 0.90 2 3 0.882 3 4 0.850 4 5 0.818 5 6 0.792 6 7 0.761 7 8 . . . . . . 0.009 142 143 0.009 143 144 0.009 144 m m m m m m m m m m < ≤ < ≤ < ≤ < ≤ < ≤ < ≤ < ≤ < ≤ < ≤ < ≤ na ts 145 0.008 145 146 0.008 146 147 0.008 147 148 0.008 148 149 0.008 149 150 0.007 150 151 m m m m m m m                        < ≤  < ≤  < ≤  < ≤  < ≤  < ≤   < ≤                           obed, et al.: the estimation of (covid-19) cases in krg 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 24-31 conclusion and recommendation conclusion during conducting the nelson allen estimator from covied-19 cases and according to the results from the practical part the following conclusions have been drawn: 1. the number of novel coronavirus cases reported was unusually high. it was in the thousands. this demonstrates how easily the virus can be spread from one person to another or from an animal to a person 2. if prevention efforts were not made in a timely manner, the number could rise to a million. in this study, it was discovered that some people may be infected with this virus without realizing it because the symptoms of infection are not visible 3. these individuals infect others without even realizing it. according to the findings, those who have recently been exposed to the disease have a better chance of surviving if medical assistance is provided without severe or timeconsuming requirements. recommendation this study suggests that those who are unsure if they have contracted the disease due to a lack of symptoms should consult a doctor right once for tests. every country’s government should establish a free testing center for such validation. making it free and easily available will encourage everyone who is sick to do a confirmatory test. this will help to slow the disease’s spread. since the virus first appeared in wuhan, this study suggests using the wuhan preventive approach to keep the infection from spreading or infecting humans. wuhan prevention concept • wash your hands/body regularly • use nose cover/mask • have your hotness/coldness of your body checked • avoid unnecessary crowd • never touch sensitive parts of your body with unclean hands/materials/equipment. references 1. d. abbas. analysis of breast cancer data using kaplan-meier survival analysis. journal of kufa for mathematics and computer, vol. 1, no. 6, pp. 7-14, 2012. 2. t. d.bluhmki. the wild bootstrap for multivariate nelson-aalen estimators. lifetime data analysis, vol. 25, no. 1, pp. 97-127, 2019. 3. r. l. cao. presmoothed kaplan-meier and nelson-aalen estimators. journal of nonparametric statistics, vol. 17, no. 1, pp. 31-56, 2005. 4. c. a. el-nouty. the presmoothed nelson-aalen estimator in the competing risk model. communications in statistics-theory and methods, vol. 33, no. 1, pp. 135-151, 2005. 5. d. a. luo. bias and mean-square error for the kaplan-meier and nelson-aalen estimators. journal of nonparametric statistics, vol. 3, no. 1, pp. 37-51, 1993. 6. r. a. jiang. a bias-corrected nelson-aalen estimator. in: iop conference series: materials science and engineering. vol. 1043. changsha: iop publishing, p. 022013, 2021. 7. a. a. winnett. adjusted nelson-aalen estimates with retrospective matching. journal of the american statistical association, vol. 97, no. 457, pp. 245-256, 2002. 8. a. s. rossa. the nelson-aalen and kaplan-meier estimators under a sequential sampling scheme. communications in statisticstheory and methods, vol. 38, no. 16-17, pp. 3077-3098, 2009. 9. h. a. zhang. on nelson-aalen type estimation in the partial koziol-green model. statistics, vol. 44, no. 5, pp. 455-465, 2010. . 37 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 research article nutritional characteristics of pregnant women and its relation with anemia during pregnancy in a sample of kurdish women/iraq rushna g. abdulwahid1, hamdia m. ahmed2* 1cardiovascular perfusionist, surgical specialty hospital/cardiac center, erbil, kurdistan region, iraq. 2basic sciences unit, college of medicine, hawler medical university, erbil, kurdistan region, iraq abstract anemia in pregnancy is a major public health problem, especially in developing countries. this study aimed to assess the nutritional characteristics of pregnant women and find out its relationship with anemia during pregnancy. a descriptive, cross-sectional study was conducted on 600 pregnant women who attended four primary health-care centers which randomly selected according to the geographical area. a  specially designed questionnaire was prepared by the researcher after extensive review of relevant literature. the severity of anemia is determined according to alene and dohe. estimation and calculation of dietary characteristics was done according to food frequency questionnaire, frequencies and percentage, mean and standard deviations, and chi-square test of association and regression analysis. there was a highly statistically significant association between anemia with eating vegetables and chicken, and a high significant relation of anemia with eating beef and eating nuts. furthermore, there was a highly significant association between severity of anemia with eating nuts, and a significant association with eating vegetables, while there was no significant association with other variables. logistic regression analysis revealed that eating less than normal of vegetables, beef, and nuts were indicated risks of anemia. eating less than normal of vegetables, beef, and nuts were indicated risks of anemia during pregnancy. keywords: anemia, diet, nutrition, pregnancy introduction anemia in pregnancy is a major public health problem, especially in developing countries. it affects 41.8% of pregnant women globally.[1] anemia in pregnancy is defined by the world health organization (who) as a hemoglobin (hb) concentration below 11 g/dl.[2] nutritional anemia is the most common type of anemia worldwide and mainly includes iron, folic acid, vitamin b 12 , and vitamin c deficiencies. iron deficiency contributes to half of the burden of anemia globally. iron deficiency affects 1.3–2.2 billion persons out of that 50 % are women of reproductive age. in ethiopia, nearly 17% of women with age of 15–49 years are anemic of these 22% are pregnant women.[3] anemia in pregnant women has severe consequences on health, social, and economic development. anemic pregnant women will be at risk of low physical activity, increased maternal morbidity and mortality, especially those with severe anemia. in addition, both pregnant women and their neonates encounter negative consequences including fetal anemia, low birth weight (lbw), preterm delivery, intrauterine growth restriction, and perinatal mortality.[1] a large number of women experience micronutrient deficiencies (of iron and vitamin a, for instance); almost half of all pregnant women in the world are thought to have anemia and 9.8 million pregnant women have night blindness. an estimated 19.1 million pregnant women (the highest proportions in africa and south-eastern asia) have low serum retinol concentrations. maternal deficiencies in micronutrients may lower infant birth weight and jeopardize development and survival: maternal iodine deficiency is associated with congenital malformations, and mental retardation in children and a link between vitamin b12 deficiency and an increased risk of diabetes has been described in india. insufficient intake of specific fatty acids, such as docosahexaenoic acid, may also impede children’s development.[4] during pregnancy, a woman needs good nutritional status for a healthy outcome. women who have a poor nutritional cihan university-erbil scientific journal (cuesj) corresponding author: hamdia m. ahmed, college of health sciences, hawler medical university, erbil, kurdistan region, iraq. e-mail: hamdia76@gmail.com received: apr 12, 2019 accepted: apr 18, 2019 published: jan 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp1-37-44 copyright © 2020 rushna g. abdulwahid, hamdia m. ahmed. this is an openaccess article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ abdulwahid and ahmed: nutritional characteristics of pregnant women 38 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 status at conception are at higher risk of disease and death; their health depends greatly on the availability of food, and they may be unable to cope with their increased nutrient needs during pregnancy in situations of food insecurity. maternal nutrition is a fundamental determinant of fetal growth, birth weight, and infant morbidity; poor nutrition often leads to long-term, irreversible, and detrimental consequences to the fetus.[4] studies have shown that the adverse consequences of maternal anemia may affect not only the neonate and infant but also increase the risk of non-communicable diseases when the child grows into an adult and the risk of lbw in the next generation.[5] low maternal hb levels are associated with increased risk of preterm delivery, lbw babies, apgar score <5 at 1 min and intrauterine fetal demise.[6] according to the results of two previous studies in erbil city regarding anemia during pregnancy, the prevalence of anemia was 55.5%, and 70% of participants had risk factors.[7,8] according to the knowledge of the researcher, no study focuses on dietary characteristics of anemic women. because the anemia during pregnancy is still a major public health problem and a high prevalence in kurdish women, therefore the researcher interested to study it concerning diet of them. therefore, this study aimed to assess the nutritional characteristics of pregnant women and find out its relationship with anemia during pregnancy. methods a descriptive, cross-sectional study was conducted to assess the nutritional characteristics of pregnant women in primary health care centers (phcc) of erbil city, kurdistan region, iraq. the study was conducted during the period october 1, 2015–november 13, 2016. the period included data collection, analysis, and interpretation. the study was conducted among pregnant women who attended antenatal care in phcc in erbil city. during time of data collection, 21 primary health centers were in erbil city which 19 of them were provided with maternal and child health care services. four phcc were randomly selected according to the geographic area from north, south, east, and west. a purposive sampling of 600 pregnant women was included in the study. the sample size was estimated using the general formula for targeted population size, allowed error 5%, prevalence (ratio of the studied phenomenon in the population in similar study) and using the 95% confidence interval. the sample size of each phcc was according to their ratio in the targeted population. the sample size from kurdistan phcc was 117, brayati was 168, mahamad bajalan was 117, and malafande was 198. mothers who had the following criteria were included in the study: pregnant women who were in 1st, 2nd, and 3rd trimester of pregnancy and woman who accept to participate in the study. pregnant women with history of chronic hypertension, diabetes, thyroid, cardiac diseases, and systematic lupus erythematosis, woman’s with hemoglobinopathies such as thalassemia and woman’s who have early and late vaginal bleeding or antepartum hemorrhage (abruption placenta and placenta previa) and hemorrhoid were excluded from the study. before data collection, the official permission was obtained from college of nursing, erbil general directorate of health, and phcc for carrying out the study in erbil city. face to face interview method and reviewing antenatal card were used for data collection after taking permission and explanation the objectives of the study to the mother’s. each interview session took approximately 15–20 min. blood hb level was measured to assess severity of anemia. the severity of anemia is determined according to alene and dohe (2014) as the following: mild (hb level between 10.0 and 11 g/dl), moderate (hb level between 7.0 and 9.9 g/dl), and sever (hb level <7 g/dl). a specially designed questionnaire was prepared by the researcher after extensive review of relevant literature, which consists of three parts: socio-demographic characteristic obstetrical characteristics, dietary characteristic of pregnant women which included how often did the mother eat the followings: fruit, vegetables, chicken, fish and seafood, beef, other meat, nuts, beans, dairy, eggs, grains, sweets, caffeinated soft drinks, coffee and tea, and what was the usual serving size for each of them. estimation and calculation of dietary characteristics was according to food frequency questionnaire (ffq). the validity of the study questionnaire was presented through panel of 14 experts of different specialties related to the field of the present study. they recommended some modifications and suggestions regarding some questions and all responses were taken into consideration in the final draft of the questionnaire format. a pilot study was conducted on 20 mothers who attended antenatal care units. according the results of the pilot study, some items of the questionnaire were removed and other was added to the questionnaire form. the proposal of the study was approved by ethical committee from the college of nursing and general directorate of phcc. informed consent was taken from study participants. data were entered into a computer using the statistical package for the science services (spss version 23) and following statistical procedures were applied: frequencies and percentage, mean and standard deviations, and chi-square test of association and regression analysis. results the highest percentage of 57.3% of the study sample aged between 25 and 36 years old. all participants of the study sample were living in urban areas. the highest percentages of 34.3% of the study sample were graduated from institute, college, and above. regarding occupation, the highest percentages of 78.7% of the study samples were unskilled workers. regarding economic status level, the highest percentage of 49.7% of the study sample had low economic status [table 1]. table 2 shows that the highest percentage of 59.2% of the study sample regarding age at the first pregnancy was in age group 13–23 years; regarding gravidity, the highest percentage of 64.3% of the study sample was multigravida; concerning parity, the highest percentage of 48.3% of the study sample was nulliparous, and the highest percentage of 71.5% of the study sample had no abortion. table 3 shows that the highest percentage of 53.2% of the study sample was in second trimester. the highest percentage of 92.3% of the study sample took iron and folic acid supplementation; the highest percentage of 87% of the study abdulwahid and ahmed: nutritional characteristics of pregnant women 39 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 sample took <30 tabs/month, the highest percentage of 97.8% of the study sample did not have anemia before pregnancy. table 4 shows the prevalence and severity of anemia of the study sample. prevalence of anemia during pregnancy was 46.2%, regarding the severity of anemia the highest percentage of 67.1% of the study sample had mild anemia, while the lowest percentage of 32.9% of the study sample had moderate anemia, and there were no cases for severe anemia. table 5 shows the dietary characteristics of the study sample for eating daily frequency, serving size, and daily amount. the highest mean for eating daily frequency was related to drinking coffee and tea 1.99, eating fruit 1.55, eating dairy 1.52, and 1.17 eating grains, while the lowest mean 0.24 eating fish and seafood, 0.31 caffeinated soft drinks, and 0.47 eating beef. regarding serving size, the highest mean was 0.98, 0.96, 0.95, and 0.93, respectively, for drinking coffee and tea, eating fruit, eating eggs, and eating dairy, while the lowest mean was 0.50 and 0.51 for eating chicken and beef and fish and seafood, respectively. regarding daily amount, drinking coffee and tea, eating fruit, dairy, and eggs had the highest mean (1.95. 1048, 1.41, and 0.84, respectively), while the lowest mean of daily amount was 0.12 for eating fish and seafood, 0.18 caffeinated soft drinks and 0.23 eating beef. table 6 shows that all of the study samples had dietary daily amounts less than normal regarding fish and seafood, beans, grains, and eggs, while fruits 92.8%, coffee and tea 83.8%, and nuts 81.5% had the highest normal dietary amounts. more than half of the study sample had normal dietary daily amounts of dairy 54.7% and beef 52.2%. furthermore, it shows that there was a highly statistically significant association between anemia with eating vegetables and chicken (p < 0.001), and a highly significant relation of anemia with eating beef (p < 0.01), and eating nuts (p < 0.05). there was no significant association of anemia with other variables (p > 0.05) such as eating fish and seafood, beans, eggs, and grains. table 1: socio-demographic characteristics of the study sample (n=600) variables f % age group/years 13–24 208 34.7 25–36 344 57.3 37–48 48 8 wife education illiterate 96 16 read and write only 24 4 primary school 106 17.7 intermediate school 85 14.2 secondary school 83 13.8 institute, college, and above 206 34.3 wife occupation high professional job 12 2 lower professional job 116 19.3 unskilled workers 472 78.7 economic status level low economic status <90 298 49.7 middle economic status 90–120 177 29.5 high economic status 121–150 125 20.8 table 2: obstetrical characteristics of the study sample (n=600) variables f % age at first pregnancy 13–23 355 59.2 24–34 229 38.2 35–45 16 2.7 gravidity primigravida 214 35.7 multigravida 386 64.3 parity nulliparous 290 48.3 primiparous 142 23.7 multiparous 96 16 grand multipara 72 12 abortion none 429 71.5 1–3 171 28.5 trimester first trimester 33 5.5 second trimester 319 53.2 third trimester 248 41.3 iron, folic acid, fero-folic supplementation yes 554 92.3 no 46 7.7 number of iron and folic acid tablet taking/month <30 522 87 ≥30 32 5.3 none 46 7.7 anemia before pregnancy yes 13 2.2 no 587 97.8 table 3: prevalence and severity of anemia among the study sample prevalence f % anemic 277 46.2 non-anemic 323 53.8 total 600 100 severity of anemia moderate 91 32.9 mild 186 67.1 total 277 100 abdulwahid and ahmed: nutritional characteristics of pregnant women 40 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 table 7 shows that there was a highly significant association between the severity of anemia with eating nuts (p < 0.01), and a significant association with eating vegetables (p < 0.05), while there was no significant association with other variables (p > 0.05). logistic regression analysis revealed that eating less than normal of vegetables, beef, and nuts were indicated risks of anemia (p < 0.05 odds ratio [or] = 0.543, 0.147, 0.611, 0.493, 0.689, 0.641, respectively). on the other hand, eating less than normal of chicken was indicated no risks of anemia (p > 0.05 or = 3.062, 1.433, 1.039, respectively) [table 8]. discussion prevalence of anemia in the present study was 46.2% which indicated severe public health problems, which was similar to the results of other studies done in bali, indonesia in 2002 which was 46.2%, in northern tanzania was 47.4% in 2011, and 48.2% in south-east asia, and 44.2% in eastern mediterranean by who 2005, in south-eastern nigeria in 2007 was 40.4%, in kakamega county, and kenya was 40% in 2014. prevalence of anemia in the present study was higher than the results of other studies done in wolayita sodo town, southern ethiopia in 2015 was 39.94%, in makkah, saudi arabia was 39% in 2012, in southeast ethiopia was 27.9% in 2014, in europe was 25.1%, and in americas 24.1% by who 2005, in nablus, palestine was 21.7% in 2007, in mekelle town was 19.7% in 2014, in the lower north of thailand was 17.5% in 2012, in kerman, and iran was 4.7% in 2010. the results of a study done by kefiyalew et al.[9] found that 55%, 32.5%, and 12.5 of the study sample had mild, moderate, and severe anemia, respectively, which was consistent with the results of the present study. dim and onah[2] reported that 90.7% had mild anemia, 9.3% had moderate anemia, and no cases of severe anemia were detected on their studies which were consistent with the results of the present study. women from lower socioeconomic groups are at higher risk of inadequate protein intake due to the associated costs. they are also more likely to choose less expensive processed foods which would put them at risk of small for gestational age babies.[10] results of the present study, there was a high significant (p < 0.001) association of anemia with eating vegetables, chicken, and beef, and a significant (p < 0.05) association with eating nuts, while there was no significant (p > 0.05) association between anemia and eating of fruits, dairy, table 4: dietary characteristics of the study sample (n=600) dietary characteristics eating daily frequency (mean) serving size (mean) daily amount (mean) fruit (apples, bananas, oranges, etc.) 1.55 0.96 1.488 vegetables (carrots, mushrooms, potatoes, etc.) 0.92 0.57 0.524 chicken (fried chicken, in soup, grilled chicken, etc.) 0.62 0.50 0.31 fish and seafood (tuna, shrimp, crab, etc.) 0.24 0.51 0.122 beef (steak, meatballs, etc.) 0.47 0.50 0.235 nuts (almonds, cashews, walnuts, etc.) 0.86 0.63 0.541 beans (tofu, chickpeas, chili, etc.) 0.82 0.56 0.459 dairy (cheese, milk, yogurt, etc.) 1.52 0.93 1.413 eggs (omelet, in salad, etc.) 0.89 0.95 0.845 grains (bread, pasta, rice, etc.) 1.17 0.63 0.737 sweets (candy, cookies, pie, etc.) 0.67 0.71 0.475 caffeinated soft drinks (cola, diet cola, energy drinks, etc.) 0.31 0.59 0.182 coffee and tea (hot coffee, iced coffee, black tea, etc.) 1.99 0.98 1.95 table 5: dietary daily amounts of the study sample (n=600) dietary daily amount less than normal normal f % f % fruit (apples, bananas, oranges, etc.) 43 7.2 557 92.8 vegetables (carrots, mushrooms, potatoes, etc.) 504 84 96 16 chicken (fried chicken, in soup, grilled chicken, etc.) 588 98 12 2 fish and seafood (tuna, shrimp, crab, etc.) 600 100 0 0 beef (steak, meatballs, etc.) 287 47.8 313 52.2 nuts (almonds, cashews, walnuts, etc.) 111 18.5 489 81.5 beans (tofu, chickpeas, chili, etc.) 600 100 0 0 dairy (cheese, milk, yogurt, etc.) 272 45.3 328 54.7 eggs (omelet, in salad, etc.) 600 100 0 0 grains (bread, pasta, rice, etc.) 600 100 0 0 sweets (candy, cookies, pie, etc.) 533 88.8 67 11.2 caffeinated soft drinks (cola, diet cola, energy drinks, etc.) 582 97 18 3 coffee and tea (hot coffee, iced coffee, black tea, etc.) 97 16.2 503 83.8 abdulwahid and ahmed: nutritional characteristics of pregnant women 41 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 sweets, and taking caffeinated soft drinks, drinking coffee, and tea. furthermore, the present study showed that there was no significant (p > 0.05) association of severity of anemia with eating beef, which was consistent to the results of aikawa et al.,[11] who found that there was no significant association of hb level with consumption of meat and meat products. results of the present study showed that eating beef less than normal had 0.688 times more likely to express risks of anemia than in nonanemic, which was consistent to the results table 6: association of dietary daily amounts with anemia during pregnancy (n=600) variables anemic non-anemic p-value chi-square test f % f % fruit (apples, bananas, oranges, etc.) 0.962 less than normal 20 46.5 23 53.5 normal 257 46.1 300 53.9 vegetables (carrots, mushrooms, potatoes, etc.) 0.002 less than normal 219 43.5 285 56.5 normal 58 60.4 38 39.6 chicken (fried chicken, in soup, grilled chicken, etc.) 0.001 less than normal 277 47.1 311 52.9 normal 0 0.0 12 100.0 fish and seafood (tuna, shrimp, crab, etc.) constant less than normal 277 46.2 323 53.8 normal 0 0.0 0 0.0 beef (steak, meatballs, etc.) 0.004 less than normal 115 40.1 172 59.9 normal 162 51.8 151 48.2 nuts (almonds, cashews, walnuts, etc.) 0.031 less than normal 41 36.9 70 63.1 normal 236 48.3 253 51.7 beans (tofu, chickpeas, chili, etc.) constant less than normal 277 46.2 323 53.8 normal 0 0.0 0 0.0 dairy (cheese, milk, yogurt, etc.) 0.060 less than normal 137 50.4 135 49.6 normal 140 42.7 188 57.3 eggs (omelet, in salad, etc.) constant less than normal 277 46.2 323 53.8 normal 0 0.0 0 0.0 grains (bread, pasta, rice, etc.) constant less than normal 277 46.2 323 53.8 normal 0 0.0 0 0.0 sweets (candy, cookies, pie, etc.) 0.425 less than normal 243 45.6 290 54.4 normal 34 50.7 33 49.3 caffeinated soft drinks (cola, diet cola, energy drinks, etc.) 0.529 less than normal 270 46.4 312 53.6 normal 7 38.9 11 61.1 coffee and tea (hot coffee, iced coffee, black tea, etc.) 0.536 less than normal 42 43.3 55 56.7 normal 235 46.7 268 53.3 abdulwahid and ahmed: nutritional characteristics of pregnant women 42 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 of abriha et al.,[12] who mentioned that consumption of meat was another factor which showed significant association with anemia in pregnant women. el-hindi[13] mentioned on their results of the study that there was a statistically significant difference (p < 0.05) between study populations with respect to nutritional, dietary status. kefiyalew et al.[9] mentioned on their results that there was no significant (p > 0.05) association of eating red meat, poultry, fish consumption, fruit and vegetable consumption with anemia, which was in contrast to the results of the present study regarding eating red meat, vegetables, and chicken, table 7: association of the severity of anemia with dietary daily amounts of the study sample (n=277) variables moderate mild p-value chi-square test f % f % fruit (apples, bananas, oranges, etc.) 0.090 less than normal 10 50.0 10 50.0 normal 81 31.5 176 68.5 vegetables (carrots, mushrooms, potatoes, etc.) 0.027 less than normal 79 36.1 140 63.9 normal 12 20.7 46 79.3 chicken (fried chicken, in soup, grilled chicken, etc.) constant less than normal 91 32.9 186 67.1 normal 0 0.0 0 0.0 fish and seafood (tuna, shrimp, crab, etc.) constant less than normal 91 32.9 186 67.1 normal 0 0.0 0 0.0 beef (steak, meatballs, etc.) 0.326 less than normal 34 29.6 81 70.4 normal 57 35.2 105 64.8 nuts (almonds, cashews, walnuts, etc.) 0.007 less than normal 6 14.6 35 85.4 normal 85 36.0 151 64.0 beans (tofu, chickpeas, chili, etc.) constant less than normal 91 32.9 186 67.1 normal 0 0.0 0 0.0 dairy (cheese, milk, yogurt, etc.) 0.797 less than normal 44 32.1 93 67.9 normal 47 33.6 93 66.4 eggs (omelet, in salad, etc.) constant less than normal 91 32.9 186 67.1 normal 0 0.0 0 0.0 grains (bread, pasta, rice, etc.) constant less than normal 91 32.9 186 67.1 normal 0 0.0 0 0.0 sweets (candy, cookies, pie, etc.) 0.217 less than normal 83 34.2 160 65.8 normal 8 23.5 26 76.5 caffeinated soft drinks (cola, diet cola, energy drinks, etc.) 0.807 less than normal 89 33.0 181 67.0 normal 2 28.6 5 71.4 coffee and tea (hot coffee, iced coffee, black tea, etc.) 0.521 less than normal 12 28.6 30 71.4 normal 79 33.6 156 66.4 abdulwahid and ahmed: nutritional characteristics of pregnant women 43 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 37-44 while it was consistent regarding eating fruits. aikawa et al.[11] found on their results that there was no significant (p > 0.05) association of anemia with eating fruit which was consistent with the results of present study, while it was inconsistent regarding eating chicken, vegetable, meat, and meat products. jufar and zewde[14] found on their results that there was no significant (p > 0.05) association of anemia with drinking tea and coffee and taking fruit, which was consistent with results of present study, while it was in contrast regarding eating animal foods and green leafy vegetables. the results of aikawa et al.[11] indicated that there was no significant (p > 0.05) association of hb level with consumption of fish, seafood, and eggs, while there was a significant (p < 0.05) relation of consumption of beans with hb levels. khapre et al.[15] mentioned that there was no significant (p > 0.05) difference of degree of anemia and was found between vegetarian and non-vegetarian type of diet. viveki et al.[16] found on their results that there was no significant association of severity of anemia with dietary habits such as vegetarian and mixed diets. clinical practice guideline nutrition for pregnancy[10] described the food pyramid guidelines as the following: starchy carbohydrates, such as whole grains and fiber-rich foods including breads, cereals potatoes, pasta, and rice, six or more servings a day from this group; where one serving is 1 bowl of cereal, 1 slice of bread or 1 medium potato, fruit and vegetables, at least five or more servings a day; 1 serving is 1 medium-sized fruit, for example, 1 apple or 3 dessert spoons of vegetables, dairy foods which includes milk, cheese, and yogurt, 3 servings a day from this group; 1 serving is 125 g yogurt, 25 g of cheese or 200 ml milk, and protein foods including meat, poultry, fish, eggs or legumes, at least 2 servings a day: where one portion is 50–75 g (2–3 oz) cooked meat, 100 g (4 oz) fish, 2 eggs or 6 dessert spoons beans. murray and mckinney[17] described the food plan and amounts per serving with recommendations intake for pregnancy as the following: food and amounts/ serving of whole grains (1 oz = 1 slice bread and ½ cup rice or pasta) for vegetables, fruits, and dairy group (1 cup milk or yogurt, 1.5 ounce hard cheese, and 2 cup cottage cheese) for protein group (1 ounce meat/poultry/fish, 1 egg, ¼ cup table 8: logistic regression analysis of the factors associated with anemia variables p value odd’s ratio 95% ci of or vegetables (carrots, mushrooms, potatoes etc.) less than normal 0.003 0.493 0.308–0.788 normal (reference category) chicken (fried chicken, in soup, grilled chicken, etc.) less than normal 0.999 1.039 1.017–1.061 normal (reference category) beef (steak, meatballs, etc.) less than normal 0.035 0.689 0.488–0.973 normal (reference category) nuts (almonds, cashews, walnuts, etc.) less than normal 0.053 0.641 0.409–1.005 normal (reference category) or: odds ratio, ci: confidence interval cooked beans, ¼ cup tofu, and 1 tablespoon peanut butter), while the recommended intakes for them as following: for whole grains 7–9 oz, vegetables 3–3.5 cup, fruits 2 cup, dairy group 3 cup, and protein group 6–6.5 oz. karaoglu et al.[18] conducted a cross-sectional study about the prevalence of nutritional anemia in pregnancy in an east anatolian province, turkey, who found that 40.2% of study sample consumed egg, 8.1% consumed red meat, poultry or fish, and 42.0% consumed fruit and vegetables daily. el-hindi (2011) concluded in a study done regarding nutrition that 55.4% of the study sample get regular nutrition, while 44.6% get not regular nutrition in anemic group, in non-anemic group and 78.3% of study sample get regular nutrition while 21.7% get irregular nutrition during pregnancy. results of a study done by karaoglu et al.[18] showed that 19.4% of study samples ate one portion of red meat, poultry, and fish consumption every day, while 27.8% of them ate less than frequent of red meat, poultry, and fish consumption. regarding eating fruit and vegetable consumption, 24.3% of participants ate one portion, while 29.1% of participants ate less frequents. these results are consistent with the results of the present study regarding eating chicken and vegetables, while they are in contrast regarding eating fruits and beef. plante et al.[19] conducted a survey on nunavik women about iron deficiency and anemia among women, they found that less than half 37.1% of the sample ate normal meat, which was consistent to the results of the present study, while it was in contrast regarding eating sweets, which indicated that less than half 39.4% of the sample ate normal sweets, on the other hand, it is also in contrast regarding drinking coffee and tea, which found that less than half 37.1% of the study sample drinking less than normal coffee and tea. conclusion eating less than normal of vegetables, beef, and nuts were indicated risks of anemia during pregnancy. references 1. l. gedefaw, a. ayele, y. asres and a. mossie a. “anemia and associated factors among pregnant women attending antenatal care clinic in wolayita sodo town, southern ethiopia”. ethiopian journal of health sciences, vol. 25, no. 2, pp. 155-162, 2015. 2. c. dim and h. onah. “the prevalence of anemia among pregnant women at booking in enugu, south eastern nigeria. medscape general medicine, vol. 9, no. 3, p. 11, 2007. 3. m. yesuf and m. wassie. “prevalence and associated factors of anemia among pregnant women of mekelle town: a cross sectional study”. bmc research notes, vol. 7, p. 888, 2014. 4. world health organization. “nutrition of women in the preconception period during pregnancy and the breastfeeding period”. provisional agenda item, meeting, 2012. 5. kalaivani, k. “prevalence and consequences of anemia in pregnancy”. the indian journal of medical research, vol. 130, pp. 627-633, 2009. 6. f. lone, r. qureshi and f. emanuel. “maternal anaemia and its impact on perinatal outcome”. tropical medicine and international health, vol. 9, no. 4, pp. 486-490, 2004. 7. a. ahmed and k. ali. “maternal anemia status among pregnant women in erbil city”, iraq. world family medicine journal, vol. 11, no. 8, p. 23, 2013. 8. t. al-khafaji, j. zangana and s. dauod. “prevalence of pregnant abdulwahid and ahmed: nutritional characteristics of pregnant 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[last accessed on 2015 mar 03]. . 42 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 research article prediction the groundwater depth using kriging method and bayesian kalman filter approach in erbil governorate kurdistan ibrahim mawlood, paree khan aabdulla omer department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq abstract the aim of this research is using the kriging method as one of geostatistics interpolation methods on the measured value of the specific part and bayesian kalman filter to identifying the depth of groundwater in erbil. geostatistics is a tool which is developed for statistical analysis of any continuous data that can be measured at any location in the space. the kalman filter is the bayesian optimum solution to the problem of estimating the unknown state of a dynamic system from noisy data and is more efficient than computing the estimate directly from the entire past observed data. the main goal of this work is to predict anew value at the unmeasured location by kriging method and bayesian kalman filter and compare these two methods. the dataset is the observed values of the (295) wells that had been taken from a known specific place which called shaqlawa – in erbil governorate. the comparison was done by calculating mean absolute error (mae) and root mean square error (rmse) for the value of the depth of groundwater in the eara of the study. the values of (mae and rmse) of each models are compared and the smaller values of them are the better interpolation as it shown in analyzing to evaluate the precision of the prediction. keywords: bayesian estimation, covariance function, gaussian random field, groundwater-surface interpolation, kalman filter, kriging interpolation method introduction the kriging is a spatial interpolation geostatistical method used for the first time in meteorology, geology, environmental sciences, agriculture, and others fields. this method is used to find the best estimator under the assumption of the second order stationarity. geostatistics is a set of tools and models that are developed for statistical analysis of any continuous data that can be measured at any location in the space. verify three data feature in statistical continuous data analysis: dependency, stationery, and distribution. with these features, you can proceed to the modeling of the geostatistical data analysis like simple kriging. in addition, the goal of this work is to predict a new value at the unmeasured location by gaussian semivariogram function (model) and compare the results of this model based on the simple kriging method and understanding their spatial variability with another approach called bayesian kalman filter. kalman filter is an optimal linear estimator which provides the estimation of signals in noise. kalman used the state transition models for the dynamic system in the estimation process. groundwater-surface interpolation in general, spatial statistics and geographic information system (gis) rely on each other in many ways. arc gis is software which can be used to create covariates for inclusion in all statistical models and to bring out the results from statistical models. the work in this study might be very important to evaluate the results from different models in simple kriging interpolation approaches. this kind of comparison presents a relevant meaning for the variability of a physical model which used as a reference to validate the interpolation results. groundwater depth of the wells of any sample point data for generates surface need to be evaluated and preprocessed before interpolation. the locations and values of sample point data will impact the interpolation result. first, all the collected data should come from the same type of wells in the same aquifer. the well information should be carefully evaluated to make sure the data reflect the dynamics of groundwater in the target aquifer, not other aquifers. second, the spatial distribution of sample point data should be carefully considered. the clustered data and sparse data in one area corresponding author: kurdistan ibrahim mawlood, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq. e-mail: kurdistan.mawlood@su.edu.krd doi: 10.24086/cuesj.v3n1y2019.pp42-49 copyright © 2019 kurdistan ibrahim mawlood, paree khan aabdulla omer. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) received: apr 27, 2019 accepted: may 29, 2019 published: jun 30, 2019 https://creativecommons.org/licenses/by-nc-nd/4.0/ mawlood and omer: prediction the groundwater depth 43 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 will cause different interpolation results. after collecting suitable data for interpolation, raster calculator in gis is used to calculate the elevation of groundwater table in each well by dividing the surface elevation (digital elevation model) with the groundwater depth.[1] gaussian random field used gaussian random field z(s) to identify the spatial correlation structure, any field of spatial is a set of random variables which parameterized by some set (d⊂rd). the simplest stochastic process form is as follows: z s s d rd( ) :{ ∈ ⊂ } (1) where: z(s): field of random spatial. s: coordinates spatial random variable. d: domain of spatial random variable. rd: d-dimensional euclidean space. any finite collection{z(s 1 ),z(s 2 ).,z(s k ).,} is multivariate normal: 1 1( ) ( ) . . ~ , ( ( ), ( )) . . ( ) ( ) i j k k z s s n cov z s z s z s s µ µ                                    if the following assumptions hold, then a spatial random field is called second-order stationary: [2] ( ( ))e z s s dµ= ∀ ∈ (2) (3) where: e: expected value. cov: covariance function of two locations. h=s 2 −s 1 : vector distance between z(s 1 ) and z(s 2 ). however, we know that the covariance function can be expressed as follows: ( ) ( ) ( )( ) [ ] 1 2 1 1 1 1 1 1 2 1 1 ( ), ( ) ( ), ( ) ( ) ( ) ( ) ( ) ( ) s s h cov z s z s cov z s z s h c h e z s z s h e z s z s h    +  = + =   = − + −  = + − (4) when the expected value is equal to zero. ( ( )) 0,e z s s d = = ∀ ∈ (5) then, cov z s z s cov z s z s h c h e z s z s h ( ), ( ) ( ), ( ) ( ) ( ) ( ) 1 2 1 1 1 1 ( ) = +( )  = = +[[ ] (6) when the mean of a second-order stationary of spatial random field is equal to zero over the d and the covariance function of the locations does not depend on s 1 and s 2 but the vector h. and for cov h cov z s z s h v z s( ) ( ), ( ) ( )= = +( )  = [ ]0 if variance or covariance function does not exist, the intrinsic hypothesis and the spatial random field are called (intrinsic stationery) if the following assumption holds as: e z s or e z s z s s d v z s z s s s s ( ( )) ( ) ( ) ( ) ( ) ( ) = −[ ] = ∀ ∈ −[ ] = − ∀ µ γ 1 2 1 2 1 2 1 0 2 ,, s d2 ∈ (7) where, v: variance, γ: semivariogram, 2γ: variogram 2 1 2 1 1 1 1 2  = −[ ] = − +[ ] = + −[ ]( ) − v z s z s v z s z s h e z s h z s e z ( ) ( ) ( ) ( ) ( ) ( ) (ss h z s e z s h z s 1 1 2 1 1 2 + −[ ]( ) = + −[ ]( ) ) ( ) ( ) ( ) (8) in addition, if the covariance function c(s 1 −s 2 )=c(h) or semivariogram γ(s 1 −s 2 )=γ(h) depends only on separation distance between s 1 and s 2 , h=||s 1− s 2 ||, then the spatial random field is called isotropic.[3] the kriging kriging is a spatial interpolation geostatistical method used for the 1st time in meteorology, geology, environmental sciences, agriculture, and other fields. this method is used to find the best estimator under the assumption of the second-order stationarity. a geological process may not be stationary in reality. in case, where the process is non-stationary, we could use non-linear functions.[4] here, the classifying geostatistical techniques are as follows [table 1]. assumptions in some of the interpolation methods especially geostatistical methods, they have their own assumptions as follow:[5] 1. stationarity 2. intrinsic hypothesis 3. isotropy and anisotropy 4. unbiased. semivariogram function and covariance function the semivariogram function is a structure of intrinsically stationary spatial random field which describes a broader class of erath phenomena. in the case of the second-order stationary spatial random processes, there is an equivalence between covariance function and semivariogram function as follows: table 1: classification of geostatistical techniques model stationary nonstationary linear ordinary/simple kriging universal kriging; kriging using irf-k nonlinear disjunctive kriging simulation simulation of irf-k irf-k: intrinsic random functions of order k ( ) ( ) ( )1 2 1 1 2 2 1 2 1 2 ( ), ( ) ( ) ( ) ( ) , s scov z s z s e z s z s c s s s s d µ µ = − − =  − ∈ mawlood and omer: prediction the groundwater depth 44 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 v z s h z s v z s h v z s cov z s h z s v z s ( ) ( ) ( ) ( ) ( ), ( ) ( ) + −[ ] = +[ ] + [ ] − +[ ] = [ 2 2 ]] − +[ ] = −[ ] = ÷ = − 2 2 0 2 2 0 cov z s h z s c c h h h c c h ( ), ( ) ( ) ( ) ( ) ( ) ( ) ( )   (10) the semivariogram function is a measure of dissimilarity between pairs of locations or observed value z(s+h) and z(s). there are three parameters of semivariogram function for the spatial second-order stationary processes: nugget effect (c 0 ), range (a), and partial sill (c) which are shown in figure 1. the sum (c 0 +c) is called the sill (marcin and marek, 2010), (sluiter, 2009). the gaussian semivariogram model the gaussian model is commonly used to represent events with a small scale spatial structure,[6] the equation for this model is similar to the normal cumulative distribution function, and it is given by [figure 2]:  z h c c h a h( ) exp ,= + − −            >0 2 0 2 1 0 (11) the simple kriging interpolation method simple kriging can deliver the value at any unmeasured location (s 0 ) using a linear estimator λ i for the measured values at locations (s 1 ,s 2 .,s n ): z s z si i i n i i n ( ) ( ),0 1 1 1= = = = ∑ ∑  (12) as the sum of all linear estimators equals one, the unbiased prediction is ensured. using the intrinsic hypothesis, the estimation variance is calculated by the formula: ( )2 20 0 0 0 0 1 1 1 ˆ( ) ( ( ) ( ( ) ( )) 2 ( ) ( ) e n n n i i i j i j i i j s var z s e z s z s s s s s       = = = = = − = − − −∑ ∑∑ (13) the semivariance (s j −s 0 ) and(s i −s 0 ) are taken from the variogram model. the goal is to minimize σ2(s) under the unbiased conditions and to find the corresponding weight. the weights are chosen to fulfill: 0 0ˆ( ( ) ( )) 0e z s z s− = according to the assumption that e[z(s 1 )−z(s 2 )]=0 which called intrinsic stationery and to solve this the lagrange multiplier (μ) is introduced, not easy solving this model by any other ways: z s s s s s so i i j i n j( ) ( ) ( ) ( )= − + = − = ∑λ γ µ γ 1 0 0 (14) the solutions to this linear equation system are the values of the linear estimators. this system is called kriging system.[7] the kriging variance is determined by: σ λ γ µk i i i n s s s2 0 1 0= − + = ∑ ( ) ( ) (15) kalman filtering the kalman filter gives unbiased, linear, and minimum variance recursive estimate to the state of a dynamic system from noisy data taken in separate real-time.[10] kf used in the formulation of a dynamic model for linear dynamical systems provides a recursive solution to the problem of the optimal linear filter. the recursive solution in every state update estimate is calculated from previous estimates and data entry; only the previous estimate requires storage. kf has been widely used in the fields of signal processing and modern control and airborne surveillance systems, radar signal processing and control, and adaptive controls. here, we will provide only the equations needed to develop discrete recursion kf, discrete state equations are given as linear dynamic for signal θ t and observation y t .[11] we have two equations: figure 1: the relationship between the γ(h) and c(h) for the second-order stationary of a spatial random variable figure 2: the gaussian semivariogram model with parameters mawlood and omer: prediction the groundwater depth 45 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 1. observation equation: y ft t t t= +θ υ 2. system equation: θ θ ωt t t tg= +−1 the initial state vector has a mean θ 0 and a covariance matrix p 0 that is: 0 0 0 0 ˆ[ ] = and [ ] = e var p   y t is the vector of noisy observation at time t, of dimension m×1. f t is the known matrix at time t, of dimension m×p relates the state vector to the observation vector y t . υt is the vector of observation error (stochastic error term) at time t, having normal distribution with zero mean and covariance v t , of dimension m×θt is the vector of unknown state parameters (signal) at time t, of dimension p×1, system states. g t is the matrix of time-varying state transition matrix dimension pxp. ωt is the system error (stochastic error term) at time t, having normal distribution with zero mean and covariance w t , of dimension p×1. where, v t and w t are variance-covariance measurement and process noise matrices, respectively. the noise vectors ω t and υ t process and measurement noise, respectively, are uncorrected white noises (have zero means) they are independent of each other and have normal probability distributions. the disturbances υ t and ω t uncorrected with the initial state that as: e( ) = 0υ θt t 0 and e( ) = 0ω θt t 0 for t=1, 2, 3,…,t bayesian estimation of dynamic linear models kalman filter is used for estimating a state of a dynamic system which gives the best-unbiased estimator using previous measurement knowledge. previous algorithms stand from the use of all the previous information to estimate the state of the system at the time of the next step. this dynamic linear model is the simple model represented as: observation equation y t =μ t +ε t system equation μ t =μ t−1+ σ μ2 where the errors ε t and ω t are mutually independent and independent of initial information probability (μ 0 |d 0 ) initial information probability (μ 0 |d 0 ) represents forecaster’s probabilistic information of about the μ 0 at time t=0. the mean m 0 and the variance c 0 are a point estimate and the associated uncertainty of μ 0 . d t consists all the information available up until time t, including d 0 , the values variances values { , : }σ σε µ 2 2 0t > , and the the observations yt,y t−1 ,…,y 1 or, the only new information becomes available at any time t is the observed value y t , where d t ={y t ,d t−1 }. here, we start expressing initial information concerning the parameter μ 0 was described in the form of a normal probability distribution with mean m 0 and variance c 0 is: 0 0 0~ ( , )n m c using a mathematical process and bayes’ theorem at the time (t−1) of the parameter μ t−1 is: 1 1 1 1( | ) ~ ( , )t t t t td n m c − − − − where, d t−1 is d yt t − −=1 1 2 2{ , , }σ σε µ . we aim now to find a final distribution for the parameter μ t at last time is the time appointed by our knowledge of all available data, or the time that we want to filter. we recompense an extremely presence of the information is: d y dt t t= −{ , }1 . then bayes’ theorem is posterior ∝ observed likelihood×prior p d p y p dt t t t t t( | ) ( | )* ( | )µ α µ µ −1 (16) where p(y t |μ t ) represents the likelihood distribution function at time t p(μt|d t−1 ) represents the prior distribution at time t both of them are distributed normally now to find all of the weighting function p(μ t |d t−1 ) and the prior probability before observation y t , using dynamic linear model and probability distribution as: e y e e e t t t t t t t t t t ( | ) ( | ) ( | ) ( | ) µ µ ε µ µ µ ε µ µ = + = + = and v y v v v t t t t t t t t t ( | ) ( | ) ( | ) ( | ) µ µ ε µ µ µ ε µ σε = + = + = 2 or ( | ) ~ ( , )y nt t tµ µ σε 2 and from the system equation: e d e e d e d m t t t t t t t t t ( | ) ( ) ( | ) ( | ) µ µ σ µ σ µ µ − − − − − = + = + = 1 1 1 1 1 and v d v v d v d c r t t t t t t t t t t t ( | ) ( ) ( | ) ( | ) µ µ σ µ σ σ µ µ µ − − − − − = + = + = + = 1 1 1 1 1 2 or 1 1( | ) ~ ( , )t t t tp d n m r − − then by substituting each of (yt|μt), p(μt|d t−1 ) in equation (2.10.1) we get mawlood and omer: prediction the groundwater depth 46 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 p t d t e y t t r t t m t ( | ) [ ( ) ( )]µ α σ ε µ µ− − + − − 1 2 1 2 1 1 (17) we can find that: p t d t e c t t m t ( | ) [ ( )]µ α µ− − 1 2 1 (18) now we get mean m t and variance ct of final distribution, respectively, are: m y r m r c r rt t t t t t t = + + = + + − 1 1 1 1 2 1 2 2 2 σ σ σ σ ε ε ε ε , and then, the final distribution for parameter μt at time t is ( | ) ~ ( , )t t t tp d n m c we can write the mean and variance of posterior probability distribution after simplifying them as: m m k y mt t t t t= + −− −1 1( ) (19) c kt t= *σε 2 (20) where k r rt t t= + −( )σε 2 1 (21) where the equation (19) called kalman filter, sometimes called for two equations (19), (20) together kalman filter, and equation (21) is kalman gain. forecasting dynamic linear models we mean by forecasting finding the h-step ahead for each distribution given the data dynamic linear model y1,y2.,yt. for a dynamic linear model, the h-step-ahead forecasting distributions, for states and observations, are obtained as a product of the kalman filter.[12,13] from the observations y 1:t to y t +h we noticed that the data y 1:t provide information about θt, which, in turn, gives information about the future state evolution up to θ t+k and consequently on y t+h .[13,14] then, the general dynamic linear model for h step is: observation equation: y f n vt h t h t h t h t t+ + + += +θ υ υ ~ [ , ]0 (22) system equation: θ θ ω ωt h t h t h t h t tg n w+ + + − += +1 0~ [ , ] (23) let a0=mt, r0=ct, then for h≥1, for forecasting at time t, the forecaster requires the h-step ahead marginal distributions, (y t+h |dt) and (θ t +h|dt), predictions start from the posterior estimate of the state, obtained from the kalman filter, on the time (t) on which the forecast is made. the mean and variance for the observation equation are: 1 1 1 ˆ ( | ) ( | ) t h t h t t t t t k t y e y d e f d fg m   + + + + + = = + = (24) q t var y d fg c g f fg w g f v t t h t h t k t t i i t t i h ( ) ( | ) ( ) ( ) = = + + + = − ∑ 0 1 (25) application, results and discussion the study area and data collection spatial variability of any regional variable is a result of complex processes which is working at the same time and over long periods of time. variation of a regional variable has never been an easy task or work. many regional random variables vary not only horizontally but also with depth such as wells of water, oil, and gas. the spatial attribute includes approximately all wells at fields that exist which spatially distributed across the study area to represent the fluctuations of levels from place to another place in whole area. the source of dataset is the observed values of the 295 wells that had been taken from the known specific place which called shaqlawa in erbil governorate. these observations are collected by gps by the ministry of agriculture, figure 3 expresses the location of each well, and each point of the dataset has its name and properties. furthermore, each point has its goal for drilling the well, some of them for drinking or irrigation and agriculture. figure 3 shows the geographical location of the data points of shaqlawa or all wells. they are shown as a surface which can be represented by the most probably prediction map and also by estimated prediction when we are applying a model of the simple kriging interpolation. the figure shows cycles that are many points closer together tend to be more alike than things that are farther apart (quantified here as spatial autocorrelation). results of groundwater-surface interpolation figure 4 can be described as continuous data and represented the random field. it shows a histogram of the observed values and how they distributed in the region. it represents the curve of the observations values, and the results after taken the suitable transformation that has a small standard deviation equal to 0.37924, the skewness of dataset is equal to −0.61614, near to zero and the kurtosis of the dataset is equal to 4.5715 not near to three. these indicated that the distributions of depth in sample data points were approximate to normal distribution. results of gaussian semivariogram simple kriging surface interpolation spatial dependency can be detected in this dataset using several tools available in geostatistical analysis exploratory spatial data analysis and geostatistical wizard in gis software. figure 3: geographical location of the dataset mawlood and omer: prediction the groundwater depth 47 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 in geostatistics semivariogram is called spatial modeling (called structural analysis or variography). table 2 shows the model called the gaussian semivariogram. this gives a quantitative description of variability about the same regional variation. the important part of the variogram is the rang which describes the distance. it shows the isotropy of the gaussian semivariogram model for (295) dataset of groundwater of wells in shaqlawa, which all points have equal directional of variability as being north-south and east-west. furthermore, the anisotropy of gaussian semivariogram model expresses the same data (554) that change with the direction which described an ellipsoid, and this ellipsoid specified by the length of two orthogonal axis (major and minor) with its orientation angle θ. figure 5 shows gaussian semivariogram model for isotropy and anisotropy surfaces, these used to predict of random field (spatial field) of groundwater well z(s 0 ) unknown value in the same area, which depends on only maximum (5) neighbors value of the measured values. table 2 contains all cross-validation of isotropy and anisotropy of simple kriging gaussian semivariogram model and the prediction value of the depth of unknown new value of well. table 3 shows all information about simple kriging by guassian semivariogram model to predict a new location of groundwater in the same area for both of isotropy and anisotropy. the prediction value of unknown is measured also depending on (5) maximum and (2) minimum value of neighbors from measured value for the depth of new well with longitude and latitude. in anisotropy gaussian semivariogram model is equal to 173.2582 and its greater than the depth of the isotropy exponential semivariogram model which equal to 132.3405. estimation the depth of groundwater using kalman filter the estimation process of parameters is made using the initial values of the depth of groundwater mean and variances are being of an estimate of it is values. equations (19), (20), and (21) are used to estimate parameters, error covariance, and kalman gain for univariate simple dynamic linear model for each observation of the groundwater depth. table 4 gives the actual value, estimated groundwater depth parameter, error covariance, and kalman gain. examining the table reveals the following: the error covariance is convergence at a time point (t=18) with the value (802.1407). kalman gain is convergence at a time point (t=16) with the value (0.383983). there is two convergences time and the last time convergence is the best estimate for the same time. a compression between groundwater depth resulting from the estimate and forecasting groundwater depth values for all observations is shown in figure 6. the results show how figure 6: difference between estimate values and forecast values for groundwater depth figure 4: the histogram of the dataset figure 5: (a and b) surfaces of the gaussian semivariogram model a b mawlood and omer: prediction the groundwater depth 48 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 closely the estimated model matches the forecasting electricity groundwater depth. the blue line is estimate values and the red line is forecasting values. evaluating accuracy (or error) measures for estimation for selections, the best approach (method) for prediction the groundwater depth two accuracy criteria were used, which are (mean absolute error [mae] and root mean square error [rmse]). they are based on the error estimate, which is the difference between the estimated and forecast groundwater depth values. the results are shown in table 5. table 3 represents that kalman filter is the better approach based on the two criteria, where each of them has a small value than simple kriging using gaussian semivariogram model for both of isotropy and anisotropy. conclusions the major results of performing the analysis of two approaches (simple kriging and kalman filter), the following main conclusions have been achieved: 1. the data follow approximately to normal distribution after taken log transformation for variability and secondorder stationary to remove the trend. 2. the variability of the depth of groundwater wells elevation in the region changes from north to south or from westnorth to south-east, this is due to the spatial dependency in the area which is one of the reasons of the depth of well. 3. for unmeasured value used simple kriging with gaussian semivariogram model, in this model the predicted value by isotropy semivariogram model is better than the anisotropy semivariogram model depending on the value of the depth of groundwater [table 2]. 4. when applied the kalman filter for estimating and forecasting the depth of groundwater, we found that the kf is suitable for estimation and yield good results, because in the presence of white gaussian noise because when the normality condition holds, the kf will give optimum and sufficient results. 5. the result shows that the bayesian kalman filter approach has the best estimation and forecasting results compared to simple kriging using the accuracy criteria (mae and rmse). table 2: results of gaussian semivariogram model simple kriging exponential semivariogram isotropy anisotropy nugget (c 0 ) 0.0714 0 rang (α) 2 2 major rang 2554.326 4998.32 minor rang 2554.326 1949.16 partial sill (c) 0.081173 0.093287 lag size (h) 416.5267 416.5267 number of lag 12 12 table 3: accuracy criteria for each approach methods mae rmse kalman filter 13.2975 18.078 simple kriging/gaussian model 29.7364 43.719 mae: mean absolute error, rmse: root mean square error table 4: results of simple kriging gaussian semivariogram model simple kriging exponential semivariogram isotropy anisotropy maximum neighbors 5 5 minimum neighbors 2 2 predicted value x 400,739 400,739 y 421,531 421,531 depth (m) 132.3405 173.2582 table 5: estimated parameters when ( ̂ 0 =126.02, p̂0 =2089.088, and w=500) convergence time actual values estimate error covariance kalman gain 1 189 126.02 2089.088 0.73123 2 201 167.5177 1156.157 0.55345 3 91 133.6806 923.7829 0.442213 4 142 137.0526 846.7026 0.405315 5 100 122.529 818.8316 0.391973 6 130 125.4203 808.4435 0.387 7 160 138.7378 804.528 0.385126 8 140 139.223 803.046 0.384416 9 50 104.9482 802.4841 0.384147 10 151 122.6342 802.271 0.384045 11 183 145.8151 802.1901 0.384007 12 100 128.2224 802.1594 0.383992 13 100 117.3854 802.1478 0.383986 14 220 156.7878 802.1434 0.383984 15 193 170.6927 802.1417 0.383984 16 180 174.2665 802.1411 0.383983 17 200 184.1478 802.1408 0.383983 18 147 169.8836 802.1407 0.383983 19 202 182.2158 802.1407 0.383983 20 151 170.2295 802.1407 0.383983 . . . . . . . . . . 291 239 164.2836 802.1407 0.383983 292 211 182.2219 802.1407 0.383983 293 73 140.2825 802.1407 0.383983 294 91 121.3589 802.1407 0.383983 295 82 106.2469 802.1407 0.383983 mawlood and omer: prediction the groundwater depth 49 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 42-49 references 1. s. i. gou. “identifying groundwater dependent ecosystems in the edwards aquifer area, zachry department of civil engineering”. texas: texas a and m university, 2010. 2. r. webster and m. olivier. “geostatistics environmental scientists”. 2nd ed. new york, usa: john wiley and sons, 2007. 3. l. marcin and k. marek. “simple spatial prediction least squares prediction, simple kriging and conditional expectation of normal vector”. poland: department of geomatics, agh university of science and technology in krakow, 2010. 4. d, sarma. “geostatistics with application in earth science”. 2nd ed. dordrecht: springer, captial publishing company, 2009. 5. r. sluiter. “interpolation methods for climate data; literature review”. knmi, r and d information and observation technology, 2009. 6. i. clark and w. v. harper. “practical geostatistics”. columbus, ohio: ecosse north america, 2000. 7. m. disse and k. amin. “review of various methods for interpolation of rainfall and their applications in hydrology, thesis of chair of hydrology and river basin management”. munich: technical university munich, 2017. 8. r. e. kalman. “a new 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(1): 1-6 research article a statistical study of the influence of covid-19 on the agricultural supply chain (vegetative) production in halabja governorate shahen m. faraj1, hazhar t. a. blbas2*, aymen i. ahmed3 1department of international business, college of law and administrative, university of halabja, iraq, 2department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region, iraq, 3independent researcher abstract it is clear that the covid-19 virus has affected human life and activities all over the world. agricultural sector is not excluded from the affection of course. importance of this issue encouraged us to carry out this statistical study aiming to show the impact of covid-19 on the agricultural supply chain (vegetative) production in halabja. three forms of self-administered questionnaire were used to collect data from three targets; farmers, wholesalers, and retailers using non-probability sampling method. descriptive analysis was used to show frequencies, percentages, and mean scores, while independent sample t-test was used to find the relationship between variables using spss software v.26. results show that there is no statistically significant relationship between the characteristic of production and residence. lockdown had a major impact on the plant production and its supplying chain in the following sequence: fewer products were delivered to the wholesaling markets by farmers, retailers bought less so consumers got less products. keywords: agricultural supply chain, farmers, wholesalers, retailers, independent sample t-test, halabja governorate, covid-19 introduction the first case of covid-19 virus attacked human was detected in china at the end of 2019,[1] the first case of the virus attacked human was detected in iraq on february 24, 2020, and in the kurdistan region on the 1st of march 2020.[2] lockdown began almost by the early cases of covid-19 attacks in iraq, so in the kurdistan region, until it reached a complete lockdown in all governorates and also the borders with neighboring countries at the end of march 2020.[3] food availability and agricultural production and how to provide food proportionate to the increasing population are one of the major issues globally.[4] especially after the occurrence of the covid-19 pandemic, and how the situation has become after the lockdown between countries and cities.[5] the effect of this, especially in non-developing countries such as iraq, in which agricultural production declined due to the wars in the eighties and early nineties of the last century, which led to the exhaustion of the state and manpower,[6] as well as a sharp declination in government support of the agricultural sector, this declination became more severe after the 2003 occupation. this sector has retarded more and more with the governments that ruled iraq until nowadays.[7] iraq has been changed from a food exporting country before the 1980s to a heavy food importer. it depends 70–80% approximately on imported agricultural products from abroad.[8] because of the lack of production or the absence of government support for the agricultural sector, the agricultural supply chain is usually short because iraqi agricultural products are often fresh from the farm to the table not from seed to table, without going through any manufacturing or storage processes.[9] this is in contrast to what was the circumstance before the 1980s, where the agriculture supply chains were longer,[10] as it started from the government side, such as distributing the necessary supplies for agricultural production, such as seeds, and fertilizer as well as the delivering of water, the distribution of lands, the purchase of some products from farmers and their introduction into many of the food industries,[11] but corresponding author: hazhar t. a. blbas, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region, iraq. e-mail: hazhar.abubaker@su.edu.krd received: november 12, 2021 accepted: december 5, 2021 published: january 30, 2022 doi: 10.24086/cuesj.v6n1y2022.pp1-6 copyright © 2022 shahen m. faraj, hazhar t. a. blbas, aymen i. ahmed. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) faraj, et al.: influence of covid-19 on the agricultural production 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 1-6 after these period hundreds of manufacturing and food processing laboratories were gradually closed due to wars, mismanagement, and corruption. therefore, it was necessary to execute a statistical study of how the agricultural supply chain (vegetative) production worked before the occurrence of the pandemic and in the period of pandemic to know its impact on both consumers and stakeholders who involved in the supply chain, including merchants, farmers, laborer, owners of wholesale and retail stores …etc., in halabja governorate, which is an agricultural governorate in kurdistan region – iraq and has abundance of surface and underground water, as well as it has fertile plains and mountainous areas that provide the possibility to cultivate different types of agricultural crops. the main hypotheses in this study are to determine whether or not there is a relationship between production characteristics and residency. materials and methods in 2014 halabja has become a province by the kurdistan region government, it is located to the east of sulaymaniyah province and nearly 90 km far from sulaymaniyah, north of iraq,[12] and it is an agricultural governorate, due to its availability of mountainous areas, as well as fertile plains and abundance of water. this is motivating to investigate this research to know how the covid-19 affect the agricultural supply chain (vegetative) production in the halabja governorate. data was collected via three forms of questionnaire including farmers, wholesalers, and retailers of halabja governorate between 1st of october, 2020 and 15th of february, 2021. non-probability sampling method used to collect the data which is snowball sampling for farmers and judgment sampling for wholesalers and retailers. the questionnaire of this study included three parts, first part was the demographic questions such as gender, age, residency location, number of experience years in the business earned by studying, and working for all of our targets (farmers, wholesalers, and retailers). the second part included some questions about production like (area of his property in donums1, yield of a product per donum in tons, and the total quantity produced) for the farmers, there were questions for retailers and some other ones for the wholesalers. the third part was a likert scale (strongly disagree = 1, disagree = 2, neutral = 3, agree = 4, strongly agree = 5) for all of our targets (farmers, wholesalers and retailers). data were analyzed using spss v.26 and descriptive analysis used to show frequency, percentage, and mean scores.[13] independent sample t-test to compare the mean between two variables,[14] it was used to find the statistically significant relationship between some characteristics of production and residency (live location) using significant level α = 0.05. the researchers selected 15 forms randomly to conducted a pilot study for all three different types of questionnaire farmers, wholesalers, and retailers before beginning the main investigation to confirm the validity of the questionnaires using cronbach’s alpha to assess the consistency of the data in which the values of cronbach’s alpha were (0.78, 0.82, 0.88) for farmers, wholesalers, and retailers respectively.[14,15] 1 1 donum=2500m2 results and discussion this research includes 74 participants which are farmers (n = 30), wholesalers (n = 10), and retailers (n = 34), all of our participants are male (100.0%), table 1 shows the table 1: descriptive statistics for demographic questions (farmers, retailers, and wholesalers) demographic questions f percent (%) age farmers 21–30 1 3.3 31–40 10 33.3 41–50 8 26.7 50+ 11 36.7 retailers 21–30 7 20.6 31–40 14 41.2 41–50 4 11.8 50+ 9 26.5 wholesalers 21–30 2 20 31–40 2 20 41–50 5 50 50+ 1 10 live location farmers inside 10 33.3 outside 20 66.7 retailers inside 13 38.2 outside 21 62.8 wholesalers inside 10 100 outside 0 0 years of working farmers 5-jan 3 10 10-jun 4 13.3 15-nov 5 16.7 16-20 6 20 21+ 12 40 retailers 5-jan 0 0 10-jun 13 38.2 15-nov 7 20.6 16-20 6 17.6 21+ 8 23.5 wholesalers 5-jan 3 30 10-jun 0 0 15-nov 2 20 16-20 4 40 21+ 1 10 faraj, et al.: influence of covid-19 on the agricultural production 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 1-6 73.3% 35.3% 30.0% 3.3% 20.6% 0.0% 3.3% 2.9% 0.0% 20.0% 41.2% 70.0% 0.0% 10.0% 20.0% 30.0% 40.0% 50.0% 60.0% 70.0% 80.0% farmers retailers wholesalers father brother friends own self figure 1: from whom did learn the profession mean and standard deviation in table 2 shows that selling less products than before the covid-19 pandemic is the first characteristic (factor) that farmers pointed because of covid19 pandemic and lockdown with mean and standard deviation (4.70, 0.837), followed by selling products cheaper than they did before to the agricultural wholesalers, people demand had less for products because of lockdown, due to covid-19, products and needs were not been able to be transported to the wholesalers markets, agricultural offices were all closed to demand their needs during lockdown, products have been left ungathered and unsold in the farmlands due to covid19, farmers haven’t been able to visit their farmlands enough during lockdown, products have been gathered but unsold in the farmlands due to covid-19, farmers haven’t been able to afford workers for their farmlands due to lockdown, products have been given more time than before the covid19 pandemic and agriculturalists weren’t been able to visit farmlands enough due to covid-19. table 3 shows the results of independent sample t-test which there is no statistically significant relationship between residence and some characteristics of production such as (area of their property in donums, yield of a product per donum in tons, tons of agricultural production sold, quantity production sold and the total quantity produced) with a p-value (0.482, 0.832, 0.849, 0.476 and 0.157) which are greater than our choosing significant level 0.05, respectively. it means, there was no difference between the mean score of inside and outside for each of the characteristics of production. figure 2 demonstrates that 100.0% of our respondents (retailers) closed their markets during lockdown so that they faced a decline during lockdown (97.1%), 5.9% of retailers said that they had delivery services to houses before lockdown while 8.8% of them said that they had delivery services to percentage of the demographic questions regarding three types of participants, about the farmers their range of age was 25–75 with an average of 48 years, about retailers the range of their age was 23–70 with an average of 42 years and the range of the wholesalers age was 24–55 with 40 years as average. furthermore, %33.3 of the farmers live inside of halabja city, %38.2 of retailers live inside halabja city, while %100 of wholesalers live inside halabja city. on the other hand, the range of the years that farmers worked was 3–50 years with an average of 22 years of working, the range of years that retailers worked was 2–35 years with an average of 33 years of working and the years that wholesalers worked was 2–23 years with an average of 14 years of working. according to figure 1, 73.3% of the farmers learned farming from their fathers, 3.3% of them learned from their brothers and friends, and 20.0% of them learned from own self. about the retailers, 35.3% of them learned the business from their fathers, 20.6% of them from their brothers, 2.9% from friends, and 41.2% from own self. furthermore, we see that exactly 30.0% of the wholesales learned the business from their fathers and 70.0% of them from own self. 100.0% 97.1% 5.9% 8.8% 14.7% 5.9% 11.8% 11.8% 0.0% 2.9% 94.1% 91.2% 85.3% 94.1% 88.2% 88.2% 0.0% 20.0% 40.0% 60.0% 80.0% 100.0% 120.0% y es n o c lo si ng m ar ke ts du rin g lo ck do w n y es n o f ac e a de cl in e du rin g lo ck do w n y es n o d el iv er y se rv ic es to ho us es b ef or e lo ck do w n y es n o d el iv er y se rv ic es to re st au ra nt s be fo re lo ck do w n y es n o d el iv er y se rv ic es to ho us es d ur in g lo ck do w n y es n o d el iv er y se rv ic es to re st au ra nt s du rin g lo ck do w n y es n o a gr ic ul tu re fo od tr ac ks a nd c ar ts ta ke n pr od uc ts be fo re lo ck do w n y es n o a gr ic ul tu re fo od tr ac ks a nd c ar ts ta ke n pr od uc ts du rin g lo ck do w n figure2: what happened in the markets before and during the time of the pandemic? faraj, et al.: influence of covid-19 on the agricultural production 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 1-6 restaurants before lockdown, sourly that means people could reach their necessaries and did not need delivery services. however, 14.7% of them said that they had delivery services to houses during lockdown and 5.9% said that they had delivery services to restaurants during lockdown, as we see delivery services were decreased during the lockdown and this belongs to the truth that people did not believe of healthy of the foods at restaurants, on the other hand, 11.8% of the retailers said that agriculture food trucks and carts taken products from them before and during lockdown. table 4 shows that with a higher percentage our respondents (retailers) said that less products are brought into markets during lockdown with mean (4.82), followed by people demand have decreased due to covid-19, products are not brought into the markets daily due to covid-19 pandemic, both of product prices have lowered due to covid19 pandemic and products were bought more expensively form wholesale markets during lockdown with the same percentage, transportation services couldn’t have been provided for the products due to covid-19, products have been sold in a wider table 3: independent sample t-test between residence (where do you live) and some characteristics of production characteristic of production n mean std. deviation t p-values how many donums of land have you farmed inside 10 13.3 4.739 -0.713 0.482 outside 20 30.53 75.673 how many tons of agricultural products have you had during the covid-19 pandemic inside 10 98.9 107.103 -0.215 0.832 outside 20 129.55 441.645 how many tons of agricultural production you sold during the covid-19 pandemic inside 10 96.9 108.625 -0.192 0.849 outside 20 124.45 442.76 from the quantity production sold how many were had as revenue inside 10 10.3 15.896 -0.723 0.476 outside 20 46 154.488 how many tons of your products have been outdated because of the covid-19 pandemic inside 10 22.5 45.822 1.455 0.157 outside 20 6.95 11.283 table 2: descriptive statistic for farmers survey questions questions strongly disagree disagree natural agree strongly agree mean std. deviation we sell our products cheaper than we did before to the agricultural wholesalers 0.00% 6.70% 0.00% 16.70% 76.70% 4.63 0.809 products have been left un gathered and unsold in the farmlands due to covid-19. 3.30% 6.70% 13.30% 33.30% 43.30% 4.07 1.081 products have been gathered but unsold in the farmlands due to covid-19. 10.00% 20.00% 13.30% 23.30% 33.30% 3.5 1.408 we sell less products than before the covid-19 pandemic. 0.00% 6.70% 3.30% 3.30% 86.70% 4.7 0.837 products have been given more time than before the covid-19 pandemic. 0.00% 30.00% 23.30% 16.70% 30.00% 3.47 1.224 people demand had less for products because of lockdown. 3.30% 3.30% 6.70% 3.30% 83.30% 4.6 1.003 we haven’t been able to afford workers for our farmlands due to lockdown. 10.00% 20.00% 13.30% 26.70% 30.00% 3.47 1.383 we haven’t been able to visit our farmlands enough during lockdown. 6.70% 13.30% 3.30% 33.30% 43.30% 3.93 1.285 due to covid-19, products and needs were not been able to be transported. 0.00% 6.70% 6.70% 23.30% 63.30% 4.43 0.898 agricultural offices were all closed to demand our needs during lockdown. 3.30% 13.30% 3.30% 26.70% 53.30% 4.13 1.196 agriculturalists were not been able to visit farmlands enough due to covid-19. 10.00% 20.00% 30.00% 13.30% 26.70% 3.27 1.337 faraj, et al.: influence of covid-19 on the agricultural production 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 1-6 table 4: descriptive statistic for retailers survey questions questions strongly disagree disagree natural agree strongly agree mean std. deviation product prices have lowered due to covid-19 pandemic. 5.90% 8.80% 5.90% 14.70% 64.70% 4.24 1.257 people demand have decreased due to covid-19. 0.00% 2.90% 0.00% 11.80% 85.30% 4.79 0.592 products have been sold in a wider range during lockdown, so that customers could have saved them later on. 32.40% 23.50% 2.90% 11.80% 29.40% 2.82 1.696 products were bought more expensively form wholesale markets during lockdown. 5.90% 8.80% 8.80% 8.80% 67.60% 4.24 1.281 less products are brought into markets during lockdown. 0.00% 2.90% 0.00% 8.80% 88.20% 4.82 0.576 products are not brought into the markets daily due to covid-19 pandemic. 5.90% 2.90% 0.00% 5.90% 85.30% 4.62 1.074 workers couldn’t have been provided during lockdown for the purpose of transportation. 38.20% 20.60% 11.80% 8.80% 20.60% 2.53 1.581 transportation services couldn’t have been provided for the products due to covid-19. 14.70% 20.60% 11.80% 14.70% 38.20% 3.41 1.54 table 5: descriptive statistic for wholesale markets survey questions questions strongly disagree disagree natural agree strongly agree mean std. deviation we have had less products in the agricultural wholesale during the covid-19 pandemic. 30.00% 0.00% 0.00% 0.00% 70.00% 3.8 1.932 we have bought products from farmers with less amount of money during the covid-19 pandemic. 40.00% 40.00% 10.00% 0.00% 10.00% 2 1.247 we have sold products cheaper than before because of the covid-19 pandemic. 20.00% 40.00% 30.00% 0.00% 10.00% 2.4 1.174 because of covid-19, products have been left unsold in the agricultural wholesale. 20.00% 40.00% 10.00% 10.00% 20.00% 2.7 1.494 we haven’t been able to afford workers for transportation during lockdown. 10.00% 30.00% 0.00% 0.00% 60.00% 3.7 1.703 transportation services couldn’t have been provided for the products due to covid-19. 60.00% 10.00% 0.00% 20.00% 10.00% 2.1 1.595 transportation problems have increased to the middle and south of iraq than before the covid-19 pandemic. 10.00% 40.00% 10.00% 20.00% 20.00% 3 1.414 during lockdown, neighbor country products such as iran, have been transported much more to the agricultural wholesale than before. 10.00% 30.00% 0.00% 0.00% 60.00% 3.7 1.703 range during lockdown, so that customers could have saved them later on and workers couldn’t have been provided during lockdown for the purpose of transportation with mean (4.79, 4.62, 4.24, 3.41, 2.82 and 2.53). as we see in figure 3, 50.0% of the wholesalers said that corona virus affected the products purchase of the people, 100.0% of them said that agriculture food trucks and carts taken food them and delivered to houses before and during covid-19 pandemic and 90.0% 0f them said that it is increased during covid-19, finally 100.0% of wholesale markets said that they have send their products to the middle or south of iraq during lockdown. table 5 shows that most of the wholesalers said that less products in the agricultural wholesale during the covid-19 pandemic have the most disadvantage to wholesale (with higher mean 3.800), followed by (they haven’t been able to afford workers for transportation during lockdown and neighbor country products such as iran, have been transported much more to the agricultural wholesale than before) with the same mean 3.700. on the other hand, transportation problems have increased to the middle and south of iraq than before the covid-19 pandemic with mean 3.000, because of covid-19, products have been left unsold in the agricultural wholesale with mean 2.700, selling products cheaper than before because of the pandemic with mean 2.400, transportation services couldn’t have been provided for the products due to covid-19 with mean 2.100 and wholesale markets have bought products from farmers with less amount of money during the covid-19 pandemic with mean 2.000. faraj, et al.: influence of covid-19 on the agricultural production 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 1-6 conclusion after studying and analyzing to know the impact of the covid-19 virus on the agriculture supply chain (vegetative) statistically, according to the responders (n = 74) we reached the following results: 1most of the farmers and retailers are aged between 31 and 40 years while most of the wholesale markets are aged between 41 and 50 years. 2majority of farmers and retailers are living outside the city while all of the wholesale are living inside the halabja city. 3most of the farmers have more than 20 years’ experience of farming, while most of the retailers have 6–10 years’ experience of marketing. on the other hand, most of the wholesale markets have 16–20 years of working. 4the highest percentage of the farmers have learned farming from of their fathers, while most of the retailers and wholesale markets learned business by themselves. 5it is clear that, selling less products by farmers (mean = 4.70), brought less products into markets (mean = 4.82) and less products in the agricultural wholesale (mean = 3.800) during the covid-19 pandemic and lockdown has the most impact on the agriculture supply chain production in halabja governorate. 6market owner reported that 100.0% of them closed their markets and 97.1% of them faced a decline during lockdown. 7with a percentage 50.0%, wholesale markets said that covid-19 effected products purchase of people, 100.0% of them said that agriculture food trucks and carts taken food of them and delivered to houses before and during lockdown and with a percentage 90.0% it is increased during covid-19. 8wholesale markets reported that with a percentage of 100.0%, they could not send products to middle and south of iraq during lockdown. 9there was no difference between the mean score of inside and outside living for each of the characteristics of production including donum of farmer’s land, having tons of agricultural products during covid-19, having tons of sold agricultural production during covid-19, revenue quantity production and outdated tons’ production because of covid-19. references 1. world health organization. novel coronavirus (2019-ncov) situation reports. geneva: world health organization, 2020. available from: https://www.who.int/emergencies/diseases/novelcoronavirus-2019/situation-reports [last accessed on 2021 sep 01]. 2. iraq: who coronavirus disease (covid-19) dashboard. n.d. available from: https://covid19.who.int/region/emro/country/ iq [last accessed on 2021 oct 05]. 3. m. k. al-malkey and m. a. al-sammak. incidence of the covid19 in iraq implications for travellers. travel medicine and infectious disease, vol. 38, p. 101739. 4. k. k. lima, a. pasqualetto, t. calil, and s. d. de castro. management of the goiás supply center (ceasa-go): its numbers and proposals for improvements. qualitas revista eletronica, vol. 18, no. 3, p. 1. 5. d. de paulo farias and f. f. f. de araújo. will covid-19 affect food supply in distribution centers of brazilian regions affected by the pandemic? trends in food science and technology, vol. 103, pp. 361-366, 2020. 6. international food policy research institute. yield gaps and potential agricultural growth in west and central africa. washington, d.c., usa: international food policy research institute, 2011. 7. faostat. available from: http://faostat.fao.org [last accessed on 2021 oct 11]. 8. r. d. schnepf and library of congress. congressional research service, iraq agriculture and food supply: background and issues. washington, dc: congressional research service library of congress, 2003. 9. s. aday and m. s. aday. impacts of covid-19 on food supply chain. food quality and safety, vol. 4, no. 4, pp. 167-180, 2020. 10. a. al-haboby, c. breisinger, d. debowicz, a. h. el-hakim, j. ferguson, r. telleria and van rheenen. the role of agriculture for economic development and gender in iraq: a computable general equilibrium model approach. the journal of developing areas, vol. 50, no. 2, pp. 431-451, 2016. 11. g. r. gibson, j. b. campbell and r. h. wynne. three decades of war and food insecurity in iraq. photogrammetric engineering and remote sensing, vol. 78, no. 8, pp. 885-895, 2012. 12. a. h. abudlqadir, b. e. hama and s. m. faraj. the effects of financial crisis on birth rates in halabja province from 2014 to 2018. economics, management and sustainability, vol. 5, no. 2, pp. 132-137, 2020. 13. k. aroian, n. uddin and h. blbas. longitudinal study of stress, social support, and depression in married arab immigrant women. health care for women international, vol. 38, no. 2, pp. 100-117, 2016. 14. h. t. a. blbas, k. f. aziz, s. h. nejad and a. a. barzinjy. phenomenon of depression and anxiety related to precautions for prevention among population during the outbreak of covid19 in kurdistan region of iraq: based on questionnaire survey. journal of public health, vol. 42, pp. 1-5, 2020. 15. h. blbas. statistical analysis for the most influential reasons for divorce between men and women in erbil-iraq. international journal of innovation, creativity and change, vol. 6, no. 2, pp. 207-216, 2019. 50.0% 100.0% 100.0% 90.0% 0.0% 50.0% 0.0% 0.0% 10.0% 100.0% 0.0% 20.0% 40.0% 60.0% 80.0% 100.0% 120.0% ye s no c o v id -1 9 ef fe ct ed p ro du ct s pu rc ha se of p eo pl e ye s no a gr ic ul tu re fo od tr uc ks , c ar ts ta ke n fo od an d de liv er ed to h ou se s be fo re c o v id -1 9 ye s no a gr ic ul tu re fo od tr uc ks , c ar ts ta ke n fo od an d de liv er ed to h ou se s du rin g c o v id -1 9 y es n o if ye s, it ha s in cr ea se d du rin g c o v id -1 9 y es n o s en di ng pr od uc ts to m id dl e an d so ut h of ir aq d ur in g lo ck do w n figure 3: how the pandemic affects wholesale markets before and after its occurs? tx_1~abs:at/add:tx_2~abs:at 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 7-11 research article apply binary logistic regression model to recognize the risk factors of diabetes through measuring glycated hemoglobin levels nozad h. mahmood1, rebaz o. yahya2, saya j. aziz1 1department of business administration, cihan university sulaimaniya, kurdistan region, iraq, 2department of business administration, cihan university-erbil, kurdistan region, iraq abstract this study aimed to identify the diabetes risk factors in the kurdistan region of iraq and explain why diabetes is rapidly spreading there, which examined some sociodemographic characteristics factors that might affect type  2 diabetes such as age, gender, alcohol consumption, and smoking, diabetes family history, and body weight. the data were collected from the hospital of diabetes named the center of diabetes in sulaymaniyah city of iraq, in which 218 diabetic cases were used for that purpose. according to the findings, some factors influence type 2 diabetes, such as gender, smoking, and body weight. for gender, females are more likely to have diabetes than males. furthermore, someone that smokes is more likely to have diabetes than those who do not smoke. furthermore, with increasing each kilogram of body weight, the diabetes degree increases as well. on the other hand, regarding the results, some factors such as age, consumption of alcoholic, and diabetes family history do not affect type 2 diabetes. depending on the findings, it is recommended that people engage in regular physical activity and consume nutritious foods to minimize weight gain, which is one of the primary causes of diabetes as well as they should quit smoking. keywords: binary logistic regression, diabetes, glycated hemoglobin, hba1c, risk factors of diabetes introduction diabetes disease is one of the most widespread infections worldwide, affecting millions of people. besides that, disease rates are increasing quickly, and it is a genuine global health emergency, imposing crucial trouble on many countries.[1] according to the who reports, it predicts that the global diabetes epidemic will nearly double by 2030. one american is killed by diabetes every 3 min, making it the leading cause of blindness, kidney disease, and even death in the united states today. regarding the international diabetes federation reports, there will be 700 million diabetics worldwide by 2045.[2] there has been an epidemic of diabetes mellitus in both developed and developing countries.[3] there are two primary varieties of diabetes. with having type 1, our body will stop making any insulin, a hormone that allows the body to utilize glucose from food sources. type 2 diabetes is a severe disease where the body cannot properly utilize food as an energy source. our bodies use glucose, or sugar, to make energy from most of the food we consume. the pancreas produces a hormone that helps glucose enter cells which is insulin. there is a lack of insulin production and an inability to utilize insulin properly in type 2 diabetes. thus, insulin sensitivity resistance and pancreatic cell failure are two characteristics of type 2 diabetes, leading to many health issues and death.[4] in general, type 2 diabetes accounts for about 95% of all diabetic cases. there has been a rise in this kind of diabetes incidence among younger people due to obesity and a family history of diabetes.[5] increased morbidity and mortality might be expected in persons with diabetes mellitus who do not control their blood sugar.[6] according to recent estimates, 50% of type 2 diabetes mellitus (t2dm) requires insulin in the first 6 years following diagnosis. insulin therapy is often necessary for many people with t2dm to achieve their desired blood sugar levels.[7] as a result of insulin’s ability to keep blood sugar levels in a healthy corresponding author: nozad h. mahmood, department of business administration, cihan university sulaimaniya, kurdistan region, iraq. e-mail: nozad.mahmood@sulicihan.edu.krd received: november 29, 2021 accepted: december 25, 2021 published: january 30, 2022 doi: 10.24086/cuesj.v6n1y2022.pp7-11 copyright © 2022. nozad h. mahmood, rebaz o. yahya, saya j. aziz. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) mahmood, et al.: regression model to recognize diabetes risk factors 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 7-11 range, it prevents a wide range of health issues that might arise from hyperglycemia. the problem statement of conducting this study was that many people in iraq’s kurdistan region were diagnosed with type 2 diabetes at various ages, and the main research questions that led this study are as follows: 1. why is diabetes rapidly increasing in the kurdistan region of iraq? 2. is there any relationship between type 2 diabetes and the factors of gender, age, consumption of alcoholic, smoking, diabetes family history, and body weight? the objectives of the study the essential goals of this research are to determine the factors that affect diabetes disease in the kurdistan region of iraq and explain the reasons for its rapidly increasing prevalence in that region. the significance of the study the significance of this research is for identifying all the factors that lead to diabetes in kurdistan so that people can be aware of the causes of the disease and better control diabetes in kurdistan. the hypothesis of the study 1there is no statistically significant difference between type 2 diabetes and gender (male or female). 2there is no statistically significant difference between type 2 diabetes and the aging of people. 3there is no statistically significant difference in type 2 diabetes based on the consumption of alcohol. 4there is no statistically significant difference between type 2 diabetes and smoking. 5there is no statistically significant difference in type 2 diabetes and diabetes family history. 6there is no statistically significant difference in type 2 diabetes and body weight. literature review glycemic control was more likely to be compromised in patients with younger age, female gender, longer disease duration, high glycated hemoglobin (hba1c) levels, obesity, and lack hypertension.[8] another study showed hba1c levels affected by sociodemographic factors in such a way that the younger and less educated participants recorded higher hba1c levels than their older and more informed counterparts.[9] the previous study revealed a significant association between smoking habits and diabetes mellitus.[10,11] the available evidence shows that the smoking habit increases insulin resistance, and it has further been associated with the risk of chronic pancreatitis and pancreatic cancer.[12] body mass index and waist circumference were the other variables that showed significant associations in the multivariable analysis. overweight people were 4.1 times more likely to have diabetes than ordinary people. this finding is consistent with research from india.[13] similarly, participants with a high waist circumference were more likely to be diabetic than their counterparts, with a prevalence of 11.2% diabetes. it has been proposed that increased abdominal fat stores interfere with insulin action by releasing free fatty acids. furthermore, fat cells secrete signaling factors such as interleukin and tumor necrosis factor, both of which influence the development of diabetes.[14] furthermore, most countries’ life expectancy has increased, with the global life expectancy at birth increasing from 65.6 years of age in 1990 to 73 years in 2017.[15] when these global aging trends are combined with the global diabetes epidemic and recent advances in diabetes patient survival, it is no surprise that older age is now recognized as an increasingly crucial diabetes risk factor.[15-18] aging raises the risk of diabetes by impairing insulin secretion and increasing insulin resistance through obesity and sarcopenia. moreover, older adults have higher rates of insufficient physical activity, increasing sedentary time, and decreasing physical activity.[19] at present, older adults (65 years and older) have the highest t2dm prevalence of any age group. diabetes cases in this age group globally are expected to increase from 122.8 million in 2017 to 253.4 million by 2045.[16] t2dm family history has been recognized as an essential non-modifiable diabetic risk factor, providing a conveniently assessed marker of the fundamental diabetic genetic component.[7] diabetic family history is a predictor of type 2 diabetes in both males and females.[20,21] notably, some research found a high correlation between diabetic family background and t2dm risk. in addition, studies from a large swedish cohort of 60-year-old men and women demonstrate that a family history of diabetes in combination with being overweight appears to be highly risky in men, raising the chance of acquiring t2dm synergistically.[21] methodology the data were collected from 218 patients that recorded their information in the hospital of diabetes in sulaymaniyah city of iraq, which included sociodemographic variables such as age, gender, weight, and clinical characteristics such as diabetes family histories. similarly, some information obtains on the patients’ smoking status and alcohol consumption. the logistic regression model was used to identify risk variables that affect type 2 diabetes. besides, testing the odds ratios, likelihood ratio, and 95% confidence intervals estimate for each factor using jmp pro, a statistical tool, version 16, was used to analyze the data. hba1c diabetes mellitus is a major epidemic affecting both developing and developed countries. the american diabetes association suggests using hba1c instead of fasting blood glucose to diagnose diabetes. hba1c is a long-term glycemic management measure that can reflect 2–3 months of glycemic history. for healthy people, hba1c levels should be <6.5%, and studies recognized that diabetic problems are controllable if the hba1c level is below 6.5%, and above 6.5% will be out of control.[22] mahmood, et al.: regression model to recognize diabetes risk factors 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 7-11 the relationship between plasma glucose and hba1c can be shown by the following formula to make sure whether the diabetes case is under control or not. then assign value 1 to diabetic cases that are controllable (hba1c < 6.5%) and value 0 to diabetic cases that are uncontrollable (hba1c ≥ 6.5%). in other words, the dependent variable (hba1c) has 1 and 0 values, that is, why a binary logistic regression model is used. +46.7 cos hba1c= 28.7 plasma glu (1) thus, the variables of the model represent as below: results and data analysis the report of table 2 provides the results of each iteration and criteria used to determine whether or not the model has converged. throughout this case, the model is a convergent gradient that was iterated 5 times. table 3 illustrates to choose the best model. according to the results shown in the table, since p < 0.001, the model is significant of the chi-squared test. in other words, the full model is better than the reduced one (model with only intercept parameter) and indicates that at least one of the parameters affects the response variable. on the other hand, a minimum akaike’s information criterion (aic) is used to select between two models in that way, the model for which aic is smallest represents the “best” with this formula. +  = − + +  − +  1 2 log 2 2 1 k aic likelihood k k n k (2) where k is equal to the number of parameters in the model and n is a sample size thus, aic=238.7672 for full model aic=256.5974 for null model therefore, the full model is the best one. furthermore, table 3 shows the r-square, which is the ratio of the negative log-likelihood values for the difference and reduced models. its ranges from 0 for no improvement to 1 for a perfect fit. a nominal model rarely has a high r-square, and it has an r-square of 1 only when all the probabilities of the events that occur are 1. in our case, r-square is 11.92%. it is now time to determine which factors affect type 2 diabetes and which do not. according to the findings in table 4, age, alcohol, and family are statistically insignificant. in other words, these three variables have no significant impact on diabetes. then, we can try rerunning the regression model without any of those variables to see how that affects prediction accuracy. the regression equation is as follows: log (odds of a control case)=−4.3217463 +1.163 (gender) + 1.053 (smoking) + 0.066 (weight) as with any regression, the positive coefficients indicate a positive relationship with the dependent variable. 1.163 is the increment to log odds of a worse outcome for females; the odds ratio e1.163=3.20 indicates that females are 3.20 times more likely to achieve a high level of diabetes (out of control) than males. for the smoking case, the odds ratio e1.053=2.87 indicates that a person with smoking is 2.886 times more likely to achieve a high level of diabetes (out of control) than one with no smoking. furthermore, with increasing 1 kg of weight body, the diabetes degree increases by e0.066=1.068. table 5 shows 95% confidence intervals for parameters (beta) and odd ratios, and we can note that confidence intervals of age, alcohol consumption, and diabetes family history have 0 between the lower and upper levels. it indicates that the coefficient of these variables might become 0, which is why age, alcohol, and family are not significant, as we explained before. when it comes to odds ratio, it is clear that log (odds ratio)=beta, and odds ratio=exp(beta) thus, regarding results in table 5, the 95% confidence interval of the odds ratio of gender, smoking, and weight (1.582827, 6.622663), (1.370512, 6.347617), and (1.030501, table 1: the variables of the model variables types of variables codes hba1c outcome (response) hba1c≥6.5%=0, hba1c<6.5%=1 gender independent male=0, female=1 age independent continuous alcohol consumption independent alcoholic=1, no alcoholic=0 smoking status independent smoker=1, no smoker=0 diabetes family histories independent yes=1, no=0 body weight independent continuous table 2: iterations of convergent gradient the model iteration objective relative gradient norm gradient 0 151.10608536 8.2627136759 4632.4497299 1 113.96878959 1.8226836288 638.86115249 2 112.15745738 0.2820389174 84.319608151 3 112.11696969 0.0077978033 2.2874541839 4 112.11693927 6.1496927e-6 0.0017917235 5 112.11693927 3.859414e-12 1.1109202e-9 table 3: whole the model model log-likelihood df chi square prob.> chi-sq. difference 15.17251 6 30.34501 <.0001* full 112.11694 reduced 127.28945 r-square (u) 0.1192 aic 238.767 observations 218 mahmood, et al.: regression model to recognize diabetes risk factors 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 7-11 1.109795), respectively, indicates that these parameters are significant because their ci does not contain 1, but the others (age, alcohol, and family) are not significant because their ci contains 1 and log (1)=0 table 6 is about wald test results, which is the same as the likelihood ratio test of table 4. here, we can note that the p-values of the variables (gender, smoking, and weight) are <0.05, and it tells us that these variables are significant, while the other variables are not significant because their p-values are more than 0.05. variance is a measure to show how much the data are spreading around the mean point, and it is equal to the square of the standard deviation. table 7 shows variance and covariance matrix for estimates of parameters, and the outcomes in the table explain that the off-diagonal elements contain the covariances of each pair of variables, and the diagonal ones of the covariance matrix represent the variances of each variable. according to the results, the covariance between each pair of (gender and alcohol), (gender and smoking), and (gender, body weight) is approximately 0.0539, 0.0312, and 0.0011, respectively, which indicates that the relationships are positive between these pairs of variables. in contrast, the covariance between gender and age is −0.0008 and between gender and family is −0.0071. the negative values indicate that there is some negative relationship between these variables. likewise, the relationships between pair variables of (age, family) and (age, body weight) are positive. on the other hand, the covariance estimates of the parameters (age, alcohol), (age, smoking), (alcohol, smoking), (alcohol, family), (alcohol, body weight), (smoking, family), (smoking and body weight), and (family, body weight) show some negative relationships between these pair of variables. conclusions the objective in this study is to illustrate the factors that affecting on the type 2 diabetes in sulaymaniyah city of iraq, according of the results, the factors of gender, smoking status, and weight body have high effecting on diabetes, in which table 4: parameter estimates term estimate (beta) std. error chi-square prob.>chi-sq. exp (beta) intercept −4.3217463 1.5420851 7.85 0.0051* −11.7478 gender 1.16270774 0.3636579 10.22 0.0014* 3.1601 age −0.0076878 0.0131161 0.34 0.5578 −.0021 alcohol 0.37083796 0.7383133 0.25 0.6155 1.0080 smoking 1.05325849 0.38882 7.34 0.0068* 2.8631 family −0.1939148 0.3332303 0.34 0.5606 −0.5271 weight 0.06587092 0.0188393 12.23 0.0005* 0.1791 table 5: 95% confidence intervals for parameters and odds ratios 95% confidence intervals parameters for parameters (beta’s) for odds ratios lower 95% upper 95% odds ratio lower 95% upper 95% gender 0.45921259 1.89049762 3.198583 1.582827 6.622663 age −0.0336268 0.01805573 0.992342 0.966932 1.01822 alcohol −0.9918519 1.98504948 1.448948 0.370889 7.279408 smoking 0.31518408 1.84807951 2.866978 1.370512 6.347617 family −0.8550631 0.45598815 0.823728 0.425256 1.577732 weight 0.030045 0.1041753 1.068089 1.030501 1.109795 table 6: effect of wald test source no. of parameters df wald chi-square prob.>chi-sq. gender 1 1 10.2224534 0.0014* age 1 1 0.34355643 0.5578 alcohol 1 1 0.25228239 0.6155 smoking 1 1 7.33791774 0.0068* family 1 1 0.33863607 0.5606 weight 1 1 12.2253152 0.0005* table 7: covariance of estimates of parameters parameters gender age alcohol smoking family weight gender 0.1322 −0.0008 0.0539 0.0312 −0.0071 0.0011 age −0.0008 0.0002 −0.0010 −0.0004 0.0003 0.0000 alcohol 0.0540 −0.0010 0.5451 −0.0619 −0.0219 −0.0001 smoking 0.0312 −0.0004 −0.0619 0.1512 −0.0075 −0.0001 family −0.0071 0.0003 −0.0219 −0.0075 0.1110 −0.0004 weight 0.0011 0.0001 −0.0009 −0.0001 −0.0004 0.0002 mahmood, et al.: regression model to recognize diabetes risk factors 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 7-11 for gender, the females have more likely to have high level of diabetes than the males. this may be due to the fact that in our society, women are more likely to do housework, but men are more likely to work outside the home, so men are more active compared to women. furthermore, for smoking case, the person with smoking is almost 3 times more likely to have diabetes than those with no smoking. furthermore, with each kilogram of body weight gained, the risk of developing diabetes increases. on the other hand, the variables of age, alcohol consumption, and diabetes family history do not have any effect on diabetes. recommendation 1it is necessary to perform additional type 2 diabetes research in all other cities in the kurdistan region of iraq to identify the fundamental risk factors that significantly influence the development of type 2 diabetes in that area. 2it is suggested that more encouragement be given to people to engage in some regular physical activities and be warned about consuming unhealthy foods that might lead to weight gain and eventually diabetes. 3men in the kurdish community have always been expected to fulfill greater responsibilities outside the home, while women have traditionally been supposed to stay home more. as a result, to avoid developing diabetes, women must engage in more physical activity and exercise. 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and women. the canadian journal of diabetes, vol. 37, no. 2, pp. 65-71, 2013. 21. c. wikner, b. gigante, m. l. hellénius, u. de faire and k. leander. the risk of type 2 diabetes in men is synergistically affected by parental history of diabetes and overweight. plos one, vol. 8, no. 4, p. e61763, 2013. 22. s. i. sherwani, h. a. khan, a. ekhzaimy, a. masood and m. k. sakharkar. significance of hba1c test in diagnosis and prognosis of diabetic patients. biomarker insights, vol. 11, pp. 95-104, 2016. tx_1~abs:at/add:tx_2~abs:at 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 review article an overview on software-defined networking and network functions virtualization security orchestration in cloud network environment israa t. aziz1*, ihsan h. abdulqadder2 1computer center, university of mosul, mosul, iraq, 2department of computer science, cihan university-erbil, kurdistan region, iraq abstract cloud networks are being used in most industries and applications in the current era. software-defined networking (sdn) has come up as an alternative tool and mechanism to follow and implement in a cloud networking environment in place of the traditional networking approaches. this paper includes the security aspects of computer networking concerning the cloud networking environment and sdn. the security risks and vulnerabilities have been listed and described in this work, and the measures that may be adapted to detect, prevent, and control the same. the use of figures, diagrams, and codes has been done as applicable. keywords: software-defined network, network functions virtualization, cloud, cryptography, distributed denial of service attacks 1. introduction cloud computing is a type of internet-based computing in which the devices communicate and are connected over a network which is usually the internet. communications and transactions take place over the network in the case of a cloud environment, leading to the introduction of many security risks and threats. software-defined networking (sdn) is a technique that is followed under computer networking that allows the network masters and administrators to make use of automated tools and applications to dynamically control and manage the networks as per the changing business requirements. orchestration of sdn is the programming of such required automated behaviors to support the business applications and services.[1] network functions virtualization (nfv), on the other hand, is a data center and network that has been specifically built and set up to manage the virtual networks and services over the cloud. nfv orchestration is the mechanism that is required to set up cloud-based applications and services over the cloud. sdn has come up as an alternative tool and mechanism to follow and implement in a cloud networking environment in traditional networking approaches. the architecture that is implemented in sdn is illustrated in figure 1. where, sdn divided into three major planes by open network foundation (onf), the architecture in sdn as follows: data plane: in this plane, forwarding devices such as switches, routers are interconnected. connection in this plane can be established by either wireless connection nor wired connection. this plane is responsible for data forwarding accordance to flow characteristics. control plane: this plane acts as brain of the network since it has the global view of entire network. this plane is responsible to program data plane elements. forwarding devices: these are hardwareor software-based data plane device. major function of these devices is to forward the data packets as per forwarding rules. here, on incoming packets, actions such as forward to specify port, drop, and forward to control plane are taken. southbound interface: this is referred as instruction set of forwarding devices that are defined by southbound application programming interfaces (apis). the interaction between control plane elements and forwarding devices is carried south through southbound interface. northbound interface: this api is offered by network operating system to application developers. this interface acts as common interface for developing new applications. management plane: this plane is often called as application plane in which set of applications that leverage the functions offered by the north interface. corresponding author: israa t. aziz, computer center, university of mosul, mosul, iraq. e-mail: israa_aziz@uomosul.edu.iq received: may 17, 2021 accepted: june 3, 2021 published: june 20, 2021 doi: 10.24086/cuesj.v5n1y2021.20-27 copyright © 2021 israa t. aziz, ihsan h. abdulqadder. this is an openaccess article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) aziz and abdulqadder: sdn/nfv security orchestration in cloud network 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 good design is the base of every software application and service. therefore, it should be made sure that the software and the networks are constructed with the standard and necessary design patterns and principles. figure 1 displays the various sets of interfaces and isolations present between the different layers of sdn that is either under evaluation or under development. for instance, the service abstraction layer (sal) in the sdn architecture has been placed to isolate the apis southbound from the controller. it would aid in the integration of the same controller with multiple apis or protocols that are southbound. according to the software design aspects of sdn, the controller is the component that should make sure that the design is as per the flow insertion constraints and the entire network invariants are fulfilled before the insertion of a new flow rule in the process. once the assurance is provided, a separate monitoring module shall be queried to retrieve the invariants.[2] figure 2 depicts the characteristics that are associated with sdn. sdn is included with some security threats in data plane as well as in control plane. vulnerable controller in sdn, most of the network functions are realized in control plane by sdn controller. however, control plane is involved with some limitation such as queuing mechanism, security, and so on. sdn controller is responsible for network information collection, network configuration, and routing collection. if sdn controller is compromised or affected by attackers, this will be a massive attack held on sdn and network performance is degraded severely. in controller, well known denial-of-service (dos), and distributed dos (ddos), target point-of-sale attack are frequently launched. risks caused by other interfaces not only controller, in sdn, all other interfaces such as switches and gateways also affected by many attacks. sdn interfaces are programmable and open natured which leads probes attackers toward it. attacks also held on application layer through northbound interfaces. moreover, each layer in the network is affected by different attacks. some attacking points in sdn are listed as follows, • sdn switch – it is a separate device that is operated over openflow (of) protocol. sdn switch is also involved with flow tables which are limited in size • link between sdn switches – this type of attack is launched in sdn data plane in which link between data plane elements are targeted by attackers. in sdn, almost all data packets are transmitted between switches in the form of plaintext. this plaintext transmission in the data plane encourages the link spoofing attacks. link between switches, controller, and switches-controller is also affected by man-in-the middle attack • link between sdn switches and sdn controller – this attack also launched as link spoofing attack in which link between controller and all switches is targeted by attacker. remaining that all flow rules are installed on switches by controller through these links. thus, this type of attack has high impact on the network • link between controllers –to support large number users, sometimes, the network is designed with multiple controllers. however, when multiple controllers are involved then attacking ratio also increased. here, communication links between controllers also affected by attackers figure 1: software-defined networking architecture figure 2: software-defined networking characteristics aziz and abdulqadder: sdn/nfv security orchestration in cloud network 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 • application software – usually, application software is built with controller on physical device. when this application software is affected by attacker, then it may inject malicious code to controller which destroys the entire network performance. thus, application software is most convenient attack point for attackers. in application layer, unauthorized access also has major impact in the security point-of-view. the primary characteristic that is associated with sdn is the logically centralized control. however, the same shall have the physically distributed controller component. a controller is a component that managed the complete network view of the associated infrastructure and also processed the entries based on the protocols as defined by the services and operations running above the same.[3] another characteristic associated with sdn is the open programmable interfaces that have the major motivation behind them to permit the networking protocols implemented in the controller to execute and evolve independently. furthermore, it makes sure that the forwarding devices are as simplified as possible. switch management protocol, third-party network services, virtualized logical networks, and centralized monitoring units are also significant characteristics associated with the sdns.[4] the rest of this paper is organized as follow, section 2 presents the attack venders including the attacks in different layers. section 3 provides the security threats and vulnerabilities in sdn. section 4 illustrates different types of security measures while section 5 concludes this paper. sdn attack vendors sdn is a networking method in which virtualization is supported by the separation of the control plane from the forwarding plane. there are usually three layers in most of the sdn architectures, which comprise a lower layer in which network devices are placed, followed by the second layer, the middle layer in which controllers are kept. the topmost layer is the higher layer which consists of the applications and services to be configured as per sdn. the sdn is still in its early phases; however, the expansion of the same is quickly increasing, leading to the emergence and development of various security attacks by malicious entities. several security threats and attacks have started to come up in association with sdn and the layers of the same. there are many attack vendors associated with these attacks, and the same has been explained as per the layers present in sdn architecture. attacks at data plane layer there are possibilities in sdn architecture that the attackers attack the network elements with the aid of the network itself. the basic technique that the attackers use in such cases is to gain unauthorized access to the network, which may be physical or virtual, and then give shape to the attacks. these attacks may be availability attacks such as denial of service attacks which have been explained in detail in the next section. in sdn architecture, the controller uses several protocols to communicate and interact with other network elements. some of the examples of such apis and protocols include of, embedded event manager, open management infrastructure, locator/id separation protocol, and various others. these protocols comprise a secure mechanism within themselves to ascertain that the communications that take place over the network are safe and secure. most of these protocols are relatively new, and there are high chances that there are security loopholes that may exist during the same implementation. these security loopholes allow the attackers to take advantage of the same and attempt to incorporate newer forms of flows in the flow table of the devices. there are spoofing attacks that are attempted to ensure that the local traffic is allowed over the network. there are malicious entities put up on the network for sniffing the same and performing unauthorized monitoring to gain relevant information through the act. such attacks are termed as man-in-the-middle attacks in which an attacker sits on the network to gain information in an unauthorized manner. the attackers perform eavesdropping on the network flows to understand the flow of traffic over the networks. data centers are the prime spot of application for most sdns. these data centers make use of several protocols in their functioning and operations, such as stateless transport tunneling, layer 2 multipath, shortest path bridging, and various others. all of these protocols comprise data packets which consist of data and information within them. the level of authentication in these protocols may not adhere to the required level to ensure secure communication. there are also instances wherein required encryption is not implemented in these packets. therefore, these protocols may emerge as the attack vendors as they may provide the attackers with the opportunity to gain access to the network and information flowing over the network. there are several data integrity and availability attacks that may be executed due to these security loopholes. attacks at controller layer after the data plane layer, the controller layer is present, and the middle layer is the target layer for the execution of several security attacks. there are several reasons behind the choice of the target as the controller layer by the attackers. if the attacker succeeds in spoofing the flows from the specific controllers, then the control of the traffic and the flows would come into the hands of the attackers. it would allow them to bypass the policies that may have been designed to enhance the security of the networks. the attacker may execute various attacks to cause damage to the controller, and one such attack is the denial of service attack. there may be unauthorized consumption of resources that may be executed in addition to the deterioration of the frequency of packet transmission across the network. such attacks would impact the network services and the flow of traffic over the network as well. the controllers deployed in sdn architecture may be done in various operating systems such as unix or linux. the vulnerabilities associated with that of the operating system become the controller’s vulnerabilities as well in such cases. there are issues with the deployment of the controllers in a production environment without strong passwords. such vulnerabilities lead to weakening the network’s security and allowing the attackers to execute several security attacks. aziz and abdulqadder: sdn/nfv security orchestration in cloud network 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 there are also instances in which the attackers create their controllers as per their set of policies and protocols and trick the network elements through the same. the network elements trust the fake controller as the original and various attacks are executed. there are malicious entries made into the flow tables which go undetected and unnoticed by the network engineers due to the nature of the controller. such attacks are the worst forms of attacks as the attackers gain complete control over the network and gain an opportunity to turn the same in whichever direction they like. attacks at sdn layer several attacks may be executed at the highest layer as the attacking vendor for the same. attackers may attempt to impact the northbound protocol and its security. many northbound apis may be used in sdn architecture such as xml, python, and java. if the attacker succeeds in breaking through the security of a northbound api, the attacker would be able to gain control over the entire network with the aid of the controller. there are also chances wherein the attackers may create their own set of policies and launch them in the network to gain control over the entire environment of sdn. there are also scenarios in which the default passwords of the apis are not changed and used. in such cases, it becomes easier for the attackers to break into the network’s security and execute various security attacks. it also allows the attackers to query the configuration associated with the sdn environment and alters the same to install their configuration in the network. security threats and vulnerabilities in sdn information breaches cloud networks deal with huge clusters of information that are passed from one source to the other. the exchange of information may involve a public cloud network, private cloud network, or likewise. the cloud network acts as a major threat agent, leading to the breaching of information by the attackers. these networks and the various access points present in the networks act as the source of entry for the attackers. they succeed in obtaining unauthorized access into the cloud networking environment.[5] the same leads to the violation of the viable properties of the information contained in the network, such as confidentiality, privacy, and the same availability.[6] information leakage and loss sdns also suffer from the loss or leakage of the data and information observed as one of the frequently occurring security attacks. when the information is passed from a source to the destination in the case of sdn, attackers get hold of the information bits and data sets which are then misused to cause damage to one or more parties associated with the same. the major threat agent, in this case, is the network and the networking protocols that are implemented. there are instances in which the information is prevented from reaching its designated destination, leading to significant damage.[7] malware attacks malware attacks are the most common form of security attacks in a computing environment, and the case is the same with sdns. the list of malware that can hurt the sdns is a long one that comprises various types such as viruses, trojan horses, worms, logic bombs, and many more. these malwares are also reproducible in most cases which makes it difficult to get rid of them. therefore, it is required to adapt the measures to prevent malware attacks to ensure the safety of the network and the information contained within the same.[8] denial of service and distributed denial of service attacks sdns introduce some advantages by decoupling the control plane from that of the data plane. however, the relationship between sdn and ddos attacks is contradictory.[9] the capabilities and properties associated with sdn allow the detection of ddos convenient and easy to execute. simultaneously, the sdn architecture in which the control place of separated from the data plane gives rise to the newer forms of ddos attacks. such a property makes sdn a target of the denial of service and distributed denial of service attacks.[10] the following is a basic ddos script that the attackers may use in an sdn environment. print “attacking mode is on! ! ! ! ! ! ! ! !” + sys.argv[1] + “! ! ! ! ! ! !” print “injecting the code “ + sys.argv[2]; def attack(): #pid = os.fork() s1 = socket.socket(socket.af_inet, socket.sock_stream) s1.connect((sys.argv[1], 80)) print “<<<<get>>>>” + sys.argv[2] + “ http/1.1” s1.send(“get/” + sys.argv[2] + “ http/1.1\r\n\r\n”) s1.send(“host: “ + sys.argv[1] + “\r\n\r\n\r\n”); s1.close() for i1 in range(1, 100000): attack() security measures ddos detection and prevention the most effective method which may be used to detect the ddos attacks in sdns is the fuzzy synthetic evaluation method. it is a method that has been developed based on fuzzy mathematics. with the aid of this method, the transformation is made from the qualitative method of evaluation to the quantitative method of evaluation. it is applicable for detecting ddos attacks as it can evaluate the uncertainties with much ease and correctness.[11] the decision-making, in this case, is based on the three factors which areas listed below. • factor set f = {f1, f2, f3……. fn} • judgment set j = {j1, j2, j3……jn} • fuzzy relationship matrix r there are also various flow checkers and flow detection mechanisms that may be used to detect ddos attacks in sdns. aziz and abdulqadder: sdn/nfv security orchestration in cloud network 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 reduction of the attack surface the control and reduction of the attack surface would prevent the security risks and attacks in the sdns. these attack surfaces are the major threat agents that allow malicious entities to enter the network and impact the security and privacy of the information.[12] the network and data masters must ensure that the information exchanges that take place over sdn make use of smaller chunks of the data instead of larger data sets and pieces. furthermore, the adaption of a secure apis in the sdn architecture would lead to the prevention of the attacks in a private networking environment.[13] reduction of the attack window along with the smaller attack surface, the network administrators must also make sure that the attack window is reduced and brought out to the minimum. the reduction in the size of the attack window would significantly reflect the depreciation of the security threats and attacks. for example, there should be the use of rotating credentials in an sdn environment adapted by the network administrators.[14] use of firewalls it is necessary to maintain the initial steps of security in sdns so that the attackers do not succeed in entering the network at all. some of the firewall rules may be used in sdn, as figure 3 described and listed below. • at a particular switch, block the traffic from a specific host such as 11.1.1.1. 00-00-02-00-00-00-00-00 block srcip=11.1.1.1 • at a particular switch, block the traffic that is coming from a specific host such as 11.1.1.2, if the particular packet say x tos has been marked with 23 and it is required to be sent to the destination for 11.1.3.1 00-00-03-00-00-00-00-00 block srcip=11.1.1.2 dstip=11.1.3.1 tos = 23 • at a particular switch, redirect traffic which is destined for 11.1.2.1 and, instead, send it to 11.1.2.2 00-00-01-00-00-00-00-00 redirect dstip=11.1.2.1 to 11.1.2.2 • at a particular switch, mark the tos field present in all the packets which are sent by 11.1.1.1 with a value of 42 00-00-04-00-00-00-00-00 mark srcip=11.1.1.1 tos 42 defense in-depth this is the security measure that shall be used in sdn to prevent security risks and attacks. the interactions that are executed in a network are used in this security measure to prevent security attacks.[15] for example, a certain part or component of the sdn may get hampered by the security attack identified by another network and security part or component present in the architecture. these interactions between various components will ensure that the security of the overall network is maintained at all times.[16] cryptography and encryption cryptography is a technique that comes under the advanced tools and techniques for preventing security attacks and enhancing information security. encryption of the information is necessary in the present times with the introduction of so many security risks and attacks.[17] cryptography is a phenomenon that makes use of public, private, or shared keys for the encryption of the information at the sender’s end, which is then transferred in the same form and is decrypted with the help of the key at the end of the receiver.[18] various encryption algorithms have been developed and may be used in the case of sdn security as well. one such algorithm is the rivest, shamir, and adleman (rsa) encryption algorithm. for instance, the text required to be sent to the destination includes the bits “sdn.” the encrypted form of the text is as specified below. o i q s i wa p h u 0 a 3 v 9 3 x x e u n + u t n 2 h x b 8 w k n l 1 1zzzj0xqaxgqkao63u4qc8dnb2fow0ls3uhrgo1vil0y wcr8mplzlxq94dsro1ek0ym0n0o5lqfumzasj914ghi i s k u k 3 m s / z i + b s 1 z e q f g a j a b + y u p o t h t q figure 3: firewall execution in software-defined networking aziz and abdulqadder: sdn/nfv security orchestration in cloud network 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 31rqhkr8zqh9pgetblpn09rbvvzppnxmi2g8g2dvug b l w s x 0 a 1 q j o v d z w 8 t 0 0 v 7 9 q h o c / o q q a 3 t r m m 6 m j n 5 w m f i s d u x n g t k s k u 7 x l v m g m s f y e c z d 6 / 9 l 7 k 2 b p r 3 l l n u z x h l n q m r l x t n l 8 g 4 q i g w m yqbe8i4pmwptqimb1kqakee9dbpxnblenteuwxw== there is a private and public key pair used to encrypt and decrypt the information while the same is exchanged between the source and the destination over the software-defined network. various other encryption algorithms can be used, such as des, triple des, and many others. one of the widely used cryptographic functions is the sha algorithm which stands for the single hash algorithm. sha-256 is one of the forms of sha that generates a unique 256bit hash for a text or data file, which is like a signature for the same.[19] for instance, the sha-256 for the text “software-defined network” is as specified as: f8551cca082c82020b1f30b07a36a68c154272f9af16b 724859ccd1c96f64ae5 access management access to the information and data transferred and exchanged over the software-defined network is often played with by the attackers. the access points are used to gain unauthorized entry to the network. therefore, it must manage access to the sdn using several administrative and technical security controls. [20] the administrative controls shall include enhanced security policies and protocols to control the access. in contrast, the technical controls shall include the use of technically advanced mechanisms such as multistep authentication, single sign-on, and likewise for the control of access.[21] advanced tools the number and frequency of data and network security attacks are on an all-time rise, and the same has led to the development of advanced tools and techniques for the detection and prevention of the same.[22] therefore, it is required for the network administrators and masters to be constantly updated with the technical changes and innovations and adapt the advanced tools available in the market to prevent security risks and attacks. various tools are now available for the management of the apis, the execution of the security controls, and the management of the security attacks associated with the sdn traffic.[23,24] network monitoring monitoring of the network at all times is also extremely important in the case of software-defined networks. such a practice will ensure that there is no illegal activity across the network and the same is identified as soon as it is attempted.[25] the networking team shall carry out administrative practices such as networking audits, reviews, inspections, and nonstop monitoring of the network. furthermore, there shall be automated intrusion detection and prevention tools that shall be implemented to ensure that the intruders do not get entry into the network.[26] network monitoring data recovery and backup strategies electronic vaulting electronic vaulting is the procedure that incorporates the legitimate official of an electronic archive by keeping up the definitive duplicate of the same and the greater part of its associated and critical exchange reports in a protected area. the same is kept at the area until the lifecycle of the report is finished and accomplished. it is done keeping in mind the end goal to ensure that the electronically marked archive remains legitimately enforceable and allowable and accordingly frames an obvious review trail. possession and consistency are resolved with this review trail as it were.[27] electronic vault does not rotate around simply the capacity of the electronic archive additionally deals with the authorized access, support, and last demolition of the record. aside from the legitimate and administrative edge, it likewise gives a protected move down to the available archive and opens electronically.[28] certain components must be available in an electronic vault with a specific end goal to make it fit for its intended purpose. electronic vault must make and give a great degree certain condition to the reports keeping in mind the end goal to guarantee the wellbeing, security, and protection of the information introduced in the archives. exchange of records starting with one area then onto the next or starting with one vault then onto the next might likewise affirm the security highlights. some archives do not permit the nonseparated duplicates of the same to live in some other area separated from the electronic vault. security of such records should be of most extreme confirmation as the loss of a unique copy will prompt losing the complete information related to the report. electronic vaults must have an incredible system to keep up and deal with the duplicates of the electronically marked archives. the duplicates must have a personality or a stamping plan that separates them from the first record. there must likewise be instruments to track the entrance to the first e-archive or the duplicates of the same.[29] point-in-time copies a point-in-time copy of an arrangement of information alludes to the copy of the information as it showed up at a specific period. it is a usable copy of the original information and has coherently happened at a specific timeframe. these copies might be perused just in nature or may give the compose capacities too. various advantages accompany this alternative to move down the information[30] safely. it is frequently observed that while creating the move down on the cloud, the server is under a gigantic load. this regularly prompts a reinforcement window, which is hard to manage to keep the financial plan and timetable constraints in thought. the same is handled through the reinforcement alternative using point-in-time copies. the issue related to the open records additionally develops during the reinforcement. clients continue updating the open documents till the last minute, and the go down of the same is not made. while creating the pointin-time copies, compose capacity of the record framework is not permitted, and consequently, the replication of the original archive or document is made. point-in-time copies do not request specialists to play out the undertaking or recover the past renditions of the record, making it simple and a savvy aziz and abdulqadder: sdn/nfv security orchestration in cloud network 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 arrangement. it is conceivable with the point-in-time copies to make the backup at constant intervals, generally impractical with the conventional techniques.[31] data deduplication information space available on the cloud is decreased with the assistance of data deduplication alongside giving the ability of information reinforcement. it ensures that the repetition related to the information is totally evacuated, and just a single duplicate or occurrence of the same is put away on the cloud. at whatever point a duplicate of information or repetitive information is discovered, it is indicated the first information. because of its productivity to expel the excess information is also named as keen pressure or single occurrence stockpiling. information deduplication joins the numerical calculations utilized as a part of the conventional pressure systems alongside delta differencing for looking at the old and new information and sparing just the bits of information that have changed. information deduplication makes the route toward reproducing fortification information practical by reducing the exchange speed and cost anticipated that would make and keep up duplicate informational indexes over frameworks. at a fundamental level, deduplication-enabled replication resembles deduplication-engaged information stores. when two photos of a support information store are made, all that is required to keep the generation or target undefined to the source are the infrequent duplicating and development of the new information divides, the general data deduplication methodology is shown in figure 4. backup options • off-site cloud backup – this is the backup option that may be used as a security measure in sdn security attacks to maintain know how of the entire data sets available on the cloud. replication of the same ifs put away on the offsite location would be a wonderful option to ensure that the data can be recovered in the occurrence of an attack or a disaster • local backup options – with the presence of more than 1 storage option for the data and information present and exchanged in an sdn environment, it would be assured that the data are safe and protected at all times. the presence of a local backup option would be an excellent way to achieve the same • scalable and easy to use – scalable backing up of the data is an easy and extremely cost-effective solution to ensure that the backing up of the data is convenient to use and execute as well • dr planning – planning is always considered an effective measure for any of the activities, and the case is the same with disaster recovery. conclusion the term sdn is a technique followed under computer networking that allows the network masters and administrators to use automated tools and applications to dynamically control and manage the networks as per the changing business requirements. orchestration of sdn is the programming of such required automated behaviors to support the business applications and services. there are many security threats and vulnerabilities associated with the cloud networking environment in the case of a softwaredefined network, such as information breaches, data leakages, ddos attacks, malware attacks, and many more. with so many security attacks, various security countermeasures are now available. they shall be implemented to ensure that the security risks and threats do not have any adverse impact and are prevented at all times. the most effective method which may be used to detect the ddos attacks in sdns is the fuzzy synthetic evaluation method. the control and reduction of the attack surface would prevent the security risks and attacks in the sdns. these attack surfaces are the major threat agents that allow malicious entities to enter the network and impact the security and privacy of the information. along with the smaller attack surface, the network administrators must also make sure that the attack window is reduced and brought out to the minimum. cryptography is a technique that comes under the advanced tools and techniques for preventing security attacks and enhancing information security. encryption of the information is a necessity in the present times with the introduction of so many security risks and attacks. the same has proved to be effective in case of security attacks associated with sdns. other security measures that may be implemented include defense-in-depth, access management, firewalls, antimalware, and many more. figure 4: data de-duplication methodology aziz and abdulqadder: sdn/nfv security orchestration in cloud network 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (1): 20-27 references 1. m. gharbaoui, b. martini, d. adami, s. giordano and p. castoldi. cloud and network orchestration in sdn data centers: design principles and performance evaluation. computer networks, vol. 108, pp. 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in software defined networking based on improved shiryaev roberts detection algorithm. journal of high speed networks, vol. 21, pp. 285-298, 2015. 31. p. mishra, d. puthal, m. tiwary and s. p. mohanty. software defined iot systems: properties, state of the art, and future research. ieee wireless communications, vol. 26, pp. 64-71, 2019. tx_1~abs:at/add:tx_2~abs:at 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 research article a comparison between brown’s and holt’s double exponential smoothing for forecasting applied generation electrical energies in kurdistan region ameera w. omer1*, hazhar t. a. blbas1, dler h. kadir1,2 1department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq, 2department of business administration, cihan university-erbil, iraq abstract the process of producing electricity from sources of energy is known as electricity production. electric also is not freely accessible in environment, thus it should be “manufactured” (i.e., converting another kinds of energy to electrical energy) by utilities with in electricity industry (transportation, distributing, and so on). moreover, the objective of this study is to compared of brown’s as well as holt’s double exponential smoothing (des) also build a best forecasting time series model among two smoothing model forecasting, as well as focuses on optimizing characteristics to use the golden section technique. this exponential smoothing approach has been one of the time series forecasting methods that would be used to forecast (generation electrical) with in kurdistan area. the issue that arises with this technique is determining the appropriate parameters to reduce predict inaccuracy. in addition, data used in this paper are (generation electrical) in kurdistan region for (132) months from 2010 to 2020. the study revealed that such data are trending modeled, indicating that a des approach from brown and holt can be used with the (stratigraphic and minitab) software. there are the same results but the result of analysis more depend on the r-program. the difference among the forecast findings acquired with optimum parameters as well as the assaying data was utilized to assess the feasibility of the forecast by completing normality and randomness tests. ultimately, the outcomes of parameterization show that the optimal value of α that in des brown is (0.22) as well as the optimal mape is 9.23616%, whereas in des holt the optimal is (0.95) as well as the optimal β is (0.05) through the optimal mape of 8.08586%. this mape of a des brown technique is greater than the mape of a des holt approach. feasibility experiments revealed that both approaches are capable of predicting. depending on the value of mape as well as evaluation process, des holt’s was recognized as the main prediction model. keywords: generation electrical, energies, double exponential smoothing parameter optimization, electricity production introduction the method of constructing electric power from various forms of energy production is known as electricity production. the kurdistan region local generation are hydro power station, gas power plant (pp), steam pp, heavy feul pp, deisel pp, erbil combined cycle pp, and suly combined cycle pp and thermal. to estimate the forecasting applied generation electrical energies in kurdistan region, a (brown’s and holt’s) double exponential smoothing (des) approaches would be created as well as contrasted to see that methodology is superior for forecasting generation electrical energies in kurdistan. furthermore, there was usually a timing lag among awareness of the imminent event or necessity also its manifestation. the fundamental reason for planning as well as predicting is to account for this advance time. there was not need for the planning when the lead time is nil or pretty limited. if indeed a lead time was considerable as well as the event’s outcome was dependent on definable variables, planning could play a major role. prediction is required in these contexts to assess while the event would happen or the need may arise, because as appropriate steps could be taken. furthermore, predicting the future is a planning method created to assist management in meeting the uncertainty of a future relied on previous data as well as predictive modeling. prediction is the science and art of forecasting the future developments by utilizing past data as well as projected this into the future with the use of a systematic method model. corresponding author: ameera w. omer, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq. e-mail: amira.omer@su.edu.krd received: october 22, 2021 accepted: november 13, 2021 published: november 30, 2021 doi: 10.24086/cuesj.v5n2y2021.pp56-63 copyright © 2021 ameera w. omer, hazhar t. a. blbas, dler h. kadir. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) omer, et al.: brown’s and holt’s double exponential smoothing 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 forecasting demand requires the application of a certain approach. many predicting approaches work on the same principle of using previous data to forecast or project future data. in other ways, predicting can be seen as a critical and necessary help to management that serves as the foundation of efficient operations. several organizations have failed in the past as a result of an ineffective forecasting approach on which the company’s whole strategy was based. forecasting effectively assists in the decrease of unneeded inventories, the improvement of product availability, and also the degree of customers’ satisfaction. the major driving factor for profit generation in today’s business world is satisfying customer’s requirements at the correct time and even in the appropriate amount. as a result, predicting with as much precision as feasible is critical to ensuring product availability at the lowest possible cost. the forecast’s reliability would be evaluated using mean absolute percentage error (mape) tool. mape displays a forecast’s reliability in the percentage form. since it was simple to read, (due to its percentage form).[1-3] a mape reliability estimate is typically more commonly employed. the smaller mape score indicates that now the prediction model performs well. table 1 shows the range of a mape score. research method (des) simple exponential smoothness does not really work well while there is a trends in the information, that is inconvenient. for such cases, various techniques have been developed that under term “ des ” or “2nd exponential smoothing,” and that is the recursion applying of the an exponentially filtering twice, hence the term “ des.” brownos des technique is indeed a prediction approach that is utilized while the data have a trend line as well as being non-stationary. this really is comparable to the terminology used for quadruple exponential smoothing, that also refers to the recursion depth. the main concept underlying des would include a component that accounts for the potential of the series displaying some kind of the trend. that slope parameter was kept up to date by using exponential smoothing. when such data reveals a trend, this approach is employed. exponential smoothing through trend is similar to simple smoothing, however that both components should be updated throughout every levelperiodic as well as its trends. this level is the smoothed estimate based on the data values at the conclusion of the each period. a tendency is a smoothed estimate of average growth at the ending of each period.[1,3-6] a basic concept of these technique was illustrated in equation (1), ( )t 1 t tz    x 1 z+ = ∝ + − ∝ (1) where z t+1 ∶ prediction one period ahead. x t : actual data at period t z t : prediction at period t ∝: smoothing parameter (0 < ∝ <1) brown’s linear exponential smoothing exponential smoothing through a tendency works similarly to basic smoothing, with the exception that two components level as well as trend should be adjusted each period. a level was the smoothed estimation of a data’s values just at ending of each period. this tendency was the smoothed estimation of average growth at every period’s conclusion. brown’s straight exponential smoothing (brown’s des) was widely used only for data with a trend line. this is utilized because when data exhibits a trending as well as to generate a linear trend. the brown’s exponential smoothing methodology is therefore appropriate for modeling time series through trend however no seasonality. that a des brown technique comprises the single parameter ∝ with a value between 0 and 1. that parameter value was being utilized to reduce the observed value (real value) in the older era exponentially.[1,4,7] des utilizing brown’s technique through the m-periodahead forecast is provided by: ( )t m ˆˆz  a b m+ = + (2) this is just a simple linear regression model. where: z t+m : ahead forecast in (m) period (â.b̂) : unknown parameters to find the m-period-ahead forecast using following steps step (1). to find coefficients of linear trend as follows: 1 2 t t tâ 2(s ) (s )= − (3) where 1t(s ) is the single -smoothing measurement and 2 t(s ) is the double-smoothing measurement. then, 1 2 t t tb ( s s )1 ˆ ∝= − − ∝ (4) where (α) is the single exponential smoothing, therefore: 1 1 t t t 1s x (1 )s −=∝ + − ∝ (5) 2 1 2 t t t 1s s (1 )s −=∝ + − ∝ (6) note. the values of 1 21 1 1(s s x )= = in period (t = 1). holt’s linear exponential smoothing holt (1957) expanded simple exponential smoothing to permit for trends predictions in data. this approach employs a prediction equation as well as 2 smoothing equations (one for each level as well as one for each trend).[4,7-10] ˆ    t m t tz l m b+ = + forecast equation (7) 1 1  (1 )( )t t t tl z l b− −=∝ + − ∝ + level equation (8) ( )1 1(1   )t t t tb l l bβ β− −= − + − trend equation (9) where: l t : denotes an estimate of the level of the series at time t. b t : denotes an estimate of the trend (slope) of the series at time t. table 1: mape score significantly mape significantly <10% excellent forecasting ability 10–20% good forecasting ability 20–50% reasonable forecasting ability >50% bad forecasting ability omer, et al.: brown’s and holt’s double exponential smoothing 58 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 ∝: smoothing constant for the data (0<∝<1) β: smoothing constant for the trend estimate (0<β<1) m: the number of periods ahead to be forecast ˆ t mz + : double forecast value of period (t+m) note: initialization level l1=z1 and trend: 4 12 1 1 2 1 4 1 (z z )(z z ) b (b b ) (b b ) or −− = − − . parameter optimization to reduce mape, the golden section approach was used to optimize the parameters. mape was determined from brown as well as holt predictions as well as contrasted to actual results. the technique of golden section was used to optimize the parameters () on des brown, while a modified version of the method of golden section was used to optimize the parameters (∝ and β) on des holt.[6] accuracy measures for predicting while choosing between various alternatives, forecasting accuracy is critical. in this context, accuracy means to predicting errors that are the difference among the actual as well as predicted values for a particular time. the forecasting error determination mape is utilized in this investigation (mape). mape is a measurement that reflects the proportion of average absolute error that has happened.[1,8] this measure is given by 1 1 1 1 * 100 ˆ * 100 n n t t t t tt t e z z mape n z n z= = − = =∑ ∑ (10) where: z t : actual demand for time period t tẑ : forecast demand for time period t n: specified number of time period t t t:e z ẑ− . results in this part, we apply the previously discussed exponential smoothing methods to analyses time series for generating electrical energies utilizing brown’s and holt’s des for the for casting applicable generation electrical energies in kurdistan area for (132) months from 2010 to 2020. trend of the generation electrical energies data on generation electrical energies were received and then plotted onto the graph to identify the data patterns (look figure 1). according to the time series plot, a (generation electrical energies) data are now on the long-term secular decrease, though at the some point the period of enhance was not important at times. a rise in certain times could be considered non-seasonal since it did not occur at the same time each month. for generation electrical energies, this method creates different table 2 and charts. the data cover 132 time periods. estimated partial autocorrelations and partial auto correlation function (acf) for generation electrical energies estimated autocorrelations for generation electrical energies the measured autocorrelations among residuals at different lags are shown in this table 2. a lag k autocorrelation coefficient assesses a relationship among residuals for time t and time t-k. the 95.0% probability limit about 0 is also indicated. unless the probability limit at a given lag does not include the calculated coefficient, there really was the statistically important correlation at a certain lag at the 95.0% level of confidence. in these situation, six of the 24 autocorrelation coefficients were statistically important at the 95.0% confidence level, suggesting that now the residuals are not entirely random (white noise). we could illustrate the autocorrelation coefficients by choosing residual autocorrelation function from of the selection of graphical choices. by using acf, a connection of a time series data through the time series itself should be computed to detect data patterns. figure 2 shows an acf plot based on the data. estimated autocorrelations for generation electrical energies the measured partial autocorrelations among the residuals for several lags are shown in this table 3. a lag k partial autocorrelation coefficient assesses the relationship among the residuals at time t and time t+k once all lower lags have been taken into consideration. it could be used to determine the order in which an autoregressive model should be constructed to match the data. the 95.0% probability limits about 0 are also indicated. unless the probability limit at a given lag does figure 1: plot a data generation electrical energies from (132) months figure 2: auto correlation function for generation electrical energies omer, et al.: brown’s and holt’s double exponential smoothing 59 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 table 2: estimated autocorrelations for generation electrical energies lag autocorrelation stnd. error lower 95.0% upper 95.0% prob. limit prob. limit 1 0.0167213 0.0873704 -0.171243 0.171243 2 −0.241384 0.0873948 −0.171291 0.171291 3 0.135039 0.092344 −0.180991 0.180991 4 −0.0592817 0.0938394 −0.183922 0.183922 5 −0.171542 0.0941248 −0.184482 0.184482 6 −0.0408148 0.0964818 −0.189101 0.189101 7 −0.0877073 0.0966135 −0.189359 0.189359 8 0.0443452 0.0972194 −0.190547 0.190547 9 0.210737 0.0973737 −0.190849 0.190849 10 −0.201387 0.100795 −0.197555 0.197555 11 −0.0940386 0.103821 −0.203486 0.203486 12 0.434518 0.104469 −0.204757 0.204757 13 −0.0416651 0.117458 −0.230214 0.230214 14 −0.19711 0.117571 −0.230435 0.230435 15 0.0534731 0.120067 −0.235327 0.235327 16 −0.0195178 0.120249 −0.235683 0.235683 17 −0.0703227 0.120273 −0.235731 0.235731 18 −0.0649174 0.120586 −0.236345 0.236345 19 −0.066637 0.120853 −0.236867 0.236867 20 0.0365189 0.121133 −0.237417 0.237417 21 0.308873 0.121217 −0.237581 0.237581 22 −0.073339 0.127083 −0.249078 0.249078 23 −0.142081 0.127406 −0.249711 0.249711 24 0.300656 0.128609 −0.25207 0.25207 not include the predicted coefficient, there seems to be a statistically important connection at which lag at the 95.0% confidence level. at the 95.0% confidence level, four of the —24 partial autocorrelation coefficients were statistically important. you could plot the partial autocorrelation coefficients via choosing partial autocorrelation function from of the listing of graphical choices. moreover, to use the pacf, a correlation of the time series data through the time series itself would be calculated in order to discover data patterns. depending on the pacf diagram is shown in figure 3. application of des using brown and holt model depending on data examination, it is recognized that the data has a trending pattern. des is the time series processing approach utilized. there really are two methods to the des technique, 1 variable linear des from brown as well as 2 parameter des from holt. a testing cases for these techniques were then evaluated utilizing parameters assigned at random. the experiment process was divided into two phases: initialization (training) utilizing training data to achieve the constituents which will be utilized to assess the forecasting as well as experimenting to make accurate predictions relying on the value of a component parts achieved from of the initializing phase as well as comparison with data test method to evaluate the value of as well as mae and mape. application of des using brown smoothing variables (α) utilized in des brown testing are chosen (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, and 0.9). the results of the mape evaluation are presented in table 4. the outcomes of these experiments revealed that the value of α = 0.2 caused the least error, because the (mape = figure 3: partial autocorrelations function for generation electrical energies omer, et al.: brown’s and holt’s double exponential smoothing 60 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 table 3: estimation partial autocorrelations for generation electrical energies lag partial stnd. error lower 95.0% upper 95.0% autocorrelation prob. limit prob. limit 1 0.0167213 0.0873704 -0.171243 0.171243 2 −0.241732 0.0873704 −0.171243 0.171243 3 0.153079 0.0873704 −0.171243 0.171243 4 −0.140071 0.0873704 −0.171243 0.171243 5 −0.0975302 0.0873704 −0.171243 0.171243 6 −0.10485 0.0873704 −0.171243 0.171243 7 −0.139082 0.0873704 −0.171243 0.171243 8 0.0528022 0.0873704 −0.171243 0.171243 9 0.155682 0.0873704 −0.171243 0.171243 10 −0.230709 0.0873704 −0.171243 0.171243 11 −0.0358917 0.0873704 −0.171243 0.171243 12 0.333182 0.0873704 −0.171243 0.171243 13 −0.0568566 0.0873704 −0.171243 0.171243 14 −0.00962131 0.0873704 −0.171243 0.171243 15 −0.082228 0.0873704 −0.171243 0.171243 16 −0.00415532 0.0873704 −0.171243 0.171243 17 0.0103137 0.0873704 −0.171243 0.171243 18 −0.0988107 0.0873704 −0.171243 0.171243 19 0.00642562 0.0873704 −0.171243 0.171243 20 −0.0551295 0.0873704 −0.171243 0.171243 21 0.218259 0.0873704 −0.171243 0.171243 22 0.00970171 0.0873704 −0.171243 0.171243 23 −0.0380208 0.0873704 −0.171243 0.171243 24 0.117758 0.0873704 −0.171243 0.171243 table 4: mape value for the des brown α 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 mape 11.2484 9.26275 9.36603 9.55898 9.67292 9.8182 9.82523 9.60284 9.4356 table 5: mape value for des holt mape β α 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 mape 0.1 10.1139 9.94129 9.93563 9.96147 10.0146 10.129 10.1839 10.0151 9.91106 mape 0.2 9.4136 9.34794 9.39981 9.44035 9.56963 9.79795 10.0164 10.2344 10.4674 mape 0.3 9.09144 9.22661 9.46055 9.79021 10.0959 10.4022 10.7455 11.1267 11.5143 mape 0.4 9.02689 9.35794 9.72886 10.1104 10.5094 10.9138 11.3833 11.867 12.3145 mape 0.5 9.00694 9.43233 9.83182 10.2438 10.7197 11.1727 11.6403 12.1686 12.6818 mape 0.6 8.90925 9.34006 9.79061 10.2269 10.6686 11.1207 11.6528 12.1753 12.6051 mape 0.7 8.79654 9.2637 9.64667 10.0507 10.5156 10.9651 11.3411 11.6038 11.7229 mape 0.8 8.62661 9.01604 9.41114 9.83381 10.1908 10.4887 10.6578 10.6803 10.6121 mape 0.9 8.38666 8.77974 9.161 9.4774 9.74875 9.89043 9.90722 9.86512 9.8109 9.26275% if the α = 0.2 which was the minimum error for forecasting, the suitable parameter in des brown is equal to α = 0.2. application of des using holt model smoothing parameters (α,β) used in des holt testing are selected (0.1, 0.1), (0.1, 0.2)…… (0.1, 0.9), (0.2, 0.1), (0.2, omer, et al.: brown’s and holt’s double exponential smoothing 61 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 0.2),…… (0.2, 0.9),……. (0.9, 0.9). the mape calculating outcomes are presented in table 5. the findings of all these trials revealed which the value of (α = 0.1 and β = 0.9) produced the smallest error. because the (mape = 8.38666% if the (α = 0.1 and β = 0.9) which was the minimum error for forecasting, the suitable parameter in des holt is equal to (α = 0.1 and β = 0.9). according to table 6 above, the holt’s des technique has the lower mape score of around 8.38666% as a result, des holt’s could well be classified as a good forecasting approach. as a result, using holt’s des as a predicting approach for the following prediction will be more appropriate. parameter optimization results for des brown parameters for optimization α des brown used the golden section approach, as well as the findings could well be found in table 7. according to the outcomes of the optimization, α = 0.22 through a mape optimal value of 9.23616%. while = 0.22, the des brown technique could be regarded to be good for prediction. the best value of mape was determined relying on the optimization outcomes. des holt’s parameter optimization outcomes the parameters optimization α and β on des holt were optimized utilizing a modified golden section technique. the outcomes of parameter optimization for des holt are presented in table 8. relying on the optimization findings, the best value of was determined α=0.95 and β=0.05 with mape optimum value of 8.08586%. as a result, the des holt technique could be considered to be good at prediction. discussion it is shown in table 9 shows the comparison between brown’s as well as holt’s des for forecasting applied generation electrical energies used in this study. furthermore, this technique will predict future generation electrical energies values. the data cover a period of (132) months. brown’s one-parameter linear exponential smoothing method is presently being used. the best prediction for future data, according to this approach, is a linear trend calculated by exponentially weighing whole past data values. even before models is fit as well as holt’s twoparameter linear exponential smoothing method was chosen, every value of generation electrical energies was changed as follows. the best prediction for future data, according to this table 8: des holt’s parameter optimization outcomes mape β α 0.05 0.06 0.07 0.08 0.09 0.1 0.11 0.12 0.13 0.14 0.15 0.85 8.31223 8.35224 8.39135 8.42957 8.46679 8.50296 8.53965 8.57689 8.61677 8.6565 8.69609 0.86 8.28947 8.32858 8.36675 8.404 8.44023 8.47538 8.51364 8.55456 8.59434 8.63298 8.67083 0.87 8.26557 8.30375 8.34097 8.37777 8.4153 8.45283 8.49215 8.53183 8.57054 8.60807 8.64593 0.88 8.242 8.28048 8.3184 8.35572 8.39245 8.43171 8.47005 8.5078 8.54588 8.58339 8.62155 0.89 8.22163 8.25912 8.29615 8.33255 8.37105 8.40933 8.44705 8.48398 8.52058 8.55875 8.59577 0.9 8.20099 8.23723 8.27313 8.31057 8.34901 8.38666 8.42346 8.45936 8.49604 8.53278 8.5736 0.91 8.17931 8.21468 8.25059 8.28874 8.32622 8.36288 8.39866 8.43469 8.47076 8.51242 8.55396 0.92 8.15662 8.19111 8.22857 8.26578 8.30228 8.33793 8.37328 8.41065 8.45232 8.49412 8.53502 0.93 8.13297 8.16867 8.20548 8.24172 8.27723 8.31264 8.35077 8.39256 8.43426 8.47501 8.51483 0.94 8.10912 8.14547 8.18134 8.217 8.25359 8.29134 8.33257 8.37418 8.41484 8.45497 8.49577 0.95 8.08586 8.12153 8.15834 8.19464 8.23179 8.27237 8.31392 8.35451 8.39493 8.43563 8.47533 table 9: the result of the mape score of parameter optimization for des brown and holt’s criteria des brown des holt’s α=0.22 α=0.95 and β=0.05 period period mape 9.23616 8.08586 forecast summary. *brown’s linear exp. smoothing was used as the prediction models α=0.22. *the forecast model has been chosen: holt’s linear exp. smoothing through α=0.95 and β=0.05 table 7: des brown’s parameter optimization outcomes α 0.15 0.16 0.17 0.18 0.19 0.2 0.21 0.22 0.23 0.24 0.25 mape 9.77084 9.59147 9.45939 9.3768 9.31287 9.26275 9.23656 9.23616 9.25097 9.26356 9.28123 table 6: the result of mape score method mape (%) des brown’s 9.26275 des holt’s 8.38666 omer, et al.: brown’s and holt’s double exponential smoothing 62 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 table 10: forecasting by using des brown and holt methods years months des brown des holt’s years months des brown des holt’s 2021 1 3063.27 3316.63 2024 1 3109.59 3971.89 2 3064.55 3334.83 2 3110.87 3990.09 3 3065.84 3353.03 3 3112.16 4008.29 4 3067.13 3371.23 4 3113.45 4026.49 5 3068.41 3389.43 5 3114.73 4044.69 6 3069.7 3407.64 6 3116.02 4062.9 7 3070.99 3425.84 7 3117.31 4081.1 8 3072.27 3444.04 8 3118.59 4099.3 9 3073.56 3462.24 9 3119.88 4117.5 10 3074.85 3480.44 10 3121.17 4135.7 11 3076.13 3498.64 11 3122.45 4153.9 12 3077.42 3516.85 12 3123.74 4172.11 2022 1 3078.71 3535.05 2025 1 3125.03 4190.31 2 3079.99 3553.25 2 3126.31 4208.51 3 3081.28 3571.45 3 3127.6 4226.71 4 3082.57 3589.65 4 3128.89 4244.91 5 3083.85 3607.85 5 3130.17 4263.11 6 3085.14 3626.06 6 3131.46 4281.32 7 3086.43 3644.26 7 3132.75 4299.52 8 3087.71 3662.46 8 3134.03 4317.72 9 3089.0 3680.66 9 3135.32 4335.92 10 3090.29 3698.86 10 3136.61 4354.12 11 3091.57 3717.06 11 3137.89 4372.32 12 3092.86 3735.27 12 3139.18 4390.53 2023 1 3094.15 3753.47 2 3095.43 3771.67 3 3096.72 3789.87 4 3098.01 3808.07 5 3099.29 3826.27 6 3100.58 3844.48 7 3101.87 3862.68 8 3103.15 3880.88 9 3104.44 3899.08 10 3105.73 3917.28 11 3107.01 3935.48 12 3108.3 3953.69 approach, is a linear trend calculated by exponentially weighing whole past data values. before fitting the model, each value of the generation electrical energies was modified as follows. as a consequence, des holt’s approach could be considered to be good at forecasting. des holt’s is chosen as the strongest forecasting model. depending on the optimization outcomes, achieved optimal value of (α=0.95 and β=0.05) through mape optimum value of 8.08586% in des holt’s technique. however, depending on the optimization outcomes in the des brown’s technique, an optimal value of α=0.22 was found, with a mape ideal value of 9.23616%. then, holt’s technique could be considered to be the most suited way for forecasting generation electrical energies data since the value of mape in des holt’s method was less than the value of mape in the des brown’s method. mape (des brown method) =9.23616% >mape (des holt’s method) =8.08586% conclusion and recommendation the contrast among brown’s and holt’s des could be established depending on the criterion of mape for omer, et al.: brown’s and holt’s double exponential smoothing 63 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2021, 5 (2): 56-63 determining the optimum data technique. with using mape as performance measurements, we concluded that the holt’s des technique is the ideal technique since the values of mape in the holt’s des would be less than the values of mape in the brown’s des. furthermore, depending on the optimization has been done α and β the optimal values achieved for des holt’s are (optimal values of α = 0.95 and β optimal value of 0.05) having mape optimal value of 8.08586%, as well as for des brown’s is (optimum value of α = 0.22 via mape optimal value of 9.23616%). a mape of the des holt’s approach was lower than that of mape des brown’s technique. future research are planned to optimize the parameters utilizing various non-linear programming approaches or to optimize by making changes to the initialization method prediction: forecasting using optimum parameter for the des brown and des holt methods this table 10 illustrates the expected values for the generation electrical energies. throughout the period while true data is obtainable; this also shows the forecasted values from either the fitting model as well as the residuals (dataforecast). it indicates 95.0% forecast limits for predictions for time period beyond the ending of a series. those limits represent where a actual data values at the chosen future time was expected to be through 95.0% confidence, providing the fitted model is adequate for the data. you could plot the predictions by choosing prediction plot from the listing of graphical choices. you could modify the level of confidence when reviewing the plot using the alternative mouse button as well as selecting pane options. choose model comparisons from of the list of tabular options to evaluate if the model appropriately matches the data. that table illustrates the predicted values for the generation electrical energies. throughout the period when real data are obtainable, this also shows the projected value from of the fitted models as well as the residuals (data-forecast). it indicates 95.0% forecast limits for predictions for periods of time well beyond ending of the series. these limits represent where the real data value is expected to be through 95.0% confidence at a given future time, providing the fitted model was adequate for the data. you could plot the predictions by choosing prediction plot from of the list of graphical choices. you could modify the level of confidence when examining the plot by using the alternative mouse button as well as selecting pane options. choose model comparisons from of the listing of tabular options to see if the model matches the data well. references 1. p. a. s. dharmawan, and i. g. a. indradewi. double exponential smoothing brown method towards sales forecasting system with a linear and non-stationary data trend. journal of physics: conference series, vol. 1810, no. 1, p. 012026, 2021. 2. m. b. hasan, and m. n. dhali. determination of optimal smoothing constants for exponential smoothing method and holt’s method. dhaka university journal of science, vol. 65, no. 1, pp. 55-59, 2017. 3. f. sidqi, and i. d. sumitra. forecasting product selling using single exponential smoothing and double exponential smoothing methods. iop conference series: materials science and engineering, vol. 662, p. 032031, 2019. 4. w. anggraeni. perbandingan metode fuzzy time series hsu dan double exponential smoothing pada peramalan nilai tukar rupiah terhadap dolar amerika. jurnal riset manajemen dan bisnis (jrmb) fakultas ekonomi uniat, vol. 1, no. 2, pp. 153-162, 2016. 5. wikipedia contributors, “exponential smoothing,” wikipedia, feb. 21, 2019. https://en.wikipedia.org/wiki/exponential_smoothing. 6. v. a. fitria. parameter optimization of single exponential smoothing using golden section method for groceries forecasting. zero: jurnal sains, matematika dan terapan, vol. 2, no. 2, p. 89, 2019. 7. a. n. aimran, and a. afthanorhan. a comparison between single exponential smoothing (ses), double exponential smoothing (des), holts (brown) and adaptive response rate exponential smoothing (arres) techniques in forecasting malaysia population. global journal of mathematical analysis, vol. 2, no. 4, p. 276, 2014. 8. f. ikhsan. forecasting of criminality problems using double exponential smoothing method. academia open, vol. 4, p. 122, 2021. 9. k. aroian, n. uddin, and h. blbas. longitudinal study of stress, social support, and depression in married arab immigrant women. health care for women international, vol. 38, no. 2, pp. 100-117, 2016. 10. r. rachmat, and s. suhartono. comparative analysis of single exponential smoothing and holt’s method for quality of hospital services forecasting in general hospital. bulletin of computer science and electrical engineering, vol. 1, no. 2, pp. 80-86, 2020. tx_1~abs:at/add:tx_2~abs:at 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 20-25 research article the full factorial design approach to determine the attitude of university lecturers toward e-learning and online teaching due to the covid-19 pandemic nozad h. mahmood1*, dler h. kadir2, hawkar q. birdawod3 1department of business administration, cihan university sulaimanyia, kurdistan region, iraq, 2,3department of statistics, salahaddin university, erbil, kurdistan region, iraq, 3department of business administration, cihan university-erbil, kurdistan region, iraq abstract the purpose of this study was to determine faculty members’ attitudes toward online learning in kurdistan region universities. the study examined the biographic and personal characteristics of the lecturer toward e-learning. the data were collected among faculty members at cihan university-sulaimaniya, and to analyze the data, a full factorial design with five main factors at two levels and no central points was applied for this specific purpose. the study’s findings indicated that there was no significant relationship between gender and lecturer attitude toward e-learning. in comparison to teachers with an msc degree, those with a phd have a more negative attitude toward e-learning. furthermore, full-time faculty members have a greater positive effect on teachers’ attitudes than part-time lecturers. likewise, the results indicate that lecturers who earned their most recent education degrees outside of iraq have a more favorable attitude. similarly, lecturers in the sciences are more favorable to e-learning than those in the arts and social sciences. in addition, the findings demonstrated that the interaction factors (gender) and (education degree) have a negative effect on lecturers’ attitudes when they are combined. besides that, the interaction of factors (country of last education degree) and (faculty member types) improves attitudes toward e-learning. based on the results, it is suggested that academic staff receive e-learning training to deepen their knowledge and understanding of such a modern teaching system. there is also a need to enhance factors related to positive attitudes toward e-learning among university lecturers. the findings of this study are necessarily significant to both teachers and educational organizations in kurdistan region universities. keywords: e-learning, attitudes, covid-19, faculty members, factorial design introduction one of the most influential sectors of human life that were stopped due to the coronavirus in most countries was the university education system. according to reports by unesco, numerous governments around the world have closed their educational institutes for the short term to control the spread of the covid-19 pandemic. approximately 1.725 billion college students globally have been suffering from the closure of colleges due to the coronavirus pandemic. in line with the unesco monitoring report, 192 countries have applied for nationwide closures.[1] staying in quarantine at home and avoiding daily physical interactions with others is one way to control the severity of this virus and prevent its rapid spread. that is why most countries and universities have decided to use online teaching as an alternative educational methodology.[2] in most developed countries, universities offer not only face-to-face instruction but also distance learning options for students who prefer to study at home rather than on campus. based on the effects of the covid-19 disease, a new education system was mandated by the ministry of higher education for the kurdistan region universities, which was a blended education system, or a mixed education system, in which the practical lectures are taught face-to-face on the campus, but the theoretical ones are online.[3,4] when compared to developed countries, developing countries such as iraq and the kurdistan region face several corresponding author: nozad h. mahmood, department of business administration, cihan university sulaimanyia, sulaimanyia, 46001, kurdistan region, iraq. e-mail: nozad.mahmood@sulicihan.edu.krd received: december 26, 2021 accepted: january 24, 2022 published: february 10, 2022 doi: 10.24086/cuesj.v6n1y2022.pp20-25 copyright © 2022 nozad h. mahmood, dler h. kadir, hawkar q. birdawod. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) mahmood, et al.: using factorial design to determine the attitude of university lecturers towards online teaching 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 20-25 obstacles when applying for online education. it might be due to the lack of having a strong internet connection and insufficient skills in the use of information technology.[5,6] likewise, learning how to use a new modern platform and an advanced application is still a weird thing, even at universities in developing countries.[7,8] the problem statement for this project is students’ dissatisfaction with the online system, as well as teachers’ performance in using it. the main research questions that guided the study are: 1. what are teachers’ attitudes toward e-learning, in general? 2. is there any association between each the factors (education degree, types of faculty members, the country of the last education’s degree, gender, stream of education), and teachers’ attitude toward e-learning? 3. is there any interaction effect between the factors towards e-learning? the objective of the study the essential goals of this research are to examine teachers’ attitudes toward e-learning in kurdistan region universities and find out the factors that influenced their attitudes toward e-learning during the covid-19 pandemic. the significance of the study the importance of this research is to increase the desire of teachers for the use of an online education system that may, in the next few years, replace the current system of on-campus education. the hypothesis of the study the first hypothesis (h1): there are no statistically significant differences in the attitudes of lecturers toward e-learning implementation, whether they are male or female. the second hypothesis (h2): there are no statistical differences in lecturers’ attitudes toward e-learning based on the type of faculty members. the third hypothesis (h3): there are no statistical differences in lecturers’ attitudes towards e-learning based on their education degrees. the fourth hypothesis (h4): there are no statistical differences in lecturers’ attitudes toward e-learning based on the last education degree in a country. the fifth hypothesis (h5): there are no statistically significant differences in the attitudes of lecturers toward e-learning implementation, whether their stream of education is art or science. literature review lecturer attitudes are extremely crucial for successfully establishing and implementing an educational method in the teaching and learning process. on the other hand, teachers are the central figures in all formal education, and their attitudes toward e-learning have a significant impact on their decisions to accept or reject the system. in other words, the success of e-learning in education is highly dependent on the lecturers’ attitudes toward it.[9,10] effectively initiating and implementing instructional innovation in e-learning processing at the university is highly dependent on lecturers’ attitudes. thus, understanding lecturers’ personal characteristics is a prerequisite for implementing an e-learning system.[11] academic staff demographic and personal characteristics, such as educational degrees, gender, educational stream, faculty type, and country of last educational degree, may influence lecturers’ favorable attitudes toward the e-learning system. several previous studies have examined the demographic and personal characteristics of teachers in relation to e-learning. there was a study that investigated the factors of gender in e-learning and discovered that gender has no influence on teachers’ attitudes. in other words, there is no statistically significant difference in attitudes regarding e-learning between men and women.[8,12] in addition, the literature links teachers’ attitudes to their personal characteristics. demographic characteristics, including gender, education level, and age, had no significant effect on their attitudes toward online learning. teachers’ nationality and marital status, on the other hand, have significant attitudes toward e-learning.[13] university teachers in the arts have negative attitudes toward e-learning, whereas lecturers in the sciences have positive attitudes.[14] methodology the general purpose of this study is to examine lecturer support and attitudes toward e-learning in terms of several personal variables and to determine the direction of their attitudes and the level of correlation between these variables. the questionnaire consists of two main sections. the first section contains the demographic profile of the participants, such as gender, education degree, types of faculty members, last education degree country, and stream of education, while the second part includes 15 statements that describe overall attitudes toward e-learning, and a five-point likert scale was used to rate each item from strongly disagree to strongly agree. then, jmp statistical software was used to analyze the data to apply a full factorial design. since there are five main factors, and each factor has two levels with a response variable, we used a 2k factorial design. which included (five main factors) indicates five main effects, and a 5 2 10 � � � � � � � ��two factor�interactions). three-factor interactions and above are very rarely significant effects.[15] description and selection of levels to conduct a 25 full factorial design, all factors must be set to two levels, the low and high levels. the factors used in the experiment and their associated levels are given in the following table 1. the planning matrix and experiment results the planning matrix and experiment results for 25 full factorial design with 32 runs, and without any center point get the following raw data. then, a randomized order was used to execute test runs of the design table 2. mahmood, et al.: using factorial design to determine the attitude of university lecturers towards online teaching 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 20-25 data analysis we need to achieve a model between the response variable (university lecturers’ attitudes toward e-learning) and the five factors to clarify their power of relationships and the effects which influenced the e-learning during the covid-19 pandemic. the results of table 3 show that r-square is 0.967806, meaning that the factors altogether explain a large amount of 96.78% of the variance in response variable (university lecturers towards e-learning), and it is evidence that the model is appropriate. furthermore, root mean square error shows a quite low 0.22727. figure 1 is a half-normal plot of parameter that employed to show the effect of main factors on the y response (university lecturers’ attitudes toward e-learning). all the factors that lie along the blue line (lenth’s pseudo standard error) are negligible. in contrast, the large effects, or significant factors are far from the blue line. the most significant effects based on this analysis show the main effects of d (last education degree’s country), e (stream of education), c (types of faculty table 1: factors and their levels factors low (-1) high (+1) a: gender female male b: education degree msc degree phd degree c: types of faculty members part-time full-time d: last education degree’s country inside (iraq) outside (other countries) e: streams of education art science table 2: planning matrix and experiment results # pattern a b c d e y 1 −−−−− −1 −1 −1 −1 −1 2.44673 2 +−−−− 1 −1 −1 −1 −1 1.98352 3 −+−−− −1 1 −1 −1 −1 1.54322 4 ++−−− 1 1 −1 −1 −1 2.05211 5 −−+−− −1 −1 1 −1 −1 3.10284 6 +−+−− 1 −1 1 −1 −1 2.19841 7 −++−− −1 1 1 −1 −1 2.273441 8 +++−− 1 1 1 −1 −1 2.31429 9 −−−+− −1 −1 −1 1 −1 3.49831 10 +−−+− 1 −1 −1 1 −1 3.40167 11 −+−+− −1 1 −1 1 −1 3.39617 12 ++−+− 1 1 −1 1 −1 3.38519 13 −−++− −1 −1 1 1 −1 3.73804 14 +−++− 1 −1 1 1 −1 3.51055 15 −+++− −1 1 1 1 −1 3.68208 16 ++++− 1 1 1 1 −1 3.44734 17 −−−−+ −1 −1 −1 −1 1 2.95147 18 +−−−+ 1 −1 −1 −1 1 2.22851 19 −+−−+ −1 1 −1 −1 1 2.11037 20 ++−−+ 1 1 −1 −1 1 2.4 21 −−+−+ −1 −1 1 −1 1 3.26823 22 +−+−+ 1 −1 1 −1 1 2.77876 23 −++−+ −1 1 1 −1 1 2.67552 24 +++−+ 1 1 1 −1 1 2.974158 25 −−−++ −1 −1 −1 1 1 4.32626 26 +−−++ 1 −1 −1 1 1 4.59298 27 −+−++ −1 1 −1 1 1 3.84214 28 ++−++ 1 1 −1 1 1 4.62367 29 −−+++ −1 −1 1 1 1 4.26693 30 +−+++ 1 −1 1 1 1 4.60214 31 −++++ −1 1 1 1 1 4.72248 32 +++++ 1 1 1 1 1 4.48045 table 3: summary of fit r-square 0.967806 root mean square error 0.22737 mean of response 3.213062 observations 32 figure 1: half-normal probability plot for the factors and their interactions table 4: parameter estimate mahmood, et al.: using factorial design to determine the attitude of university lecturers towards online teaching 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 20-25 members), and it looks like there are some interactions effects between the factors. the above figures 2 are the visualization of confidence intervals (ci) that can be applied to the f-test by using a leverage plot. the first panel shows the overall test of the model which explains the linearly relationship between actual values and the predicted values of response y. likewise, the other panels are the main factor effects for a, b, c, d, and e, respectively. each panel displays the mean as a horizontal line (blue line), a regression line (red line), and a 95% ci. since, the significance of each effect is indicated by the cis and the mean, when the ci crosses the mean line, indicating a significant effect. as the result, there are some significant effects of the main factors (b), (c), (d), and (e), whereas factor a (gender) is insignificant. y = −0.092898 (b)+ 0.1641668 (c) + 0.7567132 (d)+ 0.3396924 (e) − 0.1163893 (a*b) + 0.1226638 (d*e) (1) according to the results in table 4, the main factors of d, e, c, and b are statistically significant due to their small p-values which are less than. on the other hand, since the p-value of the main factor a (gender) is equal to 0.5082 and it is larger than, indicates that gender is statistically insignificant or shows no effect. furthermore, there is a meaningful interaction between factors d and e which shows a positive effect on the response y. likewise, there is a significant interaction between factors a and b toward a positive direction. consequently, the regression model equation becomes as below: in the linear regression equation (1), factor (b: education degree) has a negative effect on the (university lecturers’ attitudes toward e-learning), indicating that teachers with a phd degree 9.22898% have a more negative attitude towards e-learning than holders of msc degree. as well, factor (c: types of faculty members) has a positive impact on the teachers’ attitudes. in other words, full-time lecturers 16.411668% have more positive towards e-learning than part-time lecturers. furthermore, factor (d: last education degree’s country) has a positive influence on the teachers’ attitudes towards e-learning, and the results illustrated that the lecturers whose last response y a leverage plot factor names y: university lecturers’ attitudes toward e-learning a: gender b: education degree c: types of faculty members d: last education degree’s country e: stream of education b leverage plot c leverage plot d leverage plot e leverage plot figure 2: leverage plots important factors mahmood, et al.: using factorial design to determine the attitude of university lecturers towards online teaching 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 20-25 table 5: goodness-of-fit test shapiro-wilk w test w prob<w 0.983344 0.8888 ho=the data is from the normal distribution. small p values reject ho education degrees were outside of iraq 75.67132% have a more positive attitude. likewise, for the main factor (e: stream of education), the results show a positive effect on the teachers’ attitudes. to clarify, the lecturers in the scientific field 33.96924% have a more positive attitude towards e-learning than those in the art field. when it comes to interaction cases, the results demonstrate that factors (a: gender) and (b: education degree) together have a negative impact on the lecturers’ attitudes. in other words, male lecturers with a doctoral degree 11.63893% have a more negative attitude. furthermore, the interaction of factors (d: last education degree’s country) and (c: types of faculty members) together have a positive impact. in such a way, full-time faculty members that got their last degree outside the country 12.26638% have a more positive attitude towards e-learning during the covid-19 pandemic. figure 3 depicts a normal quantile plot and a histogram that is used to examine the normality of the residuals, which is one of the linear regression presumptions. since the histogram plot is like a symmetric bell curve and the points on the quantile plot close to the line, the results show a typical normal residual of the data.[16] in order to get an unquestionable normality result, the shapiro-walk test can be used. as the results in table 5, while the (prob <w) = 0.8888, and it is larger than 0.05, indicating that there is no evidence to reject the normality distribution of the data. conclusions the purpose of this study was to identify the most effective factors for increasing positive attitudes toward e-learning among faculty members at cihan university-sulaimaniya. in the full factorial designs finding the effect of main factors of (education degree), (types of faculty members), (last education degree’s country), and (stream of education) are positively significant on the e-learning attitudes. furthermore, the effect of interaction “gender and education degrees” was negatively effective. likewise, the effect of interaction “last education degree’s country and types of faculty members” is positive significant. the findings illustrated that the attitude toward e-learning increased when the faculty members were fulltime rather than part-time lecturers. it indicates that every university has its own online education system, and full-time lecturers should be more familiar with the system compared to part-time ones. similarly, faculty members with non-iraqi education degrees showed a higher level of attitude toward online learning than iraqi education degrees, and this is because most of the non-iraqi education degrees came from developed countries such as the usa, europe, and other advanced countries. next, lecturers in the scientific field have a more positive attitude towards e-learning than lecturers in the art field. the lecturers working in the scientific fields have a higher level of knowledge about computers and information technology than those working in the art and social sciences fields. on the other hand, higher level of education degrees showed a negative effect on the faculty member’s attitude. the reason could be that phd holders are older, and it is obvious that computer and information technology have been hard to obtain in our country for the last two decades. when it comes to the interaction between factors, the results showed that male lecturers with a doctoral degree have a more negative attitude, and the full-time faculty members with non-iraqi education degrees have a more positive attitude toward e-learning. recommendations based on the finding results of this study, making the following recommendations • lecturers must develop their abilities and skills to use technology to the maximum extent possible in their distance education. • conduct similar research to obtain additional information about faculty members attitude toward e-learning. • additional research is recommended to identify some different variables such as (years of experience and scientific titles) that may affect lecturer attitudes toward e-learning. • it would be beneficial to see the results from this research expanded to include lecturers and universities in order to ensure the effectiveness of e-learning. figure 3: normal quantile plot mahmood, et al.: using factorial design to determine the attitude of university lecturers towards online teaching 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 20-25 references 1. v. reddy, c. soudien and l. winnaar. the impact of school closures on education outcomes in south africa. the conversation, 2020. 2. j. collado-valero, g. rodríguez-infante, m. romero-gonzález, s. gamboa-ternero, i. navarro-soria and r. lavigne-cerván. flipped classroom: active methodology for sustainable learning in higher education during social distancing due to covid-19. sustainability, vol. 13, no. 10, p. 5336, 2021. 3. c. j. bonk and c. r. graham. the handbook of blended learning: global perspectives, local designs. new york: john wiley and sons, 2012. 4. m. p. pascual. el blended learning reduce el ahorro de la formación on-line pero gana en calidad. educaweb, vol. 69, pp. 34-49, 2003. 5. t. n. aung and s. s. khaing. challenges of implementing e-learning in developing countries: a review. in: international conference on genetic and evolutionary computing, cham: springer, pp. 405-411, 2015. 6. s. a. obed, p. a. mohammed and d. h. kadir. the estimation of (covid-19) cases in kurdistan region using nelson aalen estimator. cihan university-erbil scientific journal, vol. 5, no. 2, pp. 24-31, 2021. 7. a. andersson and å. grönlund. a conceptual framework for e-learning in developing countries: a critical review of research challenges. the electronic journal of information systems in developing countries, vol. 38, no. 1, pp. 1-16, 2009. 8. j. a. lara, s. aljawarneh and s. pamplona. special issue on the current trends in e-learning assessment. journal of computing in higher education, vol. 32, no. 1, pp. 1-8, 2020. 9. g. salmon. e-moderating: the key to online teaching and learning. milton park, abingdon-on-thames: routledge, 2012. 10. t. teo, ö. f. ursavaş and e. bahçekapili. an assessment of preservice teachers’ technology acceptance in turkey: a structural equation modeling approach. asia-pacific education researcher, vol. 21, no. 1, pp. 191-202, 2012. 11. t. teo. factors influencing teachers’ intention to use technology: model development and test. computers and education, vol. 57, no. 4, pp. 2432-2440, 2011. 12. f. martin, k. budhrani and c. wang. examining faculty perception of their readiness to teach online. online learning, vol. 23, no. 3, pp. 97-119, 2019. 13. m. s. alanazy. factors affecting faculty attitude for adopting online learning at aljouf university, saudi arabia. journal of education and learning, vol. 7, no. 1, pp. 154-162, 2018. 14. c. mercader and j. gairín. university teachers’ perception of barriers to the use of digital technologies: the importance of the academic discipline. international journal of educational technology in higher education, 17, no. 1, pp. 1-14, 2020. 15. d. c. montgomery. design and analysis of experiments. new york: john wiley and sons, 2017. 16. n. h. mahmood, s. n. fadhil n. f. ahmed. using multiple linear regression model to identify the relationship between leadership styles and job satisfaction in government organizations in sulaimani city-krg. the scientific journal of cihan university slemani, vol. 144, pp. 128, 2019. tx_1~abs:at/add:tx_2~abs:at 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 53-56 research article a genetic disaster of human cloning: ethical and scientific implications yaser a. jasim department of accounting, cihan university-erbil, kurdistan region, iraq abstract human cloning may become a viable possibility in the far future; it is no longer a spectacular science fiction wisdom. the method is accessible. the dilemma is whether it would be ethical and acceptable in society, considering any associated consequences related to human cloning. unlike a normal embryo, this one is abnormal, which has a genome derived from a combination of six sources, a human cloned genome would have one source. this procedure would undoubtedly eliminate the distinct qualities that a real child possesses. its short-term and long consequences, on the contrary, are currently unclear. cell cloning of human research potentially changes the variety of medical diseases treatment in the soon future. human cloning or non-sexual human reproduction is a multidisciplinary problem that has been studied by scholars from several perspectives, including ethical elements and science perspectives, religion, and health. because some scientific discoveries, such as human simulation, are related to human beliefs, several religious scholars have demonstrated distinctive responsivity to religious doctrine and consider such investigations to be contrary to divine verses and religious beliefs; as a result, human cloning has been prohibited. as a result, it is vital to investigate and analyze the reasoning of their contravention to demonstrate that scientific discoveries might not only not diminish but also even deepen religion. keywords: human cloning, genetic disaster, embryo, ethics, religion, and human health introduction human cloning is the technique of producing genetically identical individuals. embryo splitting or nuclear transfer can be used to accomplish this cloning process.[1] the process of separating a developing human being embryo into several parts is known as embryo splitting. every one of those components can evolve toward a blastocyst or “late embryo,” which, if implanted, might evolve into a child.[2] human cloning shares advantages and disadvantages as any developing theory and experiment, so advantages are solving infertility issues, replacing biological parts, developing genetic researches, selecting human characteristics, and developing humans. on the other hand, the disadvantages of human cloning that affects on ethics and safety are designing children, violating human right, degrading cells, identifying exclusive uniqueness, and observing psychological issues on individuals in society.[3] monozygotic twins are produced in this manner as figure 1 illustrates the process of human cloning.[4] artificial embryo splitting has been conducted successfully in cows, sheep, apes, and mice, but only up to the preimplantation phase in human beings.[5] the american society for reproductive medicine (2012) recently published a paper stating that artificial embryo splitting to expand the proportion of implanted human being blastocysts used during various infertility therapies constituted an ethical technique. embryo splitting, on the other hand, can only generate a finite number of cloned human beings since the earliest embryo can only be divided so many times and the process never generate a clone of an already living adult. the second method for creating cloned humans is nuclear transfer and does not have the disadvantages of the embryo splitting approach. most of the health problems, such as transplantation or rejection organs, can be treated by cloning the nucleus (nuclear mass/dna) in somatic cells. human cloning using somatic cell nuclear transfer is classified into two categories: reproductive cloning and therapeutic cloning.[6] due to the extreme, sociological and psychological considerations involved, human reproductive cloning remains categorically prohibited. a cloned embryo preparing for implantation into a womb must undergo comprehensive molecular testing to determine whether human cloning corresponding author: yaser a. jasim, department of accounting, cihan university-erbil, kurdistan region, iraq. e-mail: yaser.jasim@cihanuniversity.edu.iq received: december 12, 2021 accepted: march 13, 2022 published: may 30, 2022 doi: 10.24086/cuesj.v6n1y2022.pp53-56 copyright © 2022 yaser a. jasim. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) jasim: a genetic disaster of human cloning: ethical and scientific implications 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 53-56 procedure is healthy and accomplished.[7] human reproductive cloning is considered unethical according to the high risk of death. several theoretical questions, such as the origin of reproduction and human identity, have been posed, which reproductive cloning may contradict. overall, this research examines a variety of scientific and ethical challenges with embryo splitting or nuclear transfer human cloning. bioethics of human cloning bioethics is concerned with the moral implications of using humans and animals in cloning research. it is a new and multidisciplinary area that aids in the understanding and resolution of a variety of ethical dilemmas that arise in the fields of food, medicine, and life sciences.[8] it might be considered as a more effective and helpful method of protecting vulnerable species and reintroducing endangered species. even though some groups argue that goods generated from cloned animals are hazardous, they lack supporting facts.[9] one of the most significant goals of human cloning is to provide a child to a couple who cannot produce a biologically related child. one of the most important advantages of human cloning is that it aids in the elimination of illnesses that run-in families and is handed down through generations.[10] in consideration of recent advances in the field, it is critical to examine and evaluate the ethical implications of human cloning. as a result, stem cell research rules are founded on a cultural concept that provides particular protection to human embryos, but does not provide them the same moral or legal standing as fully formed persons.[11] ethical considerations about human cloning, as well as many religious references, indicate that the discipline of religious ethics is a relatively new development in islam.[12] although muslim researchers have spoken of ethical principles and created significant investigations on the refinement of human character ethics as moral behavior to guide legal judgments on human cloning.[13] various moral and ethical standards reject human cloning such standards impacts on society, children, individuals, and families. these moral behaviors that are used in a particular context and govern a consecutive method of reply based on ethics.[14] the national bioethics advisory commission contacted scientists, theologians, physicians, and others to gather data regarding ethical and moral principles as well as information concerning the safety of the organism generated through somatic cell nuclear transfer, which was the key concern.[15] due to the ethical concerns about cloning, any effort to create genetically identical embryos for clinical use in assisted reproduction, whether by embryo splitting or nuclear transfer from an existing embryo, is ultimately ruled illegal. embryo splitting human cloning approach as preliminary reports, embryo splitting was performed on the genetic abnormal human embryos in the past decades, but the split embryos succumbed after couple cell divisions. there was no further research on human embryo splitting until 2008 when scientists used blastomere biopsies to successfully develop divided human embryos to the blastocyst stage.[16] the scientists divided the cell embryos into four stages, and the four blastomeres recovered were grown separately in an empty zona pellucida. the ontological status of the embryo is a dilemma about what kind of entity embryos are. individual beings, biological humans, or individuals can all be considered embryos.[17] later, illmensee succeeded in developing blastocysts using twin human embryos.[18] recently, nothing is known about the quality of the embryos produced in this manner, and the conclusions are contradictory.[19] embryo splitting is the most standard technique of artificial cloning, in which the blastomeres of an early embryo are separated which forms two or smaller embryos. the first attempts to clone domestic beings included embryo splitting.[20] as keefer revealed that twins may be generated in sheep and cattle when cleavage stage embryos were separated; and demi embryos were transferred into donors.[21] these experiments revealed that triplets and even quadruplets were possible, but only at lower frequencies due to cellular mass loss. researchers have cloned humans by embryo splitting to improve artificial reproduction technologies, and the experiment has been approved by the american society for reproductive medicine’s ethics committee.[16] the quantity of embryos put into the womb increases the likelihood of a successful pregnancy. this technique acceptance by a reputable ethical authority implies that the issue is not necessarily the duplication of human beings.[21] most people believe that studying the causes of infertility and genetic diseases, for example, is values and moral that is likely to be achieved within existing ethical constraints. the use of human embryo models for reproduction, on the other hand, is far more difficult to justify; such applications are still a fair distance off. however, 1 day it may be possible to transplant an embryo produced from (genetically modified) stem cells to a woman’s womb to treat infertility or to avoid genetic disorders.[22] cloning through embryo splitting has been widely utilized in sheep and cattle to improve the output of progeny from genetically high-grade parents, from the 2-cell figure 1: human cloning process[4] jasim: a genetic disaster of human cloning: ethical and scientific implications 55 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 53-56 stage up to the blastocyst stage. embryo splitting is significantly less effective in mice than sheep due to the distinct pattern of early development. from a scientific standpoint, it is unlikely to be particularly successful in humans, despite the fact that monozygotic twins and greater multiples occur naturally at a low rate.[7] many religions provide numerous guidelines or ideas on how to do things correctly, on how to live. various philosophers presented several ethical principles or concepts, which can be divided into two categories: secular and spiritual. some ethical or moral concepts are classified as secular or western, whereas religion is classified as spiritual. according to islamic religion research, to grasp natural occurrences and the secret truth underlying the universe, one must eventually uncover the truth and witness evidence of allah’s majesty.[12] a theologian and neo platonic philosopher had considerable power in the early christian era. he stated that killing does not apply to animals because they do not object and are not like humans. he believes that because god created animals for the benefit of humans, people should likewise protect animals for their purposes.[9] in previous amphibian cloning experiments, a modified form of the nuclear transfer method was utilized to circumvent the limitations of embryosplitting cloning.[21] nuclear transfer human cloning approach nuclear transfer is the second to clone human beings the entire controversy began with the nuclear transfer cloning of a non-human animal, a simple technique that has created no additional issues. following dolly, scientists cloned at least cows, mice, rats, goats, pigs, bunnies, cats, horses, and dogs using the same technique, generally without chuckling. this technique implies that the issue cannot be found in the utilization of nuclear transfer as such.[16] dolly was an ovine clone who provided somatic cell nuclear transfer (scnt) to an embryo in which the nucleus was removed, and genetic material was introduced. the ethical debate had already begun before successful animal cloning, but it sparked a significant debate and calls for constitutional restrictions on human cloning. one of the major ethical problems raised by successful animal cloning is whether and under what conditions human emulation is permissible.[8] a related topic is the medical use of human cloning for stem cell research. this approach raises difficult ethical and political problems.[23] reproductive cloning is described as using scnt and other technologies to generate offspring within the original person’s shared genetic code. these same experimental approaches are used in therapeutic cloning for treatments other than reproduction such as embryonic stem cell lines or the creation of solid organs for transplant.[6] scnt for therapeutic purposes, in which embryos are not transferred for pregnancy, may provide the data necessary to make reproductive scnt safe and successful.[24] the nuclear transfer technique can be used in a strategy known as therapeutic cloning, which involves the use of embryos in therapy and research. human embryos may be produced to perform a study on anomalies, illnesses, and medication effects. this technique offers up a wide range of options for performing stem cell-related research that would otherwise be impossible to conduct on regular people. a study finding that might be utilized for therapeutic reasons could be the discovery of how to create cloned stem cells that could be used to regenerate damaged tissue in a patient.[25] human embryo research, within nuclear transfer clones, is commonly permitted for the period of 14 days following conception, subsequent culture, and scientific, and clinical usage of human embryonic stem cells is largely authorized in most nations. in terms of ethics, reproduction of humans is at the core of the cloning debate, with the concepts of design notion of enhancing individuals and advancing the human being race.[16] cloning is the process of producing genetic clones of individual organisms by nuclear transfer in animal and human biology. animal clones need advanced reproductive procedures such as microsurgery, embryo culture, and transfer into recipients (surrogate mothers), as depicted in figure 1. more specifically, a nucleus from the donor’s cell is placed in an embryo that its own nuclear dna had extracted (enucleation). this rebuilt embryo is triggered, allowing embryonic development to proceed. embryos created by this method have the potential to produce a living, genetically identical person the following transfer into a recipient, but at low efficiency.[21] future of human cloning philosophical perspectives various arguments take place over time to examine the legitimacy of animal cloning. these arguments have ended to prohibit the use of cloned animal products and to prohibit the use of animal cloning.[9] because of the significant risk of mortality, human being reproductive cloning procedure is considered unethical. numerous conceptual problems have been raised, such as the origin of reproduction and human identity, which reproductive cloning may challenge. considering the practically power growth of society and law with technology, innovation can provide us a foretaste of human cloning. because of the enormous therapeutic benefits of this approach, it is being studied all over the globe, despite the deepest ethical concerns that if recognized and perfected, this technology may be exploited for non-medical purposes as well.[26] since there is no federal legislation prohibiting human reproductive cloning in the united states; rapid progress is predicted soon, particularly in therapeutic cloning. for current therapeutic cloning research in regenerative medicine, organ transplantation is a viable medicinal and ethical justification. in the united states alone, over 114,000 individuals are waiting for organ transplants, with one candidate added every 10 min and 20 people on the waiting list dying every day.[7] nevertheless, the human race comes to whether or not to authorize human cloning, the uk government’s decision to authorize mitochondrial dna transfer was preceded by an open, early discussion including scientists, bioethicists, regulators, and the general public. conclusion overall, a variety of scientific and ethical challenges with embryo splitting or nuclear transfer human cloning emerges three aspects: (1) risk assessment, which entails performing the advantages and drawbacks study of cloning in its many jasim: a genetic disaster of human cloning: ethical and scientific implications 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 53-56 forms, as well as their possibilities. this aspect may be a not possible job, but even if it is, scientists must be aware that ethical parameters appeal to expectations and concerns rather than realistic appraisals of well-being; (2) justice, critical assessments of cloning frequently note would benefit couple individuals that it is not worth the expenditure. this aspect might be accurate. however, scientists should look into cloning in light of many ideas of justice; (3) meanings, scientists must closely review the meanings of words and occurrences, as unnaturalness. society considers the benefits of human cloning as another significant element that many people overlook, first, it allows an individual to reproduce and, second, allows experts in the area to create organs that are genetically similar and suitable for any type of transplantation. considering the hazards that human reproductive cloning can pose to the resultant person and society, human being reproductive cloning should be restricted. according to the researchers, if cloning is conducted for medical purposes, it will be deemed an ethical condition and allowed to proceed unless it poses harm to humans or animals. however, if cloning methods are used just for testing and experimentation, they must be prohibited and regarded unethical and illegal. references 1. c. hugo, m. abdulla, i. ahmed, a. assawamakin, j. bhak, s. brahmachari and g. calacal. mapping human genetic diversity in asia. science, vol. 326, no. 5959, pp. 1541-1545, 2009. 2. r. schramm and a. paprocki. strategies for the production of genetically identical monkeys by embryo splitting. reproductive biology and endocrinology, vol. 2, no. 1, pp. 1-5, 2004. 3. s. kiyani and y. munir. the issue of human cloning: a review in semitic religions’ context. al-milal: journal of religion and thought, vol. 2, no. 2, pp. 31-49, 2020. 4. h. niemann, w. kues, a. hahn and j. carnwath. somatic cloning and epigenetic reprogramming in mammals. in: principles of regenerative medicine, new york: heslin rothenberg farley and mesiti p.c. pp. 129-158, 2011. 5. b. ma, t. trieu, s. habib and x. chen. protocol for establishing mouse embryonic stem cells to study histone inheritance pattern at single-cell resolution. star protocols, vol. 1, no. 3, p.100178, 2020. 6. l. yaowei, x. wang, q. zhao, x. zhao and m. sun. robotic batch somatic cell nuclear transfer based on microfluidic groove. ieee transactions on automation science and engineering, vol. 17, no. 4, pp. 2097-2106, 2020. 7. s. shafique. scientific and ethical implications of human and animal cloning. international journal of science, technology and society, vol. 8, no. 1, pp. 9-17, 2020. 8. a. marx. the ethical and scientific debate behind human reproductive cloning by somatic cell nuclear transfer, 2018. 9. n. sinha, s. patil, u. kesigan, t. chaitanya, s. panigrahy and g. tandon. ethical concerns in animal cloning: possible risks and assessment. global bioethics enquiry, vol. 7, no. 3, pp. 128-135, 2019. 10. r. jain. human cloning: ethical issues and legal implications. indian journal of law and legal research, vol. 2, no. 1, pp. 35-46, 2021. 11. y. peng, x. huang and q. zhou. ethical and policy considerations for human embryo and stem cell research in china. cell stem cell, vol. 27, no. 4, pp. 511-514, 2020. 12. a. al delawi. role of ethics in islamic thinking in the activation of accounting information quality. utopía y praxis latinoamericana, vol. 6, pp. 179-187, 2019. 13. a. sachedina. islamic ethics and the genome question, edited by mohammed ghaly. journal of islamic ethics, vol. 1, pp. 1-4, 2021. 14. a. massoudi. reviving performance by adopting chalemleon style of leadership. journal of management and business education, vol. 5, no.1, pp. 1-19, 2022. 15. nasrullah, r. k. iqbal, s. bibi, s. muneer, s. bibi and f. n. anwar. ethical issues of human cloning. journal of medical sciences, vol. 40, no. 3, pp. 103, 2020. 16. m. häyry. ethics and cloning. british medical bulletin, vol. 128, no. 1, pp. 15-21, 2018. 17. c. okechukwu and j. emeribe. ethical implications of in vitro fertilization (ivf) and human cloning through the lens leon kass. journal of african studies and sustainable development, vol. 4, no. 1, pp. 88-104. 2021. 18. i. karl, m. levanduski, a. vidali, n. husami and v. goudas. human embryo twinning with applications in reproductive medicine. fertility and sterility, vol. 93, no. 2, pp. 423-427, 2010. 19. m. omidi, m. khalili, a. rahimi, s. nottola, f. anbari, a. faramarzi and m. palmerini. efficacy of the in vitro splitting of human preimplantation embryos from art programs. turkish journal of medical sciences, vol. 51, no. 1, pp. 68-75, 2021. 20. k. simmet, e. wolf and v. zakhartchenko. manipulating the epigenome in nuclear transfer cloning: where, when and how. international journal of molecular sciences, vol. 22, no. 1, p. 236, 2020. 21. l. keefer. artificial cloning of domestic animals. proceedings of the national academy of sciences, vol. 112, no. 29, pp. 8874-8878, 2015. 22. n. rivron, m. pera, j. rossant, a. arias, m. goetz, j. fu and s. brink. debate ethics of embryo models from stem cells. nature, vol. 564, no. 7735, pp. 183-185, 2018. 23. p. rastogi and a. kakkar. ethical issues in cloning. journal of critical reviews, vol. 7, no. 10, pp. 996-1000, 2020. 24. ethics committee of the american society for reproductive medicine. human somatic cell nuclear transfer and reproductive cloning: an ethics committee opinion. fertility and sterility, vol. 105, no. 4, pp. e1-e4, 2016. 25. t. grønning. in: cloning. in: the wiley-blackwell encyclopedia of social theory. new york: john wiley and sons, pp. 1-3, 2017. 26. m. nasr-esfahani, k. khanbeigi and a. hasannia. qur’anic views on human cloning (i): doctrinal and theological evidences. international journal of fertility and sterility, vol. 15, no. 1, pp. 73-79, 2021. tx_1~abs:at/add:tx_2~abs:at 76 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 76-80 review article fanet drone’s data applications, mobility models, and wi-fi ieee 802.11n standards for real and non-real time traffic ghassan a. qasmarrogy1, areen j. fadhil2 1department of communication and computer engineering, cihan university-erbil, kurdistan region, iraq, 2department of information system engineering, erbil technical engineering college, erbil, iraq abstract data traffic is the most important data transmission between users inside every network. this data traffic can be videos, files, voice, pictures, and many more. it is divided into two types, real-time and non-real-time traffic. most real-time traffic data have a low tolerance for delays during transmission, as they need to be quickly received between communication devices. in this paper, a comprehensive analysis was made to evaluate the two types of data transmitted through fanet drones, with different mobility models and two types of ieee 802.11 2.4 ghz and 5 ghz using the optimized link state routing protocol. metrics such as delay and throughput were measured. this paper gives an important overview of how real-time and non-real-time traffic will be handled during data transmission in fanet networks. keywords: fanet, mobility models, ieee 802.11n, non-real time traffic, real-time traffic introduction in today’s networks, most traffic transmitted between sender and receiver consists of different types of data, such as surfing the web, listening to music, making video calls, downloading files, watching youtube, and many more. this data traffic can be divided into two types of data, real-time and non-real-time traffic. the total network traffic is the total amount of moving data between the sender and the receiver. therefore, the bandwidth will be shared between both real-time and non-real-time applications. in today’s network protocols, some priority will be given to real-time applications as they have a very low tolerance for delay during transmission.[1] there are different types of ad hoc networks, such as manet, vanet, uanet, and fanet. fanet is a group of flying drones that changes its topology frequently depending on the speed and attitude of the drones while communicating using routing protocols to send and receive data between them. the data traffic transmitted between them can be recorded video, voice, or exchange information about sites by transferring files, as shown in [figure 1].[2] using different mobility models, the direction and speed of each drone will change depending on different criteria of the model chosen while exchanging data packets between the nodes, which can give different high and low performance as the routes between the drones will break due to high mobility. real-time traffic is very important to be analyzed and tested through different types of scenarios, as it is the most demanded traffic nowadays. this is the main purpose of this paper. while other non-real-time traffic will show how they can get priorities on the same link during transmission with real-time traffic. the rest of the paper will be organized as follows: section 2 will explain related work for various researchers; section 3 will explain the methodology of the paper; section 4 will give the results and analysis of the paper; and section 5 will conclude the paper. related work fanet is considered very popular these days as many researchers try to analyze its performance and enhance its ability for better properties. in[3] a survey was done for the critical points of fanet to show the performance of fanet architecture, communications, mobility models, characteristics, corresponding author: ghassan a. qasmarrogy, department of communication and computer engineering, cihan university-erbil, kurdistan region, iraq. e-mail: ghassan.qasmarrogy@cihanuniversity.edu.iq received: june 20, 2022 accepted: august 07, 2022 published: august 22, 2022 doi: 10.24086/cuesj.v6n2y2022.pp76-80 copyright © 2022 ghassan a. qasmarrogy, areen j. fadhil. this is an openaccess article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) qasmarrogy and fadhil: fanet standards for real and non-real time traffic 77 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 76-80 and fanet design. it also shows all the recommendations to help the authors work on the same and enhance these parameters. the research lists most approaches that are used to give better results in fanet. in[4] the author evaluates fanet’s different mobility models, real-time traffic, and frequency bands while measuring throughput and delay for best performance using two types of routing protocols. the result shows that 2.4 ghz gives a higher delay but more coverage than 5 ghz. in[5] a recent attempt was made to enhance and optimize the performance of video transmission in fanet with a 5 ghz frequency band while analyzing throughput, retransmission attempts, and delay. their results show that changing some metrics in wlan interface cards can give better performance to the transmission. finally, in[6] the performance of optimized link state routing (olsr) protocols was analyzed and investigated with four different data rates and scenarios while measuring the throughput, network delay, load, and retransmission attempts. the result shows that the olsr protocol was improved and gives better performance than before. fanet parameters this part will give a comprehensive explanation of all the fanet parameters calculated in this paper, such as mobility models, ieee wireless protocols, routing protocols, and data traffic types as follows: mobility models because fanet’s topology changes as it moves, it requires a specific mobility model and routing protocols to guide packets between drones to their final destination. mobility models are very important in guiding and controlling the faent movement in specific areas, as they use a mobility formation to keep all the drones in the range for transmission with a fixed direction, height, and speed.[7] the most commonly used type of mobility model is the random waypoint mobility model (rwpm). this model uses random value numbers and direction for moving and pausing, also uses random values for height and attitude. when the simulation starts, the drones start using the mobility model as these randomness values will be repeated until the end of the simulation. this model is considered very important as it simulates real-life scenarios of fanet drones moving for different life applications such as wsn wireless sensor networks, search and rescue missions, area scanning, and many more.[8] while the second type is known as the gauss–markov gmm mobility model, in this model, a drone starts moving in a specific direction with a specified speed as it reaches the endpoint, then it chooses a different direction and speed than planned. these numbers are calculated with a specific equation that depends on fanet parameters, where each drone movement is not related to other drones in the same fanet topology. [figure 2] shows the different mobility models used in this paper.[9] ieee wifi and routing protocols standards the ieee 802.11n is a wireless standard used to organize the movement of data wirelessly from the sender to the receiver, using multiple antennas to increase the data transfer rate. this protocol can support up to a 600 mb/s speed rate while operating with two different types of frequencies, 2.4 ghz and 5 ghz.[10] the multiple input/multiple output (mimo) can transmit a larger number of data packets compared to the single antenna used with the previous version of wireless standards. furthemore, mimo uses spatial division multiplexing (sdm) technology, in which multiple streams of individual data are transmitted at the same time on one single spectral channel. the technology of mimo sdm permits the bandwidth to be increased for each data stream, which helps to increase the overall speed rate. still, each data stream requires one antenna to send and one to receive; therefore, this technology uses mimo to adjust its transmission.[10] in ieee 802.11n, the 5ghz bandwidth gives more speed compared to the 2.4 ghz bandwidth as it has more nonoverlapping channels that reduce the interference between the frequencies of broadcasted devices. figure 3 shows the different types of channels between 2.4ghz and 5ghz.[11] a routing protocol is needed to guide the packets from the source to the destination. the olsr protocol is used as it gives better results in fanet scenarios to route the packets and helps them reach the destination.[12] this protocol was optimized for the old link-state protocol used in manet; [figure 4] shows the different types of fanet routing protocols. figure 1: different types of mobile ad hoc networks[2] figure 2: different mobility models[8] ba qasmarrogy and fadhil: fanet standards for real and non-real time traffic 78 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 76-80 ospf uses a group of selected multipoint relays (mprs) that forward the received broadcast messages through the flooding process. therefore, using this technique, the overhead of the network will be decreased substantially, as every drone will retransmit the received message when it receives the first message from the main source. [figure 5] shows the olsr mpr’s node.[13] the main three optimizations on olsr can be summarized as follows: 1. the information link status will only be generated by the selected mpr nodes 2. the number of control messages forwarded will decrease as only the selected mprs will broadcast the generated traffic 3. each mpr node can choose to report the main active link connected between them. therefore, these enhancements made olsr provide optimal routes between the sender and the destination, with the lowest delay and higher throughput. furthermore, made it very suitable for large and dense networks with a dynamic number of nodes.[13] network traffic there are two types of network traffic: real-time and nonreal-time traffic. real-time traffic demands are transmitted or delivered on time without any delay and with the highest quality as possible, such as videoconferencing, voip, and web browsing.[14] while non-real-time traffic has lower priority than real-time traffic, which can tolerate delays and some packet dropping, such as emails and web publishing, [figure 6] depicts the various types of data traffic transmitted in the network. results and analysis this paper examines by analyzing two types of transmission data, real and non-real-time traffic in the fanet network using a simulation area of 2000 × 2000 square meters with a 25-flying drone at a speed of 15 m/s and a height of 15 m with two types of mobility models (random waypoint mobility, or rwpm, and gauss–markov gmm). these drones will broadcast the data using wifi ieee 802.11n standards, 2 ghz and 5 ghz, with the support of the olsr routing protocol. the simulated environment was repeated 5 times while calculating the average for optimal values to calculate both throughput and end-to-end delay for both types of traffic. finally, the opnet simulator model was used to simulate the entire scenario, which is considered one of the best simulators on the market for fanet simulations. table 1 shows the simulation parameters used in this paper. the main comparison between 2.4 ghz and 5 ghz wireless protocols is about speed, bandwidth, and interference, as 2.4 ghz gives lower interference and a higher range, while 5 ghz gives more speed and bandwidth with less range coverage. in [figure 7], the throughput of both 2.4 ghz and 5 ghz traffic is shown with both real and non-real-time traffic using the olsr routing protocol, where the highest throughput was shown for video and voice as they have a lower tolerance for delay, while both http and ftp have a higher delay tolerance as they can be sent with more delay. the 5 ghz wireless protocol shows more throughput for both real and non-real-time traffic as it has higher bandwidth, but due to its lowest coverage range, its performance decreases figure 5: mpr’s nodes of olsr routing protocol[13] figure 3: different types of channels of 2.4 ghz and 5 ghz[11] figure 4: fanet routing protocols[12] figure 6: different types of data traffic[14] qasmarrogy and fadhil: fanet standards for real and non-real time traffic 79 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 76-80 in throughput for the rwpm model as many links break and more nodes randomly fly away from each other, which makes them out of range coverage. the gmm model gives better performance as the nodes move in the same direction or next to each other inside the same coverage area. figure 8 shows the delay for the same scenario of the fanet network, where 5 ghz shows lower delay in gmm as it has more bandwidth, which makes the traffic packet transmitted faster than the 2.4 ghz wireless protocol. however, the rwpm model gives higher delay as most of the links break and packet drops due to the low coverage range of the 5 ghz protocol. moreover, non-real-time traffic shows higher delays than real-time traffic as they have a longer tolerance to delays than real-time traffic for 2.4 ghz conclusion fanet is a special type of mobile ad hoc network that flies for different types of functions, such as military functions, recording, transmitting data, and many more. this data can be real-time data that has a low tolerance for delay and non-realtime data that has a higher tolerance for delay. in this paper, an evaluation has been done to measure the performance of both types of traffic using the olsr routing protocol with two types of mobility models, rwpm and gmm, that are transmitted using ieee 802.11n 2.4 ghz and 5 ghz wireless protocols. in conclusion, real-time traffic has higher throughput and the lowest delay compared to non-real-time traffic. while during retransmission attempts, the 5 ghz wireless protocol gives the best result as it covers only a short distance, while the movement of fanet drones makes the links between the nodes break more frequently and only if these nodes are close to each other. as a result, 5 ghz gives decreased performance in the rwpm model as more nodes fly away from each other, which makes them out of the coverage area. for the future work, it is recommended that more mobility models be used and more recent wireless protocols that support more bandwidth and higher speed rates, as well as more traffic. references 1. h. j. alqaysi and g. a. qasmarrogy. performance analysis of video streaming application over manets routing protocols. international journal of research in computer applications and robotics, vol. 3, pp. 22-28, 2015. 2. d. ramphull, a. mungur, s. armoogum and s. pudaruth. a review of mobile ad hoc network (manet) protocols and their applications. in: 2021 5th international conference on intelligent computing and control systems (iciccs). ieee, pp. 204-211, 2021 3. a. srivastava and j. prakash. future fanet with application and enabling techniques: anatomization and sustainability issues. computer science review, vol. 39, pp. 100359, 2021. 4. g. qasmarrogy. evaluation of flying ad hoc network topologies, mobility models, and ieee standards for different video applications. 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in: 2021 3rd international conference on computer communication and the internet (iccci). ieee, pp. 141-145, 2021. 10. s. rajbhandari, a. v. jalajakumari, h. chun, g. faulkner, k. cameron, r. henderson, d. tsonev, h. haas, e. xie, j. j. mckendry, r. ferreira, e. gu, m. d. dawson, d. o’brienand and j. herrnsdorf. a multigigabit per second integrated multipleinput multiple-output vlc demonstrator. journal of lightwave figure 7: average throughput for real-time and non-real-time traffic figure 8: average delay for real-time and non-real-time traffic table 1: simulation parameters parameters values traffic type video, voice, http, ftp area size 2000×2000 m2 fanet routing protocols olsr fanet number 25 drone mobility models rwpm, gmm simulation time 5 min node speed, height 15 m/s, 15 m wifi ieee 802.11n 2.4 ghz, 5 ghz qasmarrogy and fadhil: fanet standards for real and non-real time traffic 80 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 76-80 technology, vol. 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distributed denial of service attacks on cloud computing environment: a comprehensive review israa t. aziz1, ihsan h. abdulqadder2, thakwan a. jawad3 1computer center, university of mosul, mosul, iraq, 2department of informatics and software engineering, cihan university-erbil, kurdistan region, iraq, 3department of system and control engineering, college of electronic engineering, ninevah university, mosul 41002, iraq abstract the incorporation of numerous technologies into the traditional network makes it difficult to complete new demands such as security. the widespread conception of telecommunication technologies in the last decade has an increased in the number of more attractive security threats. however, the new technology also created many new security concerns, and the threat of distributed denial of service attacks (ddos) attack is one of the major concerns. this paper presents a comprehensive review of state-of-the-art techniques to detect ddos attacks. it aimed to describe various kinds of ddos attacks such as peer-to-peer based, web-based, and internet relay chat-based. in addition, the critical challenges against effective ddos defense mechanisms are also explored. finally, the limitations of several ddos attack detection approaches are highlighted. keywords: distributed denial of service attacks, cloud computing environment, peer-to-peer, internet relay chat introduction cloud computing has developed rapidly over recent years, and it is reshaping the information technology (it) industry. the apparent transparency, large volumes of stored data, and comparable easiness of operability of cloud computing environment (cce) make it vulnerable and easy targets for several kinds of predatory attacks, distributed denial of service (ddos) being major ones like those against cloudflare and spanhaus which are alarmingly and increasingly used for exploiting sample network management protocol (snmp). some of the major ddos attacks are flooding, spoofing, user to root, port scanning, oversized xml, coercive parsing, reflective attacks, and so forth. the ddos attacks have been reported to have significantly high chances of occurrence, first, because the tools of launching ddos are widely available, and second, because of the apparent lack of effective, timely mechanisms to defend against them. due to the frequency of attacks such as ddos, cce’s fullest gains and advantages stand highly compromised and negated. there is the need for in-depth, evidential, and research-validated studies on the topic and its many ramifications over time, ddos has evolved, and remedial actions prompted to address them and offer permanent solutions. the statement of the problem is the exact degree and magnitude of destructive powers of ddos cannot be quickly quantifiable, cce could best address these issues and find solutions are also needed. the ddos attacks on cce hold immense potential to cause significant damages and detriment, especially if uncontrolled and unremedied. in this paper, discussion of various studies carried out concerning cces is presented with more emphasis is given on the studies about ddos is done. authors in kobusińska et al.[1] aimed to determine cloud computing’s concerns and challenges, whereby they discussed grid computing and service-oriented computing, which are two related computing paradigms. they also sought to pinpoint the relationships of these two computing models with cloud computing and identified various challenges therein. on the other hand, tahirkheli et al.[2] pointed out two primary reasons that complicate the assessment of the security impact of cloud computing. according to tahirkheli et al.,[2] the current discourse on cloud computing security issues uses the terms “threat,” “risk,” and “vulnerability,” interchangeably, while they have their respective definitions. moreover, the authors claim that not every issue that’s raised is particular to cloud computing. in order to build a corresponding author: israa t. aziz, computer center, university of mosul, mosul, iraq. e-mail: israa_aziz@uomosul.edu.iq received: november 12, 2021 accepted: january 27, 2022 published: march 30, 2022 doi: 10.24086/cuesj.v6n1y2022.pp47-52 copyright © 2022 israa t. aziz, ihsan h. abdulqadder, thakwan a. jawad. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) aziz, et al.: ddos attacks on cloud 48 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 47-52 broader perspective of the threats within the cce, they delved into the meaning of cce. cloud computing is a model for enabling convenient, on-demand network access to a shared pool of configurable computing resources (e.g., networks, servers, storage, applications, and services). it can be rapidly provisioned and released with minimal management effort or service provider interaction, according to the us national institute of standards and technology.[3] within the terms used to define cloud computing and, by extension, cce, it is clear that the environment is set on on-demand self-service, implying that consumers’ resources automatically come into play, which, in itself result in fewer human interactions with the provider of the services.[4] the computing resources, in this case, include cpu time, software use, and network storage, which are engaged on a convenient, self-serve basis. second, the term “broad network access,” as used in the definition, postulates a scenario whereby various clients’ applications can use with diverse platforms access computing resources conveyed over networks.[5] common user platforms include personal digital assistants, laptops, and mobile phones, which translates to increased exposure probability heightened vulnerability. third, cce’s resource pooling aspect translates to the multi-tenancy or virtualization models.[6] these models make it possible for cces to serve multiple consumers, with the computing resources assignment varying now and then. rapid elasticity and measured service are the other terms in the definition of cces. rapid elasticity pertains to the immediate availability of computing resources instead of persistent. the outstanding issues with such a setting are that consumption can increase rapidly during peak requirement times. on the other hand, measured service brings the cces metering individual consumers’ usage even though the computing resources are combined and shared by numerous clients. in mthunzi et al.,[7] the authors analyzed the cloud computing influences each risk factor in a pool of various risks in computing. they also assessed the vulnerability issue while singling out cloud computing from the general computing domain. therefore, they asserted that cloud computing enhances some well-understood vulnerabilities and introduces new ones. akherfi et al.[8] argues that the distinctions between cloud computing, service-oriented computing, and grid computing should be investigated. as a result, they developed research topics to articulate future study problems and cloud computing objectives. more importantly, they sought to establish the benefits of the three paradigms within the context of their coexistence. the authors explain in the definition of cce to point out the risks in cloud computing; they should have depicted the strengths associated with the terms used in the definition. their dwelling on the vulnerability shows their biasness in their assessment. in al-dhuraibi et al.,[9] the authors investigate cloud computing’s advantages, and they offer the future development issues on cces, among them, the opportunity to reduce the vulnerability of cces. however, the inherent nature of the technology used in the cce implementation exposes vulnerabilities, including virtual machine escape, session riding and hijacking, and insecure or outdated cryptography. in addition to the intrinsic technologically based vulnerability, there are cloud-specific vulnerabilities.[10] such vulnerabilities can be associated with the very nature of the cloud computing setting, thus implanted in the definition of cce. according to chahal et al.,[11] the tools for launching ddos are widely available. the author further posits a lack of an effective mechanism that defends ddos attacks in a reasonable amount of time. after analyzing the various techniques available to reduce ddos attacks, they concluded that none of the studied methods met their requirement. their research sets a significant basis on the threat posed by ddos to the cces. their further assertion, based on their literature review, that ddos attacks occur more frequently than reported, is a further ramification of ddos. nevertheless, ddos remain a daunting challenge in cloud computing environments, and it requires complex simulations to cope with it. typically, ddos flood traffic into servers, systems, or networks to overwhelm the victim resources, making it difficult for legitimate users to use them. furthremore, the attacks are directed from multiple attack systems (distributed), thus complicating their detection and defending. in rossow et al.,[12] the authors closely analyzed twelve peer-to-peer (p2p) botnet variants still active. as shown in table 1, each p2p botnet has its communication protocol, message propagation technique, communication direction, command-and-control (c&c) architecture, and purpose. the major goal is written in capital letters. s = spam, t = credential theft, c = click fraud, d = ddos, m = bitcoin mining, n = network services=pay-per-install rossow et al.[12] adapted table 1. unlike p2p and internet relay chat (irc) botnets, hypertext transfer protocol (http) botnets employ a wide range of http services, making it difficult to prevent them. attacking with a trojan allows the attacker to download a zombie agent, or the trojan itself may contain one. the number of trojan attacks has increased, with most of them aimed at web servers. automated tools that exploit flaws in programs that listen for connections from remote hosts are also being used by attackers to access systems. computer security experts and law enforcement agencies can easily take down centralized botnets. as a result, botnet operators have looked for novel ways to fortify their botnets’ infrastructures. some botnet operators have reconfigured their botnets to employ p2p infrastructures to achieve this goal. because they have no single point of failure, many p2p botnets are significantly more resistant to takedown attempts than centralized botnets. p2p botnets, on the other hand, are vulnerable to specific table 1: comparison and overview of p2p botnet family protocol prop. dir. c&c propose kelihos custom gossip pull hybrid c,d,m,n,s miner custom gossip pull hybrid d,m,p nugache custom gossip pull p2p d,t sality custom gossip pull p2p d,n,p storm overnet routing pull hybrid d,s,t waledac custom gossip pull hybrid d,s,t zeroaccess custom gossip pull p2p p zeus custom gossip both hybrid d,p,t aziz, et al.: ddos attacks on cloud 49 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 47-52 types of assaults, such as node enumeration and poisoning.[13] the important outlines of this paper are providing a review for specific ddos attacks on cce, as well as the kind of detriment could cause. while the chief aims of this paper are to identify the various kinds of ddos attacks, their destructive capabilities, best these issues could be counter-attacked and resolved for the benefit of all stakeholders along the cloud continuum, preferably as permanent solutions. this paper’s structure is as follows: section 2 offered the analysis of specific ddos attacks. section 3 explained the remedies for ddos were offered previously. section 4 concludes this paper. analysis of specific ddos this section offers specific ddos attacks on cce based on published reports and other credible sources. therefore, this paper will use reports on the nature and frequency of occurrence, the extent of detriment, and reports on efforts to deal with ddos. the first step is to analyze the general architecture of ddos, which is shown in figure 1. ddos attacks are known to originate from multiple sources or multiple locations simultaneously, as evidenced by their general architecture. the participants in a ddos attack are also visible.[14] second, the importance of ddos issues can be ascribed to (1) numerous attack machines can create more attack traffic than a single attack machine, (2) several attack machines are more difficult to turn off than a single attack machine, and (3) each attack machine’s behavior can be stealthier, making it difficult to detect and shut down. therefore, the key challenge from ddos is the complexity of having a defense mechanism against them. specific ddos, based on their facilitation mechanism, plus the significance of the detriment they could cause, is as discussed here: botnet-based ddos a botnet is a network of zombie computers that have been designed to accept commands without the owner’s knowledge.[15] the infrastructure and tools currently evident in botnet launching areas are presented. notably, the critical challenges against effective ddos defense mechanisms are twofold: (1) to initiate ddos flooding attacks, a large number of zombies are used, and (2) zombies ip addresses are usually faked under the attacker’s control. thus, the attacker’s possible to add more attack machines dwindles the clients’ ability to purchase more incoming bandwidth, eventually crashing a website completely over time.[16] an attacker (master) controls a group of zombies, forming a botnet. thus, botnets consist of masters, handlers, and agents (bots) whereby the master communicates to the bots through the handlers. three categories of botnets are p2p-based, web-based, and irc-based. p2p-based these are affected through a decentralized group of malwarecompromised machines working in cooperation with an attacker without their owners’ knowledge.[17] they, therefore, lack a c&c server. these botnets avoid detection and make it hard for security researchers to access communication by being decentralized. moreover, investigators cannot take down an entire network when they detect a single bot since the p2p botnets lack a command-and-control server. while bots keep communicating, the botmaster retains commanding them, usually by digital signing. mainly, digital signing is achieved through asymmetrical encryption, which requires two keys (public and private).[18] this ensures that while one key is used to encrypt a message, the message can only be decrypted with the other key. the private key is thus kept private, while the public key is embedded in the bot. the master encrypts commands with the private key, while the bots decrypt them with the public key. bots that can accept incoming connections are called servers, while bots that cannot take incoming connections are called workers. nodes or peers are the terms used to describe the servers. to receive commands, the workers must connect to one or more peers. workers are distributed among several nodes to avoid being taken down, and they can relocate to another node if one is taken down. since this infrastructure entails bootstrapping, which offers some sort of signing; these botnets are able to prevent themselves from being hijacked by security developers. still, the bootstraps are crucial nodes, and if all of the bootstrap servers were taken down simultaneously, it would have no effect on the bots currently in the botnet, but it would prevent new infections from happening to join. while such figure 1: the general architecture of ddos aziz, et al.: ddos attacks on cloud 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 47-52 a safeguard would be tough to implement, it would also be temporary, as the botmaster would temporarily stop infecting new computers while setting up new bootstrap servers. p2p bots include sinit, phabot, spam thru, nugache, and peacomm (storm). web-based these are more recent botnets. in this setup, the commands from the botnets to the bots are communicated through the http communication protocol, making it challenging to track the control structure and the ddos command. such botnets conceal themselves within legitimate http traffic; thus, they are stealthier.[19] using the web requests, each web bots downloads instructions periodically as shown in figure 2. although it might be expected that http botnets would try to blend into regular http traffic (since they use html), a significant number of them use specially registered domain names for their purpose.[20] a proof of this claim can be shown by team-cymru.com, which studied some identified http c&cs. 46% of http botnets used a dedicated domain name, 26% used free hosting such as funpic.de and freehostia.com. based on location, for the 131 unique addresses used, for the one day, 63 of the ip addresses were located in the united states, 11 were in russia, and ten were in china.[21] an example of a web-based ddos botnet is the stacheldraht attack. these ddos tool invaders employ a layered structure and link to handlers through a client program. the client program infiltrates the handler’s systems and sends commands to the zombie agents, allowing the ddos assault. different types of internet packets, such as internet control message protocol (icmp), user datagram protocol (udp), and transmission control protocol (tcp), can be used by stacheldraht. each handler can control up to a thousand agents in some instances. the situation can even be made worse because the client can even consent to the ddos attack to become part of their machine. for example, the group anonymous used stacheldraht called operation payback. some examples of web-based bots include blackenergy, kraken, pushdo, cyber bot, and blacksun rat.[22] irc-based botnets this setup uses an online text-based instant messaging protocol to interact between servers and features a client/server structure. as a result, attackers use irc channels as handlers to deliver commands to bots utilizing legal irc ports. as a result, tracking the irc ddos’s command-and-control system is challenging. furthermore, because irc is a stand-alone software or set of scripts that connect to irc as a client, attackers do not need to keep bots on their site because they may access a list of all available bots by signing on to irc servers.[23] although irc botnets have historical operations such as game manager in games, bartenders, and puppe, these bots have evolved over time to provide special services, including access to databases, maintaining access lists, and managing channels. still, the usual large volume common to irc servers makes it easy for the attacker to hide.[24] defense against these ddos can be affected by the defender capturing their centralized command and control servers. irc botnets include sdbot, agobot, gdbot, and spybot.[25] analyzing online social sites threats reveals that irc botnets pose a significant threat. in fact, according to unpublished sources such as blogs on irc sites, it is evident that irc has continued to improve, which poses even more threat as they are building armies of bots for large-scale attacks. the worm dorkbot, also known as nrgbot, is one of the most common irc botnets, capable of stealing passwords, blocking security updates, downloading more malware, and even conducting ddos assaults using infected devices. “the bot joins a specific irc channel on an irc server and waits there for further command. this allows an attacker to remotely control this bot and use it for fun and profit. the communication between bots and their controllers is somewhat bloated; a more straightforward communication protocol would suffice. but irc offers several advantages: irc servers are freely available and are easy to set up, and many attackers have years of irc communication experience.[26] remedies and discussion in this section, we analyze the remedies for ddos offered in the previous area by looking at the reported deliberations and analyzing their success. this paper seeks to set the way towards successful contemplation on ddos. in general, defense tactics comprise a combination of attack detection, traffic classification, and response tools to block illicit traffic while allowing valid traffic.[27] application-level key completion indicators application-level ddos attacks on cces can be based on an application layer analysis to identify whether incoming traffic packets are desirable or not, allowing elasticity decisions to be activated without the negative economic consequences of figure 2: http ddos trends of prevalent based on country aziz, et al.: ddos attacks on cloud 51 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 47-52 a ddos attack.[28] upstream filtering with upstream filtering, as the name suggests, the traffic to the victim computer needs to be cleaned by passing it through digital cross-connects, proxies, direct circuits, or proxies. such upstream cleaning ensures that ddos is separated and only the “safe” traffic reaches the server.[29] applying front end hardware using front-end hardware, intelligent hardware can be deployed on the network before traffic reaches the servers. as data packets enter the system, this hardware examines them and classifies them as a priority, regular, or dangerous. ips-based prevention intrusion prevention systems (ips) can be effective if the attack has signatures linked with it. unfortunately, genuine content with malicious intent is a regular ddos trend. furthermore, intrusion-prevention systems designed for content recognition cannot detect ddos attacks based on behavior. a rate-based intrusion prevention system (rbips) is designed to analyze traffic granularly and continuously scan the traffic array for anomalies.[30] the legitimate traffic must be let pass through while the suspicious one is filtered off. switches many switches include rate-limiting and access control list (acl) capabilities. to identify and correct dos assaults, some switches include deep packet inspection, delayed binding (tcp splicing), bogon filtering, automatic traffic shaping, bogus internet protocol (ip) filtering, and system-wide rate restriction via wide area network (wan) link failover and balancing and automatic rate filtering.[31] switch treatments only if ddos attacks can be avoided while they are used. the synchronize (syn) flood attack, for example, can be mitigated by employing tcp splicing or delayed binding. still, bogon filtering can be used for attacks going to dark addresses or coming from dark addresses, while deep packet inspection can be used for content-based ddos. in addition, if both links have dos/ddos prevention mechanism, wan-link failover will work. and more still, setting the rate thresholds correctly can ensure that the automatic filtering works. blackholing and sinkhole all communication to the attacked domain name system (dns) or ip address is routed through a “black hole,” a non-existent server, or a null interface. the internet service provider can manage such a hole to be more efficacious.[32] moreover, a dns sinkhole can route traffic to a valid ip address for analysis, rejecting the bad packets and allowing the good packets through. firewalls with firewalls, a simple rule could be added, such that, based on the protocols, originating ip addresses, or ports, all incoming threat traffic is denied access.[33] however, this approach will not be able to block more complex attacks. for instance, web service; say through port 80, on experiencing an attack, it might not be possible to deny all incoming traffic through such a service port, as that would mean stopping the legitimate server’s traffic.[34] routers routers and switches are similar in that they both include ratelimiting and acl capabilities. they, like the switches, are also configured manually. however, it is essential to note that most routers are vulnerable to ddos attacks. p2p remedies almost all p2p botnets have a vulnerability in the p2p sharing mechanism. as previously stated, the nodes must keep and share a list of other nodes with the workers in order to distribute the workers around the huge number of nodes.[35] providing each node with a list of other nodes would take an excessive amount of time, if not impossible, for the botmaster to do manually.[36] as a result, the nodes do it on their own. if a new node is found to accept connections, the node to which it is linked adds it to its node list, which it then shares with other nodes. this could allow security researchers to inject hostile computers into the network through a loophole. they can introduce employees and other nodes with bogus ip addresses in this situation and share a list of other malicious nodes. if this is done and enough resources are put in place, the new rogue nodes can become a significant botnet component, thereby separating genuine nodes from workers. if the inserted malicious nodes populate the network, the workers will identify with the nodes and will have a much lower likelihood of joining the botnet. conclusion ddos remain a daunting challenge in cloud computing environments, and it requires complex simulations to cope with it. typically, ddos flood traffic into servers, systems, or networks to overwhelm the victim resources, making it difficult for legitimate users to use them. furthermore, the attacks are directed from multiple attack systems (distributed), thus complicating their detection and defending against. irc botnets pose a significant threat as they have continued to improve, which poses even more risk as they build armies of bots for large-scale attacks, the key challenges against effective ddos defense mechanisms are two-fold: (1) a large number of zombies is employed to launch ddos flooding attacks and (2) zombies ip address is typically spoofed under the control of the attacker. thus, the attacker’s possibility to add more attack machines dwindles the clients’ ability to purchase more incoming bandwidth. some remedies for ddos include application-level key completion indicators, upstream filtering, applying front-end hardware, ips-based prevention, using switches and firewalls, and blackholing and sinkholing. since ddos keeps evolving and becoming more sophisticated, this field of their remedies requires more research and development to stay at par with their development, if not to be ahead of them. in the future, this work is planned to be extended with the use of the software-defined network (sdn) and networking function virtualization (nfv) in a cloud environment to come up with new optimization algorithms to solve ddos attacks. aziz, et al.: ddos attacks on cloud 52 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 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a. welzel, c. rossow and h. bos. on measuring the impact of ddos botnets. in: proceedings of the seventh european workshop on system security, pp. 1-6, 2014. 32. p. amini, r. azmi and m. a. araghizadeh. analysis of network traffic flows for centralized botnet detection. journal of telecommunication, electronic and computer engineering (jtec), vol. 11, pp. 7-17, 2019. 33. a. k. sood, s. zeadally and r. j. enbody. an empirical study of http-based financial botnets. ieee transactions on dependable and secure computing, vol. 13, pp. 236-251, 2014. 34. k. neupane, r. haddad and l. chen. next generation firewall for network security: a survey. in southeastcon 2018, pp. 1-6, 2018. 35. l. böck, s. karuppayah, k. fong, m. mühlhäuser and e. vasilomanolakis. poster: challenges of accurately measuring churn in p2p botnets. in: proceedings of the 2019 acm sigsac conference on computer and communications security, pp. 2661-2663, 2019. 36. m. eslahi, m. rohmad, h. nilsaz, m. v. naseri, n. tahir and h. hashim. periodicity classification of http traffic to detect http botnets. in: 2015 ieee symposium on computer applications and industrial electronics (iscaie), pp. 119-123, 2015. tx_1~abs:at/add:tx_2~abs:at 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 research article an application of logistic regression modeling to predict risk factors for bypass graft diagnosis in erbil azhin m. khudhur, dler h. kadir department of statistics and informatics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq abstract in the medical world, predictive models for assessing operative risk using patient risk factors have gained appeal as a useful tool for adjusting surgical outcomes. the goal of this study was to see if there was a link between the severity of atherosclerosis as determined by angiography and changes in several key biochemical, hormonal, and hematological variables in patients who had coronary artery bypass graft (cabg) surgery. this study included 100 adult patients who had coronary angiography, as well as a standardized case– control study of acute myocardial infarction that included 60 healthy people. in addition, not all investigations of heart attack disorders were concerned with modeling; rather, they were all concerned with classification. a  family of generalized linear models called binary logistic regression was used. because most phenomena’ outcomes have only two values (alive/dead, exposed/not exposed, presence/absence, etc.), logistic regression analysis is a common method and plays an important role in health science. overall, 62.5% of individuals were grouped into surgical bypass grafts, while 37.5% were healthy people. hemoglobin a1c was wisely significant, and the odds of one unit increase led to roughly 7.488 times higher. age and body mass index had quite high and substantial effect parameters with a 1.2 times higher likelihood than those who have smaller bmi and younger. according to the study, smokers were more likely to be at risk of undergoing bypass surgery by 4.18 times. however, there was no significant link between gender, screening creatinine, cholesterol, triglycerides, high-density lipoprotein levels, lower density level, systolic blood pressure, and diastolic blood pressure with the outcome variable. keywords: bypass graft, glms, risk assessment of coronary artery bypass graft surgery, logistic regression model, explanatory analysis, odds ratio aim of the study the aim of this study is to examine factors affecting undergoing bypass surgery using multiple logistic regression. this can be done by assessing the contribution of factors that are linked to the probability of being in surgeries between patients and healthy people. research questions 1are the factors significantly correlated to bypass graft? 2what is the greatest procedure to investigate the connection between those factors and the probability of receiving surgery due to bypass graft? background to the study coronary heart disease can be defined as a disease that plaque (a waxy substance) constructs inside the coronary arteries. oxygen blood is supplied to the heart by these arteries. bypass grafting surgery is a kind of surgery that enhances the amount of blood flow to the heart. multiple coronary arteries can be bypassed by the surgeon in just one surgery.[1] from several of each specific trial or meta-analysis of various trials identifying whether cabg is better than percutaneous coronary intervention (pci) and larger result for diabetic patients with multivessel coronary artery disease (cad) at 5 years or longest follow-up, incorporating evidence from several studies, in diabetic patients with multivessel cad. the basic notion that cabg revascularization improves survival over pci is instantly supported by bayesian approaches. in diabetic patients with multivessel cad, cabg is recommended that advantage clearly exceed risk in most, if not all, diabetic corresponding author: azhin m. khudhur, department of statistics and informatics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq. e-mail: azhin.khudhur@su.edu.krd received: march 31, 2022 accepted: may 5, 2022 published: june 10, 2022 doi: 10.24086/cuesj.v6n1y2022.pp57-63 copyright © 2022 azhin m. khudhur, dler h. kadir. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) khudhur and kadir: logistic regression for predicting bypass graft 58 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 patients with multivessel cad. a suggestion of positive results in general survival in freedom declares that cabg appeared to negotiate greater survival than did pci in a single supporter of patients at an individual point in time.[2] marshall et al.[3] as used bayesian logit model for the risks associated with the coronary disease of bypass. he stated that bayesian approach is more popular nowadays among other approaches to predict mortality between patients undergoing bypass surgery. the proposed new model uses data from 12,712 patients undergoing bypass grafting procedures collected between 1987 and 1990. the authors have provided a comparative analysis between bayes theorem, logistic regression, discrimination analysis, and bayesian logit models. their data are divided into a train data set and test data set. the collected data were from various factors measured during bypass surgery and there were missing data in some variables. the data have been obtained from more than 6000 cabg surgery between 1987 and 1992 and they found outstanding risk factors that were significant. researchers gathered data at several sites across the united states depending on the number of cabg cases performed each year and surgeon experience with beatingheart procedures at each location. the study was limited to 3 years, from 1999 to 2001, to avoid any bias resulting from the centers’ early learning experiences. the database comprised patients who had pre-operative or original surgery. the authors compared various groups to determine selection criteria for mortality and morbidity. then, to eliminate selection bias, computer programming was utilized. data on patients who had heart surgery were eventually acquired and the date was then entered into a logical database. the data were evaluated using multivariate logistic regression to predict risk factors affecting mortality in 17401 isolated cabg patients. the data include 7283 (41.9%) offpump coronary arteries and 10118 (58.1%) routine cabg with cardiopulmonary bypass procedures. missing data were also considered as it accounted for more than 10%. to find predictive risk factors for mortality, researchers used the multivariate logistic regression approach. sub-groups that are almost certain to benefit were identified. the parsonnet risk model is a logistic regression model that was frequently used to characterize variation among treatment groups which can affect patient selection and results, with 47 potential risk factors investigated preoperatively to detect risk. this model is used to compare anticipated and observed death rates. with cardiopulmonary bypass as an independent variable, backward elimination techniques were employed to find significant predictors of operation mortality from a collection of 20 preoperative risk factors. a series of alternate regression models were investigated with mortality and extensive morbidity as outcome variables to discover subgroups that may benefit from the beating-heart procedure.[4] risk improvement plays an increasing role in assessing health-care quality. through the evaluation of hospitals, the implementation has become a necessary issue that needs a selection of convenient score systems. the study’s goal was to see if administrative data could be used to predict mortality in patients undergoing cabg in the hospital. between 2000 and 2001, data were gathered from the administrative database of hospital discharge abstracts in the italian region of emilia romagna. the method used for the present study was multiple logistic regression to assess the efficiency with which mortality for patients can be predicted by conforming to each various risk adjustment procedure used.[5] menard[6] as used statistical analysis on bypass graft data to investigate the relation of angiography to hematological and some biochemical variables in coronary artery patients. the study aimed to examine the relation between harshness of coronary artery disease due to angiography and alteration of some necessary biochemical, hormonal, and hematological variables in underwent coronary artery bypass graft (cabg) patients. they selected 80 adults who underwent coronary angiography patients. the present study of menard[7] used statistical modeling procedures including multivariate regression analysis on the demographic status of the patient that is used to examine the corporation of different risk factors influencing individual medical techniques and predict the effect of any risk factor on the outcome. this would commonly imply a logistic regression analysis. in 1985, the parsonnet scoring system was utilized to adjust mortality rates for the 1st time. the parsonnet scoring figure 1: logit and inverse logit functions khudhur and kadir: logistic regression for predicting bypass graft 59 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 system identified 16 risk indicators that could affect the risk of a patient undergoing heart surgery. the considered factors were gender and age of the patients. patients with a history of diabetes or blood pressure or both were separated as the independent variables. generalized linear models (glms) to make glms more relevant than general linear models, we must demonstrate some key properties, the most essential of which is that the least square estimate is no longer valid and maximum likelihood estimation is used instead. the most important elements are listed below:[8] 1random element: this element is related to the response variable’s probability distribution and is to be of the exponential family (normal, binomial, poisson, gamma, and negative binomial). each defines the response variable’s level values as well. 2systematic element: this part denotes the model’s independent (covariates) variables, which can be continuous or categorical, as well as interaction terms between predictors and polynomial functions of predictors. 3function of the link: it is generally concerned with the relationship between the random (response variable) and systematic components (predictors). the factors influence the predicted value of y: g x x x xp p i� � � � � ��� �� � � � �0 1 1 2 2 the link function g (x) and the parameters β 0 , β 1 and so on must be approximated. the three most typical link functions are shown below. 1. identity link: it is represented as g (m) = m in mathematics and is utilized in the typical linear models. 2. log link: it is represented as g (m) = log (m) in math, and it is usually utilized in log-linear models for count data (non-negative numbers). 3. logit link: it is written as g m ln m m � � � � � � � � � �1 in math. as well as for binary data (logistic regression). logistic regression model in medicine and health research, one of the most typical applications of glms is to create a responsive model between response variables with only two outcome values (e.g., alive/ dead, presence/absence, etc.) and explanatory factors, which can be any sort of data, continuous, or categorical, as from figure 1, its shape is seen as s-shape and most of the time it is recognized as s-shape. due to the structure of the response variable, there were two scenarios when glm application was required, as indicated in (binary type).[9] the first was to forecast the likelihood of survival (whether it had survived or not), and the second was to predict the likelihood of burrowing. rees and dineschandra[10] also looked at the association between the presence/absence of species in forest shrubs as a response variable versus a series of predictors including distance to nearest woodland, stand age, stand area, and so on. logistic model and parameters the logistic regression model can be expressed as follows: � � � � � � �x e e x pxp x pxp � � � � � ��� � ��� 0 1 1 0 1 11 where π(x)π(x) is the probability that y i =1 (i.e., the event is present) for a collection of specified x i ’s and βs are parameters that need to be estimated. to make the parameters easier to comprehend, we can simply convert them to odds and interpret them the same way we did in the 2 × 2 contingency table, which is the likelihood of an event occurring compared to its inverse. odds x x � � � � � � � �1 if the odds are more than one, the chance of y i =1 is greater than the reverse, and vice versa. by putting the coefficient values on an exponential function, we can simply transform logistic regression odds to odds and odd ratio. the natural logarithm can then be applied to the odds formula to make modeling easier.[11] g x ln odds ln x x � � � � � � � � � � � � � � � � � ( ) � �1 then g(x) can be controlled as follows: g x x x xp p� � � � � ���� � � �0 1 1 2 2 it is also worth noting that the maximum likelihood estimation is processed and the values are produced through computing due to their complexity. a test called the likelihood ratio test (lrt) is used to determine whether an overall model is significant, as it compares the likelihood of the null model (i.e., a model with only intercept).[12] the lrt functionality is almost similar to the f-test in linear regression and its formula is as follows: g ln l l lnl lnl0 2 0 02 2� � � ���������� where l represents the entire model’s probability and (l 0 ) represents the null model’s likelihood. the statistic (g 0 ) has a k degree of freedom (df) with an approximate two distribution (k=number of predictors in the entire model). however, we evaluate the complete and saturated model employing deviance (sse in linear regression) using a special type of lrt.[13,14] the deviance is an unexplained measure of variation in the data. d ln l l lnl lnl lnlsat full sat full full� � � � �2 2 2 ����� ����� ( ) ( ) furthermore, when it comes to determining the significance of parameters, the wald test[4] is a method for evaluating the null hypothesis. h j0 0:�� � h j1 0: � � materials and methods dataset this study is a case–control study so 100 adult patients underwent coronary angiography at erbil cardiac center khudhur and kadir: logistic regression for predicting bypass graft 60 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 and data were obtained from them. patients were followed up until the end of december 2020. data were also obtained from 60 healthy people who underwent the same coronary angiography. the obtained data were collected from both patients and healthy people. the data include gender, age, smoke, blood sugar, creatinine, weight, length, body mass index (bmi), hemoglobin a1c (hba1c), blood urea nitrogen, cholesterol, lower density level, high-density lipoprotein levels (hdl), triglycerides (tg), systolic blood pressure (sbp), and diastolic blood pressure (dbp) in erbil city. illustrating and reporting logistic regression analysis result it is preferable to construct descriptive statistics for the data and examine how the variables connect to the outcome variable. table 1 indicates that some independent factors and the dependent binary variable may have a relationship with the test value. we first demonstrate the continuous variables in accordance to control and patient groups, it is clear that age has quite an influence to get affected with the bypass graft. the result shows that the mean age for the control group is 44.87-years-old, while the patient people tend to be 59.51 and this tells us more into age, more chance to be suffering, and the p-value of the t-test can confirm that with a value <0.05. moreover, the high value of blood sugar is more likely to put an individual at risk of heart problems, as from the initial output the mean values are far away from each other among the control and patient people with 105.87 and 180.08, respectively. to support this, the driven t-test was found to be statistical significance since its p < 0.05. another interesting result is for bmi measurement, as it is recorded at 24.7 for people who are in the control group whereas a 28.61 mean value is found for the patient group. based on this, t-test reported that the mean values differed from each other statistically significantly. on the other hand, hdl and sbp were highlighted to be non-statistically significant with p-values (0.1030 and 0.6800) greater than 0.05, respectively. likewise, to assess the relationship of the response variable as being in surgery due to bypass graft definition with the categorical variables in the study, results in table 2 are found for gender as well as smoking status. the percentage of males in the patient group is about double toward of females with 64% and 36% constantly according to figure 2, while in the control group, their differences are only 13%. this leads us to confirm that males are in the position to get an operation due to bypass graft concern and the chi-square test shows that there is an association between these two variables. similarly, smoking seemed to have a quite strong influence as shown in figure 3. logistic regression analysis the dataset was subjected to a multiple logistic regression analysis to determine the likelihood of some biographic, health, and other parameters having an impact on being into surgery. rstudio version 1.4.1717 was used to conduct the analysis. we first implemented the simple logistic regression plot for each of the table 2: cross-tabulation table of chi-square for gender and smoking versus outcome variable variables control patient chi-square p-value n % n % gender male 34 56.67 64 64 0.84 0.0357 female 26 43.33 36 36 smoke yes 10 16.67 35 35 6.24 0.013 no 50 83.33 65 65 table 1: descriptive statistical analysis of the associated attributes with the outcome variable variables control patient t. test p-value mean±sd mean±sd age 44.87±14.31 59.51±8.51 −8.12 0.0000 blood sugar 105.87±14.97 180.08±99.57 −5.73 0.0000 hba1c 5.07±0.55 7.21±2.25 −7.23 0.0000 blood urea 28.13±5.44 36.88±15.33 −4.26 0.0000 creatinine 0.7±0.27 0.899±0.32 −4.05 0.0001 cho 142.62±27.19 160.61±40.78 −3.04 0.0028 tg 140.55±65.5 175.4±82.32 −2.79 0.0059 hdl 39.02±8.106 36.51±10.03 1.64 0.1030 ldl 84.38±21.6 100.57±36.38 −3.13 0.0021 weight 69.72±12.56 77.87±15.82 −3.4 0.0009 length 1.67±0.07 1.65±0.1 1.59 0.1134 bmi 24.7±3.09 28.61±5.59 −4.98 0.0000 sbp 127.85±10.61 128.99±19.86 −0.41 0.6800 dbp 74.13±6.09 78.1±12.92 −2.23 0.0271 figure 2: bar-chart demonstration between response and gender variables khudhur and kadir: logistic regression for predicting bypass graft 61 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 variables individually with the curve plot as shown in figure 4, to get a clue about the nature association among the response and covariates. it is important to spot some potential points here as seen that age, bmi, creatinine, blood sugar, and hba1c variables showed a relatively interesting link with the dependent variable. considering the descriptive statistics along with figure 4, the final model has been taken into account for further discussion. first, it was preferred to enter only the significant variables according to wald test statistics called the reduced model compared to the full model as shown in table 3. from table 4, the anova test was carried out using chisquare test to see if the model with fewer variables improved the model. consequently, it turned out that the reduced model is much better than the full model referencing aic, and another important feature is that all parameters were statistically significant. table 5’s result is perhaps the most essential in logistic regression output since it shows how the outcome variable and covariates are related. as seen, all the calculated parameters (wald statistics) were found to be statistically significant with p < 0.05. the odds ratio was calculated by exponentiation b’s value to grasp and interpret the parameter value. the most effective attribute was found to be hba1c, as per the output, it shows a significant and positive impact on the response variable holding other variables constant. the result provides us with (7.488) more chances to be selected for bypass graft operation with one unit increase in this variable in contrast to a low unit. referencing participants who smoke, the odds (4.418) indicate that according to the model, smokers have a 4.418 times higher chance of being classified as a patient than non-smokers. this is in fact enables us that people, who smoke have a higher chance to get into bypassing graft monitoring. in addition, age and bmi covariates were also found to be effective with similar strength in raising people to be considered for bypass graft surgery, and the model predicts that one unit increase in age and bmi is resulting in an increase in the odds of being suffering from bypass graft with 1.180 chance. the odds ratio for blood sugar was computed to be 1.050, with a 95% confidence interval of 0.005–0.107. a case has a 1.050 likelihood of having surgery compared to not having surgery for every one-unit rise in blood sugar, a 50% increase in the probabilities of having surgery. blood urea also played an important role to be effective in increasing risks for the response variable as with one unit increase in blood urea measurement this would lead to having odds (1.092) times chance to face bypass graft surgery compared to low blood urea unit. when comparing low blood urea units to high blood urea units, blood urea had a major role in increasing risks for the response variable, as one unit increase in blood urea measurement resulted in an odds (1.092) times likelihood of having bypass graft surgery. table 5: result of estimated parameters and odds ratio in logistic regression b s.e. wald sig. exp (b) 95% c.i.for exp (b) lower upper (intercept) −31.446 6.414 −4.903 0.000 0.000 −51.569 −17.926 age 0.169 0.038 4.399 0.000 1.184 0.087 0.294 bmi 0.167 0.081 2.049 0.040 1.182 −0.030 0.401 blood sugar 0.048 0.020 2.484 0.013 1.050 0.005 0.107 hba1c 2.013 0.691 2.912 0.004 7.488 0.567 4.180 blood urea 0.088 0.039 2.244 0.025 1.092 −0.004 0.205 factor (smoke) 2 1.486 0.714 −2.082 0.037 4.418 −3.525 0.257 table 4: chi-square test of two fitted models s. no. residual dfa. residual deviance df deviance pr(>chi) 1 153 67.871 2 145 53.833 8 14.039 0.08077 table 3: aic of two fitted models model fitted aic model 1: surgery~age+bmi+blood_ sugar+hba1c+blood urea+factor (smoke) 81.871 model 2: surgery~age+bmi+blood_ sugar+hba1c+ blood urea+creatinine+cho+tg+hdl+ldl +sbp+dbp+factor (gender) + factor (smoke) 83.833 figure 3: bar-chart demonstration between response and smoking variables khudhur and kadir: logistic regression for predicting bypass graft 62 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 table 6 shows that the full model reduces the loglikelihood value, resulting in a substantial difference between the two models. the goodness of fit test was used to evaluate the fitted model, as shown in table 7. there was no actual evidence for lack of fit because the statistical test of hosmerlemeshow was not significant, as it was for pearson chisquare or g 0 2 (deviance). furthermore, the nagelkerke r square suggests a positive impression of the model, which explained 80.8% of the uncertainty in going through bypass graft diagnosis using the factors incorporated in the model. finally, yet importantly, it is worth mentioning how the classification percentage improved after fitting the model with and without the influential factors. the overall percentage of truly predicted cases was 62.5% where there was only a constant in the model, and this had been significantly increased to 93.8%. figure 4: logit model fit of surgery on predictors khudhur and kadir: logistic regression for predicting bypass graft 63 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 57-63 conclusion and recommendation several essential criteria were discovered to be highly successful in increasing high risk among patients undergoing coronary artery bypass grafting, and they may not be the same in different places. our model would be able to properly forecast the instances with an accuracy of 80.8%, which is substantial, and it is worth noting that we arrived at the final model after eight steps of the model selection approach, which is a decent fit to explain the dataset. hba1c had a significant impact on arising the risk, and increasing one unit at hba1c increased the odds of being into surgical admission by 7.488 times, holding other variables constant. age and bmi had quite high and substantial effect parameters, with individuals who are quite an in age and had relatively high bmi led to be at risk with a 1.2 times higher likelihood than those who have smaller bmi and younger. furthermore, blood sugar and blood urea played a significant role in being effective contributors, with 1.05 and 1.09 odds ratios in favor of increasing the risk. according to the study, smokers were more likely to be at risk of undergoing bypass surgery than non-smokers, with an odds ratio of 4.418. gender, screening creatinine, cho, tg, hdl, ldl, sbp, and dbp, on the other hand, were not found to be statistically significant. it is worth noting that this study had its own limitations, such as the small number of cases. to determine how effective the covariates influenced the outcome variable, create a classification table and calculate sensitivity, specificity, and display the receiver operating characteristic curve. references 1. j. alexander and p. smith. coronary-artery bypass grafting. the new england journal of medicine, vol. 374, no. 20, pp. 1954-1964, 2016. 2. c. d. lang, y. he and j. a. bittl. bayesian inference supports the use of bypass surgery over percutaneous coronary intervention to reduce mortality in diabetic patients with multivessel coronary disease. international journal of statistics in medical research, vol. 4, no. 1, pp. 26-34., 2015. 3. g. marshall, a. l. w. shroyer, f. l. grover and k. e. hammermeister. bayesian-logit model for risk assessment in coronary artery bypass grafting. the annals of thoracic surgery, vol. 57, no. 6, pp. 1492-1500, 1994. 4. d. kadir. likelihood approach for bayesian logistic weighted model. cihan university-erbil scientific journal, vol. 4, no. 2, pp. 9-12, 2020. 5. g. q. othman, r. s. saeed, d. h. kadir and h. j. taher. relation of angiography to hematological, hormonal and some biochemical variables in coronary artery bypass graft patients. journal of physics, vol. 1294, no. 6, p. 062110, 2019. 6. s. menard. applied logistic regression analysis. vol. 106. new york: sage. 7. s. menard. coefficients of determination for multiple logistic regression analysis. the american statistician, vol. 54, no. 1, pp. 17-24, 2000. 8. m. j. mack, a. pfister, d. bachand, r. emery, m. j. magee, m. connolly and v. subramanian. comparison of coronary bypass surgery with and without cardiopulmonary bypass in patients with multivessel disease. the journal of thoracic and cardiovascular surgery, vol. 127, no. 1, pp. 167-173, 2004. 9. c. ugolini and l. nobilio. risk adjustment for coronary artery bypass graft surgery: an administrative approach versus euroscore. international journal for quality in health care, vol. 16, no. 2, pp. 157-164, 2004. 10. m. rees and j. dineschandra. risk stratification in assessing risk in coronary artery bypass surgery. in: 19th ieee symposium on computer-based medical systems (cbms’06), pp. 303-308, 2006. 11. j. hilbe. generalized additive models softwar. the american statistician, vol. 47, pp. 59-64, 1993. 12. m. beck. size-specific shelter limitation in stone crabs: a test of the demographic bottleneck hypothesis. ecology, vol. 76, pp. 968-980, 1995. 13. g. matlack. plant species migration in a mixed history forest landscape in eastern north america. ecology, vol. 75, pp. 1491-1502, 1994. 14. n. h. mahmood, r. o. yahya and s. j. aziz. apply binary logistic regression model to recognize the risk factors of diabetes through measuring glycated hemoglobin levels. cihan university-erbil scientific journal, vol. 6, no. 1, pp. 1-7, 2022. table 6: log-likelihood value of the full model and reduced model models chi-square full model log-likelihood 75.798 reduced model (constant only) log-likelihood 143.829 −2 log likelihood 67.871 p-value 0.000 table 7: goodness of fit output for the full model statistic value df p-value hosmer and lemeshow 14.337 8 0.073 nagelkerke r square 0.808 cox and snell r square 0.593 tx_1~abs:at/add:tx_2~abs:at 99 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 99-102 research article a comparison of chemical compounds between anti-diabetic drug and some medicinal plants ali j. muhialdin1, zhala b. taha2, rahman k. faraj3, ali m. hussein4, rayan s. salahalddin4, hawri a. majeed4 1department of animal production, college of agricultural engineering sciences, university of garmian, kalar, kurdistan region, iraq, 2department of forestry, college of agricultural engineering sciences, salahaddin university, erbil, kurdistan, iraq, 3department of chemistry, college of science, university of garmian, kalar, kurdistan region, iraq, 4department of biomedical sciences, cihan university-erbil, kurdistan region, iraq abstract type 2 diabetes (t2d) is a kind of diabetes marked by high blood sugar, insulin, and insulin insufficiency. adult-onset diabetes is another name for it. increased thirst, frequent urination, and unexplained weight loss are common indications and symptoms. metformin side effects include metallic taste, weakness, diarrhea, stomach upset, lactic acidosis, and vomiting. two other side effects are asthenia and a vitamin b12 deficiency. many recent studies and most health experts recommend basil seed as a better metformin substitute. the research investigates why patients who consume basil seeds have a similar response to those who take metformin. the methodology of the study consisted of two main steps, first step is to analyze basil and use hplc to determine its chemical components. the second step is to compare the broken-down components to metformin-composed materials, which is done by diluting with methanol/water (50:50 v/v) and removing the fat layer using 20 ml hexane. the findings showed that basil seed and metformin had the most similar component structure, thus the foundlings concluded that patients had the same responses without the metformin side effects, implying that basil seed stabilizes blood sugar levels. keywords: ocimum basilicum, trigonella foenum-graecum, metformin, chemical composition, type 2 diabetes introduction one of the most common metabolic disorders is diabetes mellitus. it is characterized by hyperglycemia caused by absolute or relative insulin deficiency, and it is related to long-period issues with the heart, nerves, kidneys, and eyes.[1] insulin-dependent diabetes mellitus (iddm, type 1) and non-insulin-dependent diabetes mellitus (nidm, type 2) are the two types of diabetes mellitus. the localized inflammatory reaction in the pancreas and its surrounding areas marked type 1 diabetes which is an autoimmune disease.[2] characteristics of type 2 diabetes are peripheral insulin resistance and insulin efficiency. insulin-independent sugar diabetes is substantially more common than insulin-dependent sugar diabetes among the two kinds of diabetes. in noninsulin-dependent diabetic mellitus, sulphonylureas and a few biguanides are efficient therapies for hyperglycemia, but they are unable to reestablish glucose homeostasis and normalize glucose levels.[3] the pharmacokinetic features, subsequent failure rates, and side effects of these medicines limit their use. even insulin therapy does not guarantee a long-term return to normal glucose homeostasis, and it is linked to a higher risk of cancer.[4] the advantage of medicinal plants is that they have no or few adverse effects. some of them have been utilized in traditional medical systems for 100s of years in a variety of places around the world. metformin, a drug produced from the medicinal plant galego officinalis was once used in medieval europe to treat diabetes and is still the only medical treatment that is moral for niddm patients. there is a variety of anti-diabetic plants that could be useful in the creation of medications for the treatment of sugar diabetes. an excellent source of protein in the diet for both humans and animals is fenugreek (trigonella foenum-graecum l.). fenugreek seeds corresponding author: ali m. hussein, department of biomedical sciences, cihan university-erbil, kurdistan region, iraq. e-mail: ali.m.hussein@cihanuniversity.edu.iq received: june 23, 2022 accepted: august 26, 2022 published: september 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp99-102 copyright © 2022 ali j. muhialdin, zhala b. taha, rahman k. faraj, ali m. hussein, rayan s. salahalddin, hawri a. majeed. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) muhialdin, et al.: a comparison of chemical compounds 100 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 99-102 and leaves are used to manufacture medicinal extracts and powders.[5] according to reports, it has an anti-diabetic, antiparasitic, anti-microbial, anti-infertility, anti-cancer, and hypocholesterolemic agent. it also has a lot of minerals and vitamins in it.[6] basil (ocimum basilicum l.), commonly referred to as sweet or garden basil, is widely grown in the mediterranean region.[7] diuretic, antipyretic, antispasmodic, and stomachic properties are all found in basil seeds.[8] according to various research, including medicinal and aromatic plants (map) in broiler diets resulted in, an increase in body weight, the efficiency of feed conversion, and feed cost.[9] the project’s goal is to demonstrate the chemical composition of basil and fenugreek seeds and compare it to metformin components to clarify the rationale for utilizing the seeds as a metformin substitute, which will be accomplished by completing the following objectives. materials and methods chemicals and reagents metformin hcl (mh) powder, fenugreek seeds powder, basil seeds powder, 0.1% phosphoric acid, acetonitrile, methanol/ water, hexane were used as reagents for the whole procedure. devices and instrumentation the separation was carried out using a shimadzu 10av-lc liquid chromatography system with the use of binary delivery pump model lc-10a shimadzu, and the eluted peaks were kept track of using a uv–vis 10 a– spd spectrophotometer [figure 2]. “chromatography is a physical separation method in which the components to be separated are split into two phases, one of which is stationary and the other of which moves in a specified direction.” the stationary phase is a solid, porous, and surface active substance in small particle form or a liquid deposited onto microparticulate beads on an inert solid substrate in hplc (usually silica). the mobile phase in the column is a liquid that moves across a packed bed of solid surface under pressure.[10] chromatographic conditions metformin was isolated on an flc (fast liquid chromatography) column under optimal circumstances. particle size 3 m, phenomenex c-18 column (50 2.0 mm i.d). stage of mobility: in solvent b, use 30, 70 v/v acetonitrile; in solvent a, use 0.1% phosphoric acid. uv 232 nm detection at 1.0 ml/min flow rate.[11] preparation of stock solution standard solution preparation; from the stock solution, samples of different concentration of metformin reference were produced, which was created by precisely weighing about 10 mg metformin and transferring it to a 100 ml volumetric flask, then adding methanol/water (50:50 v/v) as a diluent, shaking the sample well in an ultrasonic bath and adding diluents to bring the volume up to the desired level. this was then filtered using a membrane filter with a 0.45-mm pore size.[12] extraction of ocimum basilicum and trigonella foenum-graecum 0.5 g of sample powder was dissolved in 20 ml hexane to remove the lipid bilayer, then the aqueous layer was melted in 100 ml of 80:20 solutions (methanol: water). the distillate was supersonic at 60% duty cycles for (duty revolution for) 20 min at 25°c, followed by centrifugation at 7500 rpm for 15 min (branson conifer, usa). before evaporation under vacuum, each sample’s clear supernatant was treated with charcoal to eliminate colors (buchi rotavapor re type).[13] chemical evaluation for samples dried samples were resuspended in 1.0 ml hplc grade methanol by overtaxing. the mixture was poured through a 2.5 um disposable filter and stored for subsequent analysis at 4°c. according to optimum conditions, 20 ul of the sample was injected into the hplc system. results and discussion the figure 1 shows the standard composition of metformin analysis by hplc, the retention time for finding the chemical was 3.688 with an area 297141v and the concentration was 65.8425%. the round was stopped at 6.443. a previous study showed a similar retention time, which explains over the concentration range of 0.312–5 g/ml, the technique was linear (r2 = 0.9995). metformin had a detection limit of 0.1 g/ml and a quantitation limit of 0.3 g/ml. in the case of pharmaceutical formulations, the acquired findings revealed a good agreement with the claimed ingredients.[14] the figure 2 shows the standard composition of basil seed analysis by hplc, at the first peak, the retention time for figure 1: metformin standard composition muhialdin, et al.: a comparison of chemical compounds 101 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 99-102 figure 2: the composition of basil seeds figure 3: retention time of used extracts figure 4: peak area of used extracts finding the chemical was 1.153 with an area 59352v and the concentration was 7.868%. at the second peak, the retention time for finding the chemical was 1.642 with an area 59693v and the concentration was 7.9861%. at the third peak, the retention time for finding the chemical was 2.565 with area 48340v and the concentration was 6.4024%. however, at the fourth peak, at the time 3.81 with an area 36880v 4.874% was the most similar chemical with metformin composition. prior research found that oap-1a was a neutral heteropolysaccharide consisting of mannose (35.7%), glucose (33.32%), galactose (19.6%), and rhamnose (11.38%) which is similar with the same chemical composition of basil seeds result.[15] the figure 3 shows the standard composition of fenugreeks analysis by hplc, the retention time for finding the chemical was 1.948 with an area 218757v and the concentration 15.8732%. at the second peak, the retention time for finding the chemical was 3.15 with area 636125v and the concentration was 46.158%. at the third peak, the retention time for finding the chemical was 3.983 with area 354686v and the concentration was 25.7364%. the study provides an alternative natural strategy to treat diabetes, and we began formulating and testing these claims based on earlier studies and claims that referred to basil seeds as a drug and other studies that referred to fenugreek seeds as a medication. our data and analysis reveal that eating basil seeds as a drug will help reduce or lower blood sugar levels by a retention time of 3.81/min, indicating that it is more compatible with metformin hydrochloride, according to the study. as shown in figure 4, our findings confirmed the previous studies and researches that basil seeds are effective in treating diabetes; our study confirms those findings and adds to the evidence that basil seeds are a great alternative to metformin with few to no side effects. however, our findings suggest that fenugreek seeds are not as effective as other researchers suggest, which may call into question theories and studies that have been conducted. however, we acknowledge our limits and welcome discussion because we do not feel that our research is the ultimate and final solution, and that more and more studies in this sector, that is, comparing chemically manufactured medications to natural resources, are required. conclusion depending on the previous data and figures, each ocimum basilicum, trigonella foenum-graecum which are known as medicinally used seeds have the same chemical as metformin pills with is the official treatment used for optimizing the sugar amount in the blood, the future study will be a clinical trial which is procedure approved by fda for finding new treatment or drugs. acknowledgment this project would not have been possible without the support of cihan university-erbil by providing laboratories and equipment for researchers, all efforts and support are appreciated. muhialdin, et al.: a comparison of chemical compounds 102 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 99-102 references 1. w. h. gispen and g. j. biessels. cognition and synaptic plasticity in diabetes mellitus. trends in neurosciences, vol. 23, no. 11, pp. 542-549, 2000. 2. a. k. foulis. the pathogenesis of beta cell destruction in type i (insulin-dependent) diabetes mellitus. the journal of pathology, vol. 152, no. 3, pp. 141-148, 1987. 3. w. gepts and p. m. lecompte. the pathology of type i (juvenile) diabetes. in: the diabetic pancreas. springer us, boston, ma, pp. 337-365, 1985. 4. e. basch, c. ulbricht, g. kuo, p. szapary and m. smith. therapeutic applications of fenugreek. alternative medicine review, vol. 8, no. 1, pp. 20-27, 2003. 5. a. ahmad, s. s. alghamdi, k. mahmood and m. afzal. fenugreek a multipurpose crop: potentialities and improvements. saudi journal of biological sciences, vol. 23, no. 2, pp. 300-310, 2016. 6. m. duru, z. erdogan, a. a. duru, a. küçükgül, v. düzgüner, d. a. kaya and a. şahin. effect of seed powder of a herbal legume fenugreek (trigonella foenum-graceum l.) on growth performance, body components, digestive parts, and blood parameters of broiler chicks. pakistan journal of zoology, vol. 45, no. 4, pp. 1007-1014, 2013. 7. a. ganasoundari, s. m. zare and p. u. devi. modification of bone marrow radiosensensitivity by medicinal plant extracts. the british journal of radiology, vol. 70, no. 834, pp. 599-602, 1997. 8. r. ullah, a. s. alqahtani, o. m. a. noman, a. m. alqahtani, s. ibenmoussa and m. bourhia. a review on ethno-medicinal plants used in traditional medicine in the kingdom of saudi arabia. saudi journal of biological sciences, vol. 27, no. 10, pp. 2706-2718, 2020. 9. e. toson and m. abd ellatif. effect of using fenugreek seeds powder as a feed additive in broiler chicks’ diet on growth performance and some metabolic responses. egyptian poultry science journal, vol. 41, no. 1, pp. 31-43, 2021. 10. r. sauce, c. a. s. de oliveira pinto, c. ayala-jara, z. a. prieto, m. v. r. velasco and a. r. baby. preliminary protocol development of a hplc-tbars-evsc (ex vivo stratum corneum) assay for skin research: application in a sunscreen system. scientia pharmaceutica, vol. 89, no. 2, p. 17, 2021. 11. c. a. gulhane, w. a. panchale, j. v. manwar and r. l. bakal. liquid chromatographic method for simultaneous estimation of thiocolchicoside and etoricoxib from tablet formulation. the asian journal of pharmaceutical analysis, vol. 11, no. 2, pp. 118122, 2021. 12. e. patyra and k. kwiatek. comparison of hplc-dad and lc-ms techniques for the determination of tetracyclines in medicated feeds using one extraction protocol. chromatographia, vol. 84, no. 8, pp. 741-749, 2021. 13. s. bafadam, m. mahmoudabady, s. niazmand, s. a. rezaee and m. soukhtanloo. cardioprotective effects of fenugreek (trigonella foenum-graceum) seed extract in streptozotocin induced diabetic rats. journal of cardiovascular and thoracic research, vol. 13, no. 1, pp. 28-36, 2021. 14. k. arab, b. ghanbarzadeh, a. ayaseh and k. jahanbin. extraction, purification, physicochemical properties and antioxidant activity of a new polysaccharide from ocimum album l. seed. international journal of biological macromolecules, vol. 180, pp. 643-653, 2021. 15. h. hui, h. jin, x. li, x. yang, h. cui, a. xin, r. zhao and b. qin. purification, characterization and antioxidant activities of a polysaccharide from the roots of lilium davidii var. unicolor cotton. international journal of biological macromolecules, vol. 135, pp. 1208-1216, 2019. tx_1~abs:at/add:tx_2~abs:at 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 17-22 research article design rf frontend unit to avoid intermodulation using arduino uno adil h. mohammed, yazen s. almashhadni, ahmad n. abdulftah department of communication and computer engineering, faculty of engineering, cihan university-erbil, kurdistan region, iraq abstract designing a radiofrequency (rf) frontend is vastly realized for determining the level of integration that is required in the signal chain inside the receivers to be idealistic. the receivers are susceptible to harmful intermodulation due to nonlinear rf frontends. in this paper, intermodulation distortion is avoided by a selective prototype hardware design of rf fort end which is connected with the arduino uno for controlling the power levels or automatic level control. the measurements are tested out as a result of injecting a signals within x-band frequencies and chosen different power levels are assumed. these measurements are revealed an accepted results for the intermodulation avoidance. keywords: frontend, intermodulation, automatic level control, x-band receiver, arduino uno introduction an rf frontend is a device or module that incorporates all the circuitries between the antenna and at least one mixing stage of a receiver and possibly the power amplifier of the transmitter. it is used in a wide variety of rf products and applications such as conventional communication systems, radar systems, and electronic warfare systems (ew). rf hardware challenges for the applications of joint communication and radar sensing are studied.[1] the development of designing the frontend module has attracted in the recent years in modern communication and radar systems at x-band frequencies as a targeted this band due to its prospective opportunities for research related to new forthcoming applications, also it is consider a good candidate band for the detection of normal size objects in radar applications.[2-4] there are several approaches to combine a radar systems and communication systems. one can use an existing radar system and add communication functionalities.[5] the receiver chain in such systems is suffered from the intermodulation products which they effect receiver operation, since in a nonlinear circuit, a two mixed signals can cause a product of intermodulation distortion. the intermodulation produced when high power signal passed through an uncontrolled level gain amplifier and mixers and the generated unwanted frequencies which are confused the receiver to get real information about the target and give a false alarm, the avoidance of this undesired phenomenon is essential by proposing an achievable schemes a design of frontend circuits.[7,8] the frontend is collaborated with arduino for controlling and interfacing data as good result can be attained.[9] this paper proves that arduino uno with a digital variable attenuator in x-band can keep the input level signal with the acceptable range that is required before entering down frequency converter unit. to avoid the intermodulation problem, which is vital at high-power signal input to the mixer in rf down convertor, the automatic level control circuit is needed to be keeping the signal on a certain level. therefore, digital variable attenuator (dva) is used and it is controlled by the digital word. the digital word is generated by arduino uno according to the readout of the power level of the signal from the alarm circuit. microwave receivers in this section, the most common traditional radio receiver architectures are presented.[10] figure 1 shows microwave receiver block diagram. the major difference between electronic warfare receiver (ewr) and other conventional receivers is that their input signals are unknown. in addition to the complexity of the signal, ewr deliberately keeps some of corresponding author: yazen s. almashhadni, department of communication and computer engineering, faculty of engineering, cihan university-erbil, kurdistan region, iraq. e-mail: yazen.mahmood@cihanuniversity.edu.iq received: june 20, 2022 accepted: july 29, 2022 published: august 10, 2022 doi: 10.24086/cuesj.v6n2y2022.pp17-22 copyright © 2022 adil h. mohammed, yazen s. almashhadni, ahmad n. abdulftah. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ mohammed, et al.: design rf frontend unit to avoid intermodulation using arduino uno 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 17-22 the input signals in covert mode to inhibit detection, because it is desirable to obtain as much information as possible from the electronic order of battle (eob) in a very short period. the requirements for ewr are becoming more demanding. the receiver must have a wide input bandwidth with fine frequency resolution, high sensitivity, and dynamic range to receive as many signals as possible. the receiver must also have the capacity to process simultaneous signals. furthermore, the receiver should be able to measure the angle of arrival (aoa) of the input signal.[11] the basic function of a radio front end is to take the modulated carrier signal from the antenna, amplify and down converter the signal, select the wanted channel, and finally extract the baseband information. from this point of view, a radio receiver does not appear to be very complicated. however, when considering that the signal could be very weak or very strong depending on how far from the transmitter the receiver is, or even worse when the wanted signal is very weak but another strong interferer (blocker) is present, then the true challenges of radio receiver design are brought into the light. there exist several different ways to build a receiver that takes care of all those issues in one way or the other. intermodulation problem when a receiver provides only the frequency information and does not measure the amplitude information on the input signal, the dynamic range can be determined by its frequency measurement capability. since the frequency measurement is considering to be the primary performance of an ew receiver, the dynamic range is often determined by this capability, even the receiver measures signal amplitude, under this situation, one may quote two dynamic ranges for the same receiver. one is related to its amplitude measurement capability and the other is related to frequency measurement capability. the lower limit of the dynamic range is usually defined as the weakest signal level where the measured frequency error is within a certain predetermined range, the upper limit of the dynamic range is the strongest signal level where the measured frequency error is within a certain predetermined range. if a strong signal is received, there are some components in the receiver become work in the nonlinear region, the additional signal may appear at the output of the receiver, for example, a mixer may be generating strong intermodulation product and an amplifier may produce second harmonics when they are saturated, often, the dynamic range of a receiver is referred to as the single-signal super-free dynamic range. with this dynamic range, if one signal is present at the input of the receiver, the receiver will not generate superior signals. mathematically, the possible harmonics available at the mixer if port are given by the equation: [12] fif = nflo∓ mfrf (m and n are all integers) (1) because the only one desired output frequency (when n = 1 and m = 1), the existence of all other harmonic terms creates significant problems. elimination of these distortion products is a key goal in mixer design. the rf frontend design the frontend unit has a very important function to keep the receivers operating in the linear region of the mixer and signal amplifiers behavior without entering the non-linear regions were to avoid generating unwanted frequencies that generated by intermodulation within the intermediate frequency working range, and to ensure that it operates within its operating frequency band, and to maintain signal levels entered at a specific and controlled level. figure 2 shows the main elements of the frontend unit. protection unit the function of this unit as shown in figure 3 is to protect the receiver from high-power level frequencies emitted from very close radars. the rf switch single-pole single-throw (spst) is used to protect the system from any high-power signal, which is detected by the crystal detector (cd) after the input signal is amplified by low noise amplifier (lna) 40 db and efficiently integrating directional coupler (dc) circuitry with other circuit elements of an rf frontend, then sent to the alarm circuit passing through video amplifier (va). this circuit measures the amplitude and creates an order to open the path when high power signal input. the path can be opened as given order with high priority from the alarm circuit when it senses a high-level signal present. the total insertion loss in the protection units is stated in table 1. by the total gain of the protection, circuit can estimate the high-power signal before coming too close to our station and (switch off) disconnect the signal path to the receiver system. the order of switch-off is coming from the alarm electronic circuit which has a comparator circuit with a reference voltage figure 1: microwave receiver block diagram figure 2: main units of rf frontend block figure 3: protection unit block diagram mohammed, et al.: design rf frontend unit to avoid intermodulation using arduino uno 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 17-22 to produce logic high (1) or low (0) to switch on or off, and this circuit can be set manually to select the value of high power which consider high-power signal and must be stopped to pass to the receiver. therefore, we can consider the input signal with four different levels, as shown in table 2. table 2 shows that the signal level higher than (0 dbm) must be stopped to keep the rf components in the frontend woke in safe and not exceed input signal higher than 1 watt.[13] the alarm circuit has a function when high input power is present, the function of this circuit is to generate a digital word (3 bits), which sends the command to the rf switch (in frontend unit) to stop the running signals in the system. the input level signal will compare with the known reference level, which is representing the maximum level can the system handling with. according to that level, the order will be for a pass then no pass to the signal or the switch on then off order. the digital word has high priority and is independent whatever the mode of the scan or search in processing. so that, the output of this circuit will be connected to (or gate) and other input will be supplied from arduino uno to stop the running signal in the system. the state of the comparator output will feed arduino uno to know why the system is halt and give the order to running again later. figure 4 depicts the main block diagram of the alarm system. frequency range limiter acceding to the design consideration, the frontend work with x-band, the bandpass filter will pass only the frequencies between 8 and 12 ghz and stop or reject all frequencies out of that range, figure 5 shows the characteristic of the filter which attenuates out the band of frequencies within 70 db power range.[14] rf amplifiers the preamplifier is a one type of the amplifiers in frontend block diagram which is used to improve the signal level that it is lost part of power in the rf path due to the insertion loss of the elements and cables. this type of amplifier prefers to be a limiter amplifier. the specifications of this amplifier are shown in table 3.[15] figure 6 shows the characteristic behavior of the amplifier that can cover all the bandwidth with the gain around 30 db and noise figure around 3 db. automatic level control (alc) because the ewr deals with unknown signal and different power signals, near and far sources of the signal. alc circuit has a function, which is adding the rf path, attenuation as requested according to the level required to down frequency convertor. this circuit sends a word to the digital attenuation needed to add, which will do according to table 4. b0, b1, b2, b3, and b4 are the binary bits that must be generated from arduino uno according to the input level measuring. the alc can receive the value that needed for attenuation from the alarm circuit which sends overflow indication to the processor or direct from the processor after measuring the amplitude of the signal. then, it will send an order to control on a certain level. in this way, the close loop of alc is satisfied.[16] table 4 shows the amount of insertion loss in the attenuator according to feeding digital word. insertion gain measurement it can measure the minimum signal level and maximum signal level within the consideration of design which gives trustworthy results at the output. figure 7 demonstrates all the connected elements in the frontend with showing corresponding insertion gains. so that for low-level signal, the dva will be setting on 1.5 db as attenuation which gives the net gain (ng). ng = –0.1 + 40 – 1 – 2 + 35 – 1.5 – 1 = 69.4db (2) for high-level signal ng = –0.1 + 40 – 1 – 2 + 35 – 48 – 1 = 22.9db (3) if it considers the frequency, down converter needs around –10 dbm signal level which gives the minimum signal detection (msd) as in: [17] msd = –10 – 69.4 = –79.49db (4) the results of the estimated input signal levels in table 5 show obviously that the input level has the power greater than –30 dbm is stopped by the alarm circuit. table 1: insertion loss of protection unit element insertion loss or gain (db) 1 spst −0.1 2 lna +40 3 dc −10 4 cd −1 5 va +20 total 58.9 table 2: input levels and measured levels at cd block level no. estimate input level level at cd 1 –70 dbm –41.1 dbm 2 –50 dbm –21.1 dbm 3 –10 dbm –27.1 dbm 4 0 dbm +28.1 dbm figure 4: alarm circuit diagram mohammed, et al.: design rf frontend unit to avoid intermodulation using arduino uno 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 17-22 arduino uno connection and programming as shown in figure 8 the connection of rf frontend to arduino uno for controlling the level of the signal and protect the circuit from a high-level signal as follows: • the connection to spst by three digital pins (2, 3, and 4) as output. • the connection of video signal of alarm circuit by analog pin (a1) as input. • the connection of digital variable attenuator (dva) by four digital pins (13,12,11, and 10) as output.[18] the program was written by c++ programming with help of the arduino simulator, and the following chart of programming is shown in figure 9. the first block represents the initial setting to receive any weak signals so that pins (2, 3, and 4) must be logic high at the output to keep spst in on state, and pins (13,12,11, and 10) must be logic low to set dva as minimum attenuating value (1.5 db). the second block for reading the value of signal level and comparing with the reference level which is considered high signal level, that will be in the third block, the decision from the 4th block, if that level is high so the decision to stop receive the signal by setting pins (2, 3, and 4) high to change spst to off, and waiting for time delay and change pins (2, 3, and 4) again. the setting of pins (13, 12, 11, and 10) according to the reading of analog port a1 and changing in logic level to get accept level at frequency down converter to avoid intermodulation problem. table 3: amplifier specification specification description 1 frequency band 8–12.4 ghz 2 gain 35 db 3 noise figure <4 db 4 max. output +13 dbm 5 impedance 50 ohm 6 power supply +15 volts 300 ma figure 6: characteristics of the small-signal amplifier. figure 5: characteristics of the bandpass filter figure 8: arduino uno connection figure 7: the frontend block diagram mohammed, et al.: design rf frontend unit to avoid intermodulation using arduino uno 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 17-22 table 4: insertion loss amount based on four generated bits b4 b3 b2 b1 b0 il (db) 0 0 0 0 0 1.5 0 0 0 0 1 3 0 0 0 1 0 4.5 0 0 0 1 1 6 0 0 1 0 0 7.5 0 0 1 0 1 9 0 0 1 1 0 10.5 0 0 1 1 1 12 0 1 0 0 0 13.5 0 1 0 0 1 15 0 1 0 1 0 16.5 0 1 0 1 1 18 0 1 1 0 0 19.5 0 1 1 0 1 21 0 1 1 1 0 22.5 0 1 1 1 1 24 1 0 0 0 0 25.5 1 0 0 0 1 27 1 0 0 1 0 28.5 1 0 0 1 1 30 1 0 1 0 0 31.5 1 0 1 0 1 33 1 0 1 1 0 34.5 1 0 1 1 1 36 1 1 0 0 0 37.5 1 1 0 0 1 39 1 1 0 1 0 40.5 1 1 0 1 1 42 1 1 1 0 0 43.5 1 1 1 0 1 45 1 1 1 1 0 46.5 1 1 1 1 1 48 table 5: the output powers based on the input powers estimate input level ng require level at frequency down converter 1 –70 dbm 60 db –10 dbm 2 –50 dbm 40 db –10 dbm 3 –40 dbm 30 db –10 dbm 4 –30 dbm 22.9 db –8 dbm 5 –27dbm * * figure 11 shows the input result within (–75––32) dbm and the output result within –10.5–17.5 dbm. these results will be accepted to avoid the intermodulation problem. conclusion the measurement was done by injected signals to the frontend unit with different power levels and frequencies lies in x-band (8000 mhz–12,400 mhz). the results showed in figure 11 are illustrated the output signal level appeared with the range (–10 dbm––17.5 dbm) and it indicates a good result to avoid the intermodulation distortion problem. the fluctuation at the output range depended on steps of dva which is 1.5 db per step. furthermore, the time duration of processing gives the inversely proportional with the accuracy. the designed frontend unit can be used as an rf head for radar warning receiver (rwr) with a crystal detector. testing and results the test was done using synthesized sweepers hp 8341 a as simulating the signal was injected to frontend units and out signal was measured by spectrum analyzer hp8566b, as shown in figure 10. figure 10: the frontend laboratory testing figure 11: the injected signal and the output signal level figure 9: flowchart mohammed, et al.: design rf frontend unit to avoid intermodulation using arduino uno 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 17-22 references 1. f. bozorgi, p. sen, a. n. barreto and g. fettweis. rf frontend challenges for joint communication and radar sensing. in: 2021 1st ieee international online symposium on joint communications and sensing (jc&s.). ieee, pp. 1-6, 2021. 2. s. saha. rf front-end design for x band using 0.15 µm gan hemt technology. (doctoral dissertation, université d’ottawa/ university of ottawa, 2016. 3. a. d. stevens. design and implementation of an rf front end for the nextrad radar system. master’s thesis, university of cape town, 2017. 4. t. pongthavornkamol, a. worasutr d. worasawate, l. o. kovaisaruch and k. kaemarungsi. x-band front-end module of fmcw radar for collision avoidance application. engineering journal, vol. 25, no. 5, pp. 61-70, 2021. 5. s. dwivedi, a. n. barreto, p. sen and g. fettweis. target detection in joint frequency modulated continuous wave (fmcw) radarcommunication system. in: 2019 16th international symposium on wireless communication systems (iswcs). ieee, pp. 277-282, 2019. 6. a. v. padaki and j. h. reed. impact of intermodulation distortion on spectrum preclusion for dsa: a new figure of merit. in: 2014 ieee international symposium on dynamic spectrum access networks (dyspan). ieee, pp. 358-361, 2014. 7. y. lee, s. chang, j. kim and h. shin. a cmos rf receiver with improved resilience to ofdm-induced second-order intermodulation distortion for medradio biomedical devices and sensors. sensors, vol. 21, no. 16, p. 5303, 2021. 8. a. v. padaki, r. tandon and j. h. reed. on scalability and interference avoidance in nonlinear adjacent channel interference networks. in: 2017 ieee international conference on communications (icc). ieee, pp. 1-6, 2017. 9. i. knight. arduino to front end part ii. in: connecting arduino to the web. apress, berkeley, ca. pp. 163-202, 2018. 10. j. b. tsui and j. p. stephens. digital microwave receiver technology. ieee transactions on microwave theory and techniques, vol. 50, no. 3, pp. 699-705, 2002. 11. s. andersson. multiband lna design and rf-sampling front-ends for flexible wireless receivers. doctoral dissertation, linköping university electronic press, 2006. 12. f. marki and c. marki. mixer basics primer. marki microwave, 2010. 13. i. n. abubakar, j. tsado, u. a. dodo, e. g. ufot, j. t. zarmai, m. a. dodo and i. suleiman. development of a microcontrollerbased power transformer overload protection scheme. atbu journal of science, technology and education, vol. 8, no. 1, pp. 360-371, 2020. 14. y. lan, y. xu, t. mei, y. wu and r. xu. a 2∼ 18ghz compact microwave band-pass filter suitable for planar and threedimension flexible integration. in: 2016 46th european microwave conference (eumc). ieee, pp. 528-531, 2016. 15. j. karki. understanding operational amplifier specifications. texas instruments white paper sloa011, 1998. 16. a. h. mohammed, a. i. khoshnaw and g. a. qasmarrogy. satellite link budget calculator by using matlab/gui. in: 1st international conference of cihan university-erbil on communication engineering and computer science (cic-cocos’17). cihan university-erbil, kurdistan region-iraq. pp. 74-78, 2017. 17. t. pan and y. zhu. getting started with arduino. in: designing embedded systems with arduino. springer, singapore. pp. 3-16, 2018. 18. j. purdum and d. kidder. arduino projects for amateur radio. mcgraw-hill education, new york. 2015. tx_1~abs:at/add:tx_2~abs:at 81 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 research article study of shear transfer in modified push-off members using finite elements method: a comparative study sabih h. muhodir department of architectural engineering, cihan university-erbil, kurdistan region, iraq abstract this paper aims to investigate numerically the behavior of modified push-off specimens under the action of direct shear stress. based on the tow-dimensional finite element model developed in this research, the contribution of the aggregate interlock to resist the shear stress along the shear plane, the effect of existing of the compressive stress acting across the shear plane, the effect of the parallel reinforcement in resisting shear stress, the effect of the shear reinforcement parameter, the strains in the concrete and steel, and the actual distribution of the shear stress along the shear plane were studied. to verify the accuracy and applicability of the suggested finite element model, a comparison between the results obtained in this study and those obtained experimentally by other authors was carried out. comparison showed that the finite element results were in good agreement with the experimental results. it has been found that, for modified push-off specimens of groups without shear reinforcement across the shear plane the diagonal tension crack within the shear plane occurred at the load level which is closely to the ultimate shear strength respectively, while for specimens with both shear and parallel reinforcement, the first crack formed at about (33.7–53.0%) of the ultimate strength, also the investigation showed that the presenting of the shear reinforcement normal to the shear plane are significantly increased the shear transfer stress for all levels of loading. keywords: shear, aggregate interlock, finite element method, interface, modified push-off introduction shear transfer may be of high importance in many types of reinforced concrete members including ordinary and deep beams, slabs, corbels and brackets, shear walls and shear diaphragms and containment vessels of various types. shear transfer is generally considered as a major mechanism of load transfer along a concrete-to-concrete interface under the action of shear or under the combination effect of shear and normal force.[1-3] although the mechanism of the shear transfer, the aci-318-19 provisions[4] depends mainly on the relation between the shear transfer and the reinforcement crossing the shear plane (clamping force), as well as on the resistance generated from the friction between two sliding faces along the shear plane which is depending contact surface condition and on the coefficient of friction of the concrete used. to calculate the shear strength provided by the sear reinforcement perpendicular to the shear plane, the stress is assumed to have reached to its yield stress f y . this leads to the fact that the concrete contribution to resist the shear calculated using the aci code equations increases compared to that provided by the shear reinforcement which is expressed as v n =μa v f y, where v n = nominal shear strength, a v is the area of reinforcement crossing the assumed shear plane to resist shear, and μ is the coefficient of friction.[5,6] depending on the previously published test results hsu[7-10] developed a formula to predict the shear transfer strength of reinforced concrete members: vu = 0.822(fc /)0.406(ρfy) c (1) where: vu= unit shear strength (mpa) c = 0.159(fcc /)0.33 (2) fcc /= concrete compressive strength of 150 mm cube and taken as fc / .0 85 to this end the present study is concerned with an attempt to verify the validity of the aci shear friction provision and to investigate the influence of the direct shear stress acting parallel and transverse to the shear plane on the shear transfer strength using the finite element method (fem) and to investigate their corresponding author: sabih h. muhodir, department of architectural engineering, cihan university-erbil, kurdistan region, iraq. e-mail: sabih.alzuhairy @cihanuniversity.edu.iq received: june 05, 2022 accepted: august 03, 2022 published: august 27, 2022 doi: 10.24086/cuesj.v6n2y2022.pp81-88 copyright © 2022 sabih h. muhodir. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) muhodir: study of shear transfer in modified push-off members using finite elements method 82 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 role in the influencing the strength and deformation using the typical and modified push-off specimens. the main objectives of this study are the following: 1. study the contribution of the aggregate interlock to resist the shear stress along the shear plane 2. study the effect of existing of the compressive stress acting across the shear plane 3. study the effect of the parallel reinforcement in resisting shear stress 4. study the effect of the shear reinforcement parameter 5. investigating the strains in the concrete and steel and the actual distribution of the shear stress along the shear plane. failure criteria for formulation the failure criteria for concrete under combined states of stresses, one must agree on a proper definition of failure. such criteria as yielding, initiation of cracking, load carrying capacity, and extent of deformation have been used to define failure.[11,12] in this study, failure is defined as first crack loading and load carrying capacity of the reinforced concrete element. in general, concrete failure can be divided into tensile and compressive types. with respect to the present definition of failure, tensile failure defined by the formation of major cracks and the loss of tensile strength in concrete normal to the crack direction, while in compressive failure many small cracks develop and the concrete element loses most of its strength. the mohr-coulomb criterion in the present study was used as a load carrying capacity criteria, this is dating from 1900 and states that the failure is governed by the relation.[13] |τ| = f(σ) (3) where the limiting shearing stress τ is depending only on the normal stress (σ) in the same plane and at the same point, and where the equation (1) is the failure envelope for the corresponding mohr-circle.[9] the simplest form of equation (1) can be written as:[13] |τ|=c-σn.tan∅ (4) where: τ: the shearing stress σn: the normal stress (tensile stress is positive) c: cohesion ∅: the angle of internal friction (tan ∅ used in this study is equal to 1.4 –normal weight concrete).[14] in the principal –stress coordinate, the failure criterion (sliding criterion) given by equation (2) takes the form: [14] 1 2 1 1 2 1 01 3σ σ+( ) − −( ) − =sin sin c cos∅ ∅ ∅. (5) for σ1 ≥ σ2 ≥ σ3 [10] σ σ1 3 1 f ft c / / = = (6) in general, the mohr-coulomb criterion is a two-parameter model[13] where any combination of parameters, such as (∅, c), (fc /, ft /), experimentally observed will be adequate to characterized completely the material behavior, so it is sometimes convenient to use the parameters fc /and m, where m sin sin f f c t = + − = 1 1 ∅ ∅ / / (7) the coefficient m for concrete is considered to be 4.1,[13] then equation (3) can be written as[14,15] m. σ1–σ3 = 2.c.√m=fc / (8) where: m cos sin sin sin = − = + − [ ] ∅ ∅ ∅1 1 1 2 (9) and; f c cos sin ���������c / . .= − 2 1 ∅ ∅ (10) therefore, the value of (c) used in equation (2) can be obtained using equation (10) in terms of fc /and angle of internal friction (∅). the failure criteria given by equation (4) holds for member with shear reinforcement by adding the contribution of the shear reinforcement which can be given by the reinforcement parameter (ρ.fy) to the normal stress (σn), therefore, |τ| = c – (σn – ρfy) tan∅ (11) material and specimens characterization the dimension and reinforcement of the reinforced concrete members (modified push-of-specimens) were selected to be similar to those specimens, whose behavior being investigated experimentally by al-sharae.[15] the overall dimensions of the modified push-off-specimens that were used in this study are of length (l = 650 mm) × width (b = 400 mm) × depth (d = 150 mm). the length of the shear figure 1: typical push-off-specimen muhodir: study of shear transfer in modified push-off members using finite elements method 83 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 plane remain constant for all specimens (i.e., the cross sectional area of the shear plane 150 mm × 300 mm) and the height of the slot is constant and equal to 25 mm as shown in [figure 1]. in all specimens the shear reinforcement cross the shear plane at right angle. additional reinforcement parallel to the shear plane is provided to prevent any failure other than along the specific shear plane. the specimens are loaded by concentrated (p), without moment. when the (p) applied concentrically in the modified push-off-specimen, the shear force along the shear plane will be (p. cos ѳ) and a compressive normal force (p. sin ѳ) across the shear plane. five different values of (ѳ) was used to maintain different values of shear stress and transverse compressive stress (ѳ is the inclination of the upper point shear plane relative to the lower point). the details of specimens are summarized in [table 1]. therefore, the program of the shear transfer analysis is thus divided into the following three groups: • group sc group with plain concrete specimens. • group sp group reinforced with parallel to the shear plane reinforcement only as shown in [table 1]. • group sr (sr1, sr2, and sr3) group with shear reinforcement placed at right angle to the shear plane as well as a parallel reinforcement is provided in the critical zone, [table 1]. to study the effect of steel parameter (ρ.fy), different steel ratios were used by changing the diameter of the shear reinforcement crossing the shear plane. the detailing of all groups is summarized in [table 1]. the properties of concrete and reinforcement used in this study are tabulated in [tables 2 and 3] respectively. finite element description in the finite element formulation, the choice of a proper element is very important and effects on the accuracy of the final results of the analysis. in the current study a nine nodded table 1: details of specimens[15] spec. identity ѳo k-factor parallel rein. transvers rein. sc00 0 0 none none sc10 10 0.174 none none sc20 20 0.34 none none sc30 30 0.5 none none sc45 45 0.707 none none sp00 0 0 4φ12 none sp10 10 0.174 4φ12 none sp20 20 0.34 4φ12 none sp30 30 0.5 4φ12 none sp45 45 0.707 4φ12 none sr100 0 0 4φ12 6φ8 sr110 10 0.174 4φ12 6φ8 sr120 20 0.34 4φ12 6φ8 sr130 30 0.5 4φ12 6φ8 sr145 45 0.707 4φ12 6φ8 sr200 0 0 4φ12 6φ10 sr210 10 0.174 4φ12 6φ10 sr220 20 0.34 4φ12 6φ10 sr230 30 0.5 4φ12 6φ10 sr245 45 0.707 4φ12 6φ10 sr300 0 0 4φ12 6φ12 sr310 10 0.174 4φ12 6φ12 sr320 20 0.34 4φ12 6φ12 sr330 30 0.5 4φ12 6φ12 sr345 45 0.707 4φ12 6φ12 table 3: concrete properties[15] spec. identity cylinder compressive strength (mpa) modulus of rupture (mpa) sc00 22.9 1.78 sc10 22.9 1.78 sc20 22.1 1.73 sc30 22.1 1.73 sc45 23.2 1.82 sp00 23.4 1.85 sp10 24.6 1.83 sp20 22.5 1.89 sp30 21.8 1.89 sp45 21.8 1.89 sr100 23.4 1.74 sr110 23.4 1.61 sr120 24.2 1.61 sr130 21.8 1.84 sr145 21.8 1.84 sr200 23.4 1.87 sr210 23.4 1.61 sr220 24.2 1.61 sr230 21.8 1.84 sr245 23.4 1.87 sr300 21.8 1.61 sr310 21.8 1.61 sr320 23.4 1.84 sr330 23.4 1.84 sr345 24.2 1.87 table 2: properties of bars used[15] bar diameter φ (mm) area (mm2) f y (mpa) strain ε y 8 50.27 410 0.002 10 78.54 412 0.002 12 108.54 410 0.002 muhodir: study of shear transfer in modified push-off members using finite elements method 84 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 two-dimensional isoparametric quadrilateral elements with two degrees of freedom per node, as shown in [figure 2], were used for all constituent materials (concrete and steel) in the modified push-off specimens’ analysis. in generating the finite element, mesh certain assumptions were made to simplify the complex geometry and to reduce the size of the mesh, such as, the interface between the concrete and steel was assumed. results and discussion cracking and ultimate strength three groups of specimens (sc, sp, and sr) were analyzed using finite element model suggested in this paper to study the shear transfer strength in the modified push-off specimens. in this study, the modulus of rupture (f cr ) and the mohr-coulomb criterion were used as a first cracking and ultimate strength criteria, respectively. using the finite element model, it was found that, the group (sp) behaved to same manner to the specimens of group (sc). results of the finite element analysis are shown in [table 4]. for specimens with both shear and parallel reinforcement (groups sr), the first crack formed at about (33.7–53.0%) of the ultimate strength. depending on the first crack criterion, these cracks occurred first almost at mid-length of the shear plane approximately. as lad increased, more cracks began appear throughout the shear plane with rapid propagation away from the mid-length of the shear plane approximately. furthermore, it can be seen that the ultimate strength is increased as the external compressive stress (σ nx ) and reinforcement parameter (ρf y ) are increased. in general, characterized failure for specimens with shear and parallel reinforcement is ductile failure. al-sharae,[15] in his experimental study found that, for modified push-off specimens with (ф = 10 mm) as a main shear reinforcement, the diagonal tension crack within the shear plane occurred at about (51–55%) of the ultimate shear strength. comparison of the results obtained in this study using fem with those obtained experimentally by al-sharae[15] shows a good agreement as shown in [table 4]. for specimens of groups (sc) and (sp), the first crack formed at about (87–95.02%) and (82.4–90.5%) of the ultimate strength, respectively, as shown in [table 5]. increasing in stiffness of group (sp) as compared with that of group (sc) can be attributed of the presenting of the parallel reinforcement within the critical zone. al-sharae,[15] in his experimental study found that, for modified push-off specimens of groups (sp and sc), the diagonal tension crack within the shear plane occurred table 4: f.e.m results of the modified push-off-specimen specimen’s identity f.c.l* (kn) f.l** (kn) f.c.l/ f/l (%) sc00 143.18 154.0 93 sc10 162.1 170.4 95.36 sc20 185.13 193.0 95.28 sc30 210 227.32 93.2 sc45 267.6 307.76 87 sp00 147.58 163.05 90.5 sp10 165.32 187.6 88.12 sp20 196.3 218.0 90 sp30 219.74 250.2 87.83 sp45 273.43 331.45 82.4 sr100 122.34 256.3 47.78 sr110 145.70 318.63 45.72 sr120 183 368.0 49.73 sr130 210.8 412.02 44 sr145 254.37 480.49 53 sr200 128.5 312.84 41 sr210 160 400.0 40 sr220 225.63 469.3 48.1 sr230 247.2 538.8 46 sr245 322.35 656.81 49 sr300 136.55 406 33.7 sr310 184.84 533.0 34.6 sr320 248.9 631.7 39.4 sr330 309.3 741.63 41.71 sr345 395.34 869.7 45.5 f.c.l: first crack loading, **f.l: failure loading figure 3: load – normal compressive stress curvegroup (sc) figure 2: typical two-dimensional isoparametric element muhodir: study of shear transfer in modified push-off members using finite elements method 85 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 table 5: first crack loading and failure loading using f.e.m. and the experimental results by al-sharea[15] specimen’s identity f.l (kn) exp.[15] (1) f.l (kn) f.e.m (2) 1 2       f.c.l. (kn) exp.[15] (3 f.c.l (kn) f.e.m (4) 3 4       sc00 154.0 166.77 0.92 143.18 156.96 0.91 sc10 170.4 181.84 0.94 162.1 169.71 0.95 sc20 193.0 201.12 0.91 185.13 180.5 1.03 sc30 227.32 220.72 1.03 210 194.5 1.08 sc45 307.76 249.30 1.06 267.6 255.06 1.05 sp00 163.05 183.44 0.88 147.58 179.52 o.82 sp10 187.6 220.72 0.85 165.32 215.82 0.76 sp20 218.0 245.25 0.88 196.3 235.44 0.83 sp30 250.2 274.68 0.91 219.74 259.96 0.84 sp45 331.45 333.54 0.99 273.43 311.95 0.87 sr200 312.84 284.49 1.10 128.5 147.15 0.87 sr210 400.0 372.78 1.08 160 196.2 0.81 sr220 469.3 451.26 1.04 225.63 245.25 0.91 sr230 538.8 549.36 0.98 247.2 294.3 0.84 sr245 656.81 608.22 1.09 322.35 333.45 0.96 σ n-1 =0.0849 σ n-1 =0.095 at about (76.0–95.0 %) and (93.0–98.0%) of the ultimate shear strength respectively, and this can be attributed to the absence of the shear reinforcement across the shear plane. [table 5] shows the comparison of the results obtained by this study and those obtained experimentally by al-sharae.[15] in general, the agreement between the results is very good for some specimens and unstable for other specimens of groups (sp and sc). in general with help [tables 4 and 5], it can be conclude that, the failure load increases with increase of the normal compressive stress (σ nx ) and reinforcement parameter (ρf y ) regardless of the specimen’s group. furthermore, it can be concluded that, the reinforcement parallel to the shear plane has a little effect on the shear transfer strength. [figures 3-7] show that the cracking loads versus normal compressive stress for groups (sc, sp, sr1, sr2, and sr3), respectively. from these figures, it can be noted that the reserve in strength after the formation of cracks is increased with increasing the external compressive stress (σ nx ). furthermore, a large reserve in strength is obtained in the specimens reinforced with parallel and shear reinforcement, and is reduced a great deal when only plain concrete is present in the shear plane. the same behavior is obtained experimentally by mattock and hawkins[11] and al-sharae.[15] comparisons of [figures 3 and 4] show that the strength of specimens with the parallel reinforcement is higher than those of specimens made with the plain concrete only, but the reserve in strength is small, and in both groups failure occurred almost at a load closer to the first cracking load. furthermore, it can be observed from [figures 5-7] that the increase in strength with increase in (σ nx ) tends to stable for specimens in groups (sr). figure 4: load – normal compressive stress curvegroup (sp) figure 5: load-normal compressive stress curve group (sr1) muhodir: study of shear transfer in modified push-off members using finite elements method 86 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 in general, it can be concluded that the external compressive force will add clamping force, which resist the shear force; therefore, the shear transfer strength will increased with increasing this force. effect of the reinforcement parameter (ρ.fy) the reinforcement parameter (ρ.f y ) can be changed by varying either the reinforcement ratio (ρ), the reinforcement yield strength (f y ) or both. if the area of the shear plane constant (as in this study), the reinforcement ratio (ρ) can be changed by changing the bar size and/or spacing between the bars crossing the shear plane. mattock and hawkins[9] stated that the way in which steel ratio chanced does not affect the relationship between shear stress and the reinforcement parameter (ρ.f y ). to study the effect of reinforcement parameter (ρ.f y ), three ratios of steel reinforcement have been used (2.75, 4.92, and 6.18) using three different bar diameters. (∅8 mm, ∅10 mm, and ∅12 mm). [table 6 and figure 8] presented the results of analysis using f.e.m of specimens of groups (sr1, sr2, and sr3). these results are studied and compared to determine the effect of this parameter. it was found that, for given values of (k = sinѳ), the specimens with higher reinforcement parameter had figure 6: load-normal compressive stress curve group (sr2) figure 7: load-normal compressive stress curve-group (sr3) figure 8: effect of the reinforcement parameter (ρ.f y ) on the shear transfer stress (v u ) figure 9: max. shear stress-total normal compressive stress relationship (k=0) figure 11: max. shear stress-total normal compressive stress relationship (k=0.34) figure 10: max. shear stress-total normal compressive stress relationship (k=0.174) muhodir: study of shear transfer in modified push-off members using finite elements method 87 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 table 6: results of (vu) using finite element method specimen’s identity fc / (mpa) ρ.fy (mpa) σn (mpa) σn+ρ.fy (mpa) vu (mpa) sc00 22.9 none zero zero 3.10 sc10 22.9 none 0.592 0.592 3.41 sc20 22.1 none 1.31 1.31 3.83 sc30 22.1 none 2.25 2.25 4.49 sc45 23.2 none 4.32 4.32 6.10 sp00 23.4 none zero zero 3.85 sp10 24.6 none 0.82 0.82 4.69 sp20 22.5 none 1.86 1.86 5.43 sp30 21.8 none 3.20 3.20 6.39 sp45 21.8 none 5.41 5.41 7.65 sr100 23.4 2.75 zero 2.75 4.53 sr110 23.4 2.75 1.02 3.77 5.88 sr120 24.2 2.75 2.25 5.00 6.58 sr130 21.8 2.75 3.75 6.50 7.50 sr145 21.8 2.75 6.30 9.05 8.90 sr200 23.4 4.29 zero 4.29 5.40 sr210 23.4 4.29 1.24 5.53 7.10 sr220 24.2 4.29 2.77 7.06 8.09 sr230 21.8 4.29 4.58 8.87 9.16 sr245 23.4 4.29 7.97 12.26 11.27 sr300 21.8 6.18 zero 6.18 6.80 sr310 21.8 6.18 1.55 7.73 8.95 sr320 23.4 6.18 3.61 7.79 10.55 sr330 23.4 6.18 6.19 12.37 12.37 sr345 24.2 6.18 10.3 16.48 14.57 higher shear strength than specimens without or with lower reinforcement parameter. results presented by [figure 8 and table 6] show that the presenting of the shear reinforcement normal to the shear plane are significantly increased the shear transfer stress by (46.2%, 74.02%, and 119.4%) when groups (sr1, sr2, and sr3) are compared to the unreinforced specimens of group (sc) for k=0 (i.e., σnx =0). this increasing in the shear strength can be attributed to the clamping force which is developed in the reinforcing bars within the yield range when diagonal cracks appear. effect of total normal compressive stress (σnx+ρfy) [figures 9-13] and [table 6] show the effect of total normal compressive stress (σnx+ρfy) on the shear transfer strength. figure 13: max. shear stress -total normal compressive stress relationship, (k=0.7070) figure 12: max. shear stress-total normal compressive stress relationship (k=0.5) muhodir: study of shear transfer in modified push-off members using finite elements method 88 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 81-88 with reference to this figure, it can be noted that the presence of external normal compressive stress (σ nx ) and shear reinforcement within the shear plane enhanced dramatically the shear transfer strength of the specimens used when compared with the those without shear reinforcement or those include parallel reinforcement only. furthermore, it can be concluded that the external normal compressive stress is additive to the reinforcement parameter (ρf y ) finally it can be concluded that, if a certain shear stress is to be resisted, the area of shear reinforcement can be reduced by an amount equal to the external compressive force (σ nx ) divided by (f y ). efficiency of the fem to verify the accuracy of the finite element model suggested in this study to the shear transfer analysis, the obtained results are compared with those obtained experimentally by al-sharae.[15] the experimental results obtained by al-sharae,[15] (v u, test ) and that obtained by the finite element, (v u, cal. ) are summarized in [table 7]. the standard deviation value (σ_ (n-1)) for (v_ (u, test)/v_ (f.e.m)) is 0.102, this indicates that the predicted shear strength using fem is very effective and gave a clear picture about the behavior of the push-off specimens. conclusions based on the results presented in this study, the following conclusions can be stated. 1. the two dimensional isoparametric element used in the mesh of the fem is quite efficient in idealizing the field of displacement and the state of stresses in the modified push-off specimens with and without shear reinforcement. 2. specimens with shear reinforcement (i.e., group sr) ad a ductile failure, while the specimens without shear reinforcement (i.e., groups sc and sp) had a brittle failure. 3. the parallel reinforcement has a little or ignored effect on the shear transfer strength. 4. an externally applied compressive normal strength which acting transversely to the shear plane is additive to the reinforcement parameter (ρ.f y ) in the calculation of the shear strength. 5. shear strength is increased with increasing the total compressive stress. (σ n + ρ.f y ). 6. fem results showed a good agreement with those obtained experimentally by other authors. references 1. l. ahmed and a. ansell. direct shear strength of high strength fiber concrete. magazine of concrete research, vol. 62, no. 5, pp. 379-390, 2010. 2. p. m. d. santoa and e. n. b. julio. a state-of-the-art review on shear friction. engineering structures, vol. 45, no. 1, pp. 435-448, 2012. 3. k. h. yang, j. i. sim, j. h. kang and a. f. ashour. shear capacity of monolithic concrete joints without transverse reinforcement. magazine of concrete research, vol. 64, no. 9, pp. 767-779, 2012. 4. k. a. harries, g. zeno and b. shahrooz. toward an improved understanding of shear-friction behavior. aci structural journal, vol. 109, no. 6, pp. 835-844, 2012. 5. k. n. rahal. simplified design and capacity calculation of shear strength in reinforced concrete membrane elements. engineering structures, vol. 30, no. 10, pp. 2782-2791, 2008. 6. american concrete institute. aci committee 318, building code requirements for structural concrete (aci 318-19) and commentary. american concrete institute, farmington hills, mi, usa, 2019. 7. t. t. c. hsu. unified approach to shear analysis and design. cement and concrete composites, vol. 20, no. 89, pp. 419-435, 1998. 8. b. j. al-sulayvani and j. r. al-feel. effect of direct compressive stress on the shear transfer strength of fibrous concrete. al rafidain engineering, vol. 17, no. 2, pp. 65-75, 2009. 9. s. mahmoodreza and b. e. ross. database evaluation of interface shear transfer in reinforced concrete members. aci structural journal, vol. 114 no. 2, pp. 383-394, 2017. 10. m. soltani, b. e. ross and a. khademi. a statistical approach to refine design codes for interface shear transfer in reinforced concrete members. aci structural journal, vol. 115, no. 5, pp. 1341, 2018. 11. a. h. mattock and n. m. hawkins. shear transfer in reinforced concrete-recent research. pci journal, vol.17, no. 2, pp. 55-75, 1972. 12. r. n. khaldoun. shear-transfer strength of reinforced concrete. aci structural journal, vol. 107, no. 4, p. 346, 2010. 13. w. f. chen. plasticity in reinforced concrete. mcgraw hill, new york, 1982. 14. e. hinton and d. r. j. owen. finite element programming. vol. 90. academic press, inc., cambridge, p. 345, 1977. 15. a. j. al-sharae. experimental and analytical study of shear transfer in reinforced concrete members made with abu-ghar limestone as a coarse aggregate. m.sc. thesis, university of basrah, iraq, 1999. table 7: comparison of v u using f.e.m and v u experiment specimen’s identity v u , f.e.m. (mp) (1) v u , experim[15] (mpa) (2) 2 1       sc00 3.10 3.69 1.19 sc10 3.41 3.98 1.17 sc20 3.83 4.46 1.16 sc30 4.49 4.87 1.09 sc45 6.10 6.54 1.08 sp00 3.85 4.06 1.05 sp10 4.69 4.84 1.03 sp20 5.43 5.45 1.01 sp30 6.39 6.04 0.94 sp45 7.65 7.34 0.96 sr200 5.40 6.26 1.16 sr210 7.10 7.91 1.12 sr220 8.09 9.76 1.22 sr230 9.16 11.92 1.30 sr245 11.27 13.52 1.21 σ n-1 =0.102 tx_1~abs:at/add:tx_2~abs:at 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 26-31 research article determination of elements in indigenous vegetables using icp-ms bashdar a. sadee1,2 1department of food technology, college of agricultural engineering sciences, salahaddin university-erbil, kurdistan region-iraq, 2department of nutrition and dietetics, cihan university-erbil, kurdistan region, iraq abstract determination of elements in indigenous vegetables is important as source of dietary that are essential for human consumption. heavy metal concentrations were determined in indigenous vegetables collected in erbil city-kurdistan region of iraq. icp-ms was applied to determine total concentrations of al, co, fe, mn, ni, se and zn in malva parviflora, gundelia tournefortii, arum spp, spinacia oleracea, mentha longifolia, beta vulgaris subsp, apium graveolens, lepidium sativum l, allium kurrat. schweinf and allium fistulosum after acid digestion of the samples with the aid of microwave using nitric acid/hydrogen peroxide. the contents of heavy metals in vegetable samples varied between 40 and 451 for al, 0.04 and 0.96 for co, 53 and 247 for fe, 3.37 and 11.58 for mn, 0.26 and 5.53 for ni, 0.09 and 0.98 for se and 1.58 and 4.41 for zn on the basis of dry weight. the methodology was validated by certified reference material gbw10015-spinach and inductively coupled plasma mass spectrometry analysis. the levels of al, mn, and ni in some investigated vegetables are higher than the permissible limits for human consumption. keywords: heavy metals, icp-ms, indigenous vegetables, microwave acid digestion, validation introduction recently, more attention has been paid to cumulating of heavy metals and metalloids in vegetables. vegetables can be exposed to environmental pollution more than other food systems because of loading vegetables into the air. the accumulation and taken up of heavy metals by vegetables in eatable and non-eatable fractions at a certain quantity may lead to health concerns to both living organisms and humans.[1] food safety concerns may occur because of phytotoxicity and absorption of high levels of heavy metal by vegetation resulted from transferring high levels of these heavy metals from agriculture soils.[2,3] bioaccumulation high quantity of toxic heavy metals in vegetables can cause the deficiency of nutrients of dietary to human beings or may cause health issues for both human beings and ecosystem.[4-7] heavy metals can be categorized into vital metals (fe, mn, cu, se, zn and etc.), possibly essential (ni, v, and co) and conceivably toxic (as, cd, pb, hg, etc.). essential metals play a major role for living organisms if they exist at low or high quantity, while exposures to toxic metals may harm even at low level for long-term exposure. exposure to toxic metals can cause various serious health issues with changing degree of harshness and circumstances such as issues of kidney, neurobehavioral and developmental disarrange, hypertension, and probably cancer.[8-13] furthermore, numerous of nervous, cardiovascular, renal, neurological deterioration in addition bone illness and many other health issues may occur because of high content exposure of metals beyond maximum permissible level (mpl).[1,14] humans consume vegetables in both cooked and raw forms because they are considered as an essential part of their diet. vegetables function as acid generation throughout digestion and some available metals in vegetables are even vital biochemically and psychologically from health prospective.[15] although al is relatively low bioavailable, it has been suggested that there is a connection between aluminum and alzheimer’s disease.[16] human metabolism can be regulated via metals including, co, fe, mn, se, and zn. mn is one of the important components of many enzymes exist in human and considered an essential element functions as an activator. although co and ni are vital for human beings, higher levels than the recommended values lead to metabolic abnormal.[1] this study was conducted to evaluate the contents of heavy metals found in various edible indigenous vegetables. corresponding author: bashdar a. sadee, department of nutrition and dietetics, cihan university-erbil, kurdistan region, iraq. e-mail: bashdar.sadee@su.edu.krd received: december 26, 2021 accepted: february 16, 2022 published: march 6, 2022 doi: 10.24086/cuesj.v6n1y2022.pp26-31 copyright © 2022 bashdar a. sadee. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) cihan university-erbil scientific journal (cuesj) sadee: determination of elements in indigenous vegetables using icp-ms 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 26-31 experimental chemicals and reagents all chemical compounds utilized in this work were of high purity reagent and milli-q water (18mωcm) was used to prepare all solutions. a standards stock solution of concentration 100 µg ml−1 in 5% nitric acid (hno 3 ) for al, co, fe, mn, ni, se, and zn was obtained from cpi international, usa. hno 3 70% reagent was ordered from merck (germany) and used to digest vegetable samples to obtain extracts of al, co, fe, mn, ni, se, and zn. hydrogen peroxide (h 2 o 2 ) 37% was purchased from fischer scientific, united kingdom. a certified reference material (spinach gbw10015) was supplied by the institute of geophysical and geochemical exploration, langfang, china. all glasswares used in this work then rinsed with milli-q water followed by soaking it in hno 3 acid before use. instrumentation inductively coupled plasma mass spectrometry (icp-ms, perkin-elmer sciex elan 6100 drc ii) was applied to identify the total elements (al, co, fe, mn, ni, se, and zn) in the analyzed vegetable samples. the instrument was supplied with a sturman-masters spray chamber and a v-groove quartz concentric nebulizer for sample induction. in concise, the optimum parameters of the instrument were a radio frequency power of 1250 w with ar flow rates of 13, 0.8 and 0.95 l min-1 for cool, auxiliary, and sample gas, respectively. for the experimental results, the intensity of the following isotopes was measured: 55mn, 59co, 27al, 56fe, 60ni, 66zn, and 78se. possible interferences were eliminated using collision cell technology and also 7 % h 2 in he gaseous with average flow rates of 3.4 ml. min−1 was used. internal standards such as indium (in) and iridium (ir) were used for all vegetable samples at 10 µg l−1 (final concentration). sampling and sample preparation a vegetable samples were purchased from the local markets located in erbil, kurdistan region of iraq in 2021. the common and scientific names of analyzed vegetables are presented in table 1. all samples were rinsed with milli-q water and placed in a sealed plastic box. a freeze drier was used to dry all vegetable samples for 48 h at −40°c. a fine powder of vegetable samples with 180 µm size obtained by sieving of grounded samples using agate pestle and mortar. determination of total metals in vegetable samples 0.2g of freeze-dried vegetable samples was weighted in teflon vessels with teflon covers, 5 ml hno 3 ( 70%) and 2 ml h 2 o 2 (30%) were added. the vegetable samples were acid digested using mars x-press (cem, usa) microwave using a previous method by sadee et al.[17] the applied digestion program was as follows: vegetable samples were digested using the microwave for 43 min at 1600 w. initially, the temperature was raised up to 160°c in 15 min and stayed at this temperature for 5 min. then, the temperature ramped from 160 to 200°c over 8 min and stayed at this temperature for 15 min. the teflon vessels were left to cool at room temperature. after digestion, vegetable samples were transferred into a volumetric flask (25 ml capacity), fortified with internal standards of in and ir with an ultimate contents of 10 µg l−1 and made up to final volume (25 ml) with 2% (v/v) hno 3 using milli-q water. total metals in selected vegetables were determined utilizing icp-ms. gbw10015-spinach certified reference (crm) material was applied validation of the method. results and discussion figures of merit the slop of the applied calibration curve was used to calculate the detection limit (dl) of the icp-ms instrument (μg g−1) and 3 times standard deviation of 10 measurements of blank value. the results of dl quantities of the icp-ms are presented in table 2. validation of analytical method in order to check the quality control of used procedures, total al, co, fe, mn, ni, se, and zn were quantified in gbw10015spinach. the results for, total al, co, fe, mn, ni, se, and zn contents in the crm are tabulated in table 3. the elemental contents resulted from the experiment were in well agreement with the certified quantities (the recovery of examined elements ranged from 94% to 105%). table 1: vegetables english common name and scientific name of collected samples common english name scientific name mallow malva parviflora sunflower gundelia tournefortii arum arum spp spinach spinacia oleracea wild mint mentha longifolia chard beta vulgaris subsp celery apium graveolens garden cress lepidium sativum l leek allium kurrat schweinf spring onion allium fistulosum table 2: limit of detection for al, co, fe, mn, ni, and se (μg g−1 dry weight) measured as the mean+3 sd of the signal of method blank metals concentration μg g−1 al 0.03 co 0.03 fe 0.02 mn 0.02 ni 0.03 se 0.05 zn 0.02 sadee: determination of elements in indigenous vegetables using icp-ms 28 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 26-31 total metal determinations in vegetables the averaged results of heavy metal concentrations in selected varieties of indigenous vegetables based on dry weight are shown in table 4 and the literature reported values from various origin of the world from corresponding vegetables are listed in table 5. aluminium the concentration of al in this study ranged from 40 to 451 µg g−1 [table 4]. the lowest concentration of al in analyzed vegetables was found in malva parviflora with 40 µg g−1, while the maximum level of al was detected in spinacia oleracea with value of 451 µg g−1. it was noticed that the al contents in the vegetable samples collected were in increasing order of m. parviflora > lepidium sativum l > mentha longifolia > apium graveolens > allium kurrat schweinf > beta vulgaris subsp > gundelia tournefortii > arum spp > allium fistulosum > s. oleracea. the observed al concentration in vegetable samples is compared to the literature reported values ranged 0.160–131.640 µg g−1 [table 5]. the values of al content of each investigated vegetable of the current study higher than that reported for the corresponding vegetables by other researchers. the concentration of al in vegetables of this study except m. parviflora is higher than the maximum permissible dose limit recommended for al, 60 mg/day.[18,19] cobalt the concentrations of co in examined vegetables of this study ranged between 0.040 and 0.143 µg g−1 [table 4]. although the concentration of co in vegetables of m. parviflora, g. tournefortii and l. sativum l. were below the limit of detection of the method, the highest co content was found in a. fistulosum which was 0.143 µg g−1. the co levels in the vegetable samples collected were in increasing order of m. parviflora ≈ g. tournefortii ≈ l. sativum l > a. kurrat schweinf > a. graveolens > arum spp > b. vulgaris subsp > s. oleracea > m. longifolia > a. fistulosum. the observed literature values of co were in the range of 0.01 3.00 µg g−1 [table 5]. the values obtained for co vegetables in the current study are lower than that reported in the literature for the same vegetable species except m. longifolia, a. graveolens and a. fistulosum. the concentration of co in the vegetables of this study is well within mpl of co for adults, 50 µg/day.[8,19] iron the quantities of fe varied according to the types of vegetables concentred with quantities between 53 and 247 µg g−1 [table 4]. the minimum and maximum fe contents of the samples were found to be 53 µg g−1 in b. vulgaris subsp and 247 µg g−1 in s. oleracea, respectively. the fe concentrations in the vegetable samples collected were in increasing order table 3: total metal concentration of crm gbw10015-spinach; all experimental values are calculated in μg g−1, mean±standard deviation (n=3) metals certified value experimental value obtained extraction efficiency % al 610±60 602±40 99 co 0.220±0.03 0.229±0.016 104 fe 540±20 546±42 101 mn 41±3 42±0.59 102 ni 0.920±0.012 0.966±0.020 105 se 0.092±0.024 0.087±0.004 95 zn 35.300±1.5 33.450±0.449 95 table 4: results of analysis for total metal in the selected vegetables dry weight (mean±standard deviation) (n=3) vegetable al μg g−1 ± sd co μg g−1 ± sd fe μg g−1 ± sd mn μg g−1 ± sd ni μg g−1 ± sd se μg g−1 ± sd zn μg g−1 ± sd malva parviflora 40±0.352 <0.03 58±0.584 4.930±0.044 0.260±0.002 <0.05 4.412±0.030 gundelia tournefortii 151±1.985 <0.03 100±0.805 3.370±0.003 0.532±0.004 <0.05 3.483±0.028 arum spp 298±1.640 0.050±0.006 139±0.791 5.330±0.066 1.142±0.007 <0.05 3.092±0.021 spinacia oleracea 451±5.361 0.960±0.001 247±2.275 9.382±0.099 1.351±0.015 0.090±0.014 4.080±0.035 mentha longifolia 103±1.102 0.120±0.013 198±3.823 6.731±0.159 1.533±0.019 <0.05 3.691±0.054 beta vulgaris subsp 140±0.932 0.070±0.002 53±0.341 11.580±0.076 0.564±0.006 0.980±0.031 4.295±0.039 apium graveolens 124±1.525 0.040±0.001 81±1.020 9.622±0.115 0.812±0.008 <0.05 3.234±0.035 lepidium sativum l 65±2.191 <0.03 131±0.415 4.183±0.017 0.722±0.004 0.494±0.004 3.195±0.023 allium kurrat schweinf 132±0.633 0.040±0.001 75±0.820 10.891±0.161 0.473±0.005 <0.05 1.588±0.026 allium fistulosum 334±3.416 0.140±0.003 138±0.305 5.142±0.035 5.533±0.081 0.242±0.015 2.420±0.024 sadee: determination of elements in indigenous vegetables using icp-ms 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 26-31 of b. vulgaris subsp > m. parviflora > a. kurrat schweinf > a. graveolens > g. tournefortii > l. sativum l > a. fistulosum > arum spp > m. longifolia > s. oleracea. in the literature fe concentration has been documented in the range of 28-1952 µg g−1 [table 5]. the values of fe varied according to the vegetable species, the concentrations of fe in this study are lower than the same vegetables reported in the literature except of s. oleracea. the results of the present work with the exception of s. oleracea are well within the provisional maximum tolerable daily intake (pmtdi) of fe is 0.8 mg/kg bw.[20] manganese the levels of mn in investigated vegetables of this study ranged between 3.374 and 11.580 µg g−1 [table 4]. the lowest concentration was seen in g. tournefortii (3.374 µg g−1) sample while the highest level was 11.580 µg g−1 for b. vulgaris subsp. the fe concentrations in the vegetable samples collected were in increasing order of g. tournefortii > l. sativum l > m. parviflora > a. fistulosum > arum spp > m. longifolia > s. oleracea > a. graveolens > a. kurrat schweinf > b. vulgaris subsp. the reported literature concentrations of mn varied from 8.4 to 178.53 µg g−1 [table 5], which are higher than the values obtained for the same vegetables of this investigation. the concentration of mn in the investigated samples except m. parviflora, g. tournefortii, and l. sativum l is above the recommended intake range of mn for an adult, 2-5 mg/day.[8] nickel the calculated concentration of ni in this study was recognized in the range of 0.264–5.532 µg g−1 [table 4]. the lowest level of ni in this study was found in m. parviflora which was 0.264 µg g−1, whereas the highest content of ni was 5.532 µg g−1 for a. fistulosum. the ni contents in the vegetable samples collected were in increasing order of m. parviflora > a. kurrat schweinf > g. tournefortii > b. vulgaris subsp > l. sativum l > a. graveolens > arum spp > s. oleracea > m. longifolia > a. fistulosum. the reported literature values of ni range from 0.1 to 21.1 µg g−1 [table 5]. the contents of ni in vegetables under study were higher than those corresponding vegetables documented in the literature except g. tournefortii, s. oleracea, b. vulgaris subsp, and l. sativum l. the concentration of ni in vegetables of present work is well above the permissible limit of daily intake of ni (0.1 mg/day), which is alarming.[18,19] selenium se was lower than the dl (0.030 µg g−1) in the examined vegetable samples of the current study with the exception for s. oleracea, a. fistulosum, l. sativum l. and b. vulgaris subsp were 0.094, 0.241, 0.490 and 0.980 µg g−1, respectively [table 4]. the literature values of observed concentrations of se are varying from 0.020 to 762 µg g−1 [table 5]. except a. fistulosum, the results for this study for se in vegetables under investigation including s. oleracea, a. graveolens, and a. kurrat schweinf were lower than the literature values reported table 5: heavy metal concentration in various vegetables in different countries (literature values) vegetable concentration μg g−1 location sampling period reference al co fe mn ni se zn malva parviflora na na 296 na 0.000 na 59 ethiopia 2012 [21] gundelia tournefortii na na na na na 1.080 1952 na 408.370 na 8.400 178.530 8.400 na na na na na 820 na 20.050 iran turkey turkey not mentioned 2001 not mentioned [22] [23] [24] arum spp na 131.640 na 1.4100 na na 166.400 na na 9.900 na na 17.140 na na na na 762 9.60 na na saudi arabia iran turkey 2003 not mentioned not mentioned [25] [26] [27] spinacia oleracea na 570 0.01 na 374.700 na 131.900 na 0.000 na na 0.177 202.40 na iran india 2007 2004 [28] [29] mentha longifolia na 3 146.300 30.730 21.100 na 28.100 saudi arabia 2003 [25] beta vulgaris subsp na 70 0.010 na 257.300 na 91.770 na 0.000 na na 0.020 188.000 na iran finland 2007 2003 [28] [30] apium graveolens na 0.330 1678 164.300 3.260 na 100 iran 2007 [28] lepidium sativum l na 0.160* na 1.700 na na 118.2 na na 13.840 na na 0.100 na na na na 2.550 17.400 na na saudi arabia sweden turkey 2003 1992 2014 [25] [31] [32] allium kurrat schweinf 50 0.040 28 14 0.280 0.110 16 finland 2003 [30] *wet weight; na: not analysed sadee: determination of elements in indigenous vegetables using icp-ms 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 26-31 for the same corresponding vegetables. the content of se in s. oleracea, b. vulgaris subsp, l. sativum l, and a. fistulosum is higher than the tolerable daily intake of se which is 55 µg/day.[8] zinc the concentrations of zn in vegetables under investigation ranged between 1.588 and 4.295 µg g−1 [table 4]. the minimum level of zn in the analyzed vegetables of the current study was detected in a. kurrat schweinf which was 1.588 µg g−1, meanwhile the maximum content of zn in vegetables understudy was detected in b. vulgaris subsp (4.295 µg g−1). the zn concentrations in the vegetable samples collected were in increasing order of a. kurrat schweinf > a. fistulosum > arum spp > l. sativum l > a. graveolens > g. tournefortii > m. longifolia > s. oleracea > m. parviflora > b. vulgaris subsp. the reported literature concentrations for zn are in the range of 9.600–820 µg g−1 [table 5]. this is comparable to the obtained values of this study. the concentration of zn in investigated samples is well below the pmtdi of zn, 0.3–1.0 mg/kg bw.[8] conclusion the present study has generated data on some of heavy metal profile in common indigenous vegetables consumed in erbil city located in the kurdistan region of iraq. the contents were observed and compared with the literature values that have been reported around the world for the same vegetables. al, co, fe, mn, ni, se, and zn were measured in vegetables using hno 3 /h 2 o 2 microwave-assisted acid digestion followed by icp-ms. gbw10015-spinach was used to validate the method. vegetables of this study showed similar ability to accumulate investigated heavy metals as follows: high quantity for both al and fe; medium quantity for zn and mn and small quantity for both co and se. in addition, al was found to be at high levels in the majority of vegetables of this study, while se was below the limit of detection in the majority of investigated vegetables. more studies should be taken in order to investigate the high al contents in analyzed vegetables such measuring “plant available” for absorption or “bioavailable” of total al in soils that used to cultivate 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medicine and public health, vol. 39, p. 335, 2008. 16. g. kfle, g. asgedom, t. goje, f. abbebe, l. habtom and h. hanes. the level of heavy metal contamination in selected vegetables and animal feed grasses grown in wastewater irrigated area, around asmara, eritrea. journal of chemistry, vol. 2020, pp. 1-15, 2020. 17. b. a. sadee, m. e. foulkes and s. j. hill. obesity before, during, and after pregnancy: a review and comparison of five national guidelines. food additives and contaminants: part a, vol. 33, pp. 433-441, 2016. 18. o. d. uluozlu, m. tuzen, d. mendil and m. soylak. assessment of trace element contents of chicken products from turkey. journal of hazardous materials, vol. 163, pp. 982-987, 2009. 19. g. f. nordberg, b. a. fowler and m. nordberg. handbook on the toxicology of metals. cambridge, massachusetts: academic press, 2014. 20. world health organization. joint fao/who food standards programme (codex alimentarius commission) 39th session rome, italy, 27 june-1 july 2016 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36-40 research article the essential pheromonal elements of preputial glands of castrated and testosterone-treated mice merza h. homady1, ali a. majeed2, maysoon t. al-haideri3, mustafa t. younis4 1department of biomedical sciences, cihan university-erbil, kurdistan region-iraq, 2department of pathological analyses, faculty of sciences, kufa university, kufa, iraq, 3department of physiotherapy, cihan university-erbil, kurdistan region, iraq, 4department of general biology, cihan university-erbil, kurdistan region, iraq abstract the present study investigated the effects of both castration and treatment of castrated subjects with 0.1 ml of 50 µg/kg/day of testosterone at 3  weeks of age for 6  weeks on the pheromonal and histological structure of male mice preputial gland. the preputial gland results from the control intact group showed that six essential pheromonal compounds are present, whereas the glandular structure revealed well-developed acini with basement membrane and connective tissue around most of them. these findings also showed that castration has deleterious effects on the essential pheromonal compounds which are reduced to two compounds only (the α.-farnesene and 1- tetradecanol) as well as the histological sections from such subject appeared many atrophied vacuolated acini. on the other hand, the treatment of castrated animals with such dose of testosterone did not show any marked effects on the essential pheromonal compounds, but it was able to restore the activities of acinar development as showed by many hypertrophied alveoli. keywords: testosterone, pheromones, histology, preputial, castration introduction chemical communications are important in the social lives of many species of mammals; several of these chemicals are referred to as pheromones.[1,2] the term pheromone was created and described based on findings in insects, but whether the original definition can be applied to mammals which is still discussed.[3] when mice are stressed, they emit a specific odor into the air; because this stressrelated odor causes anxiety in the other mice, it is likely that some anxiogenic molecules are present in this odor.[2,4] pheromones have been characterized to generate either signaling activity that results in a rapid change in motor activity by the recipient, which on detection rapidly initiate behavior such as aggression or fear[5] or priming pheromones that trigger rather slower neuroendocrine or endocrine changes in the recipient.[6,7] pheromones are mainly found in the preputial glands and urine of male mice, according to numerous studies.[8] it is very well known that many of the volatile plant compounds normally found in insect natural habitats alter the perception from their own pheromone when introduced separately as a background to pheromone. olfactory communication is key for insects and rodents since it aids in the identification and location of critical resources including a food source, a mate, and threat.[9-11] chemical information exchanges are therefore not only crucial to insects or rodents but also to the functioning of the species network that makes up a community. once released into the atmosphere, these ecologically relevant signals and cues are carried by airflows, diluted, and mixed with other volatile organic compounds to form a complex and changing olfactory landscape.[12,13] the preputial glands are androgen-dependent organ; testosterone is the most well-known androgen, which is derived from dihydrotestosterone. furthermore, the activity of the preputial glands is reliant on the melanocortin 5-receptor (mc5r), that is, one of the melanocortin receptors (mcrs) are a fami1y of five g protein-coupled receptors (gpcrs; mc1r–mc5r) expressed in varied tissues, each of which serves a distinct physiological function.[14,15] because the preputial gland is a vital gland that plays an important role in the development of behavior-modifying “pheromones” in cihan university-erbil scientific journal (cuesj) corresponding author: merza h. homady, department of biomedical sciences, cihan university-erbil, kurdistan region-iraq. e-mail: merza.homady@cihanuniversity.edu.iq received: june 20, 2022 accepted: july 18, 2022 published: august 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp36-40 copyright © 2022 merza h. homady, ali a. majeed, maysoon t. al-haideri, mustafa t. younis. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). homady, et al.: pheromonal elements of mice preputial glands in relation to endocrine factors 37 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 36-40 mice, it is significantly influenced by a variety of hormones and medicinal herbs. up to our knowledge, there was no work that has done on gas chromatography and mass spectrometry (gc-ms) analysis the histological structure and major pheromonal molecules of this pheromone producing glands. the present research is aimed to investigate the effects of both castration and treatment of the castrated subjects with testosterone on the histological structure and pheromonal compounds of the preputial gland. methodology chemical (hormone) preparation all preparations administration by injection was given as “oily solution.” the oil vehicle used for these injections was a combination of one part of ethyl oleate (bhd, poole, dorset) to four parts of arachis oil (hopkin and williams, essex). almond oil was contrasted with this preparation as there are suggestions that arachis oil have estrogen action.[16] hormones were obtained in crystalline form as free alcohols from sigma ltd. (london). preparation of a compound for injection consisted of weighing the required amount of steroid, dissolving it in ethanol (analar grade from bdh, poole, dorset) and making the resultant solution up to a volume of 100 ml with oil vehicle. as 0.1 ml was generally injected, calculations were based on the quantity of material in this volume. preparation of testosterone solution: one tablet of testosterone (50 mg) (testosterone-austrian) was taken and followed by the same procedure mentioned above, to get a concentration of 50 µg/ml testosterone. injections were performed using all – tuberculin, lure filling syringe (rocket, london) all syringes and other glass were cleaned before use in decon 90. the needle was 23 × 1 gauge (arnold r. horwell ltd. london). hormonal solutions were usually injected daily for 42 days into the thigh muscle (alternating legs on consecutive days). experimental design tuck ordinary (to) strain albino mice have been bred and kept in the animal house unit of kufa university’s faculty of sciences under controlled circumstances of 12 h light: 12 h darkness. the subjects were retained in plastic cages. sawdust bedding was used, as well as food and water were provided on an ad libitum basis. animals housed in groups of comparable age (3 weeks) were assigned to the categories listed below: a. intact male mice received tap-water as a positive control, (n = 7) b. castrated animals were given tap water as experimental control, (n = 7) c. castrated subjects delivered daily with 0.1 ml of 50 µg/kg/day of testosterone from 3 weeks of age for 6 weeks. castration experimental mice were anesthesia before surgery with (500 mg ketamine + 20 mg 4/1 xylazine) for 15 min. the basic operation in these experiments was bilateral castration. after inducing general anesthesia with ketamine and xylazine and disinfecting the scrotum with 75% a1cohol, a small midline incision was made, the spermatic vessels were tied with 4.0 silk sutures, and the testes were removed. the incision was then closed with 4.0 silk sutures. mice were transformed on recovery to clean cages. assessment of pheromones before extraction, the glands are thawed at room temperature and the yellowish secretion is gathered into a clear vial by squishing, then the secretion is collected and added 1.5 ml of dichloromethane into the vial, and after 24 h, the supernatant is removed and the remaining solution stored at 0°c until gc-ms analysis within 1 week.[17] conjunction of both gc-ms was used. histological analysis mice were killed by the end of the experiment by over dose of ketamine and xylazine, and the preputial glands were used for histological investigation under light microscope. the histological methods used have been processed according to bancroft.[18] results biochemical analysis the results of table 1 showing the essential pheromonal compounds present in the preputial gland of intact control mice were six compounds, whereas these compounds are reduced in castrated animals to two only (α.-farnesene and l-tetradecanol). the treatment of castrated subjects with 0.1 ml of 50 µg/kg/day of testosterone appeared two essential compounds only (1-tetradecanol acetate and 1-hexadecanol). histology of the preputial glands the intact control group’s preputial gland was observed under the microscope and revealed normal structure [figures 1 and 2]. the gland is covered with thick connective tissue capsule, from which trabeculae protrude into the interior, partitioning the parenchymatous tissues into lobes and lobules at varied stages of growth. many acini have hypertrophied with healthy ovalshaped nuclei and numerous normal cytoplasmic organelles. the acini are bordered with basal cells that are more evident. the gland has a wide excretory duct which drains into the preputial cavity and has a broad lumen lined by stratified squamous epithelium. sections of preputial glands from castrated group [figures 3 and 4] showed atrophied acini and most of them are vacuolated with increased connective tissue capsule. in addition, the basal cells are fewer in number as compared to the control normal group. testosterone treatment of castrated animals has dramatic effects on the gland structure. such dose of testosterone was able to increase the extent of acinar development which is resulted in glandular hypertrophy with many acini at various stages of development. the parenchymatous acini were compact and many fused to form irregular masses [figures 5 and 6]. discussion the use of olfactory communication is wide-spread throughout the animal kingdom, as genetic, hormonal variations, social homady, et al.: pheromonal elements of mice preputial glands in relation to endocrine factors 38 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 36-40 imprinting, overall health, environmental factors, dietary changes, and other factors all appear to have a significant impact on how chemo signals are discharged and perceived in the sense of competition for reproductive potential.[19] at present, a different researcher uses the term “pheromone” in the sense of mammals with much less apprehension. this can be attributed, at least in part, to (i) recent groundbreaking research identifying the ubiquity of receptor proteins found in olfactory neurons, (ii) a better understanding of the “chemically selective” vomeronasal organ, and (iii) the evidence that distinct behavioral and hormonal responses in mammals can also be caused by a single chemical substance or a relatively simple synthetic mixture.[20,21] it is much easier to understand pheromone – protein interactions with a various of smaller soluble binding proteins. a number of these proteins may be involved in what are known as perireceptor events.[22] the main source of pheromones in the mice are preputial glands that are rich in wax esters, triglycerides, and table 1: the essential pheromonal compounds present in the preputial glands of: intact control mice, castrated, and castrated treated with testosterone number of essential pheromones control (intact subjects) castrated subjects castrated treated with testosterone 1 -farnesene β 2 (e)α.-farnesene and α.-farnesene α.-farnesene 3 1tetradecanol 1tetradecanol 1-tetradecanol acetate 4 1-hexadecanol 1-hexadecanol 5 1-hexadecanol, acetate 6 tetradecanol, acetate figure 1: h&e-stained cross histological section of intact mouse’s preputial glands, exposing the stroma, trabecular system, and capsule (×100) figure 2: the acini and basal cells are visible in a cross histological section of intact mice’s preputial glands stained with h&e (×400) figure 3: a cross histological section of castrated mice’s preputial glands stained with h&e displays acini depletion and that most of them are vacuolated (arrow) (×100) figure 4: transverse histological section of castrated mice’s preputial glands stained with h&e exhibits atrophied acini (arrow) and fewer basal cells (×400) homady, et al.: pheromonal elements of mice preputial glands in relation to endocrine factors 39 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 36-40 alkyl-2,3-diacylglycerols and are believed to play a role in behavioral interactions through the release of pheromones.[20] the mc5r gene is expressed in the central nervous system and in a variety of peripheral tissues, especially in the skin. the encoded protein is involved in different physiological processes, including lipid metabolism, exocrine function, and pro-inflammatory activity.[23] mc5r belongs to a group of mcrs in the melanocortin system, and each one plays a major role in the body’s physiological activities. when the mc5r gene was disrupted in transgenic mice, it disrupted their exocrine glands, resulting in a decrease in sebum production.[24,25] according to morgan and cone,[26,27] the preputial gland of rodents expresses the mc5r and secretes pheromones that encourage aggressiveness. a pheromone signal that suppresses aggression is disinhibited by mc5r deficiency. gpcrs are essential for bringing various external signals into cells and controlling a number of physiological processes. mc5r deficiency produces a preputial defect that abolishes trophic responsiveness to melanocortins.[28] it was noted that castration markedly reduced preputial gland size and sebum content. indeed, the preputial and other sex accessory organs of adult rodents seem largely dependent on continued androgenic stimulation for their differentiation and the preservation of their normal structure and function.[29] in addition, castration in the present study has a depressed effect on the essential pheromone compounds which resulted two compounds only the (α.-farnesenes and 1tetradecanol) as compared with the normal control subjects. these findings are in agreement with zhang et al.[30] of the identification of essential pheromonal compounds. the histological sections from such subject appeared atrophied preputial acini with most of them which are vacuolated. these results explanted the findings of homady[31] when he demonstrated that castration has harmful effects on the preputial glands of male mice. reducing androgens by castration, hypophysectomy, or application of antiandrogenic compounds (such as oestrogen and flutamide) results in structural involution of mice, rat ҆s sex – accessory organs, indicated by cellular atrophy, depressed size and organization of cellular organelles, disorganization and loss of granular endoplasmic reticulum, decreased numbers of free ribosomes and secretory granules, nuclear shrinkage, depleted mitochondria, loss of acid, and alkaline phosphatase activity and abolition of metabolic and functional activities.[19,32] the histochemical analysis of preputial glands from castrated mice treated with testosterone appeared two types of the essential pheromonal compounds (1-hexadecanol and 1-tetradecanol acetate). these findings suggested that such dose of testosterone did not restore the essential compounds which can be attributed to dose effect. these findings are agreed with the findings of[19,33] who demonstrated that castrated male mice treated with testosterone maintained the activity of this gland as well as sexual behavior of castrated mice. in the present findings, testosterone treatment was able to restore the preputial histology structure to limit extent which appears that it is a dose dependent. actually, testosterone appears to be a precursor to a number of other steroids and many only exert its physiological and behavioral actions after metabolic conversions. these transformations may increases the variety of candidates for behavioral control with particular pathway having differing importance in different species enabling organisms with the ability to influence two or more targets relatively independently.[34,35] up to our knowledge, there was no works were conducted using gc-ms on preputial glands of castrated or castrated treated subjects with testosterone and this study was intended to fill this gap. conclusion in the current research, six combination of essential pheromones were identified in the normal intact subjects. therefore, it seems that castration has a deleterious effect on both gland activity and essential pheromonal compounds, whereas treatment with testosterone has significant effects to restore preputial activities but did not able to restore the major essential compounds that may be contributed to the lowest dose of testosterone used. figure 6: section from castrated mice treated with testosterone having their preputial glands stained with h&e, showing a welldeveloped stroma with some hypertrophied acini (arrow) in the central part (×400) figure 5: cross-section of the preputial glands of castrated treated mice with testosterone, stained with h&e, demonstrating the restructuring of some gland regions (×100) homady, et al.: pheromonal elements of mice preputial glands in relation to endocrine factors 40 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salih, m. m. al-jubori and m. d. younus. immunohistochemical and molecular studies of p53 and kras protein and their relations to colorectal carcinoma. cihan university-erbil scientific journal, vol. 5, no. 1, pp. 28-33, 2021. 35. s. achiraman, d. s. ganesh, s. kannan, s. kamalakkannan, n. nirmala and g. archunan. response of male mice to odours of female mice in different stages of oestrous cycle: self-grooming behaviour and the effect of castration. indian journal of experimental biology, vol. 52, no. 1, pp. 30-35, 2014. tx_1~abs:at/add:tx_2~abs:at 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 research article us dollar/iq dinar currency exchange rates time series forecasting using arima model parzhin a. mohammed1, sami a. obed1, israa m. ali1, dler h. kadir1,2 1department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq., 2department of business administration, cihan university-erbil, iraq abstract the use of currency exchange estimation as a tool for economic planning is being researched as a technique for gaining economic stability. the main purpose of this study is to use the autoregressive integrated moving average (arima) model to forecast monthly us dollar against iq dinar exchange rates. the information was gathered from january 2010 to december 2020. we got the information from the website (sa.investing.com). the minimum value of root means square error and the mean absolute error are used to select the optimal model. arima (2, 1, 0) was found to be the best model for the us dollar/iq dinar series. this is the forecasted meaning for the future of this exchange rate time series, which indicates a perpetual increase continuously in the next two years. statgraphics version 15 was the statistical software package utilized to complete this project. keywords: exchange rate forecasting, us dollar/iq dinar, time-series analysis, autoregressive integrated moving average model introduction time series analysis is used in a wide variety of fields. in economics, time series are used to represent the economy’s recorded history. the consumer price index, gross national product, and unemployment, as well as the population, products, and output, are all measured in the same time series. the statistical theory of time series has sparked a lot of interest in recent years. it is generally believed that the degree of financial transactions in and out of a country depends on several factors prevailing in that country. since the advent of money, the world has always experienced the movement of money from one country to another due to economic, cultural, social, political, and other reasons. the central bank in every country usually keeps a record of such money transactions through various financial intermediaries. every country has its own money, which can only be accepted for use within its territory. hence, the establishment of foreign money became necessary. foreign money is the money of other countries of the world which serves as money in the foreign exchange market. the researcher’s study time-series data to attempt to establish a strong statistical way for forecasting a future rate of exchange of the us dollar over the iraqi dinar basis on previous data here on the exchange rate of the us dollar and the iraq dinar. using the historic date of the exchange rate of the local currency against the foreign currency, this statistical tool can be used to forecast the rate of exchange of any foreign currency. the most common technique in time series is box-jenkins modeling. the other name of it is autoregressive integrated moving average (arima) modeling. recent work on arima forecasting has revealed an apparent accuracy for forecasting. arima models have been applied for us dollar/iq dinar currency exchange rates time series forecasting. in this work, the original dataset has 132 observations. arima modeling was used to forecast two years ahead of data. the rest of the paper is structured as the following: section 2 describes the methodology of box jenkins and provides a brief introduction to the theory of arima model building. section 3 introduces the data used in this study and follows the arima procedure for building an adequate model for the forecasting exercise and discusses the results. finally, section 4 is devoted to conclusions.[1-5] corresponding author: parzhin a. mohammed, department of statistics, college of administration and economics, salahaddin university erbil, kurdistan region f.r. iraq,. e-mail: parzhin.mohammrd@su.edu.krd received: november 29, 2021 accepted: january 02, 2022 published: february 10, 2022 doi: 10.24086/cuesj.v6n1y2022.pp12-19 copyright © 2022 parzhin a. mohammed, sami a. obed, israa m. ali, dler h. kadir. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) mohammed, et al.: time series forecasting using arima model 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 methodology this section introduces some basic definitions and concepts. time series a time series is a collection of correlated observations made in a specific order across time. a discrete-time series is one in which the set t 0 of times at which observations are taken is discrete, such as when they are conducted at fixed intervals. when observations are recorded continuously across a time interval, such as when t 0 = [0, 1], a continuous-time series is obtained. if we want to emphasize that observations are recorded continuously, we’ll use the notation (t) rather than x.[6,7] stationary time series the first and second moments of a stochastic process x t are deemed stationary although they are time-invariant. to put it another way, if the first criterion is met, x t is stationary, meaning that all members of a stationary stochastic process have the same constant mean. for example, a time series generated by a stationary stochastic process must vary around a constant mean and not show a trend. the variances are often time-invariant due to the second requirement. the variance σ µ2 2x t xe x= −[ ( ) ] =0 is independent of t when k = 0. furthermore, the covariance e [(x t −μ x ) (x t−k −μ x )] = γ k depends only on the distance in time k between the two members of the process, not on t.[8,9] time series models a time series model for observed data x t is a description of the joint distributions (or, in some cases, just the means, and covariance) of a set of random variables x t , where x t is supposed to represent a realization.[10] autoregressive (ar) model a process {x t } is said to be an ar model of an order p, abbreviated ar (p), is of the form. x t =∅ 1 x t−1 + ∅ 2 x t−2 +….+ ∅ p x t−p +ϵ t (1) where it x t is stationary, ∅ 1 , ∅ 2 ,…., ∅ p , are constants (∅ p ≠0).[11] moving average (ma) model a process {x t } is called to be the ma model of order q, or ma (q) model, is identified to be x t = ϵ t −θ 1 ϵ t−1 −θ 2 ϵ t−2 −…− θ q ϵ t−q (2) where there are q lags in the ma and θ 1 , θ 2 ,….θ p (θ p ≠0) are parameters.[11] arma model the process {x t , t = 0, ± 1, ± 2.,}is said to be an arma(p, q) process if {x t } is stationary and if for all the t, x t −∅ 1 x t−1 − −∅ p x t−p = ε t + θ 1 ε t−1 + +θ q ε t−q (3) where {ϵ t } ~ n (0,σ2). we say that {x t } is an arma (p, q) process with mean μ. if x t −μ} is an arma (p, q) process.[7] arima model if the dth difference z t = ∇dx t is a stationary arma process, a time series {x t } is said to follow an integrated arma model. we state that {x t } is an arima (p, d, q) process if {z t } follows an arma (p, q) model. fortunately, we can typically take d=1 or 0 greater than for practical purposes.[9] consider an arima (p, 1, q) process. with z t = x t −x t−1 , we have z t = ∅ 1 z t−1 + ∅ 2 z t−2 +…+ ∅ p z t−p +ϵ t −θ 1 ϵ t−1 –θ 2 ϵ t−2 … −θ q ϵ t−q (4) stages of time -series model building the process of a three-step adaptive approach is used to build an arima model. first, previous data analysis helps to identify a probable model of a time series model class. second, the model’s unknown parameters are approximated. finally, diagnostics checks are carried out through residue left models to analyze the model’s appropriateness or to predict possible developments. then, we will go through each of these processes in any further context.[12] identification to make the data stable, it may have to be pre-processed. some of the other four stages below must be followed to achieve stationarity: 1start by looking at it. 2rescale it using a logarithmic or exponential transform, for example. 3take out the deterministic elements. 4and that is the distinction. to put it another way, take ∇(b)d x until you reach a stop. in most cases, d = 1,2 will enough. the autocorrelations decline to zero exponentially fast, indicating that the system is stationary. we can try to fit an arma (p, q) model when the series is stationary. the correlogram � 0  ˆ  /kkr γ γ= and partial autocorrelations ˆ kk∅ are considered. the following observations have previously been made. both the sample acf and pacf, as previously mentioned, have a standard deviation of around 1/n, where n is the length of the series. a good rule of thumb is that acf and pacf values between ±2/√n are inconsequential. for k > max (p, q), the kth order sample acf and pacf decay geometrically in an arma (p, q) process, as shown in table 1.[13,14] estimations of parameters various methods, such as the moment method, least squares, maximum likelihood, and yule-walker estimate, can also table 1: theoretical acf and pacf properties for stationary processes process acf pacf ar (p) tails off as exponential decay or damped sine wave cuts off after lag p ma (q) cuts off after lag q tails off as exponential decay or damped sine wave arma (p, q) tails off after lag (q-p) tail off after lag (q-p) mohammed, et al.: time series forecasting using arima model 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 be used to predict the values as in the tentatively defined model.[12] verification of the model the box-jenkins methodology’s confirmation procedure is rather extensive. the predicted model’s compliance with the examined data must be confirmed using various diagnostic tests.[15] forecasting[12] once a suitable time series model has been fitted, it may be used to create forecasts of observations in the future. if the current time will be is denoted by t, the forecast for x t +τ is named the τ -period-ahead forecast, then denoted by ( ) ˆ ttx τ+ . the mean squared error, which is the averaged value of the squares prediction error, is the conventional criterion to use in getting the best forecast, ( ) 2    2ˆe[( t ) ] e [  ( ) ]t ttx ex τ τ τ+ +− = is minimized. it can be seen that the best forecast in the mean square sense is the conditional expectation of x t +τ given current and previous observations, that is, x t , x t−1 ,…. x x x xt t t t� |) ( , ,...)�� + + −=τ τ(t e 1 (5) consider, for example, an arima (p, d, q) process at time t+τ (i.e., τ a period in the future): x xt i p d i t i t i q i t+ = + + − + = += + −+ ∅∑ ∑τ τ τ τδ θ 1 1   (6) and box jenkins described the flow chart shown in figure 1.[16] the dependable statistical standards to test forecasting model mean absolute error (mae) ( ) ( ) n 1t 1 1 | |e 1      n ˆ |  |ˆ n t t n t t t t t y y nmae y y n n == = − = = = − ∑∑ ∑ (7) where t = time period, n = total number of observations, and e t = (observed value forecasted value) at time t.[17] root mean square error (rmse)[1] this is simply the square root of mse: 2 1 )    ˆ( n t t t y y rmse mse n = − = = ∑ (8) application on realdata data description the dataset used in this analysis is monthly us dollar/iq dinar currency exchange rates time series forecasting using the arima model. all the data were collected from the website (sa.investing.com).data were collected for the period january 2010 to december 2020. there were overall 132 observations. the statistical analysis was computationally implemented in the stat graphics 15 software, as shown in table 2. time series plots the monthly exchange rate between the us dollar/iq dinar is depicted in the data set’s time series plot. because the variation in the size of the fluctuations with time is referred to as nonstationary in the variance, the plot reveals that the data is clear of being non-stationary in the mean and variance, seasonal, and trend patterns with an increasing variety of variance, as shown in figure 2. randomness test first of all, the randomness test of the one series should be made using the box-pierce test. the null hypotheses are the time series are random, versus the alternative hypotheses of non-randomness, to know that the data has a certain behavior than having a random behavior. as explained in table 3 for the series; as p < 0.05 to every series, therefore null hypotheses were rejected, meaning that the one-time series are not random series. the hypothesis was as follows for the exchange rate us dollar/iq dinar the series. figure 1: depicts the box-jenkins model-building methodology figure 2: time series plot of the original monthly exchange rate usd/iqd data from 1 january 2010 to 31 december 2020 mohammed, et al.: time series forecasting using arima model 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 table 2: monthly average us dollar/iq dinar currency exchange rates data for the period 1 january 2010–31 december 2020 years 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019 2020 january 1169.00 1166 1163.5 1163.25 1163.8 1164 1175.9 1181 1184.05 1186.43 1189.4 february 1165.9 1175 1163 1163 1163 1165 1196.44 1175 1184.05 1186.43 1189 march 1170 1175 1160.9 1160 1163.55 1163 1170 1166 1184 1189 1187.62 april 1170.1 1165 1160.9 1157 1162 1164 1165.7 1166 1185.24 1186.43 1187.62 may 1168 1166 1162.25 1145 1162.5 1165 1169.4 1167 1186.43 1186.43 1189 june 1168 1166 1162 1160.5 1162.9 1162 1167.2 1166 1190 1186.4 1187.62 july 1170.03 1166.05 1162.05 1162.5 1164.2 1156 1168 1167 1186.43 1186.43 1187.62 august 1171 1166 1162 1164 1161.3 1145 1162.7 1177.73 1186.43 1186.2 1187.62 september 1169.2 1166 1162 1163.8 1163.05 1131 1162.7 1166.1 1186.43 1186.43 1187.62 october 1169.05 1167 1162 1164 1162 1122 1164 1166 1186.43 1189 1189 november 1169.47 1167 1162 1160.8 1160 1108 1160.3 1182.8 1186.43 1187.62 1187.62 december 1166.5 1166.3 1163 1163.8 1142 1095 1181 1184 1186.43 1187.62 1459 figure 3: exchange rate usd/iqd series plot after non-seasonal differencing of orders one h 0 : the series is random h 1 : the series is not random data transformation after proving the non-randomness of the exchange rate us dollar/iq dinar time series, it can be concluded that all the data series have a certain behavior, so to overcome this behavior we will try to transform the data series, each according to its behavior. then, the data were transformed, for instance, a non-seasonal difference of order one for the variable. then, the series was tested again to test the existence of randomness. the hypotheses were accepted when p > 0.05 as shown in table 4. the hypothesis for the exchange rate usd/iqd series was as follows. h 0 : the series is random h 1 : the series is not random. it can be noticed that all the data series were accepted to be random after the transformations made on the series. stationary the series under consideration must satisfy the condition of being stationary; that is, the mean and variance are independent of time throughout the series. the original series needs to be differentiated to make the series stationary around meaning and to get stationary around variance for the series, we make transformations the rest original series exchange rate us dollar/iq dinar is needed non-seasonal differencing of order one transform to achieve stationary, figure 3 show the differenced series. to identify the entire order of the models for the series, we investigate the autocorrelation function (acf) and partial acf (pacf) for the original data. the acf and pacf plots are displayed in the figure 4 series. the estimated autocorrelations and pacf between corrected data values at various lags are shown in this table. the auto-correlation coefficient lag k determines between table 3: hypothesis of randomness test and p-value for data series box-pierce test p-value decision exchange rate usd/iqd 0.000 (snr) table 4: hypothesis of randomness test and p-value for transformed data series box-pierce test p-value decision exchange rate usd/iqd 1 (sr) the residuals at time t-k and time t is related and the partial autocorrelation coefficient measures the correlation between the residuals at time t and time t+k having accounted for the correlations at all lower lags 95.0% probability bounds around 0 are also indicated. there is a statistically significant association at the 95.0% confidence level if the probability bounds for a certain lag do not contain the calculated coefficient. the 95.0% probability bounds around 0 are also indicated. there is a statistically significant association at the 95.0% confidence level if the probability bounds for a certain lag do not contain the calculated coefficient. at the 95.0% confidence level, none of the 24 partial autocorrelation coefficients are statistically significant. the same procedure was conducted for the series; table 5 through table 6 contain that. mohammed, et al.: time series forecasting using arima model 16 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 table 5: estimated autocorrelations for adjusted exchange rate usd/iqd lag autocorrelation standard error. lower 95.0%probability. limit upper 95.0%probability. limit 1 0.260945 0.087038 -0.170593 0.170593 2 0.211871 0.092776 -0.181839 0.181839 3 0.189218 0.096372 -0.188886 0.188886 4 0.174333 0.099146 -0.194324 0.194324 5 0.16255 0.101442 -0.198824 0.198824 6 0.160989 0.103397 -0.202654 0.202654 7 0.16544 0.105279 -0.206343 0.206343 8 0.16592 0.10723 -0.210168 0.210168 9 0.167997 0.109158 -0.213946 0.213946 10 0.178883 0.111099 -0.217751 0.217751 11 0.187412 0.11326 -0.221986 0.221986 12 0.170011 0.115586 -0.226544 0.226544 13 0.140489 0.117465 -0.230227 0.230227 14 0.1297 0.118731 -0.232709 0.232709 15 0.127231 0.119799 -0.234803 0.234803 16 0.128438 0.120819 -0.236801 0.236801 17 0.127931 0.121849 -0.23882 0.23882 18 0.132377 0.122862 -0.240806 0.240806 19 0.131391 0.123938 -0.242914 0.242914 20 0.119234 0.124989 -0.244974 0.244974 21 0.126591 0.125847 -0.246657 0.246657 22 0.120522 0.126808 -0.24854 0.24854 23 0.10901 0.127673 -0.250235 0.250235 24 0.0940275 0.128376 -0.251614 0.251614 figure 4: the original monthly acf and pacf for the us dollar/iq dinar exchange rate data after we take the differenced series for the exchange rate, which is transformed into a stationary series of the mean and the variance, the acf and pacf plots of the differenced series show that all the lags are within the confidence limits as shown in the mentioned figure 5. choosing appropriate model after checking for stationary of the time series’ mean and variance, the model identification process is used to determine the adequate model given the acf and pacf. the appropriate model for the series is shown in table 7. at least the rmse and mae are used to determine the best models. model identification after getting on the stationary for the time series study, the acf and pacf are shown in the table accordingly for stationary data. we will examine the data in acf and pacf. the arima model for the exchange rate variable is obtained (2,1,0) 12 . estimation it was found that the best model describing the exchange rate us dollar/iq dinar series mentioned above, the parameters were estimated, as shown in table 8. mohammed, et al.: time series forecasting using arima model 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 model diagnostic checking the residuals for the fitted model should be just white noise for forecasting a good model; in other words, the chosen model has exhausted the whole behavior of the series. as a result, we compute and plot the acf and pacf of the residuals; we want to find no significant acf or pacf. furthermore, the fitted model should accurately predict the future. autocorrelation and partial autocorrelation of residual test the residual acf and pacf of the models that we develop will be checked. also, the model needs to pass the test for randomness of the residuals to prove that the model chosen has exhausted the data and that there are no significant remaining to be explained in the data. after the model diagnostics figure 5: acf and pacf of exchange rate time series after non-seasonal differencing of order one table 6: estimated partial autocorrelations for adjusted exchange rate usd/iqd lag partial autocorrelation standard. error lower 95.0%probability. limit upper 95.0%probability. limit 1 0.260945 0.0870388 -0.170593 0.170593 2 0.154285 0.0870388 -0.170593 0.170593 3 0.112343 0.0870388 -0.170593 0.170593 4 0.0880657 0.0870388 -0.170593 0.170593 5 0.0712356 0.0870388 -0.170593 0.170593 6 0.0682761 0.0870388 -0.170593 0.170593 7 0.0702673 0.0870388 -0.170593 0.170593 8 0.0654201 0.0870388 -0.170593 0.170593 9 0.0633015 0.0870388 -0.170593 0.170593 10 0.0712627 0.0870388 -0.170593 0.170593 11 0.073625 0.0870388 -0.170593 0.170593 12 0.0458525 0.0870388 -0.170593 0.170593 13 0.0129112 0.0870388 -0.170593 0.170593 14 0.00989309 0.0870388 -0.170593 0.170593 15 0.0141497 0.0870388 -0.170593 0.170593 16 0.0192469 0.0870388 -0.170593 0.170593 17 0.0192278 0.0870388 -0.170593 0.170593 18 0.0242387 0.0870388 -0.170593 0.170593 19 0.0214791 0.0870388 -0.170593 0.170593 20 0.00718463 0.0870388 -0.170593 0.170593 21 0.0199373 0.0870388 -0.170593 0.170593 22 0.0126535 0.0870388 -0.170593 0.170593 23 0.00311673 0.0870388 -0.170593 0.170593 24 -0.00737107 0.0870388 -0.170593 0.170593 table 7: evaluation of arima models no. model rmse mae 1 arima (1,1,0) (2,1,2) 12 27.219 7.557 2 arima (1,1,1) (1,0,1) 12 25.798 5.959 3 arima (1,1,2) ( 2,0,1) 12 25.717 6.122 4 arima (1,1,2) (1,0,1) 12 25.615 6.121 5 arima (2,1,2) (2,0,1) 12 25.845 6.020 6 arima (0,1,1) (1,0,1) 12 25.698 5.965 7 arima (0,1,1) 12 25.549 5.733 8 arima (1,1,1) 12 25.648 5.736 9 arima (2,1,0) 12 25.494 5.664 10 arima (1,1,0) 12 25.549 5.729 mohammed, et al.: time series forecasting using arima model 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 process, we can do further forecasts. plots are displayed in the figure 6 series. goodness of fit test it is possible to know the adequate model arima (2,1,0) for the exchange rate usd/iqd time series repeatedly using the box and pierce test. the null hypothesis is not rejected because the p-value is more than 0.05 and the model above is acceptable, implying that the residuals are also random and that the forecast will be adequate and dependable, as shown in table 9. forecasting after diagnosing the fitted model and selected as the best one, the final step comes forward forecasting. it is time to use the specified model for forecasting future values starting from january 2021 until december 2022, as shown in table 10 and figure 7. table 8: function model, parameter estimate for series model series model function model parameter estimate θ1 � θ2 � ∅1 � ∅2 � θ1 � θ2 � φ1 � φ2 � exchange rate arima (2,1,0) 12 − −∆ = ∅ ∆ + ∅ ∆ +1 1 2 1t t t tz z z e -0.28 -0.01 table 9: hypothesis and p-value for the residual series model series model hypothesis p-value exchange rate arima (2,1,0) 12 null hypothesis (h0): adequate model alternative hypothesis( h1): not adequate model 0.241 table 10: forecasting monthly average exchange rate usd/iqd series for the 24 months using arima (2,1,0) 12 model years months forecast lcl ucl 2021 january 1456.25 1405.81 1506.69 february 1379.31 1308.39 1450.24 march 1380.98 1301.65 1460.32 april 1402.79 1315.71 1489.86 may 1402.06 1306.26 1497.86 june 1395.88 1292.15 1499.62 july 1396.16 1285.46 1506.87 august 1397.91 1280.63 1515.19 september 1397.81 1274.21 1521.41 october 1397.32 1267.71 1526.93 november 1397.35 1262.02 1532.68 december 1397.49 1256.67 1538.31 2022 january 1397.48 1251.38 1543.58 february 1397.44 1246.23 1548.65 march 1397.44 1241.3 1553.58 april 1397.45 1236.53 1558.38 may 1397.45 1231.88 1563.02 june 1397.45 1227.36 1567.54 july 1397.45 1222.96 1571.94 august 1397.45 1218.67 1576.24 september 1397.45 1214.48 1580.43 october 1397.45 1210.38 1584.53 november 1397.45 1206.37 1588.54 december 1397.45 1202.44 1592.46 mohammed, et al.: time series forecasting using arima model 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (1): 12-19 conclusion the results of the analysis show that the time series data was not stationary around the mean and the variance. this means that almost all data of this kind should be different to remove any sort of trend influence on the data. because such series deal with metrology, fluctuations in the data for monthly us dollar/iq dinar currency exchange rates are to be expected. the rmse and mae are being used to select the best model. the arima model was found to be the better model for the series data arima (2,1,0) 12 . this is the forecasted means for the future of this us dollar against iq dinar time series which indicates a perpetual increase in the exchange rate. references 1. j. e. buet, g. m. barber and d. l. rigby. elementary statistics for geographers. 3rd ed. new york: the guilford press, 2009. 2. w. a. fuller. introduction to statistical time series. 2nd ed. canada: john wiley and sons, inc., 1996. 3. a. k. farhan and m. r. fakhir. forecasting the exchange rates of the iraqi dinar against the us dollar using the time series model (arima). international journal of engineering and management research, vol. 9, pp. 2394-6962, 2019. 4. e. p. clement. time series approach to forecasting rate analysis between nigeria naira and the u.s dollar. international journal of statistics and applications statistics, vol. 4, no. 1, pp. 76-84, 2014. 5. d. h. kadir. time series modeling to forecast on consuming electricity: a case study analysis of electrical consumption in erbil city from 2014 to 2018. journal of al rafidain university college, vol. 46, pp. 473-485, 2020. 6. g. e. p. box and g. m. jenkins. time series analysis forecasting and control. 4th ed. usa: library of congress catalog card number. 76-8713, 1976. 7. p. j. brockwell and r. a. davis. time series: theory and methods. 2nd ed. usa, new york: springer science and business media, llc, 233 spring street, 2006. 8. h. lutkepohul and m. kratzig. applied time series econometrics. new york: the united states of america by cambridge university press; 2004. 9. j. cryer and k. chan. time series analysis with applications. 2nd ed. usa, new york: springer science and business media, llc, 233 spring street, 10013, 2008. 10. p. j. brockwell and r. a. davis. introduction to time series and forecasting. 2nd ed. new york: springer-verlag, inc.; 2002. 11. r. s. shumway and d. s. stoffer. time series analysis and its applications with r example. 3rd ed. usa, new york: springer science and business media, llc, 233 spring street, 2011. 12. d. c. montgomery, c. l. jennings and m. kulahci. introduction to time series analysis and forecasting. 2nd ed. canada, hoboken, new jersey: john wiley and sons. inc., 2016. 13. w. wei. time series analysis: univariate and multivariate methods. 2nd ed. usa: greg tobin, 2006. 14. a. omer, h. blbas and d. kadir d. a comparison between brown’s and holt’s double exponential smoothing for forecasting applied generation electrical energies in kurdistan region. cihan university erbil scientific journal, vol. 5, no. 2, pp. 56-63, 2021. 15. t. cipra. time series in economics and finance. nature, switzerland, ag, 2020. 16. g. e. p. box, g. m. jenkins and g. c. reincel. time series analysis forecasting and control. 3rd ed. hoboken, new jersey: prenticehall, inc., 1994. 17. r. a. yaffee and m. mcgee. introduction to time series analysis and forecasting with applications of sas and spss. tokyo, toronto: academic press, inc.; 1996. figure 6: acf and pacf of residuals model figure 7: original usd/iqd series, forecasted values, and confidence limits using arima (2,1,0) 12 model for the period january 2021–december 2022, forecast (24 months) tx_1~abs:at/add:tx_2~abs:at 62 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 62-67 research article microbiological survey of ear infections in inhabitants of erbil city ameena s. m. juma, treska dh. kamil, muhsin h. ubeid department of medical microbiology, cihan university-erbil, kurdistan region, iraq abstract this study was carried out to indicate the number of infected individuals with ear infection from different genders and ages determine the antibiotic sociability for the ear swab bacterial isolates. a total of 82 ear swabs were obtained in erbil city from various hospitals and laboratories. males made up 48 (59%) of the 81 samples, while females made up 33 (41%). 0–10 years had the most bacteria isolated from ear swabs, followed by 11–20 years and then 21–40 years. pseudomonas spp. was the most prevalent bacteria found in all of the swabs (43%). pathogenic bacteria were found in 70% of the samples, while no growth was seen in 30% of the swabs. many bacterial isolates were found to be sensitive to the antibiotic amikacin, which was determined to have the best effect on 35 of them. clotrimazole was shown to be the least effective antibiotic on numerous bacterial strains, with 19 being resistant. keywords: ear infection, antibiotic sensitivity test, pseudomonas, amikacin introduction infections of the ear can affect either the inner or outer ear. ear infections are divided into three different forms, according to the centers for disease control and prevention. acute otitis media (aom), otitis media with effusion (ome), and otitis externa, sometimes known as swimmer’s ear, are the three forms. children are the ones that get ear infections the most. according to the national institutes of health (nih), three out of every four children will have at least one ear infection by the age of three (nih).[1] a bacterial or viral infection of the middle ear is known as an ear infection. inflammation and fluid accumulation in the ear’s interior canals are caused by this illness. the middle ear is a space behind the eardrum which is exposed to air. it has vibrating bones that translate sound from outside the ear into impulses that the brain can understand. ear infections are unpleasant, because he inflammation causes the eardrum to swell.[2] the most common respiratory infection in children is middle ear infection, also recognized as otitis media. middle ear infection is caused mainly by streptococcus pneumoniae (pneumococcus). pneumococcal conjugate vaccines are being used to protect young children from severe pneumococcal infections such as meningitis and pneumonia.[3] ear infections can occur as a result of poor needle technique, excessive needle retention, or the patient’s inability to properly care for his or her ear just after treatment. because the ear has a lower blood circulation than the rest of the body, once an infection occurs or involves the cartilage, it will be extremely difficult to treat. infections that are severe enough can cause the ear to swell and the cartilage to deteriorate.[4] in general, there are three types of ear infections: aom (inflammation of the middle ear). the term “ear infection” is commonly used to refer to aom. if there is fluid in the middle ear, indications or symptoms of infection, and the onset of symptoms was relatively rapid, the doctor will most likely make this diagnosis. medial otitis with effusion, where the clinician finds evidence of fluid in the middle ear if the diagnosis is ome, but no signs or symptoms have appeared. chronic otitis media with suppuration is when a doctor confirms chronic suppurative otitis media (csom), he or she discovers that a persistent ear infection caused the eardrum to tear or perforate. antibiotics are not always effective to control ear infections. what is best for the patient is determined by several of factors, including the patient’s age and the severity of his or her symptoms.[5] corresponding author: treska dh. kamil, department of medical microbiology, cihan university-erbil, kurdistan region, iraq. e-mail: treska.kamil@cihanuniversity.edu.iq received: june 20, 2022 accepted: july 11, 2022 published: august 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp62-67 copyright © 2022 ameena s. m. juma, treska dh. kamil, muhsin h. ubeid. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) juma, et al.: microbiological survey of ear infections in inhabitants of erbil city 63 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 62-67 infections of the ears are still a problem in the community. hearing loss affects 360 million individuals globally, 32 million of them are children.[6] during a year, 653,025 instances of middle ear infection were recorded in iraq, 1,737,214 cases in iran, 144,405 cases in jordan, 58,098 cases in kuwait and 463,669 cases in syria, and many other nations in the area (right diagnosis from health grades inc., 2015). otitis media was reported to affect 36.7% of the population in erbil city.[7] ear infections are a huge issue all across the world, as the below data demonstrate. in recent years, there has been some new information about this virus in the area. as a result, the following objectives were set for this research: record the number of ear infection cases in erbil city in relation to age and gender and to investigate the antibiotic sensitivity and resistance of the isolated bacteria to a variety of antibiotics. materials and methods study design this was a 4-month prospective research that was conducted from december 2017 to march 2018. in erbil city, the patients have been treated for ear infections at several hospitals and private laboratories. the samples of the study were collected from 82 patients, 48 men and 33 women. sample collection the presence of purulent material in all of the ear infections led the expert healthcare workers to conclude that they were infected. only one swab (sterilin, u.k.) was taken from each patient. each ear’s exudate was meticulously collected according to departmental regulations. to avoid the swabs drying out, the samples were delivered to the microbiology laboratory within 1 h of being collected. isolation and identification of bacteria swabs were inoculated on macconkey agar, chocolate agar, and blood agar (oxoid, u.k.) right away and incubated aerobically for 24–48 h at 37°c. due of logistical issues, anaerobic cultures were not performed. bacterial colonies on agar plates were identified using gram staining. bacterial isolates were identified and classified using biochemical techniques. standard microbiological procedures were used to characterize bacterial isolates.[8] gram stain, morphological and cultural features on agar, spore stain, motility and carbohydrate fermentation tests, nitrate reduction, catalase, hydrogen sulfide production, and indole production were among the tests that were performed. citrate utilization, gelatin liquefaction, the methyl red-voges proskaeur test, coagulase, hemolysis on blood agar, morphological and cultural characteristics on mannitol salt, and eosin-methylene blue agar are along with the other tests performed. antibiotic sensitivity test the kirby–bauer disk diffusion method was used to test antibiotic susceptibility, and the results were interpreted using the national committee for clinical laboratory standards’ recommendations. antibiotics that were applied included in the study: table 1 shows the antibiotics used in the susceptibility test on bacteria isolated from ear swabs. results swabs the total number of swabs collected in the current study was 81 ear swabs from different hospitals and private laboratories in erbil city. gender of patients from the 81 samples, 48 (59%) were male and 33 (41%) were female [figure 1]. table 1:list of antibiotics used to record the sensitivity and resistance of the isolated bacteria. abbreviation (mic*) antibiotic a (10 µg) actinomycin ak (10 µg) amikacin am (10 µg) ampicillin amc (25 µg) amoxicillin-clavulanic acid amk (30 µg) amikacin amp (10 µg) ampicillin at (10 µg) aztreonam auc (25 µg) amoxicillin-clavulanic acid az (30 µg) azlocillin c (30 µg) chloramphenicol caz (30 µg) ceftazidime cd (30 µg) cefdinir cec (30 µg) cefaclor cef (30 µg) cefazoline cep (30 µg) ceftriaxone cip (5 µg) ciprofloxacin cit (30 µg) citalopram clt (25 µg) clotrimazole cpo (5 µg) ciprofloxacin cro (30 µg) ceftriaxone cta (25 µg) closthioamide ctr (30 µg) ceftriaxone ctx (30 µg) cefaotaxime e (15 µg) erythromycin g (10 µg) penicillin gen (10 µg) gentamicin imp (30 µg) imipenem ip (30 µg) imipenem ipm (30 µg) imipenem mt (25 µg) metronidazole na (30 µg) nalidixic acid net (30 µg) netilmicin p (10 µg) penicillin pi (100 µg) piperacillin tcc (25 µg) ticarcillin-clavulanic acid ti (30 µg) ticarcillin tob (10 µg) tobramycin va (30 µg) vancomycin juma, et al.: microbiological survey of ear infections in inhabitants of erbil city 64 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 62-67 35 24 6 4 2 2 1 1 1 1 2 1 1 0 5 10 15 20 25 30 35 40 p se ud om on as s pp . n o gr ow th e sc he ric hi a co li s ta ph yl oc co cu s au re us k le bs ie lla p ne um on ia e c an di da s pp . a ci ne to ba ct er s pp . n es se ria g on or rh oe ae p ro te us s pp . p ro vi de nc ia s tu ar tii s er ra tia s pp . s ta ph yl oc oc cu s ch ro m og en es o ch ro ba ct ru m a nt hr op i figure 3: types and numbers of bacterial species cultured from ear infections 0 5 10 15 20 25 30 0-10 11-20 21-30 31-40 41-50 51-60 61-70 71-80 figure 4: ages of patients included in ear infections study bacterial isolates pseudomonas spp. was found to be the predominant bacteria isolated from ear infection, 35 (43%). no growth of bacteria represented 24 (30%) of the samples, while the other bacterial species showed a lower percentage [figures 2 and 3]. age of patients ear infections was found to be common in children, but they were found in all ages. the results showed that the children age group of (0–10) years had the highest number of cases for ear infections, compared to the other age groups [figure 4]. antibiotic sensitivity and resistance the antibiotic amikacin were found to have the best effect on many bacterial isolates, where 35 isolates were sensitive [figure 5a and b]. the antibiotic clotrimazole was found to be the least effective on many bacterial isolates, where 19 isolates were resistant [figure 6a and b]. discussion the most common cause of ear infections is a bacterial or viral infection of the middle ear, which is the air-filled space behind the eardrum that contains the tiny vibrating bones of the ear.[8] in many parts of the world, it is a public health issue.[9] another ear infection that affects the middle ear is otitis media, which is a common and major healthcare concern due to the significant economic burden it spots on the health-care system. eskimos, american indians, and australian aboriginal children, as well as dark-skin south africans, have been found to have a high prevalence. another major factor is poor living conditions and congestion. the widespread occurrence of csom in impoverished nations has been linked to poor hygiene and diet.[7] males were more affected (59%) than females, according to the findings of the present study (41%). according to other research, men are more likely than women to acquire ear infections.[10] the reason for sex dependence is still unknown.[11] ear infections are more common in children than in adults. due to the inflammation and fluid buildup in the middle ear, ear infections are typically severe. many studies in other regions, such as australia, found that indigenous children were more likely than non-indigenous children to get ear infections faster and much more severe.[12] similar results were obtained in our current study, where the highest number of cases was among the age group (0–10) followed by (11–20) years. aboriginal status was one of the most significant risk factors for otitis media in a population-based birth cohort research in canada.[13] the national longitudinal survey of children and youth in 1994/1995, on the other hand, found that 53% of newborn to three-year-old children, excluding those living on first nations reserves, had ear infections.[13] figure 1: males and females included in ear infections study 35 24 6 4 2 2 1 1 1 1 2 1 1 0 5 10 15 20 25 30 35 40 p se ud om on as s pp . n o gr ow th e sc he ric hi a co li s ta ph yl oc co cu s au re us k le bs ie lla p ne um on ia e c an di da s pp . a ci ne to ba ct er s pp . n es se ria g on or rh oe ae p ro te us s pp . p ro vi de nc ia s tu ar tii s er ra tia s pp . s ta ph yl oc oc cu s ch ro m og en es o ch ro ba ct ru m a nt hr op i figure 2: percentage of bacterial species cultured from ear infections of patients included in the study juma, et al.: microbiological survey of ear infections in inhabitants of erbil city 65 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 62-67 according to some sources, approximately 80% of children will be reported to have at least one episode of aom and between 80% and 90% will suffer at least one episode of ome before reaching school age. during an acute viral upper respiratory tract infection, aom is a complication of eustachian tube dysfunction. in 50% to 90% of instances of aom and ome, bacteria can be identified from middle ear fluid cultures.[14] furthermore, many studies have shown that breastfeeding protects against the development of otitis media.[10] this could be related to breast milk’s immunological features, which include antibodies to respiratory infections. furthermore, exposure to cigarette smoke has been linked to the development of ear infections in children, according to another study.[15] pseudomonas spp. was discovered to be the most common bacterial species (43%). pseudomonas infections are caused by gram-negative bacteria of the genus pseudomonas, particularly pseudomonas aeruginosa. they can be found in soil and water all around the planet. these bacteria can be found in wet environments such as sinks, toilets, poorly chlorinated swimming pools and hot tubs, and antiseptic treatments that are old or inactive. these bacteria are occasionally found in the armpits and genital tract area of healthy humans.[16] a high majority of cases (30%) showed no bacterial growth on the culture media used in the study. this could be due to bacteria that require media or incubation conditions to develop, or the bacteria could be anaerobic, or the cause could be fungal or viral. the antibiotic amikacin was shown to have the best effect on numerous bacterial isolates in this research, with 35 isolates being sensitive, while clotrimazole was found to be the least effective, with 19 isolates being resistant. antibiotic resistance is a regular phenomenon in most infections, with ear infections being among the most known.[17,18] in general, variations between the current study and other studies could be attributable to changes in inclusion criteria, sample sizes, microbiological approach, climate, and geographical areas, among other things.[19] conclusion a total of 81 ear swabs were collected in the present study from various hospitals and commercial laboratories in erbil city. of the 81 samples, 48 (59%) were male and 33 (41%) were female. the age group that had the highest number of bacteria isolated from ear swabs was 0–10 years followed by 11–20 years. the most common bacterial species isolated from all swabs were pseudomonas spp. (43%). most of the samples collected demonstrated the presence of pathogenic bacteria (70%), whereas 30% of the swabs demonstrated no growth. the antibiotic amikacin was found to have the best effect on many bacterial isolates, where 35 isolates were sensitive. the antibiotic clotrimazole was found to be the least effective on many bacterial isolates, where 19 isolates were resistant. figure 5 (a and b): antibiotic resistance of bacterial isolates from ear infection 0 5 10 15 20 25 30 35 40 c ip a k c n a t o b g e n n e t c r o c lt a m a t c e c t c c t i p i a m c m .t im p c t r c e f 0 2 4 6 8 10 12 14 16 c lt e c d p v a a m p c e p c t x c p o a a z a m k c t a ip m g a u c ip c a z b a juma, et al.: microbiological survey of ear infections in inhabitants of erbil city 66 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 62-67 references 1. a. bradford. ear infections: symptoms and treatment. 2014. available from: https://www.livescience.com/34720-ear-infectionssymptoms-and-treatment.html. [last accessed on 2022 may 20]. 2. s. p. jacups, d. newman, d. dean, a. richards and k. m. mcconnon. an innovative approach to improve ear, nose and throat surgical access for remote living cape york indigenous children. journal of pediatric otorhinolaryngology, vol. 100, pp. 225-231, 2017. 3. c. fortanier, r. p. venekamp, c. w. boonacker, e. hak, g. schilder, e. a. sanders and r. a. damoiseaux. pneumococcal conjugate vaccines for preventing acute otitis media in children. cochrane database of systematic reviews, vol. 2019, no. 5, pp. cd001480, 2019. 4. l. m. yin, g. h. jiang, y. wang, y. wang, y. y. liu, w. r. jin, y. d. xu, q. h. zhang and y. q. ang. use of serial analysis of gene expression to reveal the specific regulation of gene expression profile in asthmatic rats treated by acupuncture. journal of biomedical science, vol. 1606, pp. 16-46, 2009. 5. m. l. carlson. the history of otorhinolaryngology at mayo clinic. mayo clinic proceesdings, vol. 92, no. 2, pp. e25-e45, 2017. 6. b. impouma, b. n. archer, o. c. lukoya, e. l. hamblion and i. s. fall. world health organization regional office for africa weekly bulletin on outbreaks and other emergencies. emergencies, vol. 24, pp. 1392-12395, 2018. 7. f. j. khayat and l. s. dabbagh. incidence of otitis media with effusion in children with adenoid hypertrophy. zanco journal of medical sciences, vol. 15, no. 2, pp. 57-63, 2011. 8. r. d. bradford, y. g. yoo, m. golemac, e. p. pugel, s. jonjic and w. j. britt. murine cmv-induced hearing loss is associated with inner ear inflammation and loss of spiral ganglia neurons. pols pathogens, vol. 11, no. 4, p. e1004774, 2015. 9. j. j. bardy, l. sanchez and a. s. carney. otitis media in indigenous australian children: review of epidemiology and risk factors. the journal of laryngology and otology, vol. 128, no. 1, pp. s16-s27, 2013. 10. k. kong and h. l. c. coates. natural history, definitions, risk factors and burden of otitis media. the medical journal of australia, vol. 191, no. s9, pp. s34-s43, 2009. 11. r. baraibar. incidence and risk factors of acute otitis media in children. clinical microbiology and infection, vol. 3, pp. 3s13-3s22, 1997. 12. v. yiengprugsawan and a. hogan. ear infection and its associated risk factors, comorbidity, and health service use in australian children. international journal of pediatrics, vol. 2013, pp. 963132, 2013. 13. e. a. macintyre, c. j. karr, m. koehoorn, p. demers, l. tamburic, c. lencar and m. brauer. otitis media incidence and risk factors in a. paediatrics and child health, vol. 15, no. 7, pp. 437-442, 2010. 14. a. qureishi, y. lee, k. belfield, j. p. birchall and m. daniel. update on otitis media-prevention and treatment. infection and drug resistance, vol. 10, no. 7, pp. 15-24, 2014. 15. e. m. thomas. recent trends in upper respiratory infections, ear figure 6 (a and b): antibiotic sensitivity of bacterial isolates from ear infection 0 2 4 6 8 10 12 14 16 18 20 c ip a k c n a t o b g e n n e t c r o c lt a m a t c e c t c c t i p i a m c m .t im p c t r c e f 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 c lt e c d p v a a m p ce p pm i c t x cp o a a z a m k c t a ip m g c a t a u c ip n k c a z b a https://www.livescience.com/34720-ear-infections-symptoms-and-treatment.html https://www.livescience.com/34720-ear-infections-symptoms-and-treatment.html juma, et al.: microbiological survey of ear infections in inhabitants of erbil city 67 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 62-67 infections and asthma among young canadian children, health reports, vol. 21, no. 4, pp. 47-52, 2010. 16. k. bush. past and present perspectives on &#x3b2;-lactamases. antimicrobial agents and chemotherapy, vol. 62, no. 10, p. e01076-18, 2018. 17. s. s. vossoughi. clinics in laboratory medicine. elsevier health sciences, netherlands, 2021. 18. l. fiorillo, g. cervino, a. s. herford, f. lauritano, c. d’amico, r. l. giudice, l. laino, g. troiano, s. crimi and m. cicciù. interferon crevicular fluid profile and correlation with periodontal disease and wound healing: a systemic review of recent data. international journal of molecular sciences, vol. 19, no. 7, p. 11, 2018. 19. a. fares. seasonality of tuberculosis. journal of global infectious diseases, vol. 3, no. 1, pp. 46-55, 2011. . 92 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 review article social network privacy models: a systematic literature review and directions for further research abdullah abdulabbas nahi al-rabeeah1*, mohammed mahdi hashim2 1department of computer science, cihan university-erbil, iraq, 2department of computer science, auruk university, baghdad, iraq abstract privacy is a vital research field for social network (sn) sites (sns), such as facebook, twitter, and google+, where both the number of users and the number of sn applications are sharply growing. recently, there has been an exponential increase in user-generated text content, mainly in terms of posts, tweets, reviews, and messages on sn. this increase in textual information introduces many problems related to privacy. privacy is susceptible to personal behavior due to the shared online data structure of sns. therefore, this study will conduct a systematic literature review to identify and discuss the main privacy issues associated with sn, existing privacy models and the limitations and gaps in current research into sn privacy. keywords: component, privacy models, privacy rules, social network, social network privacy introduction social network (sn) has currently become a significant platform for users to share newscasts, thoughts, and post messages. as pointed out by gritzalis et al.,[1] the introduction of web 2.0 contributed to the growing number of users using sn sites (sns). users can now interact with each other, generate, or reorganize information and thoughts, and express themselves in computer-generated communities. such means of communication, along with the mutuality of user-generated content, are referred to as sns. millions of people are using sns such as google+, facebook, twitter, and instagram for daily activities. there is more than one type of sns with numerous affordances in the technological aspects, interests. moreover, their key technological features are coordinated, and the cultures that arise around sns are assorted. most sites help strangers to connect, based on shared interests, political opinions, or activities. some sites are provided to varied users, while other sites are targeted toward interested people, based on common language, or shared cultural, sexual or religious interests, or nationalitybased. sites also vary in the extent to which they incorporate new information and communication tools, such as mobile connectivity, blogging, and photo/video-sharing. in recent years, there has been an exponential increase in user-generated text content, mainly in terms of posts, tweets, reviews, and messages on sn. this increase in textual information introduces many problems related to privacy. thus, due to the need for user privacy protection in sn, many privacy tools are employed.[2] this paper conducts a systematic literature review on the privacy of sns since there are many problems not solved yet related to privacy. sn sn is a site or system of associations and connections.[3] in addition, it has another definition. these include facebook, linkedin, twitter, or content sharing systems, for example, youtube and flickr. in these (and other) sns, the user tries to find similar or perfectly matched individuals with similar interests or encounters. sn includes components of different sorts of data, for example, music, photographs, recordings, websites, connections, and outsider applications. table 1 illustrates the sn definitions from earlier researchers. the well-known sn with their main focus, default relationships, and the direction of the relationships is shown in table 2. review method this study uses a systematic review approach to answer the questions posed above. (alvarez-jimenez and alcazar-corcoles corresponding author: abdullah abdulabbas nahi al-rabeeah, department of computer science, cihan university-erbil, iraq. e-mail: abdullah.nahi92@gmail.com received: apr 18, 2019 accepted: apr 25, 2019 published: aug 20, 2019 doi: 10.24086/cuesj.v3n2y2019.pp92-101 copyright © 2019 abdullah abdulabbas nahi al-rabeeah, mohammed mahdi hashim. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ al-rabeeah and hashim: a systematic literature review and directions for further research 93 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 2014) an effective review can create a firm foundation for advancing knowledge, facilitate theory development and discover areas where the research is needed.[12] a systematic review can be defined as a process of identifying, evaluating, and interpreting all available research relevant to research questions, area of study, or a rising phenomenon of interest.[13] the reasons for conducting a systematic review are to summarize the evidence about technology or treatment, summarize the evidence of the advantages of a specific method, identify any research gaps in the existing research and provide deep understanding for new phenomena.[13] therefore, these reasons fit with the aim of our review. this study follows kitchenham and charters guidelines, where a systematic review task involves three main stages: planning the review, conducting the review, and reporting the review. each stage has certain activities. review protocol review protocol is an essential stage in performing a systematic review and specifies the methods that will be used to undertake a systematic review. figure 1 shows the review protocol stages. the goal of a review protocol is to reduce research bias.[13] the review protocol includes background, research questions, search strategy, study selection process, data extraction, and synthesis of the extracted data.[13] in this review, the research questions and background of sn are stated above. figure 1 illustrates the review protocol used for this study. inclusion and exclusion criteria the purpose of identifying inclusion and exclusion criteria is to make sure that the selected studies are relevant and related to our study. because this review focuses on understanding the sn, the consideration is only on articles from journals, conferences, workshops, book chapters, and symposia in the english language. the duration of the period of publication of the selected studies is from 2006 to 2017. the reasons for choosing this period are two-fold. first, this review is complementary to previous efforts[14] to provide a deep understanding of sn. second, the term sn has been increasingly used in various studies since 2008, and the previous major articles reviewing the state of sn research cover literature until this year, therefore, this year is required to systematically collect, analyze, and synthesize these studies for past 6 years. table 3 shows the criteria for this review. search strategy the search strategy, as depicted in figure 1, consisted of two stages: an automatic stage and manual stage. the automatic stage was to identify the primary studies of sn. based on webster and watson’s recommendations,[12] the researchers did not limit the search process to a specific set of journals; instead, and several online databases were used to cover a broad range of academic publications. the online databases used were sciencedirect, scopus, springer, ieee explorer, and web of science. these databases are considered relevant and provide high impact factor publications. to perform the automatic table 1: sn definitions from earlier researchers no. definitions reference 1. the usage of sn by an organization in communication with outside parties such as clients, sellers, and the community leonardi et al. 2013[4] 2. sns are web-based services that allow individuals to[1] construct a public or semi-public profile within a restricted system,[2] create a list of additional users with whom they share a link, and[5] view and traverse their list of connections and those made by others within the system. the nature and terminology of these connections may vary from site to site ellison and boyd 2007[3] 3. user profiles that describe their locations, interests, and education background, and provide useful information differing from links croitoru et al., 2013[6] 4. sns provide users, within their platforms, with powerful ways to interact with other users through different forms and modalities. consequently, each user can search and check the profiles of other sn members (for various reasons), exchange messages with some of them, publish videos, and post comments on shared photos, etc. hajli 2013[7] 5. sns are for finding old friends, meeting new friends, or finding people who have the same interests or problems across political, economic, and geographic borders. by creating personal information profiles that contain information such as photos, video and audio files, sns allow users to connect, post messages, send e-mails, and instant messages to each other abawajy et al., 2016[8] 6. the infrastructure of sns can support a rich variety of data analytics applications such as search, text analysis, image analysis, and sensor applications aggarwal and wang 2011[9] 7. sn technologies have opened new possibilities for sharing personal information with online networks, and millions of people routinely self-disclose personal information on snss bazarova and choi 2014[10] sns: social network sites, sn: social network table 2: well-known sn with their main focus, default relationship, and the relationships direction (gitelman 2006)[11] sn platform focus default relationship (s) relationship direction facebook general use friendship symmetrical flicker photo sharing contact and optionally friend, or family symmetrical google+ general use friends, family, acquaintances symmetrical linkedin professional business symmetrical twitter microblogging followers asymmetrical youtube video sharing subscribe-to asymmetrical sn: social network al-rabeeah and hashim: a systematic literature review and directions for further research 94 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 search, keywords were identified based on the research question of this review. the main keywords used were: “sn,” “sn privacy,” and “sn privacy issues.” the second stage was a manual search. backward and forward search methods[12,15] were used to trace the citation of the selected studies. we used the google scholar search engine to go forward and find the studies which were cited in the selected primary studies. the manual stage was used to ensure that the systematic search was comprehensive and relatively complete.[12] for managing and sorting all the studies, mendeley, a reference management tool, was used to keep all the search results and easily remove the duplicated studies. study selection process the study selection process was used to identify the studies related to the research questions of this review. using the defined keywords, the result of the initial search identified 185 studies from the automatic search. after removing the duplicated studies using mendeley, 150 remained. we then applied the inclusion/exclusion criteria, on the abstract and conclusion of each study. in this step, 90 studies were eliminated. based on kitchenham and charters’ recommendation,[13] we excluded the studies that were clearly not related to the subject of this review. full-text scanning was used for the remaining studies, with the consideration of the exclusion criteria. we also applied the manual search to the reference list of each study, to trace any missing studies. after applying the manual search, an additional 12 studies were found. thus, the final set of primary studies was 72. finally, we applied quality assessment criteria, and 12 were removed; thus, a total of 60 studies were identified as a final list of primary studies. define abbreviations and acronyms the first time they are used in the text, even after they have been defined in the abstract. abbreviations such as ieee, si, mks, cgs, sc, dc, and rms do not have to be defined. do not use abbreviations in the title or heads unless they are unavoidable. figure 1: research methodology table 3: inclusion and exclusion criteria inclusion criteria exclusion criteria full-text uncompleted studies published within the selected period (2006–2017) non-english published in the above-selected database written in english outside the selected time study manuscript in the domain of social network duplicated studies al-rabeeah and hashim: a systematic literature review and directions for further research 95 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 quality assessment applying quality assessment is considered critical to assessing the quality of the primary studies.[13] the details of quality assessment are based on quality instruments, which could be a checklist of factors or questions that need to be applied for each study.[16,17] in this review, we developed the following three quality assessment criteria to assess the quality of each study: qa1. is the topic addressed in the paper related to sn privacy? qa2. is the research problem described clearly in the paper? qa3. is the data collection method described in the paper? the three qa criteria presented above were applied to the 72 primary studies to increase our confidence in the credibility of the selected studies. the process of applying quality assessment used three levels of the quality schema (high, medium, and low),[18] in which the quality of each study depends on the loading score. for instance, studies that fulfill the criterion will be given 2; studies that partially fulfill the criterion will be given 1; and studies that do not fulfill the criterion will be given. studies that score 5 or above will be considered high quality, while if they score 4 they will be considered medium, while if they are below 4 they are considered low. after applying the qa, 12 studies were eliminated because they did not fulfill the qa criteria. the results of the qa are displayed in figure 2. research questions results rq1. what are the privacy issues in sn? the two concepts of privacy concerns and privacy attitudes are basically different. privacy concerns could be quite generic and in most cases are not bound to any specific context, while privacy attitudes refer to the assessment of exact privacy behaviors.[19] another stream of study aimed to understand self-disclosing behavior in online sn (osn), especially among young people. barnes[20] uses the term privacy paradox about the privacy behavior of young people in sn. young people tend not to realize that sm provides public space and disclose personal information that could perhaps be misused.[20] carrascal et al.,[21] led an experiment aiming to determine the monetary value of several kinds of personal information. using a web browser plugin, they prompted users to value their personal data at the time and place they were produced. in the first phase of the experiment, the browser plugin collected data about the browsing behavior of each subject. data were used to calibrate the behavior of the plugin in the second phase. in the second stage, the plugin exhibited popups as the participants were browsing the internet. popups contained two kinds of questions: questions about evaluating personal information and questions on participants’ privacy perceptions and knowledge. information valuation questions were framed as auctions.[21] for instance, one question was, “what is the questionnaire on privacy attitudes and preferences and, then, to visit an online store.” during their shopping in the store, they were engaged in a sales dialogue with an anthropomorphic three-dimensional shopping bot. participants answered most of the questions, even if these were highly personal. this indicates that even though internet users claim that privacy is a high priority, they do not behave accordingly. based on a questionnaire survey and an examination of participants’ facebook profiles, hughes-roberts 2013) 23 concluded that a general statement of user concern is not a valid indicator of privacy behavior within the network. however, he questioned the appropriateness of surveys as instruments for studying the privacy paradox.[22] lee[23] confirmed the existence of an attitude versus behavior dichotomy. they conducted a series of semi-structured in-depth interviews and an experiment to assess the influence of expected benefit and expected risk on users’ intention to share personal information. they concluded that users actively share personal information despite their concerns because they do not only consider risk but also the expected benefit of sharing.[24] reynolds et al., in their study, also found that there was little correlation between participants’ broader concern about privacy on facebook and their posting behavior.[25] in contrast to tufekci,[26] they found that the portion of posts that were visible to a large audience appeared to be independent of general privacy attitude. a web survey by taddicken also showed that privacy concerns hardly impact self-disclosure.[27] the relationship between privacy concerns and self-disclosure is moderated by various variables. perceived social relevance and the number of other social web applications used have a strong moderating effect. in this study, social relevance mainly refers to the disclosing behavior of communication partners indicating that disclosure proceeds on a quid pro quo basis, i.e., “you tell me and i tell you.” a study by zafeiropoulou et al.,[28] specifically examined location data, which are a form of personal information increasingly used by mobile applications. their survey also found evidence that supports the existence of the privacy paradox for location data. in their book liquid surveillance, bauman and lyon talk about privacy in relation to online web-based social networking administrations. bauman says: “we present our rights to privacy for butcher of our own will. or, then again maybe we simply agree to the loss of privacy as a sensible cost for the marvels offered in return”. or, then again, the weight to convey our own self-sufficiency to the slaughterhouse is so overpowering, so near the state of the rush of sheep, that exclusive of a couple especially defiant, striking, contentious and unfaltering wills are set up to make a sincere endeavor to withstand it.[29] this is an instance of the unavoidable negativity show in discourse about insurance and an instance of privacy being discussed as a generalizable thing with a lone definition figure 2: review protocol al-rabeeah and hashim: a systematic literature review and directions for further research 96 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 material to everyone. it has presented the usage of online electronic long range informal communication organizations as the opposite of assurance. bauman and lyon proceed to express their view that there has been an alteration in people’s points of view about what ought to be open and what ought to be private.[29] the authors believe that this change has been made by the coming of widespread use of online web-based sn administrations. both methodologies comprehensively characterize the philosophical utility of privacy. legal privacy, for instance, doubts about the ancient rarities of privacy, for example, the data, while social privacy is concerned with the impacts of association of people in the public eye on privacy. while there might be examples in which the two may cross, making a distinction between the two toward the start of this review is essential as it shows the wide remit of the review. even scholars with a legal background have highlighted the social aspects of privacy.[30] valerie[29] puts forward comparable assumptions, highlighting that privacy can be normally comprehended in the everyday transactions of social standards inside connections. this show how social privacy as an idea is fundamentally concerned with how people communicate with each other. in any case, given the unfathomable sorts of communications that take place between people, social privacy soon turns out to be a very wide concept. diverse ways to deal with understanding social privacy have been looked for organizations such as facebook and twitter. they also conducted a survey that revealed a strong correlation between information privacy concern and all types of responses, except for misrepresentation. contrary to previous research, blank et al.,[32] found that younger people are more likely to take action to protect their privacy than older ones. in addition, studies show a positive correlation between privacy concerns and protection behavior. lutz and strathoff[33] conducted a telephone survey in switzerland employing a questionnaire that covered several privacy-related constructs. this survey confirmed a weak but statistically significant influence of privacy concerns on protection behavior. recent surveys show that privacy concerns trigger protective responses, such as uninstalling mobile applications. a survey of smartphone users by the pew internet project[34] revealed that 54% of the mobile application users surveyed had decided to not install a cell phone application when they discovered how much personal information they would need to share to use it and 30% of the cell phone application users had uninstalled an application that was already on their cell phone because they learned it was collecting personal information that they did not wish to share. on the other hand, only 19% of cell phone owners had turned off the location tracking feature on their cell phone. table 4 shows that the studies reviewed and the methodology they used for collecting the data and how such as.[26,35-38] rq2. what are the privacy models in sn? several studies have built different models explaining mechanisms of user privacy on the internet. many of them cover issues arising when users make decisions about exposing their private information in exchange for service use, such as online shopping, internet banking, or the use of the internet in general. a limited number of studies also address issues of privacy when users communicate with other users in addition to the service users in the context of osn. nov and wattal[39] investigated privacy issues of sn. in this study, they extended earlier research on web privacy to address inquiries regarding precursors of privacy concern in social groups, and in addition, the effect of privacy concern in such groups. in the group and the group’s data sharing standards negatively affected the group members’ particular privacy concerns. they likewise found that group-specific privacy concerns not only led members to embrace more prohibitive data sharing settings but also additionally diminished the amount of data they imparted to the group. additionally, found find that data sharing was affected by system centrality. in another exploration, chai et al.[40] analyzed variables that impact web clients’ private data sharing conduct among groups of preteens and early high schoolers, which are among the most powerless gatherings on the web. this review study provides an examination system that clarifies a web client’s data conduct in ensuring privacy. two noteworthy components were found to influence web clients’ data privacy practices: (1) clients’ apparent significance attached to data privacy and (2) data privacy self-adequacy. the review, additionally, found that clients have confidence in the estimation of online data privacy and that data privacy assurance conduct differs according to gender. shin[41] analyzed privacy, trust, and privacy concern in the context of interpersonal interaction websites among purchasers, utilizing both solid scales and measures. she proposes a sms acknowledgment show table 4: studies on social network privacy study context methodology participants bamas (2011)[35] snss student survey students carrascal (2013)[21] snss survey web users hughes-roberts (2013)[23] snss survey students hau et al. (2013)[24] snss/location data experiments users of snss reynolds et al. (2011)[25] snss survey facebook tufekci (2008)[26] snss survey students taylor et al. (2013)[36] snss survey students blank et al. (2014)[32] snss survey random users boyd et al. (2010)[37] snss survey students christofides et al. (2009)[38] snss survey students lutz and strathoff (2014)[33] internet use/snss survey sample users al-rabeeah and hashim: a systematic literature review and directions for further research 97 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 by coordinating intellectual and in addition full of feeling mentalities as essential affecting components, which are driven by fundamental convictions, visual privacy, visual privacy, trust, disposition, and aim. this is the outcome of a study of sms clients which confirmed that the proposed hypothetical model clarifies and predicts client acceptance of sms significantly well. the model shows phenomenal estimation properties and builds up visual privacy, and visual privacy of sms as particular develops. the finding additionally uncovered that apparent privacy affects the impact of visual privacy on trust. nov and wattal[39] investigate privacy issues of social processing condition. in this study, they stretch out earlier research on web privacy to address inquiries regarding precursors of privacy concern in social registering groups, and in addition, the effect of privacy concern in such groups. the outcomes demonstrate that clients’ trust in other group individuals and the group’s data sharing standards negatively affect group privacy concerns. they likewise distinguish that group privacy concerns not just lead clients to embrace more prohibitive data sharing settings, additionally diminish the measure of data they impart to the group. in addition, they find that data sharing is affected by system centrality and the residency of the client in the group. shin[41] examined security, trust, and privacy concerns with regard to social networking websites among consumers using reliable scales and measures. the study proposes an sns acceptance model by integrating cognitive as well as affective attitudes as primary influencing factors, which are driven by underlying beliefs, perceived security, perceived privacy, trust, attitude, and intention. results from a survey of sns users validated that the proposed theoretical model explains and predicts user acceptance of sns substantially well. the model shows excellent measurement properties and establishes perceived privacy and perceived the security of sns as distinct constructs. the finding also reveals that perceived security moderates the effect of perceived privacy on trust. in contrast to theory-based approaches, bottom-up approaches of model construction and testing make their contribution to theory building from a selected pool of salient constructs, such as trust,[42,43] weakness,[44] or individual demeanor.[45,46] malhotra et al.[46] focus on three distinct yet closely related issues. first, drawing on social contract theory, the authors offered a theoretical framework on the dimensionality of internet users’ information privacy concerns (iuipc). the authors then attempted to operationalize the multidimensional notion of iuipc using a second-order construct and develop a scale for it. finally, they proposed and tested a causal model on the relationship between iuipc and behavioral intention toward releasing personal information at the request of a marketer. the results of this study indicate that the second-order iuipc factor, which consists of three first-order dimensions, namely, collection, control, and awareness, exhibited desirable to user’s psychometric properties in the context of online privacy. in addition, their causal model centering on iuipc fits the data satisfactorily and explains a significant amount of variance in behavioral intention, suggesting that the proposed model will serve as a useful tool for analyzing online consumers’ reactions to various privacy threats on the internet dinev and hart[44] developed and validated an instrument to measure the privacy concerns of individuals who use the internet and two antecedents, perceived vulnerability and perceived ability to control information. the results of an exploratory factor analyses supported the validity of the measures developed. in addition, the regression analysis results of a model including the three constructs provide strong support for the relationship between perceived vulnerability and privacy concerns, but only moderate support for the relationship between perceived ability to control information and privacy concerns. the authors claim that the relationship among the hypothesized and privacy concerns may be one that is more complex than is captured in the hypothesized model, considering the strong theoretical justification for the role of information control in the extant literature on information privacy. in the realm of osns, several studies have challenged the common assumption that young people do not protect their private information contrary to previous research, lutz and strathoff[33] found that younger people are more likely to act to protect their privacy than older ones. young people use a variety of protection strategies, such as using pseudonyms and giving false information,[47] restricting access to their profiles and adjusting their privacy settings,[48] limiting friendship requests, and deleting tags and photos.[49] in addition, studies show a positive correlation between privacy concerns and protection behavior aeschlimann et al.[50] conducted a telephone survey in switzerland employing a questionnaire that covered several privacy-related constructs. this survey confirmed a weak but statistically significant influence of privacy concerns on protection behavior. casalo et al.[51] investigated the impact of perceived site privacy, ease of use, and notoriety on buyer confidence with regards to web-based keeping money. the study also breaks down the trust-duty relationship, since responsibility is a key variable for building up effective long-term associations with clients. their study presents the result of privacy ease of use and reputation on buyer confidence in a site in the web-based managing an account setting. in addition, it proposes that trust positively affects purchaser responsibility, convenience and reputation have an immediate and huge impact on shopper confidence in a money-related administration site. in addition, purchaser trust is strongly identified with relationship. in addition, it is noted that trust is a key moderating element in the improvement of relationship responsibility in the webbased saving money setting. hence,[52] build-up a hypothetical system portraying the trust-based basic leadership prepares a purchaser uses when making a purchase from a given webpage, and test the proposed display utilizing a structural equation modeling strategy on web buyer purchasing behavior, with information gathered by means of a web survey. the results of the survey indicated that web buyers’ trust and perceived risk affect their purchasing choices. customer intention to trust, reputation, privacy concerns, privacy concerns, the data nature of the website, and the organization’s reputation effectively affect internet buyers’ trust in the website. bandyopadhyay et al.[53] proposed a hypothetical system to explore the elements that impact the privacy concerns of customers who utilize the internet and the results of such privacy concerns. elements al-rabeeah and hashim: a systematic literature review and directions for further research 98 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 distinguished as precursors to online privacy concerns are seen as a weakness to accumulation and abuse of individual information, perceived capacity to control information gathering and its resulting utilization, the individual’s level of internet proficiency, social awareness, and underlying social variables. the possible outcomes of online privacy concerns are the unwillingness to give individual data on the web, shunning web-based businesses, or even unwillingness to utilize the internet. the model explored depends on a correspondence theory (i.e., communication privacy management [cpm] theories). a genealogical or family tree is the main component to build their trust model. there exist many sites to manage a family tree. however, those are not popular as sns. on the other hand, many popular snss are not focused on creating a family tree. although the notion of the family tree is absent in these sites, often family members coexist in the same network as a friend or follower or fan. hence, they can easily build a family tree even if it does not exist in the architecture of the sms. using the family tree, they propose their novel approach for securing sn by calculating trust. to realize trust quantitatively, they compute two quantities for the trust baseline: a. trust value and (b) trust score: trust value captures the static component of human trust while trust score changes dynamically with human connection in sms. both of those trust indicators are based on generation circle.[54] sn users dissatisfied with the snss request for sn login credentials may also engage in public action as a form of recourse or to seek a remedy. perales et al.[55] identify two such public actions: complain to the company and complain indirectly to a third party. given the nature of interviews, individuals are not likely to complain about the requesting during them. instead, they focus on complaints to companies’ executives. indirectly complaining to a third party may include contacting the media or local politicians to seek redress. ngai et al.[56] point out that sn applications are multi-disciplinary, including such areas as marketing, knowledge sharing, customer relationship management, collaborative activities, organizational communications, education, and training. the design and development of each application system are underpinned by different personal and social behavior theories and models, including personal behavior theories (e.g., personal traits, tam, tra, and theory of planned behavior [tpb]), social behavior theories (e.g., social capital, social cognitive, and social power), and mass communication theories (e.g., psi and uses and gratifications theory), and are encompassed by a wide variety of sn tools and technologies, such as media sharing sites (e.g., youtube and instagram), blogs and microblogs (e.g., twitter and weibo), social bookmarking sites (e.g., delicious and pinterest), virtual and online communities (e.g., lonely planet and yahoo answers), sms (e.g., facebook and linkedin), and virtual worlds (e.g., second life and active world). using this model, they argue that when researchers and system developers design and develop sn applications, they should first understand the theoretical foundations of a system, and based on these foundations, choose the right tools and technologies, and process the design, development, and implementation of the system. private action by kim and ko,[52] information privacy can be undermined when internet clients lose control over how online organizations gather and handle their own data.[46] cases of such loss of control range from getting undesirable emails. the term social computing according to nov and wattal[39] alludes to applications and administrations that encourage aggregate activity and social cooperation on the web, for example, sites, wikis, interpersonal organizations, and dialog discussions. as social computing is turning into an essential part of people’s groups’ and social status, it has been receiving increasing research interest. the achievement of social networking frameworks, whose content is made altogether by client commitment, relies on the ability of the members to share. as indicated by the privacy analytics theories, people’s readiness to provide data is, thus, represented by their privacy concerns. to address this. this issue is by all accounts, especially imperative given that interpersonal organizations, for example, facebook have been confronting increasing criticism over their privacy arrangements. chai et al.[57] propose a connection between apparent significance of data privacy to web users and their conduct in ensuring their privacy on the web. as observed in past research a positive connection was demonstrated between perceived significance in a specific area and behavioral intention to carry out the activities identified with that area, they expect that web clients who put a higher importance on the significance of data privacy will exhibit an even stronger propensity to show online privacy assurance behavior than the users who place lower importance on the significance of data privacy on the web. all the factors that will affect the user privacy such as sex, age, education, etc.[58] as they point out, the iuipc will play a main role in the future. cpm theory asserts that people control their private information based on the use of personal privacy rules. through developing, learning, and negotiating rules depending on culture, gender, motivation, context, and risk/benefit ratio, people coordinate boundary linkages, boundary permeability, and boundary ownership. the theory delineates such causal relationships in a qualitative and interpretive manner, while the tpb outs forward a mechanism for the human decisionmaking process, i.e., a causal link constituting a person’s salient beliefs and evaluations, attitude toward a behavior, and behavioral intentions. table 5 illustrates the theories and research constructs for the sn privacy. rq3. what are the limitation and gaps in current research regarding sn privacy? privacy concerns exist about the potential for the collection, retention, and data mining of personal information by the federal government of the usa with respect to its use of sn for disaster recovery purposes. specifically, the use of status alerts and the creation of personal pages to establish situational awareness may raise privacy concerns.[61] others are concerned about how the information might be used, for example, would the federal government compile records after a terrorist attack to help investigate certain individuals or some concerned about the user behavior itself in the sn al-rabeeah and hashim: a systematic literature review and directions for further research 99 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 by going through all the previous studies shows the user’s attitude can be affected by many factors that will cause itself privacy.[23] a lot of research focused on the users’ behavior but did not consider all the factors that may affect the user privacy, depending on many important reasons, more importantly, the huge development happening in sn platforms and the number of users that are reaching billions of daily users using at least one sn platform.[62] smith et al.[63] point out that privacy for users is almost not available in the data or information has been posted into sns. the modern smartphones have made the using of sn very easy and accessible everywhere since the proliferation of high-speed mobile networks is enabling a culture of impulsive and carefree posting of user content. for instance, according to facebook statistics, the scale of this phenomenon per month has risen from two billion to over six billion and there were one billion active users on average at the end of 2016 on a daily basis.[64] almost most of the photos posted on sn have no relevance to privacy as from a personal perspective. from previous studies, the current models do not seem to cover all the required needs for sn users and according to marwick and boyd[65] the privacy challenges increasing day by day since the development of sns growing very fast and the number of users is around one billion for each platform. several studies have built causal models explaining mechanisms of user privacy on the internet. many of them cover issues when users make decisions of exposing their private information in exchange for service use, such as online shopping, internet banking, or the use of internet in general. however, a limited number of studies have addressed issues of privacy when users communicate with other users in addition to the service use in the context of osn. conclusion and recommendation this study provides an overview of the privacy concept in sn. to understand sn privacy, we set three research questions related to the nature of sn privacy. a systematic review approach was used to answer these questions; the review included the studies published between 2006 and 2017. after performing multiple selection processes, 60 studies were selected that focus on sn privacy. the remainder of the studies was eliminated from the review as they did not fulfill the inclusion criteria or have not reached the quality level. the study provides a clear view of sn privacy and privacy models by identifying the privacy in sn, and the main issues could be affecting the user privacy. from the data analysis, the majority of the 60 studies belong to user behavior and platforms privacy rules themes, while other areas received little attention, such as government law, business model, and security and privacy issues. in addition, the research methodologies used in these studies were identified and classified. the study found that more studies focused on the privacy of the users in sns by depending on different factors that were used previously, because of the huge number of the users in the sns. references 1. d. gritzalis, m. kandias, v. stavrou and l. mitrou. “history of information: the case of privacy and security in social media”. proceeding of the history of information conference. pp. 283-310, 2014. 2. h. zhu, c. huang, r. lu and h. li. “modelling information dissemination under privacy concerns in social media”. physica a: statistical mechanics and its applications, vol. 449, pp. 53-63, 2016. 3. n. b. ellison and d. m. boyd. “social network sites: definition, history, and scholarship”. journal of computer-mediated communication, vol. 13, pp. 210-230, 2007. 4. p. m. leonardi, m. huysman and c. steinfield. “enterprise social media: definition, history, and prospects for the study of social technologies in organizations”. journal of computer-mediated communication, vol. 19, pp. 1-19, 2013. 5. t. k. yu and g. s. wu. “determinants of internet shopping behavior: an application of reasoned behaviour theory”. international journal of management, vol. 24, p. 744, 2007. 6. a. croitoru., a. crooks, j. radzikowski and a. stefanidis. “geosocial gauge: a system prototype for knowledge discovery from social media”. international journal of geographical information science, vol. 27, pp. 2483-2508, 2013. 7. m. hajli. “a research framework for social commerce adoption”. information management and computer security, vol. 21, pp. 144154, 2013. 8. j. h. abawajy, m. i. h. ninggal and t. herawan. “privacy preserving social network data publication”. ieee communications surveys and tutorials, vol. 18, pp. 1974-1997, 2016. 9. c. c. aggarwal and h. wang, h. “text mining in social networks”. in: social network data analytics. springer, berlin, germany. pp. 353-378, 2011. 10. n. n. bazarova and y. h. choi. “self-disclosure in social media: extending the functional approach to disclosure motivations and characteristics on social network sites”. journal of communication, table 5: theories and research constructs for sn privacy study base theories research constructs lallmahamood, 2007[58] technology acceptance model perceived privacy perceived usefulness/perceived ease of use, intention to use internet banking wang et al., 2012[59] reasoned action (subjective norm/attitudes toward internet shopping), intention to shop on internet kim and ko 2011[52] reasoned action ip concern/perceived justice/social benefits from complaining/information privacy-protective responses jordan 2015[54] trust theory privacy, personal behavior, family attitude posey 2016[60] extended ethical decision-making model information privacy protection eric et al., 2015[58] personal behavior sn privacy lee 2013[23] cpm/tpb sn privacy sn: social network, cpm: communication privacy management, tpb: theory of planned behavior al-rabeeah and hashim: a systematic literature review and directions for further research 100 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 vol. 64, pp. 635-657, 2014. 11. l. gitelman. “always already new”. in: media, history, and the data of culture. the mit press, cambridge. p. 7, 2006. 12. j. webster and r. t. watson. “analyzing the past to prepare for the future: writing a literature review”. mis quarterly, vol. 26, no. 2, pp. 13-23, 2002. 13. p. brereton, b. a. kitchenham, d. budgen, m. turner and m. khalil. “lessons from applying the systematic literature review process within the software engineering domain”. journal of systems and software, vol. 80, pp. 571-583, 2007. 14. m. shanmugam and y. y. jusoh. “social commerce from the information systems perspective: a systematic literature review”. computer and information sciences (iccoins), 2014 international conference. ieee. pp. 1-6, 2014. 15. y. levy and t. j. ellis. “a systems approach to conduct an effective literature review in support of information systems research”. informing science: the international journal of an emerging transdiscipline, vol. 9, pp. 181-212, 2006. 16. w. bandara, s. miskon and e. fielt. “a systematic, tool-supported method for conducting literature reviews in information systems”. proceedings of the19th european conference on information systems (ecis 2011), 2011. 17. b. kitchenham, d. budgen, p. brereton, m. turner, s. charters and s. linkman. “large-scale software engineering questions expert opinion or empirical evidence”? iet software, vol. 1, pp. 161-171, 2007. 18. s. nidhra, m. yanamadala, w. afzal and r. torkar. “knowledge transfer challenges and mitigation strategies in global software development a systematic literature review and industrial validation”. international journal of information management, vol. 33, pp. 333-355, 2013. 19. s. kokolakis. “privacy attitudes and privacy behaviour: a review of current research on the privacy paradox phenomenon”. computers and security, vol. 64, pp. 122-134, 2017. 20. s. b. barnes. “a privacy paradox: social networking in the united states”. first monday, vol. 11, pp. 15, 2006. 21. j. p. carrascal, c. riederer, v. erramilli, m. cherubini and r. de oliveira. “your browsing behavior for a big mac: economics of personal information online”. proceedings of the 22nd international conference on world wide web. acm. pp. 189-200, 2013. 22. t. hughes-roberts. “privacy and social networks: is concern a valid indicator of intention and behaviour”? social computing (socialcom), 2013 international conference. ieee. pp. 909-912, 2013. 23. k. j. lee. “development and analyses of privacy management models in online social networks based on communication privacy management theory”. phd thesis, drexel university, 2013. 24. y. s. hau, b. kim, h. lee, y. g. kim. “the effects of individual motivations and social capital on employees’ tacit and explicit knowledge sharing intentions”. international journal of information management, vol. 33, pp. 356-366, 2013. 25. b. reynolds, j. venkatanathan, j. gonçalves and v. kostakos. “sharing ephemeral information in online social networks: privacy perceptions and behaviours”. ifip conference on humancomputer interaction. springer, berlin, pp. 204-215, 2011. 26. z. tufekci. “can you see me now? audience and disclosure regulation in online social network sites”. bulletin of science, technology and society, vol. 28, pp. 20-36, 2008. 27. m. taddicken. “the ‘privacy paradox’in the social web: the impact of privacy concerns, individual characteristics, and the perceived social relevance on different forms of self-disclosure”. journal of computer-mediated communication, vol. 19, pp. 248-273, 2014. 28. a. m. zafeiropoulou, d. e. millard, c. webber and k. o’hara. “unpicking the privacy paradox: can structuration theory help to explain location-based privacy decisions”? proceedings of the 5th annual acm web science conference. acm, pp. 463-472, 2013. 29. z. bauman and d. lyon. “liquid surveillance: a conversation”. john wiley and sons, new york, 2013. 30. j. grimmelmann. “the privacy virus”. facebook and philosophy, vol. 6, pp. 3-12, 2010. 31. d. c. locke and d. f. bailey. “increasing multicultural understanding”. sage publications, thousand oaks, 2013. 32. g. blank, g. bolsover and e. dubois. “a new privacy paradox: young people and privacy on social network sites”. prepared for the annual meeting of the american sociological association, 17 august 2014. san francisco, california, 2014. 33. c. lutz and p. strathoff. “privacy concerns and online behavior not so paradoxical after all? viewing the privacy paradox through different theoretical lenses”. papers, 2014. 34. y. liu, j. k. boyles, j. genzer and m. d. dickey. “self-folding of polymer sheets using local light absorption”. soft matter, vol. 8, pp. 1764-1769, 2012. 35. kaplan, a. “academia goes social media, mooc, spoc, smoc and ssoc: the digital transformation of higher education institutions and universities”. in: contemporary issues in social media marketing. new york: routledge, 2017. pp. 20-30. 36. i. taylor, p. walton and j. young. “the new criminology: for a social theory of deviance”. routledge, abingdon, 2013. 37. d. boyd, s. golder and g. lotan. “tweet, tweet, retweet: conversational aspects of retweeting on twitter. system sciences (hicss), 2010. 43rd hawaii international conference. ieee, pp. 1-10, 2010. 38. e. christofides, a. muise and s. desmarais. “information disclosure and control on facebook: are they two sides of the same coin or two different processes”? cyberpsychology and behavior, vol. 12, pp. 341-345, 2009. 39. o. nov and s. wattal. “social computing privacy concerns: antecedents and effects. proceedings of the sigchi conference on human factors in computing systems. acm, pp. 333-336, 2009. 40. s. chai, s. bagchi-sen, c. morrell, h. r. rao and s. j. upadhyaya. “internet and online information privacy: an exploratory study of preteens and early teens”. ieee transactions on professional communication, vol. 52, pp. 167-182, 2009. 41. d. h. shin. “the effects of trust, security and privacy in social networking: a security-based approach to understand the pattern of adoption”. interacting with computers, vol. 22, pp. 428-438, 2010. 42. l. casaló, c. flavián and m. guinalíu. “the impact of participation in virtual brand communities on consumer trust and loyalty: the case of free software”. online information review, vol. 31, pp. 775-792, 2007. 43. d. j. kim, d. l. ferrin and h. r. rao. “a trust-based consumer decision-making model in electronic commerce: the role of trust, perceived risk, and their antecedents”. decision support systems, vol. 44, pp. 544-564, 2008. 44. t. dinev and p. hart. “internet privacy concerns and their antecedents-measurement validity and a regression model”. behaviour and information technology, vol. 23, pp. 413-422, 2004. 45. m. deng, k. wuyts, r. scandariato, b. preneel and w. joosen. “a privacy threat analysis framework: supporting the elicitation and fulfillment of privacy requirements”. requirements engineering, vol. 16, pp. 3-32, 2011. 46. n. k. malhotra, s. s. kim and j. agarwal. “internet users” information privacy concerns (iuipc): the construct, the scale, and a causal model”. information systems research, vol. 15, pp. 336-355, 2004. 47. c. l. miltgen and d. peyrat-guillard. “cultural and generational influences on privacy concerns: a qualitative study in seven al-rabeeah and hashim: a systematic literature review and directions for further research 101 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (2): 92-101 european countries”. european journal of information systems, vol. 23, pp. 103-125, 2014. 48. e. hargittai and d. boyud. “facebook privacy settings: who cares”? first monday, vol. 15, no. 8, pp. 15, 2010. 49. a. l. young and a. quan-haase. “privacy protection strategies on facebook: the internet privacy paradox revisited”. information, communication and society, vol. 16, pp. 479-500, 2013. 50. l. s. aeschlimann, r. harasgama, f. kehr, c. lutz, v. milanova, s. müller, p. strathoff and a. tamò. “re-setting the stage for privacy: a multi-layered privacy interaction framework and its application”. mensch und maschine symbiose oder parasitismus? schriften der assistierenden der universität st.gallen, 2014. 51. l. v. casalo, c. flavián and m. guinalíu. “the role of security, privacy, usability and reputation in the development of online banking”. online information review, vol. 31, pp. 583-603, 2007. 52. a. j. kim and e. ko. “do social media marketing activities enhance customer equity? an empirical study of luxury fashion brand”. journal of business research, vol. 65, pp. 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conference on computational science and its applications”. springer, berlin, pp. 791-805, 2009. 58 warkentin, m., a. c. johnston, e. walden and d. w. straub. “neural correlates of protection motivation for secure it behaviors: an fmri examination”. journal of the association for information systems, vol. 17, no. 3, p. 194, 2016. 59. x. wang, c. yu and y. wei. “social media peer communication and impacts on purchase intentions: a consumer socialization framework”. journal of interactive marketing, vol. 26, pp. 198-208, 2012. 60. z. liu, q. min, q. zhai and r. smyth. “self-disclosure in chinese micro-blogging: a social exchange theory perspective”. information and management, vol. 53, pp. 53-63, 2016. 61. m. e. keim and e. noji. “emergent use of social media: a new age of opportunity for disaster resilience”. american journal of disaster medicine, vol. 6, pp. 47-54, 2011. 62. a. m. kaplan and m. haenlein. “users of the world, unite! the challenges and opportunities of social media”. business horizons, vol. 53, pp. 59-68, 2010. 63. n. dabbagh and a. kitsantas. “personal learning environments, social media, and self-regulated learning: a natural formula for connecting formal and informal learning”. the internet and higher education, vol. 15, pp. 3-8, 2012. 64. a. sarker, k. o’connor, r. ginn, m. scotch, k. smith, d. malone and g. gonzalez. “social media mining for toxicovigilance: automatic monitoring of prescription medication abuse from twitter”. drug safety, vol. 39, pp. 231-240, 2016. 65. a. e. marwick and d. boyd. “networked privacy: how teenagers negotiate context in social media”. new media and society, vol. 16, pp. 1051-1067, 2014. tx_1~abs:at/add:tx_2~abs:at 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 23-27 research article empower e-wallet payment system using secured hybrid approach of online and offline services mohammed h. shukur, reem j. ismail, laith r. flaih department of computer science, college of science, chian university-erbil,kurdistan region, iraq abstract as e-commerce expands, payment mechanisms are migrated to mobile devices, creating e-wallets. current e-wallet payment solutions are based on smartphones with internet access. e-wallets are becoming increasingly popular among individuals worldwide. despite this, a survey conducted to determine the primary factors influencing iraqis’ adoption of electronic wallets reveals a slower growth rate in iraq than in other regions. this paper proposes a design for a mobile e-wallet application that offers online-to-offline (o2o) payments to replace traditional e-wallets, credit cards, debit cards, and cash using offline connectivity and near-field communication (nfc) with sms-based payment mechanisms that do not require an internet connection. keywords: e-wallet, offline services, sms-based system, online and offline services, hybrid approach introduction the “digital revolution,” or the transition from analog to digital technology, has altered our lives and created new opportunities in a wide range of societal activities. one of these activities is an electronic commerce and money digitalization. this digitalization necessitates the development of e-wallets as the next phase of the digital revolution, as part of the transition from a physical wallet or plastic card payment system to an entirely electronic payment system.[1] e-wallet is a term that refers to a type of digital system that is used to facilitate online transactions using a smartphone. it functions similarly to a credit or debit card. an e-wallet must be linked to the user’s account through the internet to make payments.[2] an e-wallet is made up of two main parts: software and data. the information is stored in a database that includes cash, the owner’s name, contact information, shipping, or billing information, such as the customer’s address and other information used at the time of checkout on e-commerce sites, and one identification document. furthermore, users can send money using various methods, including direct sending phone numbers, quick response (qr) codes, or direct purchases.[3] in 2022, several e-wallets in iraq, including zain cash, asiahawala, fastpay, firstiraqi bank, nasspay, and zazapay, provided essentially comparable services. what differentiates them are the benefits of utilizing a certain e-wallet. zaincash is one of the most popular e-wallet payment methods in iraq at the moment. the payment system integrated with switch master card allows users to pay either through the e-wallet or mastercard to pay for services offered by the zaincash app or transfer money to a specific account using their number. zaincash is exclusively available to owners of zain phone numbers. fastpay is the most widely used e-wallet in kurdistan. fastpay enables users to send and receive money and move funds across accounts without the need to have a phone number associated with a particular network. while e-wallets are becoming more and more common among people around the world. however, due to the need for internet to complete transactions, it develops in iraq more slowly than in other places. this project focuses on study the local e-wallet and improves their cashless transaction and services by adding new features in the existing e-wallets. research problem it is anticipated that cashless and e-wallet services would account for 80% of total transaction volume between 2020 and 2025[4] but e-wallets grow slower in iraq than in other cihan university-erbil scientific journal (cuesj) corresponding author: mohammed h. shukur, department of computer science, college of science, chian university-erbil, kurdistan region, iraq. e-mail: m.kurd@hotmail.com received: june 20, 2022 accepted: july 18, 2022 published: august 05, 2022 doi: 10.24086/cuesj.v6n2y2022.pp23-27 copyright © 2022 mohammed h. shukur, reem j. ismail, laith r. flaih. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) shukur, et al.: a secure hybrid approach for e-wallet payment system 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 23-27 parts of the world. users are using e-wallets to prepaid mobile reloading, online card purchases, get some services and money transfer. iraq may transition from cash to cashless payments following an increase in cashless payments. e-payment is regarded as critical for the productivity and efficiency.[5] our research identifies the current e-wallet features, convenience, perceived trust, and customer satisfaction factors influencing consumers’ proclivity to use e-wallets through survey and application reviews. in addition, developing new features and add new function to current e-wallets will raise usability, functionality, convenience, and perceive trust. methodology this method employed the extreme programming methodology to create this project. extreme programming is an agile methodology. the main goal is to enable project changes in response to changing circumstances or customer requirements, even late in the project’s life cycle. feedback and communication are viewed as critical components of improving software development. we can connect with users by testing the application and soliciting comments on improving or adding to the software (if any). these criteria assist developers in adapting to new requirements and technologies. the extreme programming process is divided into three stages, and the following sections detail each stage in detail: fact finding: survey the survey aims to put hand on the current issue to design a new perspectives and features to local e-wallets. we conducted a survey of erbil residents. residents of various ethnicities and ages are represented, including students, employed, jobless, self-employed, and retired. the online survey, which was conducted across many platforms, attracted a total of 250 respondents. its shows 145 users have no e-wallet application, while only 105 has e-wallet application. high percentage are using their cash as they do not need cashless payment in their daily activities [figure 1]. figure 2 shows that 69% of e-wallet users use the fastpay application, according to which e-wallet is used. in response to whether, they keep money in their e-wallet to determine if they have daily transactions. about 51% of users say that they do not keep money in their e-wallet and only reload their accounts when e-payments are needed. they do not have daily transactions [figure 3]. to determine e-wallet capabilities, they respond to the type of transactions they use. table 1 shows that six out of 11 online services were used, while five transactions were not covered in e-wallet services. in response to the question of whether they had access to the internet at the time of purchase. about 59% of users report that they were not connected to the internet at the time of purchase; the highest rates of internet usage occurred at home or work. wallet owner internet location percentage home 100 work 85 malls and market 41 the survey reveals that current e-wallet issues have been identified and listed below: • large number of users who do not have access to the internet at the time of purchase figure 1: e-wallet usage figure 3: maintain money in e-wallet figure 2: e-wallet application share shukur, et al.: a secure hybrid approach for e-wallet payment system 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 23-27 • e-wallets do not support offline transaction • e-wallets do not offer touch and go payments planning we looked for related scenarios and cases that may help clarify and validate the problem assertions. to acquire information, surveys and informal interview sessions were undertaken. finally, the objectives and scopes are drawn to gain a picture of the system’s required functions. design in general, the design of the app allpay is created during this period based on the most popular e-wallet functions and adding needed screens and buttons keep the interface userfriendly and simple to learn and operate. users will feel at ease when utilizing the application because of the user experience. due to the enormous demand for offline usage of an e-wallet, sensitive user information is commonly saved on mobile devices. we suggest that this project includes optional offline device-based biometric authentication and that the most recent balance be retrieved through encrypted time-based one-time password (topt) short message service (sms), and to provide online-to-offline (o2o) services.[6] figure 4 illustrates offline login, the application uses local mobile biometric authentication employs biometrics to identify and authenticate the user seeking to access a mobile app. screen 1 shows login screen with biometrics. it may be done through fingerprint readers, facial recognition, speech recognition, and other methods. after user successful user authentication, the applications check if the devise whether connected to server or not, in case of no internet connection, the application sends an sms to the server requesting the current balance; the server will then produce rsa encrypted totp sms codes with account phone number that are refreshed every 30 s, making them more challenging to use if stolen, prevent fraud, and keeping the application. the following is a description of the login model’s execution: 1. user use biometric authentication to access the application 2. application check whether it is connected to server or not, one of below options online 1. application inquiry the balance 2. server send updated balance offline 1. application generated rsa encrypted sms contains token, using the homomorphic properties of the rsa encryption. 2. server confirm the token by decrypting it using the device’s last online token along with phone number. execute of one of the below options: a. successful authenticated, server send encrypted updated balance in totp sms b. failed authenticated, server send warning sms, and lock the account for 30 min. while logging in through offline mobile verification, the e-wallet automatically detects totp. rceive_sms in android table 1: type of transactions in e-wallets transaction type percentage online cards 33 mobile\internet prepaid cards 35 online services 10 online shopping 15 shopping 1 services 0 patrol station 0 taxi 5 tuition fees 0 bills 0 malls 0 figure 4: offline login with totp is used by app to read the encrypted totp, decrypt it with last obtained online token, and update the balance in the main screen. typically, totp detection requires the user to grant sms permissions. screen 2 illustrates that the 30 s needed to receive and update the balance. the usage of nfc in e-wallet applications is introduced in this design, allowing for more flexible and secure management, customization, and simplify the payment process.[7,8] at pressing go button on the main screen which it represents the touch and go payment. go are contactless payments that employ near-field communication (nfc) technology to securely communicate between an e-wallet and a point of sale’s (pos) terminal at time of pressing the button. this technology powers e-wallet payments shown in screen 3. the following is a description of the nfc model’s execution: 1. to make a payment, the customer must press the “go” button on their phone to enable nfc on the pos terminal. 2. the e-wallet reads the transaction information from the pos. 3. the application checks whether the user has enough credit or not using the current balance stored in the application. 4. if the user has sufficient credit, e-wallet apps send collected information to the server through online or offline (sms) communication to authorize the transaction. shukur, et al.: a secure hybrid approach for e-wallet payment system 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 23-27 5. server either authorizes the transaction or rejects the user’s request. if the user’s request was authorized, the purchase amount will be withdrawn from his account; otherwise, the user’s account will remain with the same balance. at the time of each transaction, the application checks if the amount required is within the balance; if it is not, the user will receive notification “insufficient funds to complete the payment.” in the event that the amount is equal to or less than the balance. the application will determine whether it is connected to the server online or not. in case of offline, the application sends encrypted sms to server with token and user transaction information, amount, receiver id, and other related info about the order [figure 5]. server will send back totp to user to validate the transaction. discussion the proposed e-wallet payment system, which combines online and offline services, satisfies the following standards defined: • secure authentication. additional components ensure that only authorized users may conduct online and offline transactions. biometric authentication is the first step in the login process, followed by two-factor authentication. • secured payments. in offline transactions, data are encrypted using the rsa cryptosystem’s homomorphic features; as a result, the user and any sniffer learn nothing about the data. in touch and go payment, nfc enabled at the point when the user touch go button, adding extra security to enable the payment transaction. we are not concentrating on registration at the moment, and we assume that all users have previously registered, been validated in the payment system, and have access to the internet at some point. the following procedures should be followed while modifying the online authentication method with the offline services scheme: screen 1: login, red button for biometric login screen 2: reading totp and updating user balance screen 3: touch and go payment using nfc. figure 5: online/offline transaction shukur, et al.: a secure hybrid approach for e-wallet payment system 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 23-27 1. the user chooses the biometric feature they want to employ (e.g., fingerprint and facial recognition) during the authentication procedure. 2. within the e-wallet application, a hash value is utilized with the user’s identification and approval of the user’s identity. 3. two tokens to confirm rsa encryption and decryption (server token and user token), server-generated random numbers and save it in the server database, and e-wallet at any online activity. 4. only the server accepts sms from registered and validated phone numbers then compares user tokens. 5. once linked to the internet, the e-wallet produces new tokens and verifies previously saved transactions. 6. account will be locked for 30 min if any verification fails. conclusion the solution presented in this paper includes three additional components: local biometric authentication, offline payment transactions, and touch and go nfc payment. the suggested solution applies to online and offline transactions in the banking and payment sectors. as a result, it is appropriate for countries with limited internet connections. finally, the intended contributions were successful, resulting in favorable outcomes and distinctive contributions. references 1. m. olsen, j. hedman and r. vatrapu. hci issues in mobile wallet design, in the tenth annual pre-icis workshop on hci research. china: shanghai, 2011. 2. definition of “e-wallets”. times syndication service. available from: https://www.economictimes.indiatimes.com/definition/ewallets [last accessed on 2022 mar 14]. 3. a. madan and m. i. rosca. current trends in digital marketing communication. journal of marketing research and case studies, vol. 2022, pp. 1-13, 2022. 4. v. vinatier. investment managers. available from: https://www. axa-im.com/insights/future-trends/digital-economy/why-scalematters-evolving-landscape-cashless-payments [last accessed on 2022 mar 14]. 5. g. oprescu and d. eleodor. the impact of the digital economy’s development on competition. bucharest: international conference of the institute for business administration, 2014. 6. a. dhir. v. scuotto. p. kaur and shalini talwar. barriers and paradoxical recommendation behaviour in online to offline (o 2 o) services. a convergent mixed-method study. journal of business research, vol. 131, pp. 25-39, 2021. 7. n. sabli, n. e. pforditen, k. supian, f. n. azmi and n. a. solihin. the acceptance of e-wallet in malaysia. selangor business review, vol. 6, no. 1, pp. 1-14, 2021. 8. p. pourghomi and g. ghinea. managing nfc payments applications through cloud computing. in: 7th international conference for internet technology and secured transactions (icitst), ieee, 2012. ole_link1 _goback tx_1~abs:at/add:tx_2~abs:at 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 57-61 research article serum fibroblast growth factors 23, chemerin, and vitamin d levels in patients with psoriasis shatha n. tawfeeq1, moayad m. al anzy2, entedhar r. sarhat3 1department of medical microbiology, cihan university-erbil, kurdistan region, iraq, 2department of pharmaceutical sciences, tikrit university, tikrit, iraq, 3department of basic science, faculty of medicine, tikrit university, tikrit, iraq abstract the aim of the study was to compare the levels of fibroblast growth factor 23 (fgf23) and chemerin, as well as vitamin d in psoriasis sufferers and healthy controls. from 2021 to 2022, ninety subjects were studied. the study comprised 45 individuals with psoriasis and 45 healthy controls. serum fasting fgf23 and chemerin levels of the study group were examined by sandwich enzyme-linked immunosorbent assay. the results demonstrate that the mean serum fgf23 and chemerin quantities in psoriasis patients were greater than in controls, with a statistically significant. patients with psoriasis, on the other hand, had considerably lower blood vitamin d levels than healthy controls. these findings demonstrate that psoriatic patients have lower vitamin d levels than healthy controls, and they add to the growing body of data linking fgf23, chemerin vitamin d levels, and the course of psoriasis. keywords: serum fibroblast, psoriasis, fibroblast growth factor 23, chemerin, 1,25 (oh)2d introduction psoriasis is now recognized as a chronic immune-mediated inflammatory dermatosis with an unknown etiology. it is caused by a variety of external and endogenous causes, related with a range of biochemical and immunological abnormalities,[1] affecting approximately 2% of the global population.[2] despite the fact that it does not affect just one tissue, it is linked to autoimmune reactions all over the body. sharply defined dull red spots with scales, particularly in the scalp and on extensor prominences, are used to categorize psoriasis.[3] dendritic cells and t-cells multiply in the immune system, releasing a range of inflammatory precursors cytokines and chemokines, as well as growth factors, which all have a role in the pathogenesis of psoriasis.[4] in contrast to most other fibroblast growth factors (fgfs), which do not behave as traditional hormones, the fgf23 gene is located on chromosome 12p13 and characterized by endocrine and paracrine effects.[5] it is a phosphaturic proteohormone produced mostly by osteoblasts/osteocytes in the bone that is plays a function in phosphate reabsorption and 1,25 (oh) 2d3 synthesis in the kidney. metabolism of fgf23 included the cleavage of bioactive intact fgf23 (ifgf23, 30 kd) into c-terminal (cfgf23, 12 kd) and n-terminal (18 kd) fragments. fgf23 could be cleared by kidneys through filtration and catabolism,[6,7] ffgf23 a protein with classical endocrine, but also paracrine effects. klotho, a transmembrane protein, is required for fgf23-mediated regulation of the renal phosphate transporter napiiia and cyp27b1, the major enzyme in the production of 1,25(oh)2d3.[8] adipose tissue is a metabolic and endocrine organ that is complicated, critical, and highly active. chemerin is one of the metabolically active peptide hormones, cytokines, chemokines, and adipokines released by this gland.[9-11] chemerin (16 kda), also known as tig2 or rarres2, is a newly discovered adipocyte-secreted factor that has both pro and anti-inflammatory properties. it is made as an inactive prochemerin that is transformed to its active state by serine proteases. not just in wat, it can even be found in the skin, intestines, and airways, chemerin is secreted in enormous levels. chemr23, gpr-1, and ccrl2 are the three kinds of receptors that chemerin interacts to. chemerin inhibits neutrophil transepithelial migratory process and increases apical neutrophil evacuation when chemr23 is activated.[12,13] cihan university-erbil scientific journal (cuesj) corresponding author: shatha n. tawfeeq, department of medical microbiology, cihan university-erbil, kurdistan region, iraq. e-mail: shatha.nasih@cihanuniversity.edu.iq received: june 23, 2022 accepted: july 15, 2022 published: august 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp57-61 copyright © 2022 shatha n. tawfeeq, moayad m. al anzy, entedhar r. sarhat. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). tawfeeq, et al.: role of fibroblast growth factors 23, chemerin, and vitamin d in psoriasis 58 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 57-61 vitamin d is also known as vitamin that excreted in the body after sun exposure. it refers to a group of fat-soluble a fat-soluble biomolecule, connected to bone metabolism and skeletal integrity as well as generated in the skin by the impact of uv irradiation from the sun. it has historically been known as a hormone that maintains calcium-phosphorous balance and protects the musculoskeletal system’s structure. vitamin d modulates immunity by transforming 25(oh)d (vitamin d’s hormonallyactive form) into tissue-specific, locally produced 1,25(oh)2d (vitamin d’s hormonally-active component). macrophages, t-cells, keratinocytes, and dendritic cells involved in psoriasis pathogenesis can produce and respond to 1,25 (oh)2d in a paracrine and autocrine way when 25 (oh)2d is directly accessible for cellular absorption.[14] a 25(oh)d level of <20 ng/mll indicates vitamin d insufficiency.[15] our study’s purpose is to compare the concentrations of serum fgf23, chemerin, and vitamin d in psoriasis patients to those of a healthy population of similar age and gender. materials and methods patients and controls forty-five patients of psoriasis (18 males [40%] and 27 females [60%]) with a mean age of 38.6 ± 11.2 were included in the study. forty-five ageand sex-matched normal healthy controls (16 males [35.5%] and 29 females [64.5%]) with a mean age of 37.3 ± 11.21 year as shown in table 1. patients were collected from the biochemistry department of tikrit teaching hospital in tikrit city during the period from april 18, 2021, to january 17, 2022. pregnancy, diabetes, hypertension, and inflammatory diseases other than psoriasis comprised the criterion of exclusion. the study was authorized by the local ethics committee, and all patients and volunteers in the control group gave their informed permission. measurement of fgf23, and chemerin in serum after an overnight fast, blood was drawn from patients and controls. after clotting, serum was extracted and kept in aliquots at −70°c until needed. the enzyme-linked immunosorbent assay was used to assess fgf23 and chemerin (mybiosource, usa). statistical analysis statistical analysis was done using t-test. correlation between continuous variables was estimated by pearson’s correlation coefficients. a value of p < 0.05 was considered statistically significant for each test. results the study revealed that, in comparison to the healthy control group, psoriatic cases had the highest mean of fgf23 enzyme level (109 93 pg/ml). as demonstrated in figure 1 and table 2, the difference was extremely significant (p = 0.0001). the study showed that psoriatic patients had a greater mean of serum chemerin (328.6 25.2 ng/ml) than the healthy control group, (229.1 ± 15.4 ng/ml according to the research). while s. vitamin d (21.68 ± 1.79 ng/ml) was significantly reduced in psoriatic patients (45.19 ± 4.31 pg/ml) comparing with the control group (29.92 ± 1.55 ng/ml). furthermore, parathyroid hormone (pth) was significantly higher among patients’ group (45.19 ± 4.31 pg/ml) compared to the control group (20.78 ± 1.50 pg/ml).as shown in figures 1-4 and table 2. discussion the kidneys maintain phosphorus homeostasis in healthy humans by proximal tubular active absorption or filtered phosphorus pass-through. 25-dihydroxyvitamin d (1,25 (oh)2d, parathyroid hormone and fibroblast growth factor-231 are hormones and factors that help kidneys regulate phosphorus (fgf-23).[16] the parathyroid gland’s main cells table 1 : demographic characteristics of patients (n=45) and controls (n=45) patients control age 38.6±11.2 37.3±11.21 gender male 18 16 female 27 29 figure 1: serum pgf23 in patients with psoriasis and the controls figure 2: serum chemerin in patients with psoriasis and the controls tawfeeq, et al.: role of fibroblast growth factors 23, chemerin, and vitamin d in psoriasis 59 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 57-61 manufacture and emit pth. pth is a hormone made up of peptides, released as a 115-amino-acid long pre-pro-pth at first. then, it turns into about 84 amino acids when pre-propth is degraded twice.[17] pth is a critical regulator of bone and mineral metabolic process, and its biosynthesis is influenced by changes in blood calcium levels, which trigger responses in several pathways.[18] pth levels in relation to 25(oh)d levels are often used to determine vitamin d insufficiency levels. vitamin d insufficiency appears to lower serum calcium levels, resulting in elevated pth levels. vitamin d is required for calcium absorption in the intestines in humans. furthermore, the 25(oh)d concentration, which begins to climb or no longer falls as the pth concentration lowers, may indicate vitamin d inadequacy.[19] pth is elevated when calcium levels are low, or phosphate levels are high. high levels of parathyroid hormone damage bones and cause phosphate and calcium levels in the bones to fluctuate.[9] pth-rp is only found typical skin’s basal lamina and acts as a powerful epidermal cell proliferation inhibitor. pth-rp is prevalent in keratinocytes, and 1,25 (oh)2 d3 regulates it.[20] in psoriatic lesions, pth is not present. pth-rp expression was recovered in psoriatic skin when lesions were medicated with 1,25 (oh)2d3. inflammation is one of the main causes that leads to an increase in fgf23 transcription. nuclear factor-kb, a critical component of the inflammatory signaling pathway, affects the expression of target genes, including fgf23, through transcriptional regulation. nfat was first discovered as a nuclear factor that can bind to the interleukin-2 promoter in activated t cells. nfat also has a role in innate immunological and inflammatory responses, according to research.[21-23] fgf23 promotes hepatocytes to secrete inflammatory markers including il6 and tnf, as well as fibrinogen, c-reactive protein, and vascular cell adhesion molecule-1.[24] increased fgf23 has the ability to trigger the inflammatory response by suppressing of 1,25-dihydroxyvitamin d, a strong inhibitor of inflammatory cytokine release.[25] chemerin was validated to play as an inflammatory intermediary to production of proinflammatory cytokines and encourage the relocation of immature dendritic cells and macrophages.[26] chemerin concentrations in serum samples were observed to be greater in psoriatic patients than in the control group in this investigation. in a similar manner, gisondi et al. and lora et al.[27,28] discovered elevated levels of chemerin in psoriasis patients having high disease activity. xue et al.[29] found that patients with psoriasis had considerably lower amounts of circulating chemerin than healthy controls. it is unclear what mechanism could explain these findings. we hypothesized that the mechanism was linked to chemerin’s angiogenic properties.[30] chemerin is a protein produced mostly by keratinocytes and endothelial cells, as well as white adipose tissue. il1 and tn f-both boost its production. chemerin increases angiogenesis in psoriatic skin through stimulating adipocytes and endothelial cells. in psoriasis patients’ growing skin lesions, chemerin can stimulate chemotaxis of immature myeloid dcs and plasmatoid dendritic cells.[31] chemerin may be implicated in psoriasis pathophysiological processes. chemerin expression was shown to be higher in psoriatic skin biopsies. the enhanced migration of dendritic cells and nk cells to the inflammation may be due to an increase in the concentration of this protein in mast cells, dermal fibroblasts, and endothelial cells throughout early phases of psoriasis. the stimulation of angiogenesis may also play a role in the evolution of psoriatic inflammation. chemerin’s ability to induce the development of new vasculature could indicate a role for this adipokine in psoriasis.[32] physiologically, 1,25-dihydroxyvitamin d and vitamin d receptor directly regulates the proliferation and growth of keratinocytes through vitamin d receptors. there are several pathways for the effect of vitamin d deficiency in psoriasis pathogenesis: loss of antiproliferative, anti-inflammatory, and antiangiogenic activities.[33-35] table 2: descriptive characteristics of serum fgf23, chemerin, vitamin d, and pth between between studied groups fgf23 (pg/ml) vitamin d (ng/ml) chemerin (ng/ml) pth (pg/ml) patients 106.1±15.2 21.68±1.79 328.6±25.2 45.19±4.31 control 265.7±22.1** 29.92±1.55** 229.1±15.4** 20.78±1.50** *p<0.05 significant, **p<0.0001. pth: parathyroid hormone figure 3: serum pth in patients with psoriasis and the controls figure 4: serum vitamin d in patients with psoriasis and the controls tawfeeq, et al.: role of fibroblast growth factors 23, chemerin, and vitamin d in psoriasis 60 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 57-61 psoriasis is an autoimmune inflammatory disease involving both innate and acquired immunity. it is defined as an autoimmune inflammatory sickness with th1-th17-th22 components. decreased amounts of 25(oh)d have also been linked to a higher risk of developing autoimmune pathologies mediated by th1.[36] the reason for the lower level of serum 25(oh)d in these patients may be due to inadequate 25(oh) d intake, paucity of sunlight exposure, or usage of medicines (e.g., glucocorticoids and immunosuppressive agents) that interact with 25(oh)d breakdown on a regular basis.[37] conclusion based on our observations, we have come to the conclusion that psoriasis sufferers’ serum fgf23 and chemerin levels are higher than healthy controls. patients with psoriasis, on the other hand, had considerably lower serum vitamin d levels than healthy controls. therefore, fgf23, chemerin, and vitamin d levels may be a novel diagnostic marker in psoriasis and may predict the occurrence of comorbidities in psoriasis patients. references 1. s. pal, s. sen, i. nath, a. kumar and u. k. biswas. psoriasis, an inflammatory condition associated with oxidative stress. asian journal of medical sciences, vol. 12, no. 4, pp. 24-30, 2021. 2. i. olejniczak-staruch, m. ciążyńska, d. sobolewska-sztychny, j. narbutt, m. skibińska and a. lesiak. alterations of the skin and gut microbiome in psoriasis and psoriatic arthritis. international journal of molecular sciences. vol. 22, no. 8, pp. 3998, 2021. 3. r. k. jha, a. singh, p. koundal and r. s. ambad. role of lipid and oxidative stress in psoriatic patients--a case control study. journal of evolution of medical and 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disease severity. annals of clinical laboratory science, vol. 46, no. 2, pp. 174-179, 2016. 26. e. riffat sarhat, s. j. khalaf and m. s. hamad. a study of some biochemical parameters in blood serum of patients with congestive heart failure. vol. 10, no. 5, p. 412, 2019. 27. p. gisondi, v. lora, c. bonauguri, a. russo, g. lippi and g. girolomoni. serum chemerin is increased in patients with chronic plaque psoriasis and normalizes following treatment with infliximab. british journal of dermatology, vol. 168, no. 4, tawfeeq, et al.: role of fibroblast growth factors 23, chemerin, and vitamin d in psoriasis 61 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 57-61 pp. 749-755, 2013. 28. v. lora, c. bonaguri, p. gisondi, f. sandei, l. battistelli, a. russo, a. melegari, t. trenti and g. lippi and g. girolomoni. autoantibody induction and adipokine levels in patients with psoriasis treated with infliximab. journal of immunology research, vol. 56, no. 2, pp. 382-389, 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meibodi and a. hadianfar. serum 25-oh vitamin d level in psoriatic patients and comparison with control subjects, j cutan med surg, vol. 20, no. 3, pp. 207210, 2016. 34. l. barrea, m. c. savanelli, c. di somma, m. napolitano, m. megna, a. colao and s. savastano. vitamin d and its role in psoriasis: an overview of the dermatologist and nutritionist, rev endocr metab disord, vol. 18, no. 2, pp. 195-205, 2017. 35. a. q. hamdi, e. r. sarhat, n. h. ali and t. r. sarhat. evaluation of lipocalin-2 and visfatin, and vitamin (d, c, and e) in serum of diabetic patients with chronic periodontitis. indian journal of forensic medicine and toxicolog, vol. 15, no. 2, pp. 1668-1674, 2021. 36. a. filoni, m. vestita, m. congedo, g. giudice, s. tafuri and d. j. m. bonamonte, association between psoriasis and vitamin d: duration of disease correlates with decreased vitamin d serum levels: an observational case-control study. medicine (baltimore), vol. 97, no. 25, p. e11185, 2018. 37. y. h. lee and g. g. song. association between circulating 25‐ hydroxyvitamin d levels and psoriasis, and correlation with disease severity: a meta-analysis. clin exp dermatol, vol. 43, no. 5, pp. 529-535, 2018. tx_1~abs:at/add:tx_2~abs:at 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 research article using factor analysis to determine the most important factors affecting student absenteeism at cihan university-erbil hawkar q. birdawod department of business administration, cihan university-erbil, kurdistan region, iraq abstract the review of the relevant literature reveals that the issue of absenteeism among students was not new. the researcher intended to explore the factors affecting absenteeism and to find a solution to solve this issue. for this purpose, a mixed methods research was designed. to analyze this problem, both quantitative and qualitative research methods were adopted in this study. in quantitative approach, a closeended questionnaire was developed to collect data. the sample comprised 470 students out of total population of 4892. a descriptive and inferential statistical techniques were used to analyze the data. the outcome of the study shows that common factors such as curriculum and organizational climate, transportation and recreation, administrative procedures, and psychological and economic issues all had an impact on students’ absences. likewise, the results revealed that administrative and motivational factors, including personal organization and adaptations with performance to health and second housing, are the critical factors in the student’s absences. furthermore, gender of the student plays a major role in the student’s absenteeism, and the results show that there is a significant difference between male and female students toward the absence of the students. keywords: absenteeism factors, factor analysis, component method, principle component method, cihan university-erbil introduction factor analysis is an important tool of statistical methods associated with multi-dimensional variables, that is related to a set of phenomena or attributes that help a number of variables such as people. it is a statistical method, because it depends on the first argument at the expense of correlation coefficients between these phenomena. one tries to use the factor analysis to return the causes of a group of characteristics (we call it random variables) to a smaller number of factors which can be measured entirely and can be described in a simplified and accurate manner.[1] factor analysis can also be defined as a multivariate analysis based on a set of hypotheses using the (correlation matrix) or (covariance matrix) to determine the (hypothetical factors) underlying the nature of interrelated relationships. internal relationships are among a set of variables taken for a specific phenomenon. these factors are represented by new variables but with fewer original variables. the aim of the analysis is to describe and interpret the phenomena or characteristics of the variables on the basis of few factors as possible.[1,2] one of the main important factors for the development of society and the country is the reform and improvement of the education system in universities. many studies have been conducted for this purpose. as a result, a study has been conducted to assess and understand the factors that influence a student’s academic performance.[3] another study focused on student feedback on the teaching methods used by university teachers.[4] a common topic that has been investigated to improve the education system in this region is the online education system, which has been discussed since the outbreak of the coronavirus and has become a supportive and alternative education in the universities of the kurdistan region.[5] another interesting study is to determine the factors that cause university students to hate reading books.[6] the study of absenteeism is very important for every institution and every teacher to improve the results. the reducing the rate of absenteeism is useful for making the students disciplined, punctual, and consistent.[7,8] this study aims to find different factors of absenteeism at cihan university-erbil and find a solution to reduce it. corresponding author: hawkar q. birdawod, department of business administration, cihan university-erbil, kurdistan region, iraq. e-mail: hawkar.birdawod@cihanuniversity.edu.iq received: may 20, 2022 accepted: june 18, 2022 published: july 10, 2022 doi: 10.24086/cuesj.v6n2y2022.pp1-8 copyright © 2022 hawkar q. birdawod. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) birdawod: using factor analysis to determine student absenteeism 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 the researcher’s effort was to sort out varied significant factors involved in the student’s absenteeism at the university level. it has always been a burning issue and especially nowadays since the government is concerned with improving and bringing positive changes to the educational sector. prominent level of absenteeism reported as missing 10% or more of the academic year has been a key indicator of student’s failure.[9,10] student absenteeism been given much attention today, but still it is uncontrollable issue. one thing is clear that this matter is not ignorable rather it needs greater attention and time to look into. since new college students experience many challenges physically, socially, and mentally.[10,11] research questions keeping in view the main objective of the study, the following research questions were made in light of the relevant literature. (1) what are the factors that cause absenteeism among university students? (2) do the factors of absenteeism affect boys and girls students equally? (3) are subject matter and the medium of instruction cause student’s absenteeism? (4) is there any significant difference in the perspective of students in the factors of absenteeism? students with low attendance attribute their absenteeism to the poor relationship with teachers, university administration, and class fellows. the researcher made a standard about the low attendance by fixed a cut off line which will make decision making easy about the low attendance and the accurate attendance. objectives of the study the main objectives of the study are as follows: 1. to investigate the effects of absenteeism among students of cihan university-erbil 2. to present the basic literature review regarding absenteeism among students 3. to recommend a possible solution for absenteeism after the analysis of the data. research hypothesis based on the objective and the conceptual framework of this study, null hypothesis is developed as follows: h 01 : there is no significant difference between male and female students toward student’s absenteeism. h 02 : there is no relationship between subject interest and student’s absenteeism. h 03 : there is no relationship between lecturer’s skills and student’s absenteeism. h 04 : there is no relationship between student’s attitude and student’s absenteeism. h 05 : there is no relationship between health problems and student’s absenteeism. the objectives of using the factor analysis the main objectives of factor analysis can be summarized in the following points: 1. interdependency and pattern delineation: the method of factor analysis explanation reveals the (patterns) separate of the relations between the variables and return the causes of those variables to the lowest number of factors, in which the strength of the relationship of each variable is determined by these patterns 2. parsimony or data reduction: if we have a large number of private observations of a large set of variables, these data can be concentrated in the form of a few factors that represent the variables in describing the phenomenon and clarify relationship between them 3. hypothesis testing: the factor analysis can be used to test the validity of the determinants of the type of factors affecting a set of variables, depending on the factor matrix 4. data transformation: the global analysis helps to transform data into another image in which certain conditions can be applied to some statistical methods on it 5. scaling: the method of factor analysis is useful in exploring new areas. factor analysis reduces complex relationships between a set of variables to a simple linear image 6. multicollineartiy: a factor analysis method contributes to addressing the problem of linear multiplicity, to convert variables associated with unrelated factors.[2,12-14] literature review factor model the model of the factor analysis of p (p dimensional) of the sample his size is n that it is based on a linear function of p the mean variables and m of the common factors, in which (m <p) and to (p) of the unique factors for each variable, the factor model will be as follows:[12,13] x f upx px pxm mx px1 1 1 1� � �� � (1) where: x: represents the random vector of observed variables. µ: represents the vector of variables. a: the loading factors matrix represents the changes. f: is the random vector of the common factors selected from pvariables. u: represents the random vector of the only variables of the variables. basic assumption of factor analysis the basic hypothesis of factor analysis is based on the existence of correlations between a set of variables unstudied and that these correlations are the result of the existence of interrelated factors, in which the factor analysis seeks to explain these linkages with the least number of independent factors among them, this hypothesis assumes the standard value (standard value) of the variables, which may be distributed in natural distribution with a mean of zero well as for the elimination of variables.[15,16] based on this hypothesis, the vector of the media and the variance of the variables will be zero vectors, var x i( ) = and e x o( ) � �� birdawod: using factor analysis to determine student absenteeism 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 in this case, the factor model takes the following form: z f up p m m p* * * *1 1 1� �� (2) for the covariance matrix for each of the common factor drivers (f) and the only factors (u) assuming that, they are independent: � � � � � � �f u f u ff fu uf uu m m m� � � � � � � �� � � �� � �� � �� � �� � � � � � � � � � � �nn n m n m� �� � � � � � � whereas: φ: represents the heterogeneity matrix of the common factors f� � . ψ: the country matrix represents single factor variance. as for the matrix of variance of the variables x� � and (σ) denoted by: e x x p p( ') *�� so (σ) that it is a symmetric positive definite and full rank matrix. moreover, since the goal in factor analysis is to estimate the parameters of the model, it was necessary to express the model (2) in a more appropriate form, since: � � � � � �� e x x e af u af u( ) ( )( ) � � �� � � � �� �ae ff a ae fu e uf a e uu( ) ( ) ( ) ( ) the structural matrix (ω), which represents the covariance between observed variables and factors, they are: � � ��p m x f� � � �( ) these hypotheses divide the total variation into three variations. common variance represents a part of the total variance that is associated with the rest of the variables and is calculated from the coefficients of the general factors and symbolized by the symbol hj 2� � .[13,17] h a a aj j j jm 2 1 2 2 2 2� � � �... (3) specific variance it represents a part of the total variance that is not associated with the other variables, but is associated with the same variable, which is part of the only factor variation that equals u b ej j j 2 2 2� � (4) where: uj 2 : is the only factor variation. bj 2 : variance of the variable j . ej 2 : error variance. error variance which is not explained by the common factors that can be described as residual or error, is the variation resulting from errors in the sample drag or measurement or any changes that lead to data instability and symbolizes ej 2� � .[13,16] e h bj j j 2 2 21� � �( ) (5) it is worth mentioning that factor analysis cannot be used if the sample size is less than 50 observations, and it is preferably when the sample size contains 100 observations or more. the commonalities the standard value is the sum of the squares of the saturations of the factors of that variable, and it represents the common variance that is explained by the common factors extracted from the correlation matrix analysis, that is, it represents the extent of interference between the variables and the common factors: [13,17] h a a aj j j jm 2 1 2 2 2 2� � � �... where j=1,2,3,………, p i=1,2,3,……, m or h aj ij i m 2 2 1 � � � many of the factor analysis methods are needed when applying the factor analysis to an initial estimate of the quantities of communion hj 2� � and placing them in the diagonal elements of the correlation matrix (r) instead of the units. these methods are the method of analysis of the main factor used in our research, which needed to estimate these values and place them in the main diameter of the correlation matrix.[17-19] many of the factor analysis methods, including the main factor analysis method, need when used to a preliminary estimate of the quantities of commonness and put them in the diagonal elements of the correlation matrix instead of the units; then, it is called the reduced correlation matrix, and one of the characteristics of the common values is that they are always positive values, its value lies between zero and one (because it is part of the total variance, which is equal to one), that is, it: [14,16,19] 0 12≤ ≤hj factor analysis methods there are several ways to estimate the matrix of factor loading for the first solution, the following is the most common: 1. principal components method 2. factor method principal 3. method maximum – likelihood 4. image method 5. unweighted least squares 6. generalized least squares 7. alpha method birdawod: using factor analysis to determine student absenteeism 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 8. the centered method 9. rao method. we will explain the main vehicle method, for adoption in the applied side, because it is more accurate in estimation.[15,20,21] principle component method the basic idea of this method is to find the main universes, the number of which is equal to the number of studied variables (x j ) where (j=1,2,3,……,p) that follow the multivariate normal distribution with the aver mean of the population ( )µ and the covariance and covariance matrix (σ), meaning that ( , )� � � x and then choose the first component of the main components, which is modified to find the following model:[16,19,22,23] x l f l f l f u x l f l f m m p p p 1 11 1 12 2 1 1 1 1 2 2 � � � � � � � � ............... .................� � � � � � �l f upm m p (6) where: f 1… f m : represent the unrelated general factors that were extracted from (p) the variables. l ji : represents the factor loading (i) of the variable (j). u 1 …u p : are the only factors methodology the results of this study will be based on the literature review and the responses collected from students through questionnaires. the response rate was appropriate to analyze the data. data were analyzed in percentage response of respondents and using factor analysis. the sample size is 470 students out of total population of 4892. the sample was calculated based on (5%) margin of error. the respondent was divided into 240 female students a 51.06% and 230 of the respondents were male 48.94%. the study was conducted on the basis of a sample of 470 students selected randomly. the data were collected by survey questionnaire as a mean tool. after classifying the data from the forms of the sample, each statement was calculated as a variable given symbols, using the statistical package (spss25). the data include a set of variables to explain the reasons for absence students. results and discussion the purpose of this study is to determine the factors influencing students’ absences, to show the most important variables and their sequence in terms of their effect on the studied phenomenon for the method of factor analysis, the principal components were used to analyze the correlation matrix of the studied variables, and based on the matrix of the coefficients of the circulating factors in a manner (varimax) and show the importance of each variable based on the relationship of this variable with other variables. the number of moral factors was selected according to (kaiser scale) based on the characteristic values (the factor variation), keeping any factor with eigen value greater than one. the significance of the loadings for each factor was determined on the basis of the comparison between factor loadings, where the largest load value was selected in each factor, and then, it is compared with other factor loads for same variables. the (varimax) matrix was applied on all students and it was taken into consideration for the purpose of knowledge and distinguishing the reasons for absenteeism. the principal components (p.c): the first stage for all students in table 1, we present the results obtained in this way for 470 students and that represent the size of the sample (males and females). it is noted from the table that the total number of factor variances is equal to 24, which represents the total variance of all the variables studied (the standard values). the shaded parts indicate the presence of the nine main significant factors effect on the reasons for absence of both genders together, table 1: analysis using the main compounds method for all students (male and female) factor factor variance (distinguished value) variance ratio factor cumulative percentage of variance 1 4.295 17.897 17.897 2 2.321 9.67 27.568 3 1.887 7.861 35.428 4 1.556 6.481 41.91 5 1.379 5.745 47.655 6 1.266 5.276 52.931 7 1.197 4.987 57.918 8 1.139 4.748 62.666 9 1.064 4.433 67.099 10 0.865 3.605 70.704 11 0.85 3.543 74.247 12 0.821 3.419 77.666 13 0.703 2.931 80.597 14 0.652 2.716 83.313 15 0.571 2.38 85.693 16 0.55 2.294 87.986 17 0.496 2.067 90.053 18 0.455 1.896 91.949 19 0.41 1.707 93.656 20 0.382 1.59 95.246 21 0.344 1.434 96.681 22 0.328 1.365 98.046 23 0.267 1.113 99.158 24 0.202 0.842 100 total 24 100 birdawod: using factor analysis to determine student absenteeism 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 which represent the number of factors whose value exceeds one. these nine factors explain a percentage (67.099%) of the total variance of variables explained by these nine factors. although they explain different percentages of variation, but they important in determining the variables affecting the absence of students and these nine factors are explained, respectively, 17.897%, 9.670%, 7.861%, 6.481%, 5.745%, 5.276%, 4.987%, 4.748%, and 4.443% of the total variance. first factor this factor is important in influencing the causes of absence, where it explains (17.897%) of the total variance. this factor is saturated with significant saturation for the following variables and according in sequence: depending on the book only to explaining the academic subjects x21 and by amount of 0.696 and the value of communality is 0.573, difficulty of a number of subjects x22 by amount of 0.674 and the value of communality is 0.625, incompatibility between students x17 by amount of 0.614 and the value of communality is 0.539, the non-encouraging way of teachers in presenting the course material x2 by amount of 0.570 and the value of communality is 0.706, the weakness of the humanitarian relations between teachers and students x1 by amount of 0.551 and the value of communality is 0.714, student unwillingness to education x7 by amount of 0.511, and the value of communality 0.612. the weakness of academic subjects to motivate student x20 by amount of 0.485 and the value of communality is 0.644 within the saturation of this factor and in both genders and the quantities shown in the shaded parts of the table. this factor can be called (scientific) factor which is an important indicator to determine the reasons for absence of students in both genders. this factor can be called the curriculum factor and the social and organizational climate. it can be said that the depending on the book in the commentary may be difficult to understand the student and that the lack of the introduction of examples and evidence may lead the student to a state of helplessness and lack of desire to lecture and then evade attending lectures on this side, and on the other hand, it can be said that the lack of an adequate social climate in the college reflects the negative effects on harmony which leads the student to search for other groups with them and, thus, was always absent. on the other hand, there is a lack of interest in the humanitarian aspect in dealing with students and weakness in consideration about their feelings, desire, and circumstances. in addition, the limited number of teachers makes it difficult to deal with both academic as well as the human side of the student. therefore, understanding the students’ circumstances can be a helpful tool to decrease absenteeism. in particular, the modern literature in the field of education and psychology refers to the importance of using psychological methods to link the subject to the needs of students and their interests and tendencies, all this leads to the departure of students from the study schedule and then their absence from lectures. second factor this factor is very important and comes in second place in terms of importance, which it interprets (9.670%) of the total variance and this factor satisfies a significant saturation of the following variables and in sequence, unhealthy conditions in the accommodation x24 by amount of 0.845 and the value of communality is 0.775, failure to provide appropriate classroom environment x8 by amount of 0.845% and the value of communality 0.755, and also poor living conditions of university students x6 by amount of 0.626 and the value of communality is 0.612. the weakness of the moral imprints of this factor in both genders are called the second administrative and scientific factor. this factor can be called accommodation factor and economy first. the problem of student stability is a fundamental factor affecting the extent of his commitment to the continuation, especially in the beginning of the academic course. lack of good health conditions in the accommodation of the heating and furniture and other conditions can be leading to infection diseases. hence, it can be counted among the reasons for the absence of students from this permanence on the one hand, and on the other hand, a number of students from outside the center of the province maybe delay in obtaining accommodation due to the frequent procedures and complexities of routine as well as the concern of providing accommodation that burdens the students. this will result in the inability of students to connect to the school, and on the other hand, the nature of the economic conditions experienced by the student, which reflected the impact especially on the students’ ability to maintain their commitment due to the high wages of travel compared to the potential of students, especially those who suffer from poor living conditions, constant absences. third factor this factor comes in third place in terms of importance, it explains 7.861% of the total variance and saturated this factor by amount of 0.639, 0.795 and the value of communality is 0.745, 0.676 of the variables lack of transportation facilities x5, and lack of comfort in university campuses x16. this factor can be called (transportation and recreational) factor. the lack of means of transport and the inability of the student to pay the transportation fees may result in a lack of commitment. on the other hand, the absence of students may sometimes refer to psychological reasons related to the student’s need for resting, so the lack of comfort causes the student to feel a sense of distress and tension and resort to other places away from the college, which leads the student to the absence of permanence. fourth factor this factor comes in fourth place in terms of importance, it explains 6.481% of the total variance and saturated this factor by amount of 0.691, 0.805 and the value of communality is 0.599, 0.744 of the variables unclear vision of future career for college graduates x12 and management delay announcement of results x10. this factor can be called (future and administrative) factor. as we say, this factor can be referred to the acceptance controls, as well as the lack of the future, which affects in some way the motivation of the students negatively and their desire for continuous work, so he suffers from a birdawod: using factor analysis to determine student absenteeism 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 table 2 : (loading matrix) moral factors and common quantities of all the factors produced by the main compound method after rotation for all students (male and female) variable first factor second factor third factor forth factor fifth factor sixth factor seventh factor eighth factor ninth factor dispersion x1 0.551 −0.09 0.113 −0.06 0.538 −0.15 −0.2 −0.171 0.08 0.714 x2 0.57 −0.252 −0.06 0.04 0.119 0.281 −0.14 −0.137 0.424 0.706 x3 0.102 0.204 0.122 0.193 0.005 0.045 0.759 0.051 −0.07 0.69 x4 0.081 0.413 0.057 0.067 0.583 0.168 0.145 0.232 −0.11 0.639 x5 0.045 0.131 0.759 −0.17 0.183 0.138 0.03 0.243 0.085 0.745 x6 −0.018 0.626 0.375 0.113 0.181 −0.06 0.135 −0.117 0.049 0.616 x7 0.511 0.42 0.359 0.254 0.415 0.139 −0.079 0.057 −0.14 0.612 x8 0.099 0.845 −0.02 −0.09 −0.03 0.108 −0.015 0.015 0.105 0.755 x9 −0.101 0.221 0.143 −0.02 0.023 0.015 −0.023 0.038 0.811 0.74 x10 0.074 −0.034 −0.12 0.691 0.153 −0.04 0.215 −0.049 −0.17 0.599 x11 0.422 0.225 0.265 0.188 −0.01 0.154 −0.433 0.145 −0.3 0.657 x12 0.07 0.001 0.164 0.805 −0 −0.16 −0.073 0.14 0.121 0.744 x13 0.113 0.269 0.328 0.027 −0.05 0.721 0.034 −0.149 −0.07 0.745 x14 0.387 −0.043 0.333 −0.35 0.346 −0 0.461 0.053 −0.03 0.724 x15 −0.103 0.055 0.085 0.072 0.713 0.055 0.039 0.017 0.064 0.543 x16 0.241 0.003 0.639 0.364 0.067 −0.05 0.201 −0.065 0.155 0.676 x17 0.614 0.179 0.117 0.152 0.042 0.026 0.257 −0.082 −0.01 0.539 x18 −0.036 −0.04 0.002 −0.24 0.056 0.713 −0.078 −0.105 0.033 0.589 x19 −0.049 −0.107 −0.32 0.052 0.288 0.556 0.219 0.263 0.108 0.641 x20 0.485 0.132 −0.05 0.196 −0.18 −0.06 0.183 0.439 0.299 0.644 x21 0.696 0.137 0.21 −0.1 −0.07 −0.07 −0.028 0.054 −0.06 0.573 x22 0.674 0.048 −0.16 0.169 0.002 0.049 0.088 0.234 −0.22 0.625 x23 −0.052 0.109 −0.13 −0.03 −0.11 0.119 0.016 −0.867 0.024 0.812 x24 0.099 0.859 −0 −0.03 0.083 −0.04 0.09 −0.053 0.081 0.775 %variance ratio factor 17.9 9.67 7.861 6.481 5.745 5.276 4.987 4.748 4.433 psychological condition that is directly make him/her unstable in his time and presence. fifth factor this factor comes in fifth place in terms of importance, it explains (5.745%) of the total variance and saturated this factor by amount of 0.713, 0.583 and the value of communality is 0.639, 0.543 of the variables various personal and family problems x15 and students work during or after university x4. this factor can be called (social) factor. many personal and family problems add to the sense of increasing the burden of work, which enables them to maintain their commitment to sustainability. on the other hand, the poor economic situation of the student reflected its effects, especially to the potential of the students and the extent of their commitment to full time, some of them to work despite being a student to improve economic level. sixth factor this factor comes in sixth place in terms of importance, it explains 5.745% of the total variance. this factor is saturated with a significant saturation of the following variables and in sequence: a number of students were preoccupied with relationships with others x13 by amount of 0.721 and the value of communality is 0.745. there are also two variables poor follow-up by the administration to students’ absences x18 and strong friendship bonds among students x19 by amount of 0.713, 0.756 and the value of communality is 0.589, 0641. respectively and sequentially, this factor may be referred to the (first administrative) or the motivational. the weak follow-up of the administration of the students’ absences affects the motivation of the students negatively and their desire for continue as well as the existence of a range of psychological and social needs are not saturated students, birdawod: using factor analysis to determine student absenteeism 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 some of them are personal needs and their interests and need any affiliation to others, so their motivation is an important issue in terms of students to stay away from the study, because they do not agree and their needs and tendencies which leads to their departure from permanence. seventh factor this factor comes in seventh place in terms of importance, it explains %4.987 of the total variance and this factor is saturated with a significant saturation of the following variables and in sequence: poor organization of the study schedule x3 by amount of 0.759 and the value of communality is 0.690, explanation of the subject by the teacher in a language far from the language of the student x14 and frequent daily examinations by a number of teachers x11 by amounts of 0.461, 0.433 and the value of communality is 0.657, 0.724. the weakness of the moral imprints of this factor in both genders are called the second administrative and scientific factor. these three variables are called the second administrative and scientific factor. eighth factor this factor comes in seventh place in terms of importance, it explains 4.748% of the total variance and this factor is saturated with a significant saturation of the following variables and in sequence: the health situation of the students x23 by amount of 0.867 and the value of communality is 0.812. only within the moral parameters of this factor can be called (health) factor. since the absence of a number of students is always attributed to the suffering of health, as the infection of some diseases, especially cold diseases and the associated drugs, due to the siege leads to the student’s immaturity to physical and psychological rest and avoid it from work so can be counted among the reasons for the student’s absence. ninth factor this factor is ranked ninth and most important in terms of importance. it explains 4.433% of the total variance, and this factor satisfies a significant saturation of the variant of long distance from the university x9 by amount of 0.811, and the value of communality is 0.740. the ninth factor can be called (accommodation) conclusions after addressing the applicable aspects of the study, and obtaining the results of the implementation of the principal of components method, the researcher came with the following conclusions: 1. during the work on the factor analysis for all students, the researcher found that there are nine main moral factors that allow us to determine the main effective reasons (variables) that lead to student’s absence for both genders and each of them individually 2. the study revealed that the explained ratio of total variance is the nine factors in cases (all students, only females, only males) equal (67.009%, 74.014%, and 72.585%), respectively, this means that the explained ratio of total variance analysis that been done separately for each gender has increased comparing to the explained ratio in the analysis of both genders, and that assures our assumptions of importance of doing analysis of each gender individually, to get a better result 3. the study shows that the most important variables that have moral effects on students’ absence are curriculum and organizational climate, transportation and recreation, administrative procedures, and psychological and economic issues. administrative and motivational factors including: personal organization and adaptations with performance to health and second housing. for females, living and economy, personal adaptation to the course and curriculum, admission and graduation regulations, psychological, health and administrative, transport and recreation, acceptance procedures, organizational and physiological climate, motivation, for males, curriculum climate and human relations, housing, economics, administrative management, admission controls and administrative procedures, motivation, recreation, personal adjustments, and permanence and health 4. differential results of the factor analysis among both genders means that the order or sequence of the important variables that effect the female absence are different from the order or sequence of male’s absence variable. thus, that assure our assumptions of doing the analysis of each gender individually, the purpose is to determine the reasons that lead to absence of gender individually 5. not giving the real reasons of absence by students may prevent the desired results from being obtained. recommendations the following recommendations can be made in light of the study conclusion: 1. the study assists the determination of main factors that affect students’ absence (and for each gender individually); hence, the study recommends the importance of doing researches and studies on the same topic, and take other variables (reasons) in consideration, and for greater number of students to come up with a valid recommendations and that’s for spotting the important reasons to reduce the phenomenon of absence at the university 2. the importance of taking the principles of humanitarian relationships into account for interrelationship between teachers with students 3. use interesting and affective teaching methods, in a way that enriches class environment with fruitful communications among teachers, and students on the other hand. furthermore, encouraging students to study and find other academic resources to overcome the difficulties related to certain subjects 4. accommodate students for their necessities, especially providing transportation, provide suitable job opportunities for students, particularly those in need, weather inside the university or in coordination with organizations, as well as providing financial support to facilitate their affairs 5. the need to apply instructions and guidelines, especially in regard to students’ absence by assigning part of the passing grade according to attendance 6. minimize the routine procedures regarding registration of birdawod: using factor analysis to determine student absenteeism 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 1-8 students in the university and taking in consideration the importance of students desires to do that 7. care for the cleanliness of student’s accommodation and the provision of health condition to avoid the possibility students to be infected by diseases. references 1. i. ajzen. attitudinal vs. normative messages: an investigation of the differential effects of persuasive communications on behavior. sociometry, vol. 34, no. 2, pp. 263-280, 1971. 2. k. v. 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absenteeism on the academic performance of pupils of ebom-bomfa jhs. unpublished dissertation. university of cape coast, 2017. 19. j. a. al-mahdawi. an analytical statistical study on the factors affecting juvenile delinquency, a master’s thesis submitted to the department of statistics, college of administration and economics. iraq: university of baghdad, 1987. 20. n. wadesango and s. machingambi. causes and structural effects of student absenteeism: a case study of three south african universities. journal of social sciences, vol. 26, no. 2, pp. 89-97, 2011. 21. a. al-sabawi. organizational knowledge and its impact on determining the entrances to the formulation of strategy: a field study of the opinions of senior management in a sample of joint stock companies in the city of mosul. rafidain development journal, college of administration and economics university of mosul, 1999. 22. g. pehlivan. the school absenteeism among high school students: contributing factors. educational sciences: theory and practice, vol. 16, no. 6, pp. 5-17, 2016. 23. h. q. pirdawd. transformed and solving multi-objective linear programming problems to single-objective by using correlation technique. cihan international journal of social science, vol. 1, pp. 30-36, 2017. tx_1~abs:at/add:tx_2~abs:at 29 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 29-34 research article inhibition of rac 1 protect against platelet-induced liver and kidney injury in diabetes mellitus rundk a. hwaiz1, helen j. sabri2 1department of clinical biochemistry, hawler medical university, kurdistan region, iraq, 2department of medical biochemical analysis, cihan university-erbil, kurdistan region, iraq abstract diabetes mellitus (dm) both (type  1 and type  2) are one of the common causes for activation of platelet. inflammation-induced abnormal platelet function contributes to chronic complications, which are the leading causes of death and morbidity among diabetics. rac1 has been shown to regulate a variety of platelet functions; predicted rac1could regulate platelet release of cxcl4, which leads to kidney injury in dm. dm’ effect on rac1 activation, a 21kd g-protein implicated in platelet activation, was investigated and platelet induced inflammation and kidney injury. swiss albino male mice were pretreated with 5 mg/kg of a specific rac1 inhibitor nsc23766 and injected with (45 mg/kg body wt.) streptozotocin, twice for 5 days. moreover, the concentration of serum chemokines cxcl4 was assayed using elisa and histology score for kidney was examined. our results showed that dm was induced in mice by streptozotocin. in addition, platelet chemokines (cxcl4) were markedly higher in diabetic mice when compared to the sham (control) group. moreover, pre-treatment with nsc23766 decreased liver and kidney injury assessed by histology score, p < 0.05. our study reveals that rac1 has a critical role in platelet chemokines secretion due to diabetes-induced inflammation in the liver and kidneys, targeting rac1 could be a target for innovative treatment to control inflammation in diabetic individual. targeting platelets involved in inflammatory pathways could be part of a strategy to control and manage diabetes and its consequences. keywords: rac1, platelet, cxcl4, diabetes mellitus, liver and kidney injury introduction diabetes mellitus (dm) is a state in which set to be failure of the langerhans islet of the pancreas to produce sufficient insulin, or for the insulin to be resistant.[1] the figures of people with diabetes are expected to soar by 550 million by the end of the year 2035.[2] diabetes has detrimental consequences that are mainly because of it is micro and macroangiopathy which include several debilitating neurological complications and nephropathies in which serious glomerular complications develop leading to dropping the level of glomerular filtration rate (gfr) and eventually renal failure.[3,4] determining factors for the increased levels of gfr in early dm have been observed utilizing different physiological experiments commencing factors and pathophysiology routes are particularly ambiguous to determine as the relation between form and function tense to be complicated microangiopathy complications in diabetes were found as renal lesions in type 2 diabetes where hyperfiltration did not sound to occur while.[5,6] liver is a crucial organ in the regulation of lipid, glucose, and protein metabolism and is one of the numerous organs affected by this metabolic illness diabetes. the most specific markers of hepatic injury are alanine aminotransferase (alt), and aspartate aminotransferase (ast), which are found in the hepatocellular mitochondria and cytosol. diabetes is frequently accompanied by elevated serum levels of (alt), (ast), and gamma glutamyltransferase.[7] excess free fatty acids are directly toxic to liver cells in the insulin-resistant state, t2dm is thought to be a contributing factor in the development of liver diseases such as cirrhosis, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis, hepatocellular carcinoma will eventually develop.[8] pre-diabetic individuals or impaired glucose tolerance subjects have demonstrated lower levels of mean platelet corresponding author: rundk a. hwaiz, department of clinical biochemistry, hawler medical university, kurdistan region, iraq. e-mail: rundk.hwaiz@hmu.edu.krd received: june 14, 2022 accepted: january 27, 2023 published: march 01, 2023 doi: 10.24086/cuesj.v7n1y2023.pp29-34 copyright © 2023 rundk a. hwaiz, helen j. sabri. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) hwaiz and sabri: role of rac1 in diabetes mellitus 30 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 29-34 volume (mpv) than diabetic people in several studies.[9] higher platelet activity is frequently found to be more in diabetic patients, platelet glycation proteins promotion are directly link to platelet reactivity.[10] diabetic patients are doomed to develop higher platelet reactivity, as both insulin resistance and deficiency levels up platelet reactivity.[11] chronic diabetic patients in general are not an exclusion of the vicious circle in which hyperglycemia leads to high platelet activation: the latter is thought to be contributing in the producing the reactive oxygen species (ros) in endothelium since this have been showed in several studies. other than this ros production in endothelium happens directly due to glucose metabolism and autooxidation indirect ros production may also occur through the development of pro-inflammatory cytokine receptors for advanced glycation end products (rage).[12] cytosol of platelets embed pro-inflammatory derivatives like cxcl4 which is sent to be important mainly produced from rna and the fate is to be spilled on platelet activations. this pro-inflammatory derivative is released upon platelet activation during cecal ligation and puncture mice which in its turn leads to expression of cxcl4 and this may cause aggregation of other immune cells and exaggerate inflammation during abdominal sepsis.[13,14] rac1a small guanine nucleotidebinding proteins (g protein) is a key signaling molecule that coordinates intracellular transduction pathways that lead to nadph oxidase activation.[15] similar to other small gtpases, rac1 must be isoprenylated to move from cytosol to the plasma membrane, where its presence facilitates the assembly of some nadph oxidase subunits.[16,17] this assemblage is a very important step for the enzymatic activation of the rosproducing nadph oxidase system. as a result, rac1 activation may be a key mechanism in hyperglycemia-induced vascular oxidative stress. rac1 a small gtpase protein playing a crucial role in dm as a signaling molecule.[18] furthermore, platelet secretion of cxcl4 in sepsis is also controlled by rac1.[13,14,19] numerous theses have emphasized the principal contribution of rac1 in sepsis, lamellipodia formation, as well as the activation of phospholipase, granule secretion, and clot retraction in platelets. so far, the molecular and cellular mechanisms contribute to the inflammation and complication effects of diabetes are only partially recognized; hence, the aim of the present study was to investigate the rac1 role in releasing cxcl4 from platelets in dm and the inhibition of rac1 to protect against liver and kidney damage due to dm. materials and methods animals swiss albino male mice were used in all experiments 8–9 weeks of age (weight: 20–25 g) the department of pharmacy hawler medical university, iraq, is in accordance with animal welfare standards legislation, and the regional animal experimentation ethical committee has given its approval. the animals were housed in a pathogen-free environment with a 12–12-h light–dark cycle. water and food were provided twice daily clean and fresh drinking water provided specific nipple used to access ad libitum. the trials were conducted after a 7-day acclimatization period. animals were sub-grouped in to three groups sham (injected with saline only), vehicle (injected with streptozotocin), and treatment group which pretreated with nsc23366 and streptozotocin (nsc+stz), each group contains five mice. with environment enrichment, the mice were maintained in cages with no more than five mice per cage in each group. materials streptozotocin (stz; glentham life science. ltd., u.k.) is the chemical that uses for induction of dm.[20] naoh the buffer that uses for preparation of streptozotocin was made by dissolving (10.7 g) of sodium citrate in (200 ml) of distilled water and (9.6 g) of citric acid added and the volume completed to l000 ml with distilled water. by adding (naoh) to the solvent, the ph of the solution was adjusted to (4.5). nsc23766 (n6‐[2‐[[4‐(diethylamino) ‐1‐methylbutyl] amino] ‐6-methyl‐ 4‐pyrimidinyl] ‐2 methyl‐‐4, 6‐quinolinediamine trihydrochloride, chem cruz, santa cruz biotechnology, california). the accu-chek active blood glucose meter was used to monitor the blood glucose level in the study. animal experiments (5 mg/kg) of rac1 inhibitor (nsc23766) was administered intraperitoneally to the animals based on past research, this dose of nsc23766 was chosen.[19,21,22] after 30 min, the animals were treated using multiple (i.p.) stz (45 mg/kg body weight) was injected intraperitoneally into the experimental mice to produce dm. stz was dissolved in a buffer of 0.01 m sodium citrate (ph = 4.5) and given to mice for 5 days in a row. this dose of stz is selected on the base on the previous study.[23] to minimize hypoglycemia produced by stz, after the injection, the animals were given a glucose solution (5% w/v) to drink overnight. sham mice were given an equivalent dose of vehicle (citrate buffer) only. for 5 days, stz and nsc+stz treated mice were housed in normal settings after dosing was completed. in fact, after these time period mice developed diabetes by measuring fasting blood glucose levels were approximately 11.1 mmol/l. blood samples from the tail vein of nsc+stz and stztreated mice were taken after a 12-h fast to assess blood glucose levels. diabetes was defined as fasting blood glucose levels higher than 11.1 mmol/l in diabetic mice.[23] moreover, hence, they were selected for further studies. intravenously, sedation was achieved by administering 75 mg ketamine hydrochloride (hoffmanla roche, basel, switzerland) and 25 mg xylazine (janssen pharmaceutica, beerse, belgium) kg-1 body weight. the animals were harvested and blood took from vena cava. the serum was allowed to coagulate at room temperature for 10–20 min. before being centrifuged for 20 min at 2000–3000 rpm. the serum was extracted from the supernatant and kept at 80°c for use in an elisa test later. for histopathology, the kidney was preserved in formaldehyde. biochemical determination before the start of the procedure, all experimental animals’ blood glucose levels were monitored. fasting blood glucose levels were checked on a regular basis until diabetes was confirmed. mice with a fasting blood glucose level of 11.11 mmol/l or above were classified as diabetic. blood was collected from the tail veins of all experimental animals (2–3 µl). an accu-chek active blood glucose meter was used to monitor the blood glucose levels. hwaiz and sabri: role of rac1 in diabetes mellitus 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 29-34 elisa serum cxcl4 level measured successfully in all groups of mice and samples assessed by enzyme-linked immunosorbent assay (elisa) using the (mouse platelet factor 4 elisa kit, bt lab cat. no e0686mo). following the manufacturer’s directions, at 450 nm absorbance was measured, a standard curve had been set on each microplate by diluting a known concentration standard. using a logistic curve-fitting technique, the mean absorbance for the wells was used to calculate the chemokine concentration for each sample. the linear section of the standard curve contained all of the absorbance values. the data were represented as ng/ml. histopathology kidney tissue was fixed overnight in a 10% formaldehyde phosphate buffer before being dried and paraffin embedded. hematoxylin and eosin were used to stain four micrometer sections. a modified scoring system was used to quantify kidney injury in a blinded manner.[24] including mixed inflammatory cells, necrosis (reversible injury), apoptosis, fibrosis, vascular congestion and edema, and degeneration (reversible injury) infiltration is evaluated on a scale of 0–4, zero represents (absence) and four represents (extensive). the mean value was calculated after assessing five random locations in each tissue sample. the sum of all six criteria determines the histopathology score. statistics the data were provided as mean values with standard error (se). nonparametric tests were used to do statistical analyses (mann–whitney). n is the total number of mice in each group, and p < 0.05 was considered significant. statistical analysis was performed by use of spss (ibm corp., armonk, n.y., usa). results streptozotocin-iinduced dm in mice injection of (45 mg/kg body wt.) of stz to mice significantly increased the fasting blood sugar of mice compared to sham group, p < 0.05. however, the group that treated with rac1 inhibitor markedly reduced the high blood sugar induced by streptozotocin. pre-treatment of mice with 5 mg/kg of the rac1 inhibitor (nsc23766), reduced the fasting blood sugar from 556.20 ± 26.65 to 375.20 ± 20.7 with p < 0.05 [figure 1]. hence, attenuating rac1 activation by nsc23766 prevents high blood glucose which might induce platelet activation. rac1 regulates platelet secretion of cxcl4 in dm dm increased plasma levels of cxcl4 from 6.40 ± 0.4 ng/ml in sham mice up to 13.60 ± 1.32 ng/ml, corresponding to a 2.12-fold increase [figure 2]. the results showed induction of dm by injecting (45 mg/kg body wt.) szt in plasma [figure 2] suggesting that dm induces the platelet chemokine secretion cxcl4 in diabetic mice. notably, (nsc23766) a rac1 inhibitor, significantly reduced dm-induced platelet aggregation and figure 1: blood glucose concentration. fasting blood glucose levels were measured on day of harvest after the mice were induced with stz (vehicle) for 5 days and pretreated with rac1 inhibitor (nsc23766). data represent mean ± se (sham = 5, vehicle = 5). *p < 0.05 versus sham figure 2: activated platelets secrete chemokines in diabetic mice. elisa was use d to quantify the levels of cxcl 4 in the diabetic mice plasma. data represent mean ± se (sham = 5, vehicle = 5). *p < 0.05 versus sham chemokines secretion in platelets [figure 2], showing that nsc23766 which is an effective inhibitor of rac1 activation, also inhibit the increased level of platelet chemokines. pretreatment with nsc23766 attenuate serum levels of cxcl4 in diabetic mice from 13.60 ± 1.32 ng/ml to 7.20 ± 0.8 ng/ml, equating to a drop of more than 80% [figure 2]. histopathology alterations of kidney in diabetic mice histopathological change of kidney regarding normal control showed normal kidney histology architecture, while treated group (induced diabetes) showed mild vacuolar degenerations, scattered chronic inflammatory cells infiltration. while after giving treatment, there will be reduction in the inflammatory cells and reduction in degeneration [figure 3b]. the induction of dm by injection of streptozotocin, significantly induced hwaiz and sabri: role of rac1 in diabetes mellitus 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 29-34 kidney injury in diabetic mice compared to the sham group (1.13 ± 0.08) with p < 0.05 [figure 3a]. however, administration of nsc23766 to the mice reduced the histological score to 0.26 ± 0.21 with p < 0.05 [figure 3a]. histopathology alterations of liver in diabetic mice histopathological change of liver normal control showed normal liver histology architecture, while treated group (induced diabetes) showed vascular congestion, degenerative hepatocyte (reversible injury), and scattered chronic inflammatory cell infiltration (lymphocyte). while in the pre-treated group with nsc23766, there was only mild vascular congestion [figure 4b]. our findings revealed significant liver damage in the diabetes induced animals. streptozotocin injections used to induce diabetes in mice dramatically increased liver damage compared to the control group 1.4 ± 0.08 with p < 0.05 [figure 4]. nsc23766, on the other hand, reduced the histological score in mice to 0.73 ± 0.06, with p < 0.05 [figure 4a]. discussion the current findings suggest that rac1-mediated platelet activation and cxcl4 secretion from platelets play an important role in dm. these data imply that platelets play a critical role in diabetes, and that reducing rac1 signaling and/or cxcl4 function could be effective dm therapy options and its complication associated with platelet activation. platelets are important for wound healing and thrombosis, but they also contribute to the host’s response to bacterial invasion by performing a variety of pro-inflammatory actions.[25] platelets, for example, govern a variety of features of responses of leukocytes to severe infections, according to research.[13,14,19] platelets have been demonstrated to play an important role in the development of dm in the previous investigations; however, in this study, we found that the injection of mice with streptozotocin (45 mg/kg body wt.) significantly induce the dm compare to sham group p value <0.05. similar to our study, the previous studies showed the elevation of glucose after induction of streptozotocin.[23,26-28] however, the role of rac1in regulation of platelet chemokines in the dm not figure 4: rac1 regulates liver damage in dm. (a) rac1 regulates liver damage in dm. histology score in the liver. (b) rac1 regulates liver damage in dm. representative (h&e ×100) sections of the liver are shown. (a) normal histology of the liver of normal control group. (b) vascular congestion, and degenerative hepatocyte as shown in the (arrow on the vehicle), and scattered chronic inflammatory cell infiltration. (c) treatment group showed only mild vascular congestion a b figure 3: rac1 regulates kidney damage in dm. (a) histopathology score in the kidney. (b) rac1 regulates kidney damage in dm. representative (h&e ×100) sections of the kidney are shown. (a) normal histology of the kidney of normal control group. (b) scattered chronic inflammatory cells as shown in (arrow 1), and vacuolar degeneration (arrow 2). (c) treatment group showed reduction in inflammatory cells and vacuolar degeneration ba hwaiz and sabri: role of rac1 in diabetes mellitus 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 29-34 been studied earlier. induction of streptozotocin-induced dm, significantly increased platelet chemokine secretion. however, the induction of dm by streptozotocin was abolished by the pretreatment with rac1 inhibitor (5 mg/kg), by more than 80% reduction [figure 2]. this emphasizes the role of platelets and rac1 in the dm molecular process. moreover, histological study showed the kidney destruction in diabetic mice which induced by injection of streptozotocin. pre-treatment with rac1 inhibitor (5 mg/kg) significantly reduced the kidney damage induced by dm. the role of platelets in renal damage was demonstrated in the previous studies.[29-32] talat et al. showed that platelet count significantly increased in the diabetic patients.[33] the glomerulus and tubules basement membrane thickens and this leads to recruitment of inflammatory cells. gene expression and protein production of extracellular matrix components such as collagen iv, laminin, and fibronectin increase when extracellular matrix components accumulate. the weight of the kidneys was reported to increase as a result of these changes.[34] our result showed pathological changes in the liver produced in diabetic mice induced by (45 mg/kg body wt.) streptozotocin. a recent study has shown liver injury induced by stz.[35] furthermore, another study revealed that the occurrence and progression of diabetic liver injury are influenced by oxidative stress.[36] in this study, we documented a significant reduction of mixed inflammatory cells and apoptosis of pre-treatment hepatocytes with rac1 inhibitor in mice. the rac1 inhibitor (5 mg/kg) administration significantly reduced necrosis, fibrosis, vascular congestion and edema, an extreme change shown between diabetic mice and pre-treated rac1 inhibition. furthermore, the latest study showed that nsc23766 treatment significantly alleviated the hepatic ri in mice as well as reducing ros production and oxidative stress.[37] pro-inflammatory cytokines were observed to be acutely induced in the liver and kidneys of rats exposed to gnps in several studies.[38-40] in platelets, cxcl4 is one of the most prevalent chemokines.[13,14] numerious studies have found that patients with metabolic syndrome had higher levels of (il-6), (tnf), cxcl16, and (crp) than healthy people. furthermore, metabolic syndrome has been linked to an increase in the number of immune cells such as leukocytes, monocytes, and platelets.[41-43] interestingly, the level of cxcl4 in diabetic mice’s serum was higher than in sham animals, according to this study findings [figure 2]. rac1 has been implicated in the regulation of platelet chemokine production in the previous investigations.[13,14,19] furthermore, it is well known that high blood glucose levels assist in the development of ros on the endothelium lining of arteries.[44,45] ros, a signaling molecule produced by nadph, is vital in the advancement of inflammation and vascular damage in diabetes.[46] rac1, a small g protein, is a key signaling molecule that connects intracellular signaling pathways to nadph oxidase activity.[47] conclusion in the present study, we showed that rac1 has a function in platelet activation and that the chemokine cxcl4 is overexpressed in diabetic mice. in addition, morphological change associated with dm in mice as a consequences of platelet activation. rac1 inhibition may thus be a therapeutic drug to regulate dm through attenuation of chemokine, which is a signaling molecule in aggravating inflammation, as well as chemokine production liver and kidney damage by activated platelets in dm. aknowledgment i would like to express my heartfelt gratitude to my college, hawler medical university, and salahadin university research center, for providing me with the 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service attacks in internet of things using software-defined networking and ensemble learning shima rashidi1, adil h. mohammed2, yusra. a. salih3 1department of computer science, college of science and technology, university of human development, sulaymaniyah, kurdistan region, iraq, 2department of communication and computer engineering, faculty of engineering, cihan university-erbil, kurdistan region, iraq, 3department of database technology, college of informatics, sulaimani polytechnic university, sulaymaniyah, kurdistan region, iraq abstract the internet of things (iot) is a novel approach to automate connections between smart devices without involving humans. the utilization of this structure is growing, and its application range is continually expanding. we confront additional issues as the usage of these networks grows, such as the presence of attackers and combating their attacks. these networks’ performance may be improved, and their development can be accelerated, with new solutions to these difficulties. a new method for improving iot security is proposed in this research, which is based on software-based network and collaborative learning. the suggested solution divides the network domain into numerous subdomains, each with its own controller for exchanging security rules with other subdomains. all of a subnet’s node traffic are routed through the subnet’s control node in this topology. as a result, each controller node employs an integrated learning model to continually evaluate network traffic data and detect assaults. this learning model incorporates an artificial neural network, a decision tree, and a new biz model that uses statistical information gathered from each data stream to identify the likely existence of assaults. nslkdd database data were utilized to assess the proposed method’s performance, and its accuracy in identifying denial of service attacks was compared to earlier approaches. keywords: denial of service attacks, intrusion detection system, cumulative learning, software-based network, internet of things introduction the internet of things (iot) is a notion that was first presented in the late 20th century and refers to the intelligent interaction between machines and humans over a large communication platform such as the internet. all smart gadgets in this communication system have a unique identity and the capacity to interact with other devices. each device that can be connected to the structure of the iot is called an object, and the result of the integration of all objects in a global information network will form the iot. in general, the interactions of objects in this communication structure are beyond the traditional machine-to-machine communication and can lead to huge changes in areas such as transportation, industry, treatment network, and so on. despite these benefits, several of the network’s fundamental properties, such as its breadth, variety of objects, and usage of numerous protocols, have posed management issues.[1] a big network, in most cases, cannot function properly without some form of organization. this is due to issues such as user authentication and network architectural scalability. some of the existing difficulties can be overcome in a cost-effective manner using software defined networks (sdn).[2] a software-centric network is a novel communication network design that has been proposed to achieve aims such as more efficient network dynamics confrontation, improved network flexibility, and improved network manageability. the control layer is separated from the data layer in this design, which is made up of two primary components: forwarding elements and sdn controllers. delivery equipment’s role is to exchange packets through proprietary hardware or software. the controller, on the other hand, is a piece of software that operates on a hardware platform and is designed to carry out specific functions. to authenticate users on the network, the suggested solution cihan university-erbil scientific journal (cuesj) corresponding author: adil h. mohammed, department of communication and computer engineering, faculty of engineering, cihan university-erbil, kurdistan region, iraq. e-mail: adil.mohammed@cihanuniversity.edu.iq received: june 17, 2022 accepted: july 29, 2022 published: august 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp49-56 copyright © 2022 shima rashidi, adil h. mohammed, yusra a. salih. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). rashidi, et al.: detecting denial of service attacks in internet of things 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 employs an sdn-based architecture. it separates the network’s domain into sdn subdomains, with each subdomain in charge of authenticating users in its own domain. on the other side, the iot ecosystem gives greater opportunity for attackers to conduct threats such as denial of service (dos) assaults due to the exponential growth in the number of connected devices and apps. application layer dos attacks are a type of malicious attack that targets the osi model’s top layer (where common internet requests such as http get and http post occur). identifying the origins of attacks and security risks are another topic to consider while constructing the proposed model in this context. to this purpose, the controller nodes in each sdn subnet manage the network traffic pattern and identify the existence of probable attacks on their subnet using a model based on ensemble learning techniques.[3] several learning models (such as decision tree, neural network, and new biz) are utilized in the proposed cumulative learning model to identify the origins of assaults, and the final diagnosis is determined by voting among the output findings of the learning models. the following is the order in which this article will be continued: in the second section, we’ll look at works that are linked to the research topic. the proposed method is described in detail in the third section, and the results of evaluating its performance in a simulated environment are discussed in the fourth section. finally, in the research’s fifth section, the findings are summarized and recommendations for further research are provided. related studies perrone et al.[4] examined the message queuing telemetry transport (mqtt) protocol’s security and defined the different security requirements for establishing the iot and safeguarding devices from application layer attacks using the mqtt protocol. andy et al.[5] looked at certain iot attack scenarios and assessed iot security to mitigate these attacks in another study. the authors’ study focuses on iot messaging protocol security and attack scenarios against open authentication servers; they’ve described it. although the viability of such attacks is currently debatable, the open authentication functionality is deactivated in most deployments of message servers in industrial contexts to decrease the danger of unauthorized access and associated assaults.[6] firdous et al.[7] investigated a model of syn flood denial attack and its influence on message mediators to better understand security vulnerabilities in iot application layer protocols. when inserting fuzzy data between the user and the server, their suggested technique examines the behavior of applications at the application layer. they employed a fuzzy proxy mechanism combined with a non-standard closed variable header data pattern to evaluate both the broker and the user’s behavior when presented with unexpected data. experiments suggest that this method may uncover application layer vulnerabilities in messaging intermediates to some extent.[8] cipla has released f-secure mqtt-fuzz, a comparable tool that has been designed to assess application layer vulnerabilities in the mqtt protocol for commercial reasons.[9] moustafa et al.[10] suggested a technique for identifying iot-based threats that rely on characteristics derived from tcp protocol analysis. a set of attributes characterizing the communication protocol between users is utilized as input to learning models for this purpose, and the sort of attack is detected by categorizing these features. the fundamental difficulty with this study is that it uses restricted information as input to learning models to detect assault types. accurate attack detection at the iot application layer necessitates access to a large amount of data. syed et al.[11] suggested a machine learning-based method to identify application-layer assaults on the iot to overcome this problem. to determine the sort of attack, this method additionally leverages statistical information from the exchanged packets, as well as aspects of the communication protocol between the objects. the effectiveness of artificial neural networks and decision trees in accurately diagnosing the types of assaults has been investigated in this study, and the results reveal that the decision tree is superior in correctly diagnosing the types of attacks. kharkongor et al.[12] suggested an iot routing protocol that takes into account the energy consumption of heterogeneous network devices. this solution proposes an sdn controller that serves as a management center for network security and preventing hostile nodes from gaining access to the network. in this paper, first, routing algorithms are classified, then the proposed method for secure routing in the iot is presented. the routing algorithm proposed in this paper consists of six steps, which are: registration of nodes by the controller; monitor the entire network by the controller; receive neighbor information by each node in the network; calculate the remaining energy of neighbors; select and send data based on the remaining energy of the nodes to the neighbor; and block malicious nodes and prevent them from reaccessing the network using the controller. a machine learning-based approach for identifying dos assaults on grid intelligent networks was proposed by zhe et al.[13] preprocessing procedures, feature extraction, and classification are all part of their suggested strategy. principal component analysis was utilized to minimize the feature dimensions in the feature extraction stage, and a support vector machine (svm) was employed to identify dos assaults in the classification step. the simulation results suggest that svm outperforms categorization methods such as decision trees and new bay. in[14] dong and sarm suggested two ways for identifying software-based dos attacks the first one is adopts the degree of ddos attack to identify the ddos attack , the second on is improved k-nearest neighbors (knn) algorithm based on machine learning (ml) to discover the ddos attack. suggested methods in this part, we’ll go through the suggested software-based network and aggregate learning approach for detecting service denial threats in the iot framework in detail. to provide a safe communication platform between network objects, the suggested solution employs a software-centric network. the network topology is separated into subnets in this situation. an sdn controller node is assigned the responsibility of authentication and communication management of the members of each subnet in this topology. in addition to this communication structure, network traffic is monitored using an integrated learning model based on neural networks, decision trees, and the niobiz model. as a result, each controller node on its subnet uses this learning model to detect assaults and security concerns. it is required to first define some of the terms that will be used in the following sections; let’s pay. figure 1: block diagram of the proposed method figure 2: an example of the process of communication between objects in the proposed method rashidi, et al.: detecting denial of service attacks in internet of things 51 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 first definition: an active node is a wireless node that is connected to one of the network’s active nodes. otherwise, we refer to it as an inactive node. definition 2: subnet: a portion of the imagined global network that is administered centrally by an sdn controller node, and whose members can connect with other subnets through the controller node’s administration and rules. network nodes in wireless networks have varying communication qualities due to different manufacturing processes for radio equipment. as a result, the network that is assumed is heterogeneous. each software-driven network controller node is equipped with a learning model that can record and interpret the data traffic that passes through it. this is a learning model that combines artificial neural networks, decision trees, and the new bayes model; it is used to identify security assaults and threats in the network that corresponds to the controller node. figure 1 depicts the suggested method’s steps in detail as a diagram (1). the sdn domain is separated into numerous subdomains in the first phase of the proposed technique, and the responsibility of monitoring each subdomain is assigned to a controller who may exchange security rules with other subdomains. each controller will deliver a list of certified users relevant to its section to other controllers in the suggested way. if communication between two users is required, the user’s trustworthiness is determined by exchanging messages between the controllers. the data exchange activity will take occur if at least one controller has authenticated each of the two participants to the connection. according to the software-centric software structure suggested in this study, all communications between nodes in a subnet are routed through the subnet’s control node. as a result, each controller node employs a learning model to continually evaluate network traffic data and detect assaults. this model is an integrated learning system that uses an artificial neural network, a decision tree, and new biz to identify the presence of dos assaults using statistical data gathered from each traffic flow. each of these steps will be described below. sdn-based network communication structure the network domain is separated into multiple subdomains in the first phase of the proposed technique, and the responsibility of monitoring each subdomain is assigned to a controller who exchanges security rules with other subdomains. the position information of items in the network may be used to segment them in a pattern. as a result, each subnet is treated as a cluster. each cluster node on the network only communicates with its sdn controller directly (it will not even communicate with its neighbors). the goal is to have network users authenticate through the sdn controller to avoid security threats both inside and outside the clusters. furthermore, using this structure, each node is forced to communicate its traffic with others through the controller node, making it possible to monitor and identify attacks for all information transmitted in the network using the learning model. the controllers of each subdomain must communicate information about their members for data to be securely exchanged across mobile nodes in the network. when a source node wants to communicate data to another node, it sends the target node id to its subdomain controller first. if the source and destination nodes are on the same subdomain, the two nodes are connected by sending a reply message to the source node. otherwise, the message supplied from the source is transmitted to the central controller c by the controller node. following receipt of this message, the c t node sends packets to the controllers of the other subdomains with the target node id. a confirmation message is sent to the source node by the controller that has the destination node in its subdomain through the central c t node. a link will be formed between the two nodes in this manner. figure 1 shows an example of the suggested algorithm’s communication process between objects (2). figure 2 assumes that a node in subdomain 1 such as a wants to connect with a node in subdomain 3 such as b. in this scenario, node a sends a message to controller c1 specifying the target node id. this controller transfers the received packet to the central controller ct since node b is not in the c1 domain. this message is also sent to other controllers by this controller (c2 and c3). because the destination node b is in figure 4: neural network structure to determine the type of attacks in the aggregate model of each controller node figure 3: how controller nodes work in identifying attacks based on cumulative learning rashidi, et al.: detecting denial of service attacks in internet of things 52 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 the subdomain of node c3, this subdomain sends a response packet to the source node. finally, using the discovered route, the data packet is transmitted between the two nodes. the suggested aggregation approach is used to analyze traffic information and detect intrusions while data are being routed by controller nodes. the structure of this learning model will be described in the following sections. detection of attacks based on cumulative learning as mentioned earlier, each controller node in the softwaredriven network is equipped with an integrated learning model that is capable of recording and processing the data traffic flowing through it. this learning model consists of the following learning models: 1. artificial neural network 2. decision tree 3. new biz model. each of the above learning models analyzes the traffic patterns that travel through their respective controllers, and then classifies them using the voting mechanism. the learning model based on these controller nodes will solely evaluate traffic received by its subnet nodes to lessen the complexity and computational strain imposed on them. by doing so, malicious malware may be avoided from infecting network equipment and routers from the start of the transmitting process, and the rogue node can be easily recognized. figure 1 shows an example of this procedure (3). on figure 3, it is assumed that the two nodes are in the same subnet to keep things simple. node a is a malicious node, whereas node b is a regular node. assume that each of these nodes wants to send each other a message. as previously stated, all nodes in the network exchange data through their subnet controller, which processes all messages supplied by subnet members using the integrated system’s learning models. when node a transmits a malicious message to the controller, the communication features are retrieved and categorized by three models of artificial neural networks, decision trees, and the nivebase model before each operation. the aggregate system’s ultimate output is then decided using the voting process, depending on regular communication or the existence of an assault. the connection will be stopped and the message will be erased if the integrated system qualifies the collected characteristics. this happened in the case of a hypothetical message sent from node a to node b. the message sent by node b, on the other hand, is noticed by the integrated system in the regular controller and forwarded to node a. the process of recognizing assaults using the integrated learning system will be described in the following sections. data processing the first step in the process of identifying dos attacks by the proposed aggregation system is data preprocessing. data preprocessing is done through the following steps: • numerically quantify the nominal characteristics of the traffic flow being processed. for example, the “connection type” attribute can have one of the icmp, udp, and tcp modes, which are replaced by numbers 1–3. the numerical properties obtained for the traffic flow are normalized using the following equation. − = −   i min i max min n n n n n (1) the input property vector for normalization is represented by n i and the minimum and maximum values in the n i property vector are represented by n min and n max , respectively. thus, the traffic flow characteristics are translated into a numerical representation in the range,[0,1] and these characteristics are employed as input to learning models in the proposed method’s subsequent phases. classification of features based on artificial neural network the first learning model used in the proposed integrated system for detecting dos attacks is the artificial neural network. this neural network is a prosthetic network with a hidden layer. the secret layer of this network has 10 neurons and its transmission function is determined by logarithmic sigmoid type. furthermore, the number of input layer neurons is equal to the number of traffic flow characteristics, and the number of output layer neurons is equal to the number of types of attacks. the amount of output from these neurons determines the type of attack determined by the neural network. the structure of this network is shown in figure 4. the artificial neural network is the initial learning model utilized in the proposed integrated system for detecting dos threats. this neural network is a hidden layer prosthetic network. this network’s hidden layer comprises 10 neurons, and its transmission function is of the logarithmic sigmoid type. rashidi, et al.: detecting denial of service attacks in internet of things 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 classification of features based on artificial neural network in addition, the number of input layer neurons equals the number of traffic flow characteristics, whereas the number of output layer neurons equals the number of attack kinds. the sort of assault decided by the neural network is dictated by the amount of output from these neurons. figure 1 depicts the network’s structure (4). in addition, the number of input layer neurons equals the number of traffic flow characteristics, whereas the number of output layer neurons equals the number of attack kinds. the sort of assault decided by the neural network is dictated by the amount of output from these neurons. figure 1 depicts the network’s structure (4). the neural network learning algorithm for training based on the input data is as follows: 1. at the beginning of the neural network training, the initial values of the weights for the w c vector are set equal to random numbers and t = 0 is considered 2. for each layer in the neural network, the input vector di is applied to the neurons of the current layer of the neural network and based on the weight vector of the neurons in the current iteration and the logarithmic sigmoid activation function, the output d is calculated 3. the values of neural network weights in the w c vector are updated using the following relation:[15] ( ) ( ) µ+ = + × ×1j jw t w t e x (2) in the above relation, μ is the learning rate, e is the difference between the actual output and output of the neuron, and x is the input data. steps 2–4 are repeated until the neural network error is less than the threshold value or t < t max , increasing by t one value. otherwise, the neural network training will end. classification of properties based on the decision tree the decision tree is the second learning model utilized in the proposed integrated system for identifying assaults. the decision tree algorithm is a data mining approach that, despite the fact that it does not involve sophisticated computations and is simple to grasp, has comparable accuracy to other classification methods. as a result, it may be used to solve a variety of categorization problems. as a result, a structure based on decision tree and regression classification and regression trees are described in this part to identify assaults. the decision tree created by the division and regression tree attempts to anticipate and classify future observations. the goal of this strategy is to eliminate contaminants in each of the categories. when all of the items in a subset belong to the same goal category, a node is totally free of impurities. intervals and categories are two sorts of predictive and objective adjectives. all divisions will be binary, meaning that each node will have only two subgroups. instead of employing stop laws, the decision and regression tree creates a succession of subtrees by first creating a big tree and then pruning it until only the root node remains. the classification cost of each subtree is then estimated using cross-validation. finally, the decision tree model is built by selecting the subtree with the lowest projected cost. classification of properties based on new biz the root node and other parent nodes in the final tree define the branch, while the leaf nodes define the target classes. the tree traversal begins at the root node and continues until it reaches a leaf to classify the input data. the class that the data reach by scrolling the tree branches will be the decision tree’s output for the input data. the proposed integrated system employs a new base model as the third learning model for detecting dos threats in controller nodes. the bayesian technique is basically a way of categorizing events based on their likelihood of occurring or not occurring. the chance of an event occurring in the future may be calculated in the new business categorization by looking at prior events. bayesian classification is used for problems in which each instance of x is selected by a set of attribute values and the objective function f (x) from a set such as v. the bayesian mechanism for classifying a new sample is to identify the most likely class or target value of v map by having the attribute values <a 1 , a 2 …, a n > that describe the new sample:[15] ∈= …1 2üüüjüüüv argmax p v a a a (3) using bayes’ theorem, the above statement can be rewritten as follows:[15] ( ) ( ) ( )∈ ∈ … = … = … 1 2 1 2 1 2 | , , ,   , , ,     ( , , , | ) ( ) j j j n j map v v n v v n j j üüü üüü p a a a argmax p a a a v p v (4) now, using educational data, we try to estimate the two sentences of the above equation. it is easy to calculate from educational data how much vj is repeated in the data. but calculating different sentences p (a 1 , a 2 ,…, a n /v j ) will not be acceptable in this way unless we have a large amount of educational data. therefore, we have to look at each sample several times to get a good estimate of it. the bayesian method of classification is based on the premise that attribute values are conditionally independent of having objective function values. in other words, this assumption implies that provided that the output of the objective function is observed, the probability of observing the attributes a 1 , a 2 …, a n is equal to multiplying the probabilities of each attribute separately. if we replace this concept with equation 4, the bayesian classification method results:[15] ( )∈= ∏  ( ) |jüüü i v argmax p v p a v (5) vnb is the output of bayesian classification for the objective function. the number of p sentences (a i /v j ) to be calculated in this method is equal to the number of attributes multiplied by the number of output sets for the objective function, which is much less than the number of p sentences (a 1 , a 2 ,…, a n /v j ). detection of attacks in a cumulative model based on voting technique the voting approach is the final stage in detecting dos threats in the proposed aggregate system. the goal of the voting procedure is to increase the accuracy of algorithm rashidi, et al.: detecting denial of service attacks in internet of things 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 classification when compared to when each algorithm is employed individually. this is known as aggregation-based learning or voting. each of the classification algorithms may have errors in classifying some samples; therefore, the purpose of voting-based techniques is to reduce the resulting error and increase the accuracy of sampling samples. it has been theoretically proven that the use of voting techniques can improve the results. simulation and results as a consequence, in the final stage of the proposed technique, the neural network, new biz, and decision tree classification models each do the classification operations of the test samples individually, and the final output of the system is decided by voting on the results of all three models. the suggested model was evaluated using data from the nslkdd database[16] for this purpose. more than 25,000 data records in the field of information shared in the network are contained in the database used to evaluate the performance of the proposed model. packet data sent during various forms of network incursions are stored in these records. table 1 shows the data from the nslkdd database (1). table 1 shows that around 80% of the data in the database are allocated to assaults. this database comprises 42 statistical aspects of traffic flow characteristics in each data record. during the simulation, it is assumed that each node in the network exchanges information with other nodes depending on one of the records in this database. the data flow traffic information is categorized by the learning models in the controller node integration system throughout the data exchange process, and the existence of assaults is identified. the performance of the suggested aggregate model in distinguishing regular traffic flows from streams associated with dos assaults will be examined in the following sections. to ensure the validity of the results, the simulation operation was repeated 10 times. during this process, in each iteration, 90% of the database samples are used as learning model samples and the remaining 10% are used as test samples. thus, after 10 repetitions of the experiment, the exchange of all samples belonging to normal categories and dos attacks in the nslkdd database will be simulated by network nodes. furthermore, the performance of the proposed aggregate model in detecting dos attacks is compared with two svm learning models in zhe et al.[13] and the nearest neighbor k algorithm knn in.[14] the accuracy results of each of the compared algorithms in detecting network attacks are shown in figure 5. during several iterations, figure 5a depicts the accuracy of the proposed technique and the methods compared to it. the average accuracy of the various approaches is also shown in figure 5b. as shown in figure 5, the attack detection operation may be done with 99.6% accuracy employing a combination of artificial neural network, decision tree, and new biz in the suggested aggregate system, which is a little improvement. it’s 2% more precise than the approaches described in zhe et al.[13] and dong and sarem.[14] the disruption matrix will be examined in greater depth to analyze the efficacy of the suggested aggregation mechanism in identifying assaults. the clutter matrix resulting from the detection of dos assaults during 10 replication simulations for the proposed aggregation system is compared to the clutter matrix coming from the svm[13] and knn[14] approaches in figure 6. the value 35,201 in the first row and column of the perturbation matrix shown in figure 6a reflects the number of normal connections examined that were accurately recognized as normal by the suggested aggregation method. the clutter matrix identifies this number as tn. the number 92 in the second row and first column represents the number of regular communications that the suggested dos attack model mistakenly detected. in the clutter matrix, this quantity is designated as fp. the clutter matrix’s second row and second column, which reflect the number 40,068, identify service denial assaults that the proposed aggregate system accurately classifies as tp occurrences. in addition, the number 219 in the first row and second column represents dos attempts that the table 1: types of information available in the nslkdd database type of attacks percentage of available data normal (no attack) 19.48 dos attacks 73.9 u2r attacks 1.34 r2l attacks 5.2 probe attacks 0.07 figure 5: accuracy of learning algorithms in detecting network attacks by controller node. (a) accuracy in different iterations and (b) average accuracy b a figure 6: disruption matrix (a) proposed aggregate system, (b) support vector machine, and (c) k-nearest neighbor in attack detection c ba rashidi, et al.: detecting denial of service attacks in internet of things 55 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 suggested aggregating system mistook for regular. the clutter matrix identifies this number as fn. the suggested method’s improved efficiency in detecting dos assaults is confirmed by a comparison of its matrix with the examined approaches. table 2 compares the results of different learning models with the test results of the proposed integrated system for detecting dos assaults in the network. the sensitivity and specificity criteria are compared in this table. the ratio of total assaults accurately recognized by the learning model is called sensitivity, and it’s determined as follows: = + tp sensitivity tp fn (6) in this relation, tp is the number of attack streams that have been correctly detected and fn is the number of attack traffic streams that have been erroneously identified as normal streams. the property criterion is used to measure normal streams that are correctly classified. this criterion is calculated as follows: = + tn specificity tn fp (7) tn is the number of accurately recognized normal traffic streams, whereas fp denotes the number of normal traffic streams that have been identified as attack traffic streams. when the efficiency of the suggested approach is compared to the efficiency of the compared methods, it is clear that the provided solution may improve the criteria of accuracy, sensitivity, and specificity in the process of detecting dos assaults. conclusion a new technique for detecting dos attacks in the structure of iot is provided in this research, which is based on software-based network and machine learning. the suggested solution divides the sdn domain into numerous subdomains, each with its own controller for exchanging security rules with other subdomains. each controller will deliver a list of certified users relevant to its section to other controllers in the suggested way. in this example, if two users need to interact, the user’s credentials are passed back and forth between the controllers. the data exchange activity will take occur if at least one controller has authenticated each of the two participants to the connection. in the topology suggested in this study, all traffics between nodes in a subnet are routed through the subnet’s control node. as a result, each controller node employs an integrated learning model to continually evaluate network traffic data and detect assaults. this learning model incorporates an artificial neural network, a decision tree, and a new biz model that uses statistical information gathered from each data stream to identify the likely existence of assaults. matlab software was used to implement and assess the suggested approach, and the results of simulating the new method’s performance were compared to earlier methods. the data from the nslkdd database were utilized in the tests, and the suggested learning model’s accuracy in identifying various forms of network assaults was assessed. the results of these tests revealed that the suggested method’s cumulative learning model can identify dos attacks in network traffic flows with accuracy of 99.6%. other machine learning models, such as probabilistic neural networks, deep learning approaches, and others, might be used in the future to identify assaults. it appears that optimization algorithms may be used to rank learning models and then establish a weight value for each learning model in the integrated system based on that ranking. as a result, the future research might focus on resolving this problem. references 1. j. li, m. siddula, x. cheng, w. cheng, z. tian and y. li. approximate data aggregation in sensor equipped iot networks. tsinghua science and technology, vol. 25, no. 1, pp. 44-55, 2019. 2. j. marietta and b. mohan. a review on routing in internet of things. wireless personal communications, vol. 111, no. 1, pp. 209-233, 2020. 3. x. dong, z. yu, w. cao, y. shi, and q. ma, x. a survey on ensemble learning. frontiers of computer science, vol. 14, no. 2, pp. 241258, 2020. 4. g. perrone, m. vecchio, r. pecori, and r. giaffreda. the day after mirai: a survey on mqtt security solutions after the largest table 2: comparison of the performance of the proposed integrated system with other learning models specificity percentage percentage of sensitivity percentage accuracy title 96.9221 98.4955 97.6568 svm[13] 98.4660 96.7359 97.6581 k-nearest neighbor[14] 99.4564 99.7393 99.5885 proposed integration system rashidi, et al.: detecting denial of service attacks in internet of things 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 49-56 cyber-attack carried out through an army of iot devices. in: 2nd international conference on internet of things, big data and security, iotbds, pp. 246-25, 2017. 5. s. andy, b. rahardjo and b. hanindhito. attack scenarios and security analysis of mqtt communication protocol in iot system. in: 2017 4th international conference on electrical engineering, computer science and informatics (eecsi), pp. 1-6, 2017. 6. x. liu, t. zhang, n. hu, p. zhang and y. zhang. the method of internet of things access and network communication based on mqtt. computer communications, vol. 153, pp. 169-176, 2020. 7. s. n. firdous, z. baig and a. ibrahim. modelling and evaluation of malicious attacks against the iot mqtt protocol. 2017 ieee international conference on internet of things (ithings) and ieee green computing and communications (greencom) and ieee cyber, physical and social computing (cpscom) and ieee smart data (smartdata), pp. 748-755, 2017. 8. h. ramos, s. villalba, r and lacuesta. mqtt security: a novel fuzzing approach. wireless communications and mobile computing, 2018. 9. a. vähä-sipilä, “mqtt fuzz”., 2015. available from: https:// github.com/f-secure/mqtt_fuzz 10. n. moustafa, b. turnbull and k. choo. an ensemble intrusion detection technique based on proposed statistical flow features for protecting network traffic of internet of things. ieee internet of things journal, vol. 6, no. 3, pp. 4815-4830, 2018. 11. n. f. syed, z. baig, a. ibrahim and c. valli. denial of service attack detection through machine learning for the iot. journal of information and telecommunication, vol. 4, no. 4, pp. 482-503, 2020. 12. c. kharkongor, t. chithralekha and r. varghese. a sdn controller with energy efficient routing in the internet of things (iot). procedia computer science, vol. 89, pp. 218-227, 2016. 13. w. zhe, c. wei and l. chunlin. dos attack detection model of smart grid based on machine learning method. in: 2020 ieee international conference on power, intelligent computing and systems (icpics), pp. 735-738, 2020. 14. s. dong and m. sarem. ddos attack detection method based on improved knn with the degree of ddos attack in softwaredefined networks. ieee access, vol. 8, pp. 5039-5048, 2019. 15. m. bharati and m. ramageri. data mining techniques and applications. indian journal of computer science and engineering, vol. 1, no. 4, pp. 301-305, 2010. 16. r. bala and r. nagpal. a review on kdd cup99 and nsl nsl-kdd dataset. international journal of advanced research in computer science, vol. 10, no. 2, pp. 64-67, 2010. tx_1~abs:at/add:tx_2~abs:at 147 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 147-152 research article relationship between sera vitamin d, calcium, and magnesium levels in pregnant women with and without postpartum depression hataw a. mohammed1, hero h. muhammed saed1, bushra m. jarallah2 1department of chemistry, college of education, garmian university, kalar, iraq, 2department of medicine, college of medicine, garmian university, kalar, iraq abstract postpartum depression (ppd) is a significant mental health case after childbirth, distinguished by behavioral changes and emotional disturbance. ppd is a significant public issue estimated to affect 10–15% of women worldwide. the study’s objective was to investigate whether low sera concentrations of vitamin d, calcium, and magnesium during the third trimester of pregnancy were associated with ppd. the study population included 80 pregnant women between the ages of 25 and 35 years. in this study, we followed up the pregnant women until birth. after childbirth, those women were divided into two groups: group 1 (40 healthy women as a control group) and group 2 (40 unhealthy women with ppd disorder). when the blood samples were taken, vitamin d level was determined using an elisa device. while, calcium and magnesium were measured using the spectrophotometric method. a  health questionnaire structure was used for screening depression after birth. after measuring pregnancy serum concentrations of vitamin d of both two groups, we found that there was no significant relationship between vitamin d concentrations in pregnant women and ppd (p = 0.6497). a significant difference in sera calcium (p = 0.0003) and magnesium levels (p = 0.0005) between the control group and depressed group was observed. our study detected that vitamin d concentrations in the serum of women during their last trimester of pregnancy had no statistical link with their risk of ppd. whereas direct association in calcium and magnesium deficiency in the third trimester of pregnancy and happening of ppd were reported in this study. keywords: calcium, magnesium, postpartum depression, third trimester of pregnancy, vitamin d introduction postpartum depression (ppd) is one of the most common and public health problems of childbirth.[1] approximately one-fifth of women present depression during pregnancy and after birth,[2] yearly affecting about 10–15% of postnatal women.[3] it sets within a few months to a year after birth. however, some evidence indicated postpartum depression 4 years after birth.[4] it is characterized by low appetite, fatigue, the feeling of guilt, and sleep problems.[5] untreated maternal illness is potentially associated with adverse consequences for the mother, her infant, and her family.[6] the etiology remains unclear and complex.[7] most commonly risk factors such as having a history of anxiety and depression during pregnancy, stressful life events, biological factors, and low social support are involved in the progression of postpartum depression.[8] furthermore, there are many researches about the contribution of nutritional status of minerals and vitamins, which are associated with several body’s metabolic regulations for depression patients.[9] for instance, vitamin d is one of the nutritional factors linked with cognitive decline in mental health. it has been hypothesized that a low concentration of vitamin d may take part in depression.[10] research supports the connect between depression and vitamin d in three ways. first menon et al.[11], vitamin d plays as a neuroactive hormone. sundry sources have demonstrated that receptors of vitamin d are largely distributed throughout the brain of the human and play a key role in mood regulation.[12] second, vitamin d has a role in regulating the immune system and reducing the release of inflammatory substances.[13] finally, vitamin d as well has a defensive influence versus a low concentration of specific hormones such as serotonin and dopamine.[14] cihan university-erbil scientific journal (cuesj) corresponding author: hataw a. mohammed, department of chemistry, college of education, garmian university, kalar, iraq. e-mail: hataw.adil@garmian.edu.krd received: june 16, 2022 accepted: october 23, 2022 published: december 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp147-152 copyright © 2022 hataw a. mohammed, hero h.m. saed, bushra m. jarallah. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). mohammed, et al.: vitamin d and minerals with postpartum depression 148 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 147-152 concerning macromolecules, calcium (ca) and magnesium (mg) are the most predominant intracellular cation. certain studies have found elevation of ca and mg among depressed pregnant women. in contrast, others show a decrease in levels of ca and mg.[15] calcium has several vital functions in the human’s body.[16] more than 99% of total ca is present in the skeleton, supporting their framework. while, the residual 1% is set in the blood, muscle, and the fluid between cells which is necessary for contraction of muscle, expansion and contraction of blood vessel, hormones and enzymes excretion, and posting messages through the nervous system.[17] in human’s body, magnesium is one of the fundamental minerals and a cofactor of over 300 enzymatic reactions.[18] deficiency of mg has been reported in depression. it is a natural calcium channel blocker, and it is needed for nerve function and appropriate relaxation. ca enhances the nerves and mg calms them down. all these systems have been associated with the depression’s pathophysiology. mg as well dampens the calcium ion-protein kinase c-linked neurotransmission and catalyzes the na-katpase.[19] when mg is declining, it is possible to notice an increase in the activity of the n-methyl-d-aspartate receptor (nmda receptor), as well as substance p, which may cause increased pain and elevated hormones related to stress.[20] according to our investigation, this study was the first trial held to measure vitamin d, ca, and mg levels together in the serum of pregnant women in their last trimester of pregnancy and studying their relation with postpartum depression. the primary goal of this study was to illustrate if low levels of vitamin d, ca, and mg in the sera of pregnant women in their third (last trimester) of pregnancy were linked with their affecting by postpartum depression or not in the iraqi population/garmian/kalar. methods and clinical measurements clinical measurements the population of this study was conducted on the sera of 80 healthy pregnant women aged 25–35 years. we measured sera vitamin d, calcium, and magnesium for those pregnant women in their third trimester of their pregnancy. afterbirth, we followed their cases. if they did not suffer from postpartum depression, we considered them as group 1; healthy women or control group (40 women). on the other hand, the women who suffered from postpartum depression, we regarded them as group 2; unhealthy women or depressed women (40 women). the pregnant donors did not have any severe diseases in the third trimester of pregnancy. for example, they didn’t suffer from hypertension, liver disorders, congenital heart diseases, renal dysfunction, cancers, kidney disorder, rheumatoid arthritis, blood diseases, lung disease, and diabetes or any other diseases. we excluded those pregnancies that took vitamins, minerals, and any other drugs. in addition, group 2 (depressed women) and group 1 (healthy women) provided their written, informed approval to take part in our study. the blood samples of pregnant women were collected from shahed sheray naqeb hospital in kalar which is the center of garmian region from september (2020) to february (2021). approximately 5 ml of blood samples were gained from pregnant donors, then centrifuged at 1500 rpm for about 10 min to obtain the serum, after that divided into tiny fractions, and kept frozen at −30°c for analysis. estimation of vitamin d enzyme immunoassay was applied for the fixing of 25(oh)vitamin concentration in sera of the third trimester of pregnant women using of monobind kit,[21] using an elisa device. estimation of calcium serum calcium level was estimated using a colorimetric method using bilbao diagnostic kit.[22] estimation of magnesium serum magnesium ion was determined spectrophotometrically using bilbao diagnostic kit.[23] statistical analysis statistical analysis was achieved by graphpad prism. the outcomes were expressed as mean ± standard deviation. the p-value was considered significant if it was < 0.05. results the comparison of the quantitative variables between group 1 and group 2 is described in table 1. the ages of both groups were between 25 and 35 years. bmi for group 1 was 28.55 ± 4.419 kg/m2, while for group 2 was between 27.96 ± 4.014 kg/m2. the table 1 illustrates that there was a significant statistical association between age and group 2 (26.97 ± 5.623), as well as with healthy group 1 (30.09 ± 5.933) and p = 0.0274. box plots of age between two subjects are shown in figure 1a. for bmi, there was non-significant interaction with both two groups (p = 0.5987), that shown in figure 1b. the normal ranges of sera kits which used in this study were ca; 8–10 mg/dl and mg; 1.7–2.2 mg/dl, whereas vitamin d was found in the normal sera ranges (v. d; 30–50 ng/ml). as described in the table, there was also a slight decrease in sera vitamin d concentrations in group 2 compared with group 1 (34.10 ± 4.254 and 34.88 ± 6.495), respectively. however, there was no significant connection between them (p = 0.6497). box plot of vitamin d is shown in figure 1c. besides, group 2 or unhealthy women with ppd had lower sera levels of ca and mg than group 1 or healthy women (8.673 ± 0.5503, 1.568 ± 0.3292) and (9.136 ± 0.4401, 1.876 ± 0.2454), sequentially, with a significant statically link between them (p = 0.0003 and 0.0005), respectively, as shown in figure 1d and e, sequentially. discussion postpartum depression is known as an episode of major depression that is temporally linked with childbirth.[24] ppd is a severe threat to the child, mother, and other members of the family.[19] the findings of our study provide that there was a statistically different in ages between both groups were p = 0.0274, as shown in table 1. furthermore, there was no mohammed, et al.: vitamin d and minerals with postpartum depression 149 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 147-152 statistical link among the bmi (the patients set and the healthy subjects) in which p = 0.5987 was considered, as shown in table 1. many factors are placed a woman at danger of developing ppd, such as age, type of insurance, marital status, whether or not the pregnancy was planned, race/ethnicity, type of feeding, and level of educational.[25-29] in addition, studies have shown the beneficial impact of dietary factors during pregnancy on depressive symptoms. one of those dietary factors is vitamin d which has been proposed to usefully influence gloom in adults. it has been supposed that vitamin d may work as a neuroactive hormone.[12] a lot of studies confirm the role of vitamin d in conception, placentation, improvement of pregnancy, and pregnancy outcomes inclusive the offspring’s health. altogether, vitamin d supports placental outgrowth and task by regulating placental calcium transport, and by exerting immunomodulatory effects, critical for pregnancy maintenance.[30] many researchers have found that receptors of vitamin d are primarily doled out throughout the brain of human, and its lack mutates neurotransmitters which are familiar to be included in of depression’s symptoms.[12] although the results of our study indicated a slight decrease in vitamin d concentrations among the pregnant women who afterbirth affected by ppd; we found that the sera vitamin d levels had no significant link between women with ppd and those with non-ppd (p = 0.6497), as shown in table 1. this decrease in vitamin d levels agrees with those gotten by abedi et al., aghajafari et al., murphy et al., accortt et al., ribamar et al., and lin et al.,[5,12,31-34] they also found sera vitamin d concentrations were significantly lower in depressed women than in the healthy subject. similarly, the study wang et al.[35] did not record a considerable connect between depressive symptoms and low concentrations of vitamin d through pregnancy. in addition, robinson et al.[36] showed that at 18 weeks gestation, a significantly low level of vitamin d in sera was linked with a major danger of the symptoms of ppd. furthermore, in the study of fu et al.,[37] the researchers reported that sera vitamin d concentrations after delivery were significantly lower in women with ppd than those without ppd. moreover, research of fallah et al.[14] at 18 weeks gestation measured concentration of vitamin d sera, table 1: comparisons of numerical variables for group 1 and group 2 parameters group 1 (healthy) means±sd group 2 (depression) means±sd p number 40 40 age (years) 30.09±5.933 26.97±5.623 0.0274 s bmi (kg/m2) 28.55±4.419 27.96±4.014 0.5987 ns vitamin d (ng/ml) 34.88±6.495 34.10±4.254 0.6497 ns ca (mg/dl) 9.136±0.4401 8.673±0.5503 0.0003 s mg (mg/dl) 1.876±0.2454 1.568±0.3292 0.0005 s data are shown as the mean ± standard deviation (means±sd). sd: standard deviation, bmi: body mass index, kg/m2: kilogram per meter square, ng/ml: nanogram per milliliter, mg/dl: milligram per deciliter, p value is the probability, significant p < 0.05, s: significant, ns: not significant, student’s t-test figure 1: box plot of the results, (a) age was significantly lower in group 2 in compared with the group 1, (b) body mass index was not significant in both group 1 and group 2, (c) vitamin d was not significant among both groups, (d) ca was significantly lower in group 2 in compared to group 1 (e). mg was significantly lower in the second group in compared to the first group. group 1: 40 healthy women without postpartum depression (ppd) as a control group. group 2: 40 unhealthy women with ppd disorder as depressed women d cba e mohammed, et al.: vitamin d and minerals with postpartum depression 150 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 147-152 where they detected a significant association between quality of the sleep through pregnancy and ppd with deficiency of vitamin d. on the other hand, our finding did not agree with the study of nielsen et al.[1]; where their outcomes did not find a connection between danger of ppd and low vitamin d levels through pregnancy. in contrast, an increased danger of postpartum depression was detected among women with the highest levels of vitamin d. similarly, tan et al.[38] found contrast data to our results. they reported that high blood sera of vitamin d concentrations have a preventive impact on maternal depression. notably, when blood concentration of vitamin d was raised, the maternal depression risk was the lowest. in addition, in a study gould et al.,[39] concentrations of vitamin d were measured at delivery, and they did not reveal a consistent link between vitamin d concentrations and ppd. finally, a study of williams et al.[40] found that levels of vitamin d were not linked with diagnoses of generalized anxiety trouble or major depressive turmoil. further, vitamin d has been increasingly attached to mental health cognitive decline, and it has been hypothesized that hypovitaminosis d may give a share to depression.[10] electrolyte dysfunction has been reported with depressive disorders for the past six decades. the most predominant intracellular cation is calcium.[15] dysregulation of ca is profound in disorders of the nervous system such as depression and dementia.[9] in the biosynthetic pathway, ca activates tryptophan hydroxylase, leading to the synthesis of serotonin. furthermore, the calcium/calmodulin-dependent system may promote the synthesis of dopamine in the brain. in the immune system cells, ca acts as a signal as well. in the extracellular, changes in ca level may impact the neuromuscular tissues excitability contributed in regulation of emotional.[41] despite that, findings from this study suggested that sera ca levels were considerably lower in group 2 (women with ppd) in comparing to group 1 (healthy women without ppd) (p = 0.0003). this finding was in agreement with the study done by paoletti et al.[42]; when their donors’ group was randomized to calcium/vitamin d treatment, they reported a decrease in the degree of depression among healthy puerperal women. however, our study did not agree with a study that was done by sharma et al.[15] which reported that there was no correlation in the depressive symptoms severity with sera levels of ca. further, a previous study done by li et al.[41]; detected no important link between dietary ca consumption and depression risk. mg also plays a role in the entry of ca into the neuron. magnesium is probable to keep the neuron versus the cell’s death by regularizing this entrance and may be linked with the depression danger through preserving the nervous system. consumption of magnesium dietary may be connected with depression by safeguarding the nervous system.[41] mg extends antidepressive action over a set of mechanisms that in the end, encourages a drooping of the glutamatergic pathway, and a raise of brain-derived neurotrophic factor (bdnf) and serotonin.[42] regarding the trace element mg, similarly to ca results, our data showed that levels sera of mg were statically lower in group 2 (depressed subject) than group 1 (undepressed donors) (p = 0.0005). this finding was in agreement with studies done by nechifor and nechifor;[43,44] where they record a reduction of erythrocyte mg in adult patients with medium, major, and severe depression. furthermore, concerning mg supplementation[41,45] recorded an opposite connection among community-dwelling adults between mg consumption and symptoms of depressive. on the other hand, these grades do not agree with those gotten by wojcik et al.[46]. their results did not elucidate a relevance between decreased sera mg concentration and severity of depressive symptoms in a specific affective disorder, postpartum depression. furthermore, this study fard et al.[47] concluded that mg did not minimize depressive symptoms and postpartum anxiety. in addition, the study of ram et al.[15] indicated no correlation in sera concentrations of mg and severity of depressive symptoms in patients with a major disorder of depressive. similarly widmer et al.[48] have reported raised mg in erythrocytes and plasma of moderate and severely depressed patients. deficiency of mg could lead to numerous psychiatric symptoms involving tremors, headaches, behavior disturbances, irritability and psychotic behavior, generalized tonic–clonic, focal seizures, vertigo, and depression.[19] it has also been suggested that the transmit of huge quantities of this mineral from the blood of mother to the fetus with another nutrient may give a share in the appearance of ppd.[18] finally, it can be said that vitamin favorably modifies functions of the brain, antagonizing the evolution of ppd.[42] conclusion so far, low vitamin d, ca, and mg concentrations may be a neglected danger factor for ppd. however, to our knowledge, this is the first trial that has been carried out to clarify the impact of vitamin d, ca, and mg insufficiencies together on the occurrence of ppd. we have recorded that lower concentrations of vitamin d, ca, and mg in the third trimester of pregnancy are connected to a danger for announcing symptoms of postnatal depressive in the 1st days following delivery. therefore, ensuring appropriate consumption of vitamin d, ca, and mg through pregnancy may be one way of conserving versus postpartum mood disturbance in mothers. finally, health policymakers must pay awareness to the reality that measuring vitamin d, ca, and mg concentrations must be considered one of the elementary tests for pregnant women to be treated accordingly. references 1. n. o. nielsen, m. strøm, h. a. boyd, e. w. andersen, j. wohlfarht, m. lundqvist, a. cohen, d. m. hougaard, and m. melbye. vitamin d status during pregnancy and the risk of subsequent postpartum 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penninx. the association between low vitamin d and depressive disorders. molecular psychiatry, vol. 19, no. 4, pp. 444-451, 2014. 11. v. menon, s. k. kar, n. suthar, and n. nebhinani. vitamin d and depression: a critical appraisal of the evidence and future directions. indian journal of psychological medicine, vol. 42, no. 1, pp. 11-21, 2020. 12. f. aghajafari, n. letourneau, n. mahinpey, n. cosic, and g. giesbrecht. vitamin d deficiency and antenatal and postpartum depression: a systematic review. nutrients, vol. 10, no. 4, p. 478, 2018. 13. g. sağlam, g. pektaş, s. karakullukçu, a. pektaş, and d. s. aykut. vitamin d deficiency is associated with depression, anxiety and sleep disturbance in pregnant women. journal of academic research in medicine, vol. 11, no. 1, pp. 51-55, 2021. 14. m. fallah, g. askari, and z. asemi. is vitamin d status associated with depression, anxiety and sleep quality in pregnancy: a systematic review. advanced biomedical research, vol. 9, p. 32, 2020. 15. a. s. ram, r. r. abhijit, and b. s. sangeeta. serum calcium and magnesium levels do not correlate with severity of major depressive disorder. indian journal of neurosciences, vol. 2, no. 1, pp. 6-10, 2016. 16. g. cormick and j. m. belizan. calcium ıntake and health. nutrients, vol. 11, no. 7, p. 1606, 2019. 17. b. hanna. the role of calcium correction during normal pregnancy at third trimester in mosul. oman medical journal, vol. 24, no. 3, pp. 188-194, 2009. 18. a. serefko, a. szopa, p. wlaź, g. nowak, m. radziwoń-zaleska, m. skalski, and e. poleszak. magnesium in depression. pharmacological reports, vol. 65, no. 3, pp. 547-554, 2013. 19. s. etebary, s. nikseresht, h. r. sadeghipour, and m. r. zarrindast. postpartum depression and role of serum trace elements. iranian journal of psychiatry, vol. 5, no. 2, pp. 40-46, 2010. 20. a. andretta, e. d. batista, m. e. schieferdecker, r. r. petterle, c. l. boguszewski, and e. dos santos paiva. relation between magnesium and calcium and parameters of pain, quality of life and depression in women with fibromyalgia. advances in rheumatology, vol. 59, no. 1, p.55, 2019. 21. s. k. sawhney and r. singh. introductory practical biochemistry. 2nd ed. narosa publishing house, new delhi. p. 361, 2009. 22. w. l. clark, e. s. baginski, s. s. marie, and b. zak. spectrophotometric study of a direct determination of serum calcium. microchemical journal, vol. 20, no. 1, pp. 22-32, 1975. 23. e. m. gindler and d. a. heth. colorimetric determination with bound calmagite of magnesium in human blood serum. clinical chemistry, vol. 17, no. 7, p. 662, 1971. 24. c. guille, r. newman, l. d. fryml, c. k. lifton, and c. n. epperson. management of postpartum depression. journal of midwifery and women’s health, vol. 58, no. 6, pp. 643-653, 2013. 25. m. righetti-veltema, e. conne-perreard, a. bousquet, and j. manzano. risk factors and predictive signs of postpartum depression. journal of affective disorders, vol. 49, no. 3, pp. 167-180, 1998. 26. c. t. beck and r. k. gable. comparative analysis of the performance of the postpartum depression screening scale with two other depression instruments. nursing research, vol. 50, no. 4, pp. 242-250, 2001. 27. centers for disease control and prevention. pregnancy risk assessment monitoring system (prams): prams and postpartum depression. centers for disease control and prevention, united states. 2004. 28. c. l. dennis and e. hodnett. psychosocial and psychological interventions for treating postpartum depression. cochrane database of systematic reviews, vol. 4, p.cd006116, 2007. 29. c. t. beck. state of the science on postpartum depression: what nurse researchers have contributed-part 1. mcn: the american journal of maternal/child nursing, vol. 33, no.2, pp. 121-126, 2008. 30. f. cyprian, e. lefkou, k. varoudi, and g. girardi. immunomodulatory effects of vitamin d in pregnancy and beyond. frontiers in immunology, vol. 10, no. 10, p. 2739, 2019. 31. p. k. murphy, m. mueller, t. c. hulsey, m. d. ebeling, and c. l. wagner. an exploratory study of postpartum depression and vitamin d. journal of the american psychiatric nurses association, vol. 16, no. 3, pp. 170-177, 2010. 32. e. e. accortt, c. d. schetter, r. m. peters, and a. e. cassidybushrow. lower prenatal vitamin d status and postpartum depressive symptomatology in african american women: preliminary evidence for moderation by inflammatory cytokines. archives of women’s mental health, vol. 19, no. 2, pp. 373-383, 2016. 33. a. ribamar, b. almeida, a. soares, b. peniche, p. c. jesus, s. cruz, and a. ramalho. relationship between vitamin d deficiency and both gestational and postpartum depression. nutrición hospitalaria, vol. 37, no. 6, pp. 1238-1245, 2020. 34. d. lin, j. zhang, y. bian, m. pan, and x. liu. association between circulating concentrations of vitamin d and risk of antenatal, postnatal depression: a meta-analysis. archives of clinical psychiatry, vol. 48, no. 4, pp. 186-190, 2021. 35. j. wang, n. liu, w. sun, d. chen, j. zhao, and w. zhang. association between vitamin d deficiency and antepartum and postpartum depression: a systematic review and meta-analysis of longitudinal studies. archives of gynecology and obstetrics, vol. 298, no. 6, pp. 1045-1059, 2018. 36. m. robinson, a. j. whitehouse, j. p. newnham, s. gorman, p. jacoby, b. j. holt, m. serralha, j. e. tearne, p. g. holt, p. h. hart, and m. m. kusel. low maternal serum vitamin d during pregnancy and the risk for postpartum depression symptoms. archives of women’s mental health, vol. 17, no. 3, pp. 213-219, 2014. 37. c. w. fu, j. t. liu, w. j. tu, j. q. yang, and y. cao. association between serum 25-hydroxyvitamin d levels measured 24 hours after delivery and postpartum depression. an international journal of obstetrics and gynaecology, vol. 122, no. 12, pp. 1688-1694, 2015. 38. q. tan, s. liu, and d. chen. poor vitamin d status and the risk of maternal depression: a dose-response meta-analysis of observational studies. public health nutrition, vol. 24, no. 8, mohammed, et al.: vitamin d and minerals with postpartum depression 152 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 147-152 pp. 2161-2170, 2021. 39. j. f. gould, a. j. anderson, l. n. yelland, l. g. smithers, c. m. skeaff, r. a. gibson, and m. makrides. association of cord blood vitamin d at delivery with postpartum depression in australian women. australian and new zealand journal of obstetrics and gynaecology, vol. 55, no. 5, pp. 446-452, 2015. 40. j. a. williams, v. c. romero, c. m. clinton, d. m. vazquez, s. m. marcus, j. l. chilimigras, s. e. hamilton, l. j. allbaugh, a. m. vahratian, r. m. schrader, and e. l. mozurkewich. vitamin d levels and perinatal depressive symptoms in women at risk: a secondary analysis of the mothers, omega-3, and mental health study. bmc pregnancy and childbirth, vol. 16, no. 1, p. 203, 2016. 41. b. li, j. lv, w. wang, and d. zhang. dietary magnesium and calcium intake and risk of depression in the general population: a meta-analysis. australian and new zealand journal of psychiatry, vol. 51, no. 3, pp. 219-229, 2017. 42. a. m. paoletti, m. m. orru, m. f. marotto, m. pilloni, p. zedda, m. f. fais, b. piras, c. piano, s. pala, s. lello, f. coghe, r. sorge, and g. b. melis. observational study on the efficacy of the supplementation with a preparation with several minerals and vitamins in improving mood and behaviour of healthy puerperal women. gynecological endocrinology, vol. 29, no. 8, pp. 779-783, 2013. 43. m. nechifor. interactions between magnesium and psychotropic drugs. magnesium research, vol. 21, no. 2, pp. 97-100, 2008. 44. m. nechifor. magnesium in major depression. magnesium research, vol. 22, no. 3, pp. 163-166, 2009. 45. f. n. jacka, s. overland, r. stewart, g. s. tell, i. bjelland, and a. mykletun. association between magnesium intake and depression and anxiety in community-dwelling adults: the hordaland health study. australian and new zealand journal of psychiatry, vol. 43, no. 1, pp. 45-52, 2009. 46. j. wojcik, d. dudek, m. schlegel-zawadzka, m. grabowska, a. marcinek, e. florek, w. piekoszewski, r. j. nowak, w. opoka, and g. nowak. antepartum/postpartum depressive symptoms and serum zinc and magnesium levels. pharmacological reports, vol. 58, no. 4, pp. 571-576, 2006. 47. f. e. fard, m. mirghafourvand, s. mohammad-alizadeh charandabi, a. farshbaf-khalili, y. javadzadeh, and h. asgharian. effects of zinc and magnesium supplements on postpartum depression and anxiety: a randomized controlled clinical trial, women and health, vol. 57, no. 9, pp. 1115-1128, 2017. 48. j. widmer, j. g. henrotte, y. raffin, p. bovier, h. hilleret, and j. m. gaillard. relationship between erythrocyte magnesium, plasma electrolytes and cortisol, and intensity of symptoms in major depressed patients. journal of affective disorders, vol. 34, no. 3, pp. 201-209, 1995. tx_1~abs:at/add:tx_2~abs:at 112 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 research article different concentrations of honey’s antimicrobial activity against staphylococcus aureus by hplc determination raid d. thanoon1, abdalmuhaymen s. alshaeen2, hasnaa r. abdullah2, laith a. abdulhussain2, wurood j. mohammed2, abdullah b. hassan2 1department of medical biochemical analysis, cihan university-erbil, kurdistan region, iraq, 2department of general biology, cihan university-erbil, kurdistan region, iraq abstract honey has been used in ancient times as a treatment. it is used for healing wounds and also can be used as antiseptic to kill bacteria and other microorganisms. the aim of this study was to examine the activity of honey as a bactericidal and bacteriostatic and to measure the most active compounds of honey with highly bacterial inhibition zone using high-performance liquid chromatography (hplc) technique. in this experiment, the honey was used to check whether the honey can be used as an antimicrobial agent or not. first by well diffusion method is done by adding different concentrations of honey (25, 75, 100, 125, 150, and 200  ml). the results showed a little inhibition zone for the three types of honey but in different size for each type. the industrial honey showed the largest effect. different concentrations were used (10%, 30%, and 50%). the antibiotic sensitivity was applied, the result showed only two resistant antibiotics (azithromycin and erythromycin). the concentration of catalase, amylase, invertase, and glucose oxidase in honey tested by hplc and showed the concentration of each substance conc. of catalase = (32877 ÷ 170,253) × 20 = 3.862  u/ml; conc. of amylase  =  (136,985  ÷  180,849) ×  20  =  15.149  u/ml; conc. of invertase = (58,466 ÷ 193,624) × 20 = 6.039  u/ml; and conc. of glucose peroxide = (105,204 ÷ 163,245) × 20 =12.889 u/ml. moreover, the presence of gluconic acid “organic acid” gives the honey its acidic characteristic which is about 3.2 ph. keywords: honey, staphylococcus aureus, antimicrobial activity, antibiotics, high-performance liquid chromatography test introduction honey could be a good health-care product in addition to its previously use as a part of the traditional medicine in the treatment of several wound’s types.[1] in the recent decade, the microbial resistance is becoming one of the serious concerns due to the extensive use of antimicrobial agents. as a part of the traditional antimicrobial preparations, honey has gotten more and more consideration as there is an increase in the investigations on its antimicrobial properties but is the honey have an effect on bacteria?[2] phytochemicals are already presents as they can thus affect the antimicrobial capacity.[3] there are numerous in vitro and in vivo studies which were illustrated the antibacterial, antifungal, and even the antiviral activity of honey.[4-6] however, the ability of honey to kill bacteria “bactericidal effect” is due to the diversity of honey composition which includes about 181 constituents.[7] one of the major compositions of honey is carbohydrates in addition to the vitamins, minerals, enzymes, amino acids, and water by 15–18%. moreover, ripened honey consists of 80% sugars; mainly glucose and fructose as well as some sucrose and maltose along with 18% water. the high concentration of sugars combined with a low moisture content creates osmotic stress, which prevents spoilage of honey by microorganisms. only slight dilution of honey can result in yeast growth, but the sugar content of honey is sufficient to retain antibacterial activity of honey when diluted to approximately 30–40%. at higher dilution rates, the antibacterial activity could be due to the other compounds other than sugars.[8,9] staphylococcus aureus is gram-positive pathogenic bacteria which causes a wide variety of diseases. many methods were developed to treat s. aureus-related diseases but the treatment of this bacteria is still incomplete due to the appearance of different strains which are resistant to antibacterial agents corresponding author: raid d. thanoon, department of medical biochemical analysis, cihan university-erbil, kurdistan region, iraq. e-mail: raid.duraid@cihanuniversity.edu.iq received: june 24, 2022 accepted: august 12, 2022 published: september 25, 2022 doi: 10.24086/cuesj.v6n2y2022.pp112-118 copyright © 2022 raid d. thanoon, abdalmuhaymen s. alshaeen, hasnaa r. abdullah, laith a. abdulhussain, wurood j. mohammed, abdullah b. hassan. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) thanoon, et al.: antimicrobial activity against staphylococcus aureus by hplc determination 113 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 such as “methicillin-resistant s. aureus.”[10] in addition, it can be found as a microbial flora on the mucosal membrane and skin.[11] some are aerobic and few are anaerobic and it can survive in high concentration of salt reaching about 10%.[12] honey is could be produced from various sources, and its antimicrobial activity varies significantly in response to its origin. the antibacterial activity of honey has been attributed by physical factors such as osmolarity, acidity, and chemical factors. studies were reported that commercial honey had antimicrobial effects against s. aureus. the use of honey as an antimicrobial agent was started in 2000 bc as it showed some different results against different bacteria and depending on the honey that was used. the added value of honey has attracted the clinicians and scientists to concentrate on it and how to be used extensively.[13] the second major advantage of the honey is that bacteria have not developed the same rate of resistance against the natural products as the one for the antibacterial agents.[14,15] furthermore, the bacterial resistance to many antibiotics can cause different chronic types of wounds which can sometimes progress to amputation or even death.[13,16] the aims of study were to examine the activity of honey as a bacterial inhibitor and to measure the most active compounds of honey with highly bacterial inhibition zone using high-performance liquid chromatography (hplc). methodology collection and preparation of honey test solutions there were three common honey samples collected. the honey samples were stored at 37°c for 24 h before the preparation process of different concentrations of each honey type. in addition, the honey types were kept in dark bottles to guarantee that they will be away from the sunlight.[17] materials 1. two types of agar were used • nutrient agar • mueller-hinton agar 2. three types of honey • black honey • industrial honey • natural honey 3. pure culture of s. aureus 4. ten different antibiotics: (cloxacillin, trimethoprim, azithromycin, ciprofloxacin, penicillin, rifampin, methicillin, erythromycin, clindamycin, and gentamicin) 5. swabs and petri dishes 6. needle and loop for culturing bacteria 7. bunsen burner 8. sterilizers (alcohol). isolation of bacteria inactive pure culture of s. aureus was present, so the bacteria could be reactivated by making a new subculture on nutrient broth by transferring a sample from the inactive old culture to the new one using swabs to be subjected then for incubation at 37°c for 24 h.[18] preparation of nutrient broth a 28 g of nutrient broth powder was mixed with 1 l of distilled water and the required number of media is about 20 ml, to obtain the needed 20 ml, 0.56 g of nutrient broth powder was prepared following the ratio and proportion equation. addition of honey on the cultured s. aureus mueller-hinton agar was prepared and then poured on the petri dishes. first, about 18 petri dishes were required with about 20 ml of media for each as the total media volume that will be needed is 360 ml. in terms of the mueller-hinton agar composition, 38 g of the powder should be mixed with 1 l of distilled water. accordingly, to have 18 petri dishes with a total media volume of 360 ml; 13.68 g of mueller-hinton powder was required following the ratio and proportion equation. finally, 13.68 g of powdered mueller-hinton was added to 360 ml of distilled water to a 500 ml flask followed by boiling on a bunsen burner. for more sterilization, the flask was kept in the autoclave.[19] agar well diffusion a 20 ml of the media was poured into each petri dish and remained for solidification to form the wells. all materials that were used had been sterilized in the autoclave for 15 min. punching force was applied using a sterile tip with 5 mm width to develop on the plate to form wells with the same diameter. in addition, the excess agar was removed and the plates were subjected to incubation for 24 h at 37°c. culturing s. aureus the bacteria were cultured using swaps by dipping them in the previously cultured test tubes. furtherly, the bacteria were spread on the petri dishes following the spread plate technique instructions to accomplish the culturing process. addition of different honey concentration before adding various honey concentrations, the petri dishes were labeled with the type and concentration of the honey to be added as follows: s1: industrial honey, s2: black honey, and s3: natural honey. each type of honey was tested in terms of activity in different concentrations as these concentrations were, respectively; 25, 75, 100, 125, 150, and 200 in mcl. furthermore, micropipette was used to add the different concentrations in the center of the wells and further incubation for 24 h. dilution method in the current method, the three types of honey were tested in different concentration using the nutrient broth and then cultured overnight in the tubes following a specific procedure.[20] sterilization of test tubes heat proof glass test tubes were used to be subjected for high temperatures to guarantee a complete sterilization process using the autoclave at 121°c for about 15 min. thanoon, et al.: antimicrobial activity against staphylococcus aureus by hplc determination 114 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 preparation of nutrient broth fifteen sterilized test tubes were needed for the whole three types of honey in addition to 150 ml of media as each test tube requires 10 ml of nutrient broth. to prepare 150 ml of nutrient broth media; following the previously discussed ration proportion equation, 4.2 g of the powder will be added to 150 ml of distilled water in a flask. in addition, the mixture was boiled using a bunsen burner and then autoclaved for 15 min. dilution of honey the three types of honey were tested by taking 10%, 30%, and 50% concentrations respectively from each. concentrations were expressed as percentage and weight based on the density of 1.42 g/ml as the 10% of honey equals 0.142 g/ml, 20% equals 0.284 g/ml, and 50% equals 0.701 g/ml of the honey. culturing of s. aureus a swab from pure culture of s. aureus was obtained and then cultured in 10 ml of nutrient broth as the step was repeated for each test tube. furthermore, different concentrations of honey were added to the cultured test tubes as the samples were incubated for 24 h and the results were checked. sterilization of test tubes heat proof glass test tubes were used to be subjected for high temperatures to guarantee a complete sterilization process using the autoclave at 121°c for about 30–60 min. preparation of nutrient broth fifteen test tubes were needed for the whole three types of honey in addition to 150 ml of media as each test tube requires 10 ml of nutrient broth. to prepare 150 ml of nutrient broth media; following the previously discussed ration proportion equation, 4.2 g of the powder will be added to 150 ml of distilled water in a flask. in addition, the mixture was boiled using bunsen burner and then autoclaved for 30–60 min. dilution of honey the three types of honey were tested by taking 10%, 20%, and 50% concentrations, respectively, from each. concentrations were expressed as percentage and weight based on the density of 1.42 g/ml as the 10% of honey equals 0.142 g/ml, 20% equals 0.284 g/ml, and 50% equals 0.701 g/ml of the honey. culturing s. aureus a swap from pure culture of s. aureus was obtained and then cultured in 10 ml of nutrient broth as the step was repeated for each test tube. furthermore, different concentrations of honey were added to the cultured test tubes as the samples were incubated for 24 h and the results were checked. antibiotic sensitivity test the effect of 10 types of antibiotics on s. aureus was tested after being cultured “s. aureus” on mueller-hinton agar. the reason behind using mueller-hinton agar is that it is inert; does not have effect on the antibiotics. this test was done to show whether s. aureus is strong bacteria or not and compare the results the other tests that honey used as antimicrobial agent. there were two petri dished used as five disks of antibiotics were put in each and then incubated for 24 h. in addition, the results were checked whether resistant or sensitive following the size of inhibition zone measurement. one factor at a time was used to improve the process parameters and the ideal circumstances for each type. inhibition zone measurement the transparent area that surrounds the disks was measured using a ruler as the area refers to the sensitivity of bacteria to the antibiotics along with the measurement of the diameter of inhibition zone that has been subtracted from the disks. hplc test preparation a 10 g of honey was shipped to baghdad to be homogenized in 50 ml of 100 mm sodium sulfate buffer (ph 7.0), containing 1 mm ascorbic acid and 0.5% (w/v) polyvinylpyrrolidone, for 5 min at 4°c. the homogenate was filtered through three layers of cheesecloth, and the filtrate was then run at 5000× g for 15 min, and the superannuate was collected. sample residue was then resuspended in 1.0 ml hplc grade methanol by vortexing, and the mixture was then run through a disposable 2.5 um filter. this was followed by storage at 4°c for further analysis, and finally, 20 ul of the sample was injected into the hplc.[21] equipment shimadzu 10 a v-lc with a binary delivery pump model lc-10 a was used for the separation, and a uv–vis 10 a-spd spectrophotometer was used to track the peak elution. the sequence of the eluted material of the standard mentioned in table 1. hplc determination of enzymes in honey the extract was separated using an fast liquid chromatographic column with a 7 um particle size, a nuclosit 4000-7 pei, anion exchange for protein and peptides (125 × 4.0 mm i.d) column, mobile phase, linear gradients from 0% b to 100% b in 10 min, and solvents a and b with different concentrations of tris-acetate at different ph levels. uv detection is set to 280 nm with a 1.5 ml/min flow rate. results agar well diffusion assay 24 h of incubation were illustrated that the bacteria were resistant to all samples “s 1, s2, and s3” at 25 µl concentration. table 1: the sequence of the eluted material of the standard seq. subjects retention time minute area 1 catalase, cat 2.02 170,253 2 amylase 3.19 180,849 3 invertase 4.10 193,624 4 glucose oxidase 5.19 1632 thanoon, et al.: antimicrobial activity against staphylococcus aureus by hplc determination 115 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 the first sample (artificial honey) started its activity against bacteria at the concentrations 75 µl and 100 µl as the inhibition zone was 0.2 mm while at the same concentrations of the rest two samples, s. aureus was resistant. in addition, at 125 µl, the inhibition zone of s. aureus for the first sample was about 0.3 mm in the same time of being resistant in the rest two samples. at 150 µl, samples 2 and 3 (black and natural honey) started their activity against bacteria as the inhibition zone for the second sample was 0.2 mm while for the third sample was 0.1 mm as per the figures 1-4.[22] dilution method the first honey sample (industrial honey) on a certain concentration was tested and then mixed it with nutrient broth and s. aureus as well as the same concentration from the other two samples as they were subjected for incubation for 24 h. the results showed that the bacteria were resistant based on the observed growth in the three test tubes. when the concentration of all honey samples was increased, the bacterial growth was detected but in lower rates. the lowest concentration of the honey in clear test tube was considered as the minimum inhibitory concentration “mic,” table 2. aureus resistance against antibiotics the effect of 10 types of antibiotics on s. aureus was tested after being cultured ‗s. aureus on mueller-hinton agar. the reason behind using mueller-hinton agar is that it is inert; does not have effect on the antibiotics. the resistance and sensitivity of s. aureus were tested to some of the antibiotics including; rifampin, clindamycin, gentamicin, methicillin, ciprofloxacin, penicillin, cloxacillin, trimethoprim, azithromycin, and erythromycin as the results are illustrated in table 3. hplc test of honey the concentration of catalase, amylase, invertase, and glucose oxidase in honey tested by hplc and by following the equation below shows the concentration of each substance: conc. of sample = (area of sample ÷ area of standard) × cons. of standard × dilution factor: conc. of catalase = (32877 ÷ 170253) × 20 = 3.862 u/ml conc. of amylase = (136985 ÷ 180849) × 20 = 15.149 u/ml conc. of invertase = (58466 ÷ 193624) × 20 = 6.039 u/ml conc. of glucose peroxide = (105204 ÷ 163245) × 20 =12.889 u/ml discussion the findings showed that the three different honey samples have antibacterial action against the gram-positive bacterium s. aureus. the possible reason could be that honey consists mainly of carbohydrates (approximately 82%), water and other minor components including; proteins, minerals, and phytochemicals.[23] the water activity of honey ranges from 0.5 to 0.6 which could be low enough to prevent bacterial growth. furthermore, the high sugar content of honey can induce the osmotic pressure on the bacterial cells the thing that will initiate the process of water flow to the outside of the cell, leading to the dehydration and shrinkage of the cell thus the inability to survive.[24] figure 2: effect of different concentrations of natural honey on staphylococcus aureus figure 1: effect of different concentrations of industrial honey “s 1” on staphylococcus aureus figure 4: concentration of enzymes found in industrial honeyfigure 3: antibiotic sensitivity test thanoon, et al.: antimicrobial activity against staphylococcus aureus by hplc determination 116 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 moreover, the presence of gluconic acid “organic acid” gives the honey its acidic characteristic which is about 3.2 ph the thing that is not optimal for further bacterial growth which is provided by neutral ph.[25] the degree of antibacterial action changed agreeing according to the type of bacteria and honey. the observed variations in the behavior of distinct honeys types at vary concentrations as well as the nature of honey production’s impact such an effective pattern on their inhibitory activities that were directly proportional to the increased honey concentrations respectively.[24] it has been detected that these phytochemicals are the ones responsible for medical and biological activities of honey within the treatment of infections, burns, wounds, and ulcers. using cybow 11 test strips, the ph was determined for all varieties of honey and their concentrations, and it was found to be 5, indicating that the honey is acidic. in addition, honey’s significant osmotic impact (approximately 80% wt/vol of concentrated sugars) and microorganism susceptibility to hydrogen peroxide act as inhibitors of bacterial development.[26] in this research, agar well diffusion and the dilution method were both employed to assess the antibacterial activity of honey. acidity, non-hydrogen peroxide activity, high osmotic effect, and the presence of phytochemical components, which are helpful in regulating bacterial dispersal, are the main actions of honey. because honey has a significant amount of sugar, bacteria cannot grow in it because the ph is low enough to prevent many types of bacteria from growing in it. honey’s primary antibacterial component, hydrogen peroxide, is produced when the enzyme glucose oxidase is triggered by honey dilutions. furthermore, other components found in honey and give it the ability to kill or inhibit the growth of bacteria.[27,28] all of the investigated honeys’ mic and inhibitory zone values against s. aureus demonstrated bactericidal and bacteriostatic activity. our findings are comparable to those of other studies.[29] the bacteria were resistant to the honey sample if any zone had a diameter of <3 mm. the bacterium appeared to be sensitive to the honey sample, though, as shown by a zone diameter higher than 7 mm.[30] these honey samples might be used topically to heal wounds because the bacteria are known to infect wounds and are sensitive to the honey samples. it is known that honey may be used to cure an infected wound.[31] it was shown that 50% (v/v) of the three samples’ concentration of honey that prevented the growth of which 90% of staphylococcus aureus accordingly. the findings of earlier studies reflect our ideals. these variations in honey’s antibacterial action may result from its diverse bee sources or species.[32] in addition, samples of honey included alkaloids, tannins, and flavonoids, all of which are known to have antimicrobial properties.[19] data from both the agar diffusion and broth dilution methods indicated that bacteria were more resistant to the second and third samples more than industrial honey, despite the fact that this result is supernatural, according to previous research, the natural honey has greater antibacterial activity than modified honey. this may be due to fraud by the companies exporting these two samples and selling them as a natural honey. however, in our studied if we take into account that the two samples are natural, the second sample was having greater antibacterial activity than the third, the reason is due to the different nectar source of the bees or the change in acidity and water rate in both samples. the industrial honey is tampered with honey. it started off as pure honey, but later other ingredients – glucose, dextrose, molasses, sugar syrup, flour, corn syrup, starch, or any other product – were added to boost its volume or alter its qualities. inverted sucrose from beet or cane sugar is artificial honey. to mimic genuine honey, it has been altered in look, flavor, and odor.[33] there are various researches studied the chemical components of industrial honey in general, regardless of the company that manufactured it to distinguish between natural honey and artificial honey. a total of 800 samples of industrial honey were examined for this reason. since the designs of honey with 0 and 7 and 0 and 0 and c-4 sugar content (percent) were so similar, it was possible to determine that honey with 7 and c-4 sugar content (percent) was also free of c-4 sugars. the 13c protein levels of both honey groups and the 13c honey values show a very good association. a helpful characteristic for contaminated honey with a c-4 sugar content (percent) 7 is that the c-4 sugar content (percent) 7 group had lower (p 0.05) values (16.30) for the 18o value compared to other honey. table 2: mics of honey against staphylococcus aureus growth solution % mic result s1 10 no mic negative 20 no mic negative 50 mic positive s2 10 no mic negative 20 no mic negative 50 mic positive s3 10 no mic negative 20 no mic negative 50 mic positive mic: minimum inhibitory concentration table 3: antibacterial activity of antibiotics against staphylococcus aureus antibiotic (mcg) sensitivity inhibition zone cloxacillin (10 mcg) sensitive 8.1 cm trimethoprim (10 mcg) sensitive 6.2 cm azithromycin (15 mcg) resistant no inhibition ciprofloxacin (10 mcg) sensitive 10.4 cm penicillin g (10 mcg) sensitive 5.3 cm rifampin (5 mcg) sensitive 8.1 cm methicillin (10 mcg) sensitive 3.5 cm gentamicin (10 mcg) sensitive 7.8 cm clindamycin (10 mcg) sensitive 6.5cm erythromycin (10 mcg) resistant no inhibition thanoon, et al.: antimicrobial activity against staphylococcus aureus by hplc determination 117 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 a hplc test was carried out to determine the components of industrial honey and their concentrations to determine why industrial honey had a greater impact on honey. this test’s results revealed that industrial honey is primarily composed of carbohydrates, catalase, and invertase, and that its low water content also plays a role in the concentration-dependent killing of bacteria. hplc test showed that the most common type of enzymes present in the honey is organic acidic and is explained the acidity of honey. it also showed a high amount of glucose oxidase which plays an important role in killing bacteria by oxidation of glucose to h 2 o 2 .[7] many researches have employed the earlier techniques while using the agar diffusion method to measure the activity of honey. when researchers researched honey or the natural antibacterial components of honey, they discovered a number of issues with the agar diffusion method. a few modifications in the experimental settings, such as agar volume, inoculation concentration, and incubation conditions, may cause significantly different findings. these include insensitivity, wherein modest levels of antimicrobial activity are not always evident.[34] finally, the data acquired by the dilution techniques and time-kill assay did not confirm the results of the agar diffusion obtained in the current investigation, which indicated limited activity. it was obvious that there was no direct correlation between mic and zone size. this shows that results from the agar diffusion technique may not always be an accurate reflection of overall antibacterial activity. this research also studied the effect of some antibiotics on bacteria to see if s. aureus is resistant or sensitive to it. we have noticed that s. aureus was sensitive to some antibiotics, including rifampin, clindamycin, gentamicin, methicillin, ciprofloxacin, penicillin, cloxacillin, and trimethoprim, and it was resistant to azithromycin and erythromycin. conclusion all tubes that were cultured with addition of honey showed a level of turbidity except 10% diluted tubes which was a clear tube (no turbidity) that refers to mic. in the well diffusion method, the three types of honey had different effects at the same concentration of honey. low concentration had not any effect and high concentration of honey showed a different effect. although the industrial honey had more effect (larger inhibition zone), we cannot consider it as an antimicrobial agent because the inhibition zone was not large enough compared to the concentration of honey and other antibiotics that were used. references 1. n. a. albaridi. antibacterial potency of honey. international journal of microbiology, vol. 2019, pp. 1-10, 2019. 2. p. mcloone, m. warnock and l. fyfe. honey: a realistic antimicrobial for disorders of the skin. microbiology and immunology, vol. 49, pp. 161-167, 2016. 3. c. v. nolan, j. harrison and j. a. g. cox. dissecting the antimicrobial composition of honey. antibiotics, vol. 8, p. 251, 2019. 4. k. s. hossain, m. g. hossain, a. moni, m. m. rahman, u. h. rahman, m. alam, s. kundu, m. m. rahman, m. a. hannan and m. j. uddin. prospects of honey in fighting against covid-19: pharmacological insights and therapeutic promises. heliyon, vol. 6, p. e5798, 2020. 5. s. anand, m. deighton, g. livanos, e. c. k. pang and n. mantri. agastache honey has superior antifungal activity in comparison with important commercial honeys. scientific reports, vol. 9, p. 18197, 2019. 6. o. escuredo, l. r. silva, p. valentão, m. c. seijo and p. b. andrade. assessing rubus honey value: pollen and phenolic compounds content and antibacterial capacity. food chemistry, vol. 130, pp. 671-678, 2012. 7. s. bogdanov and p. martin. honey authenticity mitteilungen aus dem gebiete der. lebensmitteluntersuchung und hygiene, vol. 93, pp. 232-254, 2002. 8. j. t. seil and t. j. webster. antimicrobial applications of nanotechnology: methods and literature. international journal of nanomedicine, vol. 7, pp. 2767, 2012. 9. a. f. henriques, r. e. jenkins, n. f. burton and r. a. cooper. the effect of manuka honey on the structure of pseudomonas aeruginosa. european journal of clinical microbiology and infectious diseases, vol. 30, pp. 167-171, 2011. 10. m. a. mundo, o. i. padilla-zakour and r. w. worobo. growth inhibition of foodborne pathogens and food spoilage organisms by select raw honey. international journal of food microbiology, vol. 97, no. 1, pp. 1-8, 2004. 11. h. goossens, m. ferech, v. r. stichele and m. elseviers. outpatient antibiotic use in europe and association with resistance: a crossnational database study. lancet, vol. 365, no. 9459, pp. 579-587, 2005. 12. e. hołderna-kędzia and b. kędzia. miody odmianowe i ich znaczenie lecznicze. wydawnictwo duszpaster stwa rolników, włocławku, poland, 2002. 13. j. cooke. when antibiotics can be avoided in skin inflammation and bacterial colonization? a review of topical treatments. current opinion in infectious diseases, vol. 27, no. 2, pp. 125-129, 2014. 14. s. mandal, m. deb mandal and n. k. pal. antimicrobial resistance pattern of salmonella typhi isolates in kolkata, india during 1991-2001: a retrospective study. japanese journal of infectious diseases, vol. 55, pp. 58-59, 2002. 15. a. mansour, k. enayat, n. m. s. neda, a. behzad. antibacterial effect and physicochemical properties of essential oil of zataria multiflora boiss. asian pacific journal of tropical medicine, vol. 3, pp. 439-442, 2010. 16. b. joshua and n. d. keshu. comparing the antibacterial and functional properties of cameroonian and manuka honeys for potential wound healing-have we come full cycle in dealing with antibiotic resistance? molecules, vol. 20, no. 9, pp. 1606816084, 2015. 17. v. m. french, r. a. cooper and p. c. molan. the antibacterial activity of honey against coagulase-negative staphylococci. journal of antimicrobial chemotherapy, vol. 56, pp. 228-231, 2005. 18. o. o. akanbi and b. o. omafuvbe. microbiological and physicochemical properties of some commercial nigerian honey. african journal of microbiology research, vol. 3, no. 12, pp. 891-896, 2009. 19. a. simon, k. traynor, k. santos, g. blaser, u. bode and p. molan. medical honey for wound care still the “latest resort”. evidence based complementary and alternative medicine, vol. 6, no. 2, pp. 165-173, 2009. 20. d. dinkov. perspection of royal jelly and bee honey as new antibacterial therapy agents of hospital infections. journal of clinical pathology and laboratory medicine, vol. 1, no. 1, pp. 5-8, 2017. 21. k. j. abhishek, v. ravichandran, s. madhvi and r. k. agrawal. synthesis and antibacterial evaluation of 2-substituted-4,5diphenyl-n-alkyl imidazole derivatives. asian pacific journal of tropical medicine, vol. 3, no. 6, pp. 472-474, 2010. thanoon, et al.: antimicrobial activity against staphylococcus aureus by hplc determination 118 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 112-118 22. v. p. shenoy, m. ballal, p. g. shivananda and i. bairy. honey as an antimicrobial agent against pseudomonas aeruginosa isolated from infected wounds. journal of global infectious diseases, vol. 4, no. 2, p. 102, 2012. 23. f. g. p. moumbe, f. zambou and m. kaktcham. antimicrobial activity of probiotic strain lactobacillus plantarum isolated from ‘‘sha0a” and assement of its viability in local honey. the journal of microbiology, biotechnology and food sciences, vol. 3, pp. 226231, 2013. 24. d. m. manisha and m. shyamapada. honey: its medicinal property and antibacterial activity. asian pacific journal of tropical biomedicine, vol. 1, no. 2, pp. 154-160, 2011. 25. s. natarajan, d. williamson, j. grey, k. g. harding and r. a. cooper. healing of an mrsa-colonized hydroxyurea-induced leg ulcer with honey. journal of dermatological treatment, vol. 12, pp. 33-36, 2001. 26. p. l. miorin, n. c. levy, a. r. custodio, w. a. bretz and m. c. marcucci. antibacterial activity of honey and propolis from apis mellifera and tetragonisca angustula against staphylococcus aureus. journal of applied microbiology, vol. 95, pp. 913-920, 2003. 27. n. al-waili, k. salom and a. a. al-ghamdi. honey for wound healing, ulcers, and burns; data supporting its use in clinical practice. the scientific world journal, vol. 11, pp. 766-787, 2011. 28. m. diez. characterization of uniflora honey by their physical characteristics. food chemistry, vol. 79, pp. 373-379, 2002. 29. m. ahmed, s. sahile and c. subramanian. evaluation of antibacterial potential of honey against some common human pathogens in north gondar zone of ethiopia. international journal of pure and applied zoology, vol. 2, no. 4, pp. 286-295, 2000. 30. o. e. agbagwa and n. frank-peterside. effect of raw commercial honeys from nigeria on selected pathogenic bacteria. african journal of microbiology research, vol. 4, no. 16, pp. 1801-1803, 2010. 31. a. kingsley. the use of honey in the treatment of infected wound. the british journal of nursing, vol. 10, no. 22, pp. 13-16, 2001. 32. r. zintchem, g. o. fankem, c. fokunang, p. mkounga, e. t. achick, r. kamgang, d. gatsing and t. dimo. in vitro evaluation of the antimicrobial properties of mallotus oppositifolium decoction leaf extracts and fractions. international journal of biological and chemical sciences, vol. 9, no. 2, pp. 815-824, 2015. 33. n. prica, m. živkov-baloš, s. jakšid, z. mihaljev, b. kartalovid, j. babid and s. savid. moisture and acidity as indicators of the quality of honey originating from vojvodina region. arhiv veterinarske medicine, vol. 7, no. 2, pp. 99-109, 2014. 34. p. h. s. kwakman, j. p.c. van den akker, a. güçlü, h. aslami, j. m. binnekade, l. de boer, l. boszhard, f. paulus, p. middelhoek, a. a. te velde, c. m. j. vandenbroucke-grauls, m. j. schultz and s. a. j. zaat. medical-grade honey kills antibiotic-resistant bacteria in vitro and eradicates skin colonization. clinical infectious diseases, vol. 46, no. 11, pp. 1677-1682, 2008. tx_1~abs:at/add:tx_2~abs:at 161 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 161-165 research article grid to ground solutions in construction projects shamal f. ahmed department of surveying and geomatics, faculty of engineering, tishk international university-erbil, kurdistan region, iraq abstract nowadays, global navigation satellite system (gnss) techniques are considered the cornerstones in the field of surveying and civil engineering industry. at present, a plethora of companies is employing gnss in their daily activities. the engineering construction projects are designed based on ground distances although rapid static works on grid coordinate system, where the distances between points vary from the ground. the established control points should be compatible with the design by converting distances to ground. this study investigates the effect of the scale factor (sf) to reduce the linear distortion when grid is converted to ground. for this purpose, eight different distances from 64.858 m to 887.974 m were taken with rapid static method. the results reveal that whereas the distances are increased, the difference between the grid and ground is proportional, and for the smallest distance, the difference was 54 mm and for the largest was 398 mm. besides, the grid distances were converted to ground distances using the combined sfs that works as a divider; the average difference was decreased from 175 mm to 45 mm, and the maximum difference was decreased from 398 mm to 85 mm. keywords: global navigation satellite system, total station, grid ground distance, scale factor, conversion introduction now in many construction projects, different instruments are used for surveying purposes such as global navigation satellite system (gnss) and total station (ts). gnss is a multi-constellation satellites that can be utilized to determine the receiver’s position on the earth by sending signals from space that contains navigational message to the gnss receivers. one of the most useful methods in gnss positioning, for a small distance and in millimeter accuracy, is the rapid static method. it is a relative positioning that uses the carrier phase to determine the coordinates of unknown point (rover). on the other hand, the other receiver (base) is occupied on a point with known coordinates which should be tracked the same satellites.[1-3] details on rapid static, operational considerations, data transmission, ambiguity resolution, and processing can be found in.[4] the occupation time is planned according to the length of the baseline, the numbers of satellites, the geometric dilution of precision, and the type of the instruments reference[5] stated that the integer ambiguity can be fixed more reliable and faster when the occupation time is adequately long. the accuracy of this method is about 3–5 mm.[1] on the other hand, ts instruments can carry out all the functions that transits and theodolites are capable of doing, with more efficiency. in addition to that, they have the ability to observe distances quickly and accurately with the angle and can compute some mathematical calculations by a microprocessor that is fixed in the instrument and display the results in real-time.[2] according to references,[5,6] gnss works on projected (grid) coordinates, whereas the ts works on ground coordinates. when these two instruments are combined and used together in a project, linear distortion occurs. this distortion cannot be eliminated, but it can be minimized.[7,8] in this paper, eight different baselines will be taken in different lengths by both gnss (rapid static method) and ts to present the effect of scale factor (sf) to minimize linear distortion when these types of instruments are combined. grid to ground solutions there are three different surfaces of the earth for per project area, as illustrated in figure 1, the ground (ts distances), the ellipsoid, and the grid (gnss distances) plane. ground distances are distances between objects on the topographic surface of the earth. it can be measured with chains or calibrated wheels with odometers and are needed whenever an analysis depends on actual distances on the earth’s surface. this can be corresponding author: shamal f. ahmed, department of surveying and geomatics, faculty of engineering, tishk international university-erbil, kurdistan region, iraq. e-mail: shamal.fatah@tiu.edu.iq received: september 13, 2022 accepted: october 30, 2022 published: december 30, 2022 doi: 10.24086/cuesj.v6n2y2022.pp161-165 copyright © 2022 shamal f. ahmed. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) ahmed: grid to ground solutions in construction projects 162 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 161-165 the case for highway construction, hydrology, and meteorology, for example, mappers and surveyors seldom measure ground distances, because most maps are compiled with horizontal or reduced distances.[9] the distance measured by an electronic distance measurement (edm) is called an edm distance. in general, they are not straight-line distances, because the light emitted by the instrument is refracted by the earth’s atmosphere. a euclidean straight-line distance is called a spatial distance. one can attempt to remove the atmosphere’s effect on edm distances by measuring atmospheric temperature, humidity, and pressure.[1] the projected (grid) coordinates are derived from ellipsoidal coordinates in a particular geodetic datum and coordinate reference system after applying a projection like universal transverse mercator projection (utm).[10] when points are moved from one surface to another, as shown in figure 1, the distance between those points varies, resulting in a change in scale owing to the distortion. when intending to depict a curved surface on a plane, it is impossible to do so without introducing some degree of distortion. it indicates that there is a shift in the “real” relationship that exists between points that are situated on the surface of the earth and the representation of that relationship on a plane. because distortion cannot be completely removed, the most that can be done is to minimize its impact. one of the types of distortions is known as linear distortion which is the difference in distance between a set of projected (map grid) coordinates and the actual horizontal distance between those points (ground), represented by “δ”. to transform points from the grid to the ground, first an ellipsoid transformation must be performed, and then, a ground transformation must follow. each step has its own scale shift. scale factor the scale factor (sf) is the value which specifies the distanced points, such as between a and b on the grid plane (grid distance (dgrid)) and those matching two locations on the ellipsoid (ellipsoid or geodetic distance (dellip)). the cause of a scale difference is due to the transformation that occurs from a flat (grid plane) to a curved surface (ellipsoid). equation (1) calculates the sf at each point: ° °= + 2 2 sf 2 e k r (1) where: k: central meridian’s sf r: earth’s radius e: difference between false easting at the central meridian and easting of the point first, by calculating the grid distance between the two points and then multiplying that result by the effective sf (effsf), it is possible to get the approximate ellipsoidal distance between the two points (a and b). this will yield the value of the approximate ellipsoidal distance. the effective sf (effsf) can be calculated using (2), which is as follows: + + = 4 effsf 6 a ab bsf sf sf (2) where: sf a : stands for sf point a sf b : stands for sf point b sf ab : stands for sf point between sf a and sf b . using equation (3) the grid distance (dgrid) can be altered to an ellipsoidal distance (dellip). = dgri dellip d effsf (3) the scale value that defines the distance between two points on the ellipsoid (geodetic distance, or dellip) and those same two points on the earth (ground distance, or dground) is known as the elevation factor (ef). equation (4) can be used to obtain the ef: = + ef r r h (4) where: h: mean ellipsoidal height between a and b by applying the ef in (5), the ground distance (dground) can be calculated from the geodetic distance (dellip) = d dg el round lip ef (5) the most frequent transformation is that between points on the grid plane and points on the ground. the combination factor (cf) is the scale value that will permit a direct figure 1: different types of surfaces ahmed: grid to ground solutions in construction projects 163 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 161-165 transformation from grid to ground. equation (6) illustrates how to compute the combination factor by multiplying the sf by the ef: cf = effsf × ef (6) using the combination factor, one can determine the ground distance (dground) from the grid distance (dgrid), using (7): = d dg gr round id cf (7) data and field observations eight points were marked on a satellite image in iraq-erbil at latitude 43.993° e and longitude 36.1901°n with different distances. reconnaissance was done for points to check the intervisibility of points, as shown in figure 2. the procedure commenced by measuring coordinates of the eight points using gnss receiver (leica gs16) by the rapid static method, because the baselines between base and the rover (at points) were small. subsequently, the points were measured using ts by the traverse method. points were randomly distributed to investigate the different distances between gnss and ts instrument [figure 3]. coordinates were measured by rapid static method, starting from point (t1) to (t8) and the occupation time for each point was equal to 15 min. the collected data were processed by leica infinity software (version 3.0.1) to get the coordinates of the points. on the other hand, the ts (topcon 255) was used to observe the points by traverse method which started and terminated on t1 which was a common point. results and discussion the collected raw data by rapid static method were post processed with the selected parameters which were: cutoff angle was 15°, ephemeris type was precise, gnss types were gps, glonass, galilleo, minimum duration for float solution [static] was 5’ and sampling rate was 1 s. as illustrated by table 1, the grid distances (gnsss) are the smallest, whereas the ts distances are the biggest. in each figure 6: the differences between grid and by total station figure 2: study area figure 4: the differences from grid to ellipsoid figure 3: different distances figure 5: the differences from ellipsoid and ground ahmed: grid to ground solutions in construction projects 164 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 161-165 table 1: distances (m) from–to grid ellipsoid ground ts t1–t2 560.730 560.893 560.927 560.979 t2–t3 152.641 152.686 152.695 152.730 t3–t4 195.873 195.930 195.942 195.983 t4–t5 300.378 300.466 300.484 300.541 t5–t6 64.858 64.877 64.881 64.912 t6–t7 241.178 241.249 241.263 241.291 t7–t8 548.874 549.035 549.068 549.102 t8–t1 887.974 888.233 888.287 888.372 table 2: differences (m) from-to grid ellipsoid ellipsoid ground grid-ts ground-ts t1–t2 0.164 0.034 0.250 0.052 t2–t3 0.045 0.009 0.089 0.035 t3–t4 0.057 0.012 0.110 0.041 t4–t5 0.088 0.018 0.163 0.056 t5–t6 0.019 0.004 0.054 0.031 t6–t7 0.071 0.015 0.113 0.028 t7–t8 0.161 0.033 0.228 0.034 t8–t1 0.259 0.053 0.398 0.085 surface, the distances are increased by a rate close to the ts distances. from figures 4-8 and table 2, it seems obvious that the difference between grid and ts increased proportionally. due to the impact of the effective sf (effsf), the grid distances were increased considerably by approximately 79% and the maximum difference was 25.9 cm, minimum difference was 4.5 cm, and the average was 10.8 cm. the distances were converted to ground by the ef, and they were slightly increased. the maximum difference was 5.3 cm, minimum difference was 0.9 cm, and the average was 2.2 cm. after applying the combined sf to get the ground distances close to the ts distances, the maximum difference was 8.5 cm and the minimum was 2.8 cm. conclusion to sum up, various instruments, such as the gnss and the ts, are utilized in modern construction projects for surveying. an avoidable linear distortion occurs when both of them are used in a project. this is due to the fact that gnss works on grid and ts works on ground coordinate system. in this study, to investigate the impact of the sf to reduce linear distortion, gnss (rapid static method) and ts have been used to determine the accuracy of the factor. eight different distances were taken, the differences between gnss and ts distances were increased proportionally. due to the influence of the effective sf, the grid distances have been significantly increased by about 79%; the maximum difference was 25.9 cm, the minimum difference was 4.5 cm, and the average difference was 10.8 cm. the ef was used to convert the distances to ground; consequently, they have been slightly increased. the greatest difference was 5.3 cm, whereas the smallest difference was 0.9 cm, and the average difference was 2.2 cm. by applying the combined sfs that worked as a divider the grid distances were converted to ground distances, the average difference was reduced from 17.5 cm to 4.5 cm whereas the maximum difference from 39.8 cm to 8.5 cm. it is an indisputable fact that sources of error like the instrumental, personal, and environmental affect the observations. moreover, if more accurate ts was used with high precision the result might be more accurate. references 1. c. d. ghilani and p. r. wolf. elementary surveying: an introduction to geomatics. prentice hall, new jersey, 2012. 2. b. f. kavanagh and d. k. slattery. surveying: with construction applications. prentice hall, new jersey, 2010. 3. j. van sickle. gps for land surveyors. crc press, florida, 2008. 4. s. f. ahmed and r. a. abbak. effect of different network geometry on gnss results. selcuk university journal of engineering sciences, vol. 17, no. 1, pp. 1-18, 2018. 5. a. el-rabbany. introduction to gps: the global positioning system. artech house, london, 2002. 6. a. el-mowafy. surveying with gps for construction works using the national rtk reference network and precise geoid models. in: presented at the 1st fig international symposium on engineering surveys for construction works and structural engineering, pp. 1-14, 2004. 7. i. alil. an introduction to map projections and low distortion projections (ldp). 2020. available from: https://www. conradblucherinstitute.org/workshops [last accessed on 2022 aug 10]. figure 8: all differences figure 7: the differences between ground and by total station ahmed: grid to ground solutions in construction projects 165 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 161-165 8. m. l. dennis. ground truth-design and documentation of low distortion projections for surveying and gis. in: arizona professional land surveyors workshop, vol. 11, pp. 40-45, 2008. 9. t. h. meyer. grid, ground, and globe: distances in the gps era. surveying and land information science, vol. 62, no. 3, pp. 179-202, 2002. 10. v. janssen. understanding coordinate systems, datums and transformations in australia. in: proceedings of the surveying and spatial sciences institute biennial international conference, pp. 697-715, 2009. tx_1~abs:at/add:tx_2~abs:at 31 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 31-34 research article electromagnetic field pollution level measurement for mobile phone networks at cihan university-erbil adil h. mohammed department of communication and computer engineering, faculty of engineering, cihan university-erbil, kurdistan region, iraq abstract the health effects of the impact of radiofrequency (rf) electromagnetic fields (emf) have become of increasing concern. we conducted a specific absorption rate (sar) study and measurements of wireless and mobile phone networks to evaluate the hazard of rf radiation limit. our measurements have been carried out at different locations on the campus of cihan university-erbil. the rf strength level measurement of a base station on site was done by using a spectrum analyzer with a dipole antenna and electromagnetic field detector (emf) application in the handsets for mobile networks, then comparing our measurements with the international slandered sar levels. keywords: antenna, electromagnetic fields pollution, mobile network, specific absorption rate limit, specific absorption rate introduction for more than many years, most people have been concerned about the possible health consequences of exposure to radiofrequency radiation (rf and mobile devices). excessive levels of rf fields are known to induce various physical effects on the human body. today, after the wide variety boom in public use of wireless devices, especially cell phone networks, it has emerged as obligatory to ensure that these products no longer divulge their users to potentially harmful degrees. at the frequencies at which the maximum of these devices operate, the acknowledged health outcomes are middle on or around tissue heating. specific absorption rate (sar) and sar limits sar is referred to as the specific absorption rate and is the time byproduct of the incremental energy (dw) absorbed or dissipated in the incremental mass (dm) contained in an amount (dv) of a given density (ρ):[1] sar d dt dw dm d dt dw pdv � � � � � � � � � � � � � �              (1) sar should be considered an “absorbed dose rate” and is related to electric fields at a point by: sar e � � � 2 � � (2) where: σ = conductivity of the tissue (s/m) ρ = mass density of the tissue (kg/m3) e = root mean square (rms) of electric field strength (v/m). the bounds, which apply in widespread for cell phones and similar apparatus, there are two limits in every one of them a low rate for exposure averaged over all frames and a higher rate relevant to nearby publicity to components of the body. this partial-frame sar is averaged over a quantity of tissue defined as a tissue volume within the form of dice. american necessities differ from the global requirements of their call for lower spatial joint restriction, which is the typical limit over a smaller quantity (1 g of tissue as opposed to 10 g). in addition, they require an extended time over which the sar is to be averaged. still, for the reason that it is miles assumed that a person with the portable tool will be uncovered to the maximum power available from the device during the required average time, the required time averaging of the output all through sar size does not require practice. the ansi/ieee and icnirp published rf publicity limits measures that must no longer be handed to human health. the ansi/ieee limits the spatial-average special absorption rate (sar) to at least one. 6 w/kg over 1 g of tissue over halfhour.[2] corresponding author: adil h. mohammed,department of communication and computer engineering, faculty of engineering, cihan university of erbil, kurdistan region, iraq. e-mail: adil.mohammed@cihanuniversity.edu.iq received: january 07, 2023 accepted: june 04, 2023 published: august 25, 2023 doi: 10.24086/cuesj.v7n2y2023.pp31-34 copyright © 2023 adil h. mohammed. this is an open-access article distributed under the creative commons attribution license (ccby-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) mohammed: electromagnetic field pollution level measurement for mobile phone networks 32 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 31-34 in addition, the icnirp limits the sar to 2-four w/kg.[3] mobile telephones commonly have a time-averaged energy output of 600 mwatt or much less and yield numerically modeled mind sars which might now and again exceed 1.6 w/kg restriction; however, which commonly lie inside the ansi/ieee “managed surroundings” condition of 8w/kg averaged over 6 min. the following table 1 represents the international fashionable used for measuring sar restriction. mobile phone networks the cellular smartphone era consists of the essential components: the provider and the consumer. the bottom stations (bs) are on the provider side, “the antenna distribution on the towers which speak with the mobile phones.” on the consumer side is cell smartphone handsets. base stations bs emit electromagnetic radiation continuously and at some distance extra strength than cell phones which emit electromagnetic radiation continuously simplest all through calls or makes use of information. in the rest of the instances, that is, idle mode, mobile phones emit a regular pulse of electromagnetic radiation for base stations to continuously preserve track of the geographic role of the telephones in their “cell community.” modern-day 2nd era (2g) bs antennas are associated with transmitter powers of 20 – a hundred watt, despite the fact that the ultra-modern 3rd era (3g) bs antenna use much less electricity, on average 3 watt in city regions. in rural areas, the bs power output is much better due to the many regions that need to be included between base stations positioned in far-off places. by implementing urban sprawl strategies, technology has advanced and the data demands of cellular networks have increased, the number of towers has increased significantly, but companies have made little effort to share such towers. although the number of antennas in our urban environment is low, the increased number of antennas has improved cell phone reception in moving elevators, large building centers and basements, and other “dead” locations. the ability of a mobile phone user to be instantly “discovered” by a base station almost anywhere on the planet is astonishing, and the widespread current should show nearly prevalent generation and propagation. network,[5] and a new generation of mobile networks evolved node b (enb).[6] mobile antennas and antenna arrays antennas generally have a specific directivity. an omnidirectional antenna radiates in all directions (horizontally viewed), whereas a sector antenna effectively radiates in only one (horizontal) sector [figure 1]. this allows more spectrum reuse due to less interference. therefore, most base stations in congested areas such as cities are sectoral. the recommended gain for sector antennas is 10–20 dbi. this means that the radiated power in the direction of interest can be 10–100 times stronger than an omni antenna but correspondingly weaker in other laws.[7] massive multi input multi output (mimo) antenna arrays massive mimo antenna array is a recent application in 5th mobile networks (5g) and beyond, which supports the sub 6 ghz frequency as well as millimeter wave (mmwave) radio frequencies,[8] nowadays mmwave massive mimo antennas serving frequencies below 100 ghz but it can reach up to 300 ghz or even terra hertz frequency band shortly. the main advantage of massive mimo is the ability to serve multiple users and multiple devices simultaneously within a condensed area by very narrow beam by deploying beamforming technology[8] figure 2; due to the high gain of these antennas, a meager power can be transmitted from the base stations compare to table 1: sar limit guideline[4] guideline sar limit head, neck, and trunk health canada safety code 1.6 w/kg averaged over 1 g of tissue europe 2.0 w/kg average over any 10 g of tissue australia 1.6 w/kg averaged over 1 g of tissue us 1.6 w/kg averaged over 1 g of tissue india 1.6 w/kg averaged over 1 g of tissue icnirp 2.0 w/kg average over any 10 g of tissue sar: specific absorption rate figure 2: massive multi input multi output beamforming figure 1: antenna radiating for mobile base station mohammed: electromagnetic field pollution level measurement for mobile phone networks 33 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 31-34 conventional omnidirectional antennas, furthermore only the mobile users will be reached with bs transmission. at the same time, they are active users (in call or data stream use), which could be a significant improvement to reduce emf pollution and lead to reduce sar in public areas, see figure 3, on the other hand, massive mimo has become an exciting candidate to improve the mobile handsets[9,10] in the same way to reduce transmission power directed to the bs with compare to omnidirectional antennas which transmitting almost in all directions, so overall massive mimo technology is an improvement to reduce sar at both bs and mobile handsets. figure 4: site of cihan university-erbil measuring sar level in cihan university-erbil at the cihan university campus, there are nine buildings as shown in figure 4. the mobile base station is allocated on the top of building 5, which has four floors with an additional room on the top; buildings 1, 2, 3, and 4 have three floors, and building 8 and 9 have four floors. the measurement has been taken using a tektronix spectrum analyzer covering a frequency band up to 1ghz, connected to a dipole antenna that matches the downlink frequency to measure the power level in dbm at several locations. to duplicate and confirm, our measurements a mobile application been used to indicate the power levels as well. the application which called mobile network emr application was used to measure that strength level from designated base station. table 2 shows the power levels measured in dbm and sar calculated values at different locations (buildings) in the university. using equation (2) to calculate sar, it is assumed the average parameters for that conductivity of the tissue σ = 150 (s/m), and mass density of the tissue ρ = 1250 (kg/m3), the electric field intensity e is determined from (3),[11] where η is the free space characteristic impedance = 120 π ω ≈ 377 ω and s is power density e s2 � . �� (3) according to the measurements and calculation the highest sar level at building 8 and research center, which is about 7.17 × 10-9 w/kg, and it is considered too small comparing to the international standard. conclusion the work shows that there are many factors effect on the sar level, it can be summarized by two main parts, first on the base station parameters (power transmit, antenna types, antenna installing angles, and the distance), and the second part is the body mass and time can be explosion from that radiation. figure 3: antenna radiation pattern for mobile handsets, (a) omnidirectional antenna, (b) massive multi input multi output antenna beamforming toward the bottom stations b a table 2: strength measurement and sar calculated at cihan university-erbil location power (dbm) sar building 1 -76 1.13638e-09 building 2 -76 1.13638e-09 building 3 -86 1.13638e-10 building 4 -80 4.524e-10 building 5 -85 1.43061e-10 building 6 -71 3.59354e-09 cafeteria -66 1.13638e-08 building 8 -68 7.17006e-09 building 9 -74 1.80104e-09 research center -63 2.26737e-08 mohammed: electromagnetic field pollution level measurement for mobile phone networks 34 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 31-34 at cihan university-erbil campus, the measurement proves that the sar level for the mobile base station operating onsite is below the standard sar limit value and it will not be considered as hazard. meanwhile; we believe the radiation from mobile handsets results higher sar level to the users when they are in call or data usages comparing to the sar level from base station at cihan university. references 1. d. seabury and e. t. s. lindgren. an update on sar standards and the basic requirements for sar assessment. available from: https://www.conformity.com [last accessed on 2023 jan 01]. 2. ieee standard for safety levels with respect to human exposure to radio-frequency electromagnetic fields, 3 khz to 300 ghz (ieee c95.1-1991). the institute of electrical and electronics engineers, inc., new york, 1992. 3. international commission on non-ionizing radiation protection. guidelines for limiting exposure to time-varying electric, magnetic, and electromagnetic fields (up to 300 ghz). international commission on non-ionizing radiation protection. health physics, vol. 74, pp. 494-522, 1998. 4. available from: https://en.wikipedia.org/wiki/specific_ absorption_rate [last accessed on 2023 jan 11]. 5. v. g. khurana. mobile phones and brain tumours-a public health concern. surgical neurology, vol. 77, p. 205-214, 2009. 6. s. park, y. shin and d. kim. design and implementation of protocols related to radio network layer over enb in 3gpp lte system 2007 ieee 66th vehicular technology conference, 2007. 7. p. a. ramsdale and a. wiener. cellular phone base stations: technology and exposures. radiation protection dosimetry, vol. 83, no. 1-2, pp. 125-130, 1999. 8. s. sun, t. s. rappaport, r. w. heath, a. nix and s. rangan. mimo for millimeter-wave wireless communications: beamforming, spatial multiplexing. ieee communications magazine, new york, 2014. 9. g. p. kumar, n. agarwal, p. kranthi and s. s. babu. design considerations to calculate sar in multiband mimo antenna for mobile handsets ieee wispnet 2016 conference, 2016. 10. g. p. kumar and s. s. babu. design of multiband mimo antenna in compact mobile handsets and sar distribution, ieee iccsp 2015 conference, 2015. 11. j. k. raines. simple formula relating dbm and dbu. vol. 18, no. 2. ieee broadcast technology society newsletter, new york, pp. 27-29, summer 2010. tx_1~abs:at/add:tx_2~abs:at 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 research article proteinase inhibitors of pigeonpea cv. bsmr 736: characterization and bioefficacy against helicoverpa armigera(hübner) faiyaz k. shaikh1,2, sarwan w. bradosty3, prafull p. gadge2, manohar v. padul4, bhimrao v. jaiwal5, ajit b. patil5 1centre for biotechnology, pravara institutes of medical sciences (du), loni, maharashtra, india, 2department of biochemistry, dr. babasaheb ambedkar marathwada university, aurangabad, maharashtra, india, 3department of community health, college of health technology, cihan university-erbil, kurdistan region, iraq, 4department of biochemistry, institute of science, mumbai, maharashtra, india, 5department of biochemistry, ghulam nabi azad art, commerce and science college barshitakli, akola, maharashtra, india abstract pigeonpea is an agriculturally important leguminous crop with high vulnerability to insect pest attack specifically, helicoverpa armigera. the proteinase inhibitors (pis) mediated host plant resistance against insect pests is a promising sustainable agricultural research practice. the current study was carried out to perceive biochemical characterization of proteinase inhibitors named ppti in the pigeonpea (cv. bsmr 736). the purification of ppti from crude protein seed extract was achieved by acetone precipitation, n-lp-ief, and trypsin affinity chromatography. it was found to inhibit bovine trypsin and hagps in vitro. the optimal conditions for inhibition were ph  8 and temperature 40ºc. the ppti showed four isoinhibitors bands on native, non-reducing and reducing sds-page in the range of 26.7–19.3 kda. on resolution on two-dimensional gel electrophoresis (2-de), ppti produced nine pi variant spots having isoelectric point (pi) 6.6, 6.6, 6.3, 6.1, 5.9, 5.8, 5.7, 5.6, and 5.6. an artificial diet containing ppti reduced the h. armigera larval weight about 69%, with 25% mortality. for eco-friendly sustainable agricultural practices, natural compounds like ppti could be expressed in transgenic crops to prevent the invasion of h. armigera in pigeonpea. keywords: proteinase inhibitors, pigeonpea, ppti, helicoverpa armigera, hagps introduction proteinase inhibitors (pis) are plant proteins regulating endogenous proteinases and acting as defense molecules against biotic stresses.[1,2] they occur in tubers, seeds, leaves, and flowers.[3] with constitutive occurrence, enhanced expression was observed upon insect and pathogen attacks.[4] they are effective in the treatment of cancer, blood clotting, hemorrhage, and inflammation.[5] they are also useful as markers in studies of evolution and systematics.[6,7] pigeonpea (cajanus cajan l.) is a leguminous crop cultivated by farmers of subtropical and tropical regions. it is a major crop of proteinrich diet among others for the vegetarian population of the world. the range of biotic factors constrains the productivity of this crop. it is host to around 200 species of insects, among them helicoverpa armigera is vivacious.[8] helicoverpa armigera belongs to lepidoptera species in the family noctuidae. it is also named as cotton bollworm or corn earworm or scarce bordered straw.[9] the larvae of h. armigera feed on a wide range of plants, including significant no. of cultivated crops. the pest account about us$ 317 million loss in semiarid tropics in pigeonpea crop whereas above us$ 2 billion in other crops.[10] farmers resort to the use of environmentally aggressive and ineffective pesticides to overcome these losses.[11] hence, the probing and improving crop cultivars displaying resistance to h. armigera has huge potential in pest management.[12] host plant resistance could be attained through the use of pis and is promising sustainable agricultural research practices.[13,14] earlier insect in vivo feeding assays using artificial diet incorporated pis and transgenic plants with corresponding author: faiyaz k. shaikh, centre for biotechnology, pravara institutes of medical sciences (du), loni 413 736, maharashtra, india. e-mail: faizbiochem@gmail.com received: june 20, 2022 accepted: july 7, 2022 published: august 01, 2022 doi: 10.24086/cuesj.v6n2y2022.pp9-16 copyright © 2022 faiyaz k. shaikh, sarwan w. bradosty, prafull p. gadge, manohar v. padul, bhimrao v. jaiwal, ajit b. patil. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) shaikh, et al.: proteinase inhibitors of pigeonpea cv. bsmr 736 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 pis expressions have shown promising results.[15] the pis are known to interfere with the digestive process of insects by inhibiting gut proteolytic enzymes. the arresting proteolytic activity causes limitation in the amino acids indispensable for the stages of development of insects.[16] several pis have been identified from seed extracts of pigeonpea.[17] pigeonpea encompasses predominantly pis of the bowman-birk pis (bbis) family.[18] bbis were found to be ineffectual against hagps.[19] the probable cause of feebleness lies in the dynamic gut protease expression system of h. armigera. the gut is found to express proteases that are pis insensitive or degrading pis.[20,21] alongside bbis, kunitz pis are also reported from pigeonpea.[22] henceforth, the design of suitable pis based defense strategies against h. armigera requires a broad understanding of various classes of pis in pigeonpea as well as their effectiveness against h. armigera. the current study was commenced to perceive biochemical nature of pis in the pigeonpea (cv. bsmr 736). materials and methods chemicals and accessories immobilized trypsin agarose and porcine skin gelatin from porcine were procured from sigma-aldrich, usa. protein ladders were procured from genei laboratories pvt. ltd, banglore, india. gelatin coated x-ray films were purchased from fuji film, usa, and selvas photographic ltd., silvassa. ipg strips (11 cm, ph gradients 3-11), reagents and buffers required for 2d electrophoresis were obtained from ge healthcare bio-sciences corp., usa. the chemicals required for solution assays and 1d electrophoresis were procured from sisco research laboratories pvt. ltd. (srl), india. all chemicals and accessories utilized in this research were of the utmost purity available. pigeonpea seeds sample the pigeonpea seeds (cv. bsmr 736) were acquired from the agricultural research station, badnapur (ms) india. procurement of insects the larvae (2nd and 4th instar) of h. armigera were obtained from chickpea and pigeonpea meadows at maliwada, aurangabad maharashtra, india. in vivo insect feeding bioassays were performed with a chickpea flour-based synthetic diet.[23] extraction of ppti from pigeonpea the seeds of pigeonpea were finely powdered in a mixer grinder. the solvents acetone and hexane were used for depigmentation and defatting respectively. the resulting dry defatted (fat removed) powder was adjourned in 1:6 (w/v) 100 mm tris-hcl buffer with ph 7.8 comprising 1% polyvinylpyrrolidone with stirring conditions for 120 min. the resulting homogenate was further spin at 6,000 revolutions per minute (rpm) for 20 min at 4°c. the clear solution obtained by centrifugation was used for solvent precipitation of crude proteins by cold acetone. acetone precipitated crude proteins were dialyzed against distilled water. the resulting precipitate was freeze-dried. helicoverpa armigera gut proteinases (hagps) extraction the h. armigera larvae were dissected at a lower temperature and stowed at –20°c. the gut is ruptured in alkaline buffer (100 mm glycine-naoh buffer, ph 9.6) at 10°c. the resulting lysate was centrifuged at 10,000 rpm for 20 min at 4°c. the clear solution obtained was used as crude hagps.[3] native liquid-phase isoelectric focusing (n-lp-ief) of ppti the crude proteins extract was loaded on n-lp-ief by mini rotofortm cells (bio-rad laboratories india pvt. ltd.). about 18 ml crude proteins extracts (0.4 g of proteins) were mixed along with 0.5% triton x-100 and 20% glycerol. the 2% amphoteric molecules (40% ampholytes, biolyte®, bio-rad) were selected/2000 µg of protein. partitioning of crude proteins was carried out using ph 3–10 gradients at a continuous voltage of 12 w. the fractions obtained were further assessed for the presence of pis using reveres zymography and solution assays. the fraction enriched with pis activity was concentrated in 1m nacl through an ultrafiltration membrane (amicon ultracell®10k, millipore). the resulting concentrated fraction (ppti) was subjected to affinity chromatography. affinity chromatography of ppti the concentrated protein fraction (no. 13) acquired by n-lp-ief was subjected to affinity chromatography using immobilized trypsin agarose column (1.5 ml). the suspension contain (16) units/ml trypsin packed gel. the column was equilibrated with 100 mm tris-hcl buffer with ph 7.8. repeated elution of fractions of ppti was eluted in 0.1n hcl with ph 3.0. the obtained fractions condensed and dialyzed using an ultrafiltration membrane. detection of ppti by gxcp technique and reverse zymography pigeonpea crude pis and affinity chromatography purified ppti were qualitatively analyzed by gel-x ray-film contact print (gxcp) technique[24] plus reverse zymography. pis were applied on 1-dimensional native page (polyacrylamide gel electrophoresis).[25] the pis were electrophoresed at a constant current (20 ma) until the tracing bpb (bromophenol blue) dye touched the end of the gel. for the gxcp technique, the gel was submerged in 100 mm tris-hcl buffer, ph 7.8, and then immersed in 100 µg/ml trypsin for 10 min. after removing excess trypsin, the gel overlapped on the x-ray film for 10 min at 37°c. after incubation, hydrolyzed gelatin from x-ray film was removed by washing with distilled water. the pis bands were seen as unhydrolyzed gelatin. the x-ray film was then photographed. the resulting gel, after washing with water stained for protein bands using coomassie brilliant blue r 250 (cbb r 250) dye. the identical protocol was employed for detecting pis of hagps using 100 mm glycine-naoh, 0.3 m cacl 2 buffer, ph 9.6. the experimentation was repeated thrice with 3 repeats. to analyze pis by reverse zymography, the native page gel was immersed in 100 mm tris–hcl buffer, ph 7.8 for 10 min and further incubated in 1% casein (made in 100 mm tris–hcl buffer with ph 7.8) for 20 min. subsequently, shaikh, et al.: proteinase inhibitors of pigeonpea cv. bsmr 736 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 the gel was immersed in trypsin (0.1%) solution for 10 min at 37°c. afterward, the gel was washed with distilled water and stained with cbb r 250 dye. the polypeptide content of ppti was explored by 12% reducing sodium dodecyl sulfate– polyacrylamide gel electrophoresis (sds-page).[26] ppti visualization by 2-de (2-dimensional electrophoresis) pigeonpea crude pis and affinity chromatography purified ppti were further analyzed by 2-de and 2-d reverse zymography using 7 cm ipg strip with 3–10 ph gradients (bio-rad, usa). rehydration of ipg strips was carried out using buffer comprising 2% chaps (w/v), 0.2% (v/v) ipg buffer, 2m thiourea, 7m urea, pigeonpea pis, and trace of bpb dye. the ief (isoelectric focusing) was carried out at 20ºc on the protean® ief system (bio-rad, usa) according to manufactures protocol (catalog # 163-2099). after ief, strip was equilibrize for 30 minutes in equilibration buffer (2% sds, 30% glycerol), and 50 mm tris–hcl buffer with ph 8.8 at 37°c. the ipg-strip was further exposed to 12% sds page and the gel subsequently was subjected to reverse zymography by the following methodology mentioned above. the resulting protein spots were visualized by cbb r-250 and silver stain. using 2d-sds page standards, isoelectric points (pis) of ppti were determined. solution assay of pis pis activity was assessed using method reported earlier by kakade et al.[27] the reaction mixture comprising pis and trypsin in hcl and 100 mm tris–hcl buffer, ph 7.8, 300 mm cacl 2 were incubated for 10 min. to this 300 µl, bapna (1 mm in dimethyl sulfoxide) was added. the reaction mixture was adjusted to 1.2 ml by the buffer. after end of reaction time, that is, 30 min, 300 µl 30% acetic acid was added to terminate the reaction. the blank and enzyme control tubes were prepared concurrently. the optical density was computed at 410 nm. the pis activity against hagps was calculated by mixing the pis with hagps for 20 min at 37°c in 100 mm glycine-naoh buffer with ph 9.6 containing 300 mm cacl 2 . one unit enzyme activity is defined as the release of 1.0 µm of p-nitroaniline per minute under standard conditions whereas one pis unit could be inhibition of one enzyme unit. effects of ph on ppti the buffer systems, that is, 100 mm glycine-hcl (ph 2), 100 mm sodium citrate (ph 4 and 6), 100 mm tris-hcl (ph 8), and 100 mm glycine-naoh (ph 10, 12) were used to study effect of h+ ion concentrations on ppti activity using standard assay parameters. effects of temperatures on ppti temperatures range 20°c–80°c (at 10°c intervals) in dry water bath was used to study effect of temperature on ppti activity using standard assay parameters. insect bioassay of ppti the larvae of h. armigera (2nd instar) were nurtured on a synthetic chemical diet comprising altered dosages of ppti maintaining 28 ± 2°c and 16:8 light/dark day length conditions. the experiment was performed in three replicates using a 100 ml synthetic chemical diet.[23] the pis was integrated into the synthetic chemical diet at 3 altered dosages, that is, 5, 10, and 15 µg/g of food, w/w). the diet deprived of pi fraction was considered as the control diet. all the experimental food was solidify at 4°c for 24 h before offered to the h. armigera. all larvae were distributed in individual vials for feeding experiment and nourished daily. the food in vials were changed daily and not permitted to dry or be fully consumed. the weights of h. armigera were noted at identical time daily. protein determination the total protein was quantified using method devised by lowry et al. using bovine serum albumin (bsa) as the standard protein.[28] statistical analysis the experimental data reported in the present research were performed in triplicate. the statistical analysis, mean ± s.d, and anova (one-way analysis of variance) were performed in microsoft excel 2010. the statistical significance examined with p ≤ 0.001. results and discussion pis act as the plant’s endogenous arsenals to counterbalance insect pest activities. overexpression of pis in the most vulnerable target tissues and/or raising transgenic plants expressing pis is the current strategy against insect pests.[29,30] the exploration of pis based h. armigera resistance in pigeonpea may prove a vital component in ipm (integrated pest management). the current investigation was aimed at exploring the biochemical nature and characteristics of ppti from pigeonpea cv. bsmr 736. pigeonpea pis extraction and detection as per the literature review, most of the pigeonpea cultivars including bsmr 736 exhibits monomorphic pis proteins pattern on the electrophoretic gel.[19] here in accordance, we are also reporting monomorphic pis bands from the crude proteins extract of pigeonpea cv. bsmr 736 by gxcp technique against bovine trypsin [figure 1]. the gxcp facilitates simple and rapid analysis of the proteinases and their inhibitors from seeds and other samples. prefractionation of pis by n-lp-ief several research groups employed various strategies for the prefractionation of proteins before electrophoretic characterization and preparative isolation of desired protein and n-lp-ief was found to be convenient.[31,32] the n-lp-ief allows the separation of protein even if it is present in low abundance in the sample based on their isoelectric point (pi). conventional fractionation strategies such as (nh 4 ) 2 so 4 precipitation, subsists traces of salts in the sample which creates trouble in further purification and identification of molecules. hence, to overcome these issues, we applied acetone precipitated protein onto n-lp-ief. about 20 fractions shaikh, et al.: proteinase inhibitors of pigeonpea cv. bsmr 736 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 were collected after n-lp-ief ranging in ph from 3 to 10. the prefractionation by n-lp-ief revealed that pis was restricted to acidic to neutral fractions with maximum at pis 6.6, 6.6, 6.3, 6.1, 5.9, 5.8, 5.7, 5.6, and 5.6. (fraction no.13). earlier pis showing activity in the acidic to neutral pis were reported in the pigeonpea cv. abhaya and icp 7118.[18,33] this prefractionation also showed the presence of more number of pi bands in fraction 13 on x-ray films suggesting active fractionation of low abundant pis. figure 2 displays the sds-page stained gel map of the total fractionated pis (20 fractions) collected from the n-lpief with equal volume loaded on the gel. it indicates the distribution of pis in different fractions, and the relative numbers of pis of each fraction are also different. however, it was observed that most of the pis had diffused into several neighboring fractions. this may be due to the diffusion of pis in the solution since it is not isoelectrically focused as it would be in a gel.[34] fraction 13, showed more activity, and numbers of pis was used for further purification purpose. trypsin-agarose affinity chromatography the n-lp-ief fraction was further applied to the trypsinagarose affinity column. a trypsin-agarose resin is selectively purifying an inhibitor of trypsin. the pis bound to the matrix were eluted in 0.1n hcl with ph 3.0. these pis were collected, condensed and dialyzed using membrane ultrafiltration. the unbound fraction did not show any ti activity. the concentrated fraction was eluted from the trypsin-agarose column showing ti activity at 280 nm was named as ppti and used in further characterization study. one dimensional electrophoretic characterization of ppti on affinity purification, the ppti displayed separation of 4 pis bands, namely, pi1, pi2, pi3, and pi4 on native as well as nonreducing sds-page as compared to pi pool present in the crude seed extract. this might be due to the self-association tendency of pis during purification stages. earlier such a phenomenon of self-association was observed in bbi like pis.[35] such pis were also reported from two cultivars of pigeonpea.[20,21] the ppti showed inhibitory activity against bovine pancreatic trypsin and hagps checked by native reverse zymography [figure 3]. treatment of ppti with β-mercaptoethanol slightly alters the migration of the pi4 band on the gel indicating its reduction [figure 4]. the four pis on non-reducing and reducing sdspage with β-mercaptoethanol exhibit molecular weights in the range of 26.7–19.3 kda. this finding is in agreement with earlier studies that reported high molecular weights of pis from various legume seed extracts.[36] the self-association of pi molecules might result in the overestimation of molecular mass as reported for several legumes.[37,38] furthermore, the occurrence of ppti in a group of four isoinhibitors may be attributed to the particularly individual narrow range masses and high molecular weights of these pis may also be associated with the different degrees of glycosylation.[39] other possibilities like oxidation of cysteine residues have been reported earlier.[40] similar findings of purification of ti with a group of three isoinhibitors were reported from acacia victoriae bentham seeds.[41] the functional role of occurrence of many isoinhibitors is not clear; it seems to be a part of approaches adopted by the host plant to ensure its existence. inhibition assay of ppti the trypsin and hagps activities were reduced in the presence of crude pis as well as an affinity-purified ppti in vitro [figure 5]. previously, moderate to low inhibition potential against hagps was reported from several pigeonpea cultivars by in vitro and in vivo bioassays.[19,23] in agreement with the above studies, our finding advocates that pigeonpea (cv. bsmr 736) could be a potent source of pis. optimum ph of ppti in general, legumes pis are identified to be stable under extremes of ph. the ppti was found to be active at a varied range of phs 3–12 with optimum inhibition at ph 8 [figure 6]. this finding is in agreement with earlier studies where optimum pis activity was retained between ph 4 and 10.[16,36,37] the optimum activity of pi from other legumes such as mungbean (v. radiate l. wilczek) seeds was also observed at ph 8.0.[41] the gut content of h. armigera is highly alkaline figure 2: isoelectric point (pi) based fractionation of acetone precipitated protein n-lp-ief was carried out between ph 3-10. an equal volume of each fraction was loaded and visualized by sds-page gxcp. arrow (↓) indicates fraction (13) used for subsequent analysis figure 1: detection of tis of pigeonpea by gxcp technique. the sample (100 µg) was loaded in each lane (i-iv) on 10% native page, pis activity bands were detected by immersing the gel in bovine trypsin and overlapping gel on the x-ray film shaikh, et al.: proteinase inhibitors of pigeonpea cv. bsmr 736 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 figure 3: electrophoretic analysis (10% native page reverse zymography) of ppti. lanes i-iv crude protein extract (100 µg), the active fraction (fraction-13) from n-lp-ief (50 µg), affinity column pooled fraction (15 µg), and membrane filtration fraction (10 µg; 10 kda cut-off), respectively with maximum hagps activity at ph 10.0–11.0.[42] this result with ppti indicated that it is suitable against hagps, which work in the alkaline gut of h. armigera. optimum temperature of ppti the optimum temperature of ppti was found to be 40°c and showed considerable activity even at 80°c [figure 7], suggesting that ppti is cysteine-rich which imparts high stability and conformational rigidity at high temperatures.[43] this study is in agreement with earlier finding where pis with stability up to a temperature of 80°c was reported from pigeonpea.[16,37] analysis of pi variant isoforms of ppti the isoinhibitors complexity of affinity-purified ppti was further resolved by 2-de. figure 8 illustrates 2-de analyses figure 4: 12% sds-polyacrylamide electrophoresis of ppti. lane m, molecular weight standards; lane i crude extract, lane ii n-lp-ief fraction no. 13, lane iii ppti with βmercaptoethanol; proteins were detected by dye cbb r-250 figure 6: effect of ph on ppti activity. an assay was executed in altered buffer solutions, as revealed in the methodology section. the results are mentioned as mean ± sd, n = 3 figure 7: effect of temperature on ppti. an assay was executed at altered temperatures for 10 min as mentioned in the methodology section. the results were presented as mean ± sd, n = 3 figure 5: trypsin and hagps inhibition assays by (a) crude seed protein, (b) acetone ppt. protein, (c) n-lp-ief fraction, and (d) ppti. the enzyme activity was assayed using bapna. the results were reported as mean ± sd, n = 3 a b dc shaikh, et al.: proteinase inhibitors of pigeonpea cv. bsmr 736 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 of pigeonpea seed proteins. the pigeonpea cv. bsmr 736 is a rich source of proteins hence several proteins with varied mw and pi were seen on the gel [figure 8a] when stained with cbb r-250. subsequently, staining of the gel with silver stain increased the quantity of proteins bands on the gel corroborating the sensitivity of strain [figure 8b]. when the gel was digested with trypsin in reverse zymography, most of the proteins disappeared due to the hydrolytic action of trypsin [figure 8c]. several trypsin inhibitors or trypsinresistant proteins could be detected on the lower and upper panels of the gel. the lower panel of the gel is shown by the circle corresponding to the affinity purified ppti. the 2-de analysis showed 9 pi variant isoforms of ppti with pi range 5.6–6.6, matching their isoinhibitors migration on the gel [figure 9]. for instance, they were collected from alkaline regions earlier only very slight pi dependent variations were observed among them with molecular weights nearly in same the range as observed in 1-d sds page with β-mercaptoethanol [figure 9a]. the ppti remained active after exposure to several detergents and urea used in 2-d gels assessed by reverse zymography [figure 9b]. the pi variant isoforms showing identity toward bbi like pis were reported from the seeds of pigeonpea cv. abhaya and cv. icp 7118.[16,19] the advent of different pis classes in the same plants with the potential to inhibit the same enzyme is due to convergent evolution.[44] bioefficacy of ppti the kunitz, bbi, cysteine, knottin pis, and lectins are the most studied pis showing future potential to use them in insect resistance strategies.[45,46] the faith of the pi strategy can largely depend on the nature of the insect response to the ingested pis. hence, the effectiveness of affinitypurified ppti against hagps was assessed by integrating 3 altered dosages (i.e., 5, 10 and 15 µg/gm w/w of diet). here artificial diet containing ppti reduced the h. armigera larval weight by 69%, with 25% mortality. the average larval weight was reduced in the control from 689 ± 17.45 mg to 214 ± 12.45 mg when fed a 15 µg/gm diet, at the end of the feeding assay period (10 days), indicating a substantial decline in the growth and development of h. armigera with p ≤ 0.001 [figure 10]. conclusions in a nutshell, the above results indicate that the ppti is responsible for the reduction in the growth of h. armigera with low mortality. further research in understanding the proteomic nature of isoinhibitors variant isoforms of ppti is required. comprehensive knowledge of the occurrence of isoinhibitors of ppti in host plants like pigeonpea might provide insect resistance modules with resistance against h. armigera invasion. acknowledgment dr. faiyaz khudaboddin shaikh is thankful to university grant commission (ugc) and ministry of minority affairs, government of india, new delhi for maulana azad national fellowship (manf). references 1. p. r. shewry. tuber storage proteins. annals of botany, vol. 91, pp. 755-769, 2003. figure 10: effect of ppti on the development of h. armigera larvae. the larvae were permitted to feed on a synthetic chemical diet with ppti and without ppti. larval weight was noted every 24 h. each mean signifies three replicates ± sd, and a difference among analyses of p ≤ 0.001 was considered significant figure 9: two-dimensional electrophoretic analyses of ppti isoforms. (a) ppti protein spots (b) reverse zymogram. un-digested proteins signify pis. black arrow specify the location of pis. lane i, protein ladder in kda; lane ii, purified pis (ppti) from affinity column. the phs range is shown on the gel a b figure 8: two-dimensional electrophoretic analyses of pigeonpea (bsmr 736) seed proteins. (a) cbb r-250, (b) silver staining, and (c) pis activity visualization by reverse zymography b ca shaikh, et al.: proteinase inhibitors of pigeonpea cv. bsmr 736 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 9-16 2. m. clemente, m. g. corigliano, s. a. pariani, e. f. sánchez-lópez, v. a. sander and v. a. ramos-duarte. plant serine protease inhibitors: biotechnology application in agriculture and molecular farming. international journal of molecular sciences, vol. 20, no. 6, pp. 1345, 2019. 3. m. v. padul, r. d. tak and m. s. kachole. protease inhibitor (pi) mediated defense in leaves and flowers of pigeonpea (protease inhibitor mediated defense in pigeonpea). plant physiology and biochemistry, vol. 52, pp. 77-82, 2012. 4. f. d. leo and plant-pis: a database for plant protease inhibitors and their genes. nucleic acids research, vol. 30, no. 1, pp. 347-348, 2002. 5. g. c. mello, i. a. desouza, s. marangoni, j. c. novello, e. antunes and m. l. macedo. 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ling, h. y. qi and c. w. chi. protein, cdna, and genomic dna sequences of the towel gourd trypsin inhibitor. a squash family inhibitor. journal of biological chemistry, vol. 268, no. 2, pp. 810-814, 1993. 44. b. prakash and m. r. murthy. source and target enzyme signature in serine protease inhibitor active site sequences. journal of biosciences, vol. 22, no. 5, pp. 555-565, 1997. 45. f. k. shaikh, s. w. hamad, s. w. hamad and a. a. shinde. in vitro screening of seed extracts of medicinal plants for protease inhibitory activity. cihan university-erbil scientific journal, vol. 3, no. 1, pp. 61-65, 2019. 46. a. a. shinde, f. k. shaikh and m. s. kachole. exploration of amylases producing competency of helicoverpa armigera gut bacterial strain, bacillus subtilis rtsba6 6.00. cihan universityerbil scientific journal, vol. 3, no. 1, pp. 75-79, 2019. tx_1:abs~at/tx_2:abs~at 85 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 85-89 research article effect of isatis spp. extraction on the growth of aspergillus niger and candida albicans zehra odabaş-serin1*, ali h. salim agha2, zhala b. taha2 1department of forest, kahramanmaraş sütçü imam university, faculty of forestry, 46040, kahramanmaraş, turkey, 2department of bioengineering and science, kahramanmaraş sütçü i̇mam university, faculty bioengineering and science, 46040, kahramanmaraş, turkey and cihan university-erbil, department of biology, iraq abstract in this study, the crude extraction of isatis spp. (isatis tinctoria, isatis busichaina, and isatis lusitanica) was investigated for their antifungal activity. each of methanol, ethanol, and h2o was used as a solvent. due to find the most effective part of each species, the plant parts were used separately. the aim of this research was to determine the natural products effect on two common pathogenic fungi aspergillus niger and candida albicans. the results show that most of used plants have significant effect on both used fungal species. extracted each of flower, i. tinctoria was showed the best results comparing to the other used species. extracted leaf and flower of i. tinctoria by methanol were showed the best result on a. niger and c. albicans. the growth zone was around 90 mm for control and 61 mm at 75% concentration of methanolic and ethanolic extraction. the flower was for followed by the stem. depend on the results, the methanol was showed highest number, then ethanol and the lowest inhibition zone area was for h2o extraction. keywords: antifungal activity, aspergillus niger and candida albicans, isatis spp., pathogenic fungi introduction a latest study presented that tertiary care clinics are consuming antimicrobial and antifungal agents improperly. this rising trend is an ever-increasing threat to the health of humans who are currently at risk of dying from previously fixable infections. unluckily, drug resistance (adaptation of the drug target site, drug modification, or restricted drug penetration) improves at a pace quicker than new drug development.[1] this sensation is also happening in the pathogenic microorganism candida albicans, an otherwise commensal kinds found in and on the human body.[2] modern antifungal agents are less active against c. albicans now than when they were first introduced because candida species have developed resistance to these agents. in addition, research by the major pharmaceutical companies in the development of antifungal agents has been on the decline due to low coming back on investment.[3] materials and methods plant extracts three methods were used to extract water, ethanolic, and methanolic, according to several studies, with some modification as needed and as follows:[4] aqueous extraction hot water extraction was performed by mixing 40 g of plant form with 160 ml of distilled water twice (i.e., 1:4 weight:volume), heated to 80–90°c, then stirred and left to cool down and mixed together then placed in refrigerator for 24 h, which was then administered through several layers of medical gauze, and again administered by buchner funnel using filter papers whatman no.1 to dispose of the nonpowdered parts and fibers to obtain the raw liquid extract and then put it in the rotary evaporator device at a temperature not exceeding 40°c, working on the basis of evaporation under pressure and then put the extracted in the shaker incubator at a temperature of 30–35°c. after the drying of the extract, corresponding author: zehra odabaş-serin, kahramanmaraş sütçü imam university, faculty of forestry, 46040, kahramanmaraş, turkey. phone: +90 3443001780. fax: +90 3443001712. e-mail: zehra@ksu.edu.tr received: jan 16, 2020 accepted: apr 27, 2020 published: jun 20, 2020 doi: 10.24086/cuesj.v4n1y2020.pp85-89 copyright © 2020 zehra odabaş-serin, ali h. salim agha, zhala b. taha. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) mailto:zehra@ksu.edu.tr https://creativecommons.org/licenses/by-nc-nd/4.0/ odabaş-serin, et al.: antifungal activity of isatis spp 86 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 85-89 it was kept in a sealed containers in the freezer (−4°c) and wrote the information on each until use. ethanolic extraction prepare the ethanol extract by mixing 20 g of plant powder in 200 ml of absolute ethanol with stirring (i.e., 1:10 weight:volume) and then leave the mixture in the refrigerator for 24 h. then, the extract was filtered through several layers of gauze and then filtered for 2nd time using whatman no.1 filter papers to remove the non-powdered plant parts and the remaining fibers, then put in the rotary evaporator device at 40°c and after evaporation of the alcohol obtained a thick layer of the extract and then placed in the shaker incubator shaker incubator at 25–30°c. after drying the extract, it was kept in a sealed container in a freezer with writing the information on each until use. methanolic extraction prepare the methanol extract by mixing 20 g of plant powder in 200 ml of absolute methanol with stirring (i.e., 1:10 weight:volume) and then leave the mixture in the refrigerator for 24 h to soak. then, the extract was filtered through several layers of gauze and then filtered for 2nd time using whatman no.1 filter papers to remove the non-powdered plant parts and the remaining fibers, then put in the rotary evaporator device at 40°c and after evaporation of the methanol obtained a thick layer of the extract and then placed in the shaker incubator shaker incubator at 25–30°c. after drying the extract, it was kept in a sealed container in a freezer with writing the information on each until use. media preparation the above studied media were prepared according to the information on the package by the manufacturer company[5] then sterilized at 121°c and 15 lb/kg2 for 15 min. the medium was distributed in petri dishes (diameter 9 cm), and the dishes were left under the temperature of the laboratory until the hardening of the medium in sterilizer hood. a quantity of distilled water was also sterilized for use as a substitute for extracts in control transactions (ddh 2 o). fungi two types of fungus obtained from the media center in erbiliraq for diagnosing the disease with mention the strain type were used in this study. • aspergillus niger atcc 16404 • c. albicans atcc 10231 preparation of petri dishes to test extracts against fungus for this study, concentration method was used by mixing with the medium. the fungal isolates were planted in petri dishes on the potato dextrose agar media, mixed with three different concentrations of plant extracts that mentioned in previously, then incubated the dishes in the incubator at 25 ± 2°c and 37 ± 2°c. for example, candida species need a temperature of between 25 ± 2°c and 37 ± 2°c for optimal growth according to the nature of their growth to fungus or semi-yeast. the process of incubation continued for 10 days. the readings were taken every 3 days according to replicates, influenced by concentrations of plant extracts.[6,7] statistical analysis glm-univariate analysis was applied to the arithmetic means obtained in spss with 95% confidence interval. if the difference was significant (p ≤ 0.05), the homogeneity groups were determined by tukey b test. the analytical test was done separately for each solvent type (hot water, ethanol, and methanol) to show the effect of the samples part (flower, leaf, and stem) and extract concentration (25%, 50%, and 75%). results and discussion the antifungal compound is one of the vital substances in pharmaceutical sciences that can find in the natural medicinal plants. this needs to identify the activity of the samples compound by different parameters. in this study, the antifungal activities of extracts obtained with solvents that vary from nonpolar to polar were investigated. the results were took after the growth was rich the optimum point of two types of used fungi in optimum incubation condition. the growth of the control petri dishes was 90 mm for both. the following table is showed the effect of isatis tinctoria on a. niger atcc 16404 and c. albicans atcc 10231 growth. the best results were with flower extraction by methanol with 75% concentration against a. niger atcc 16404, following by the stem with methanolic extraction 75% against a. niger atcc 16404 too. in this table, all of the methanolic extraction had significant effect on both fungi a. niger atcc 16404 and c. albicans atcc 10231 [table 1]. the second table is showed the effect of isatis busichaina on a. niger atcc 16404 and c. albicans atcc 10231 growth. the best results were with leaf extraction by methanol with 75% concentration against c. albicans atcc 10231, following by the flower with methanolic extraction 75% against a. niger atcc 16404. in this table, all of the methanolic extraction had significant effect on both fungi a. niger atcc 16404 and c. albicans atcc 10231 [table 2]. however, table 3 shows the effect of isatis lusitanica on a. niger atcc 16404 and c. albicans atcc 10231 growth. the best results were with flower extraction by methanol with 75% concentration against a. niger atcc 16404, following by the flower with methanolic extraction 75% against c. albicans atcc 10231. in this table, all of the methanolic extractions had significant effect on both fungi a. niger atcc 16404 and c. albicans atcc 10231 [table 3]. depend on the results, the significant results were obtained from the extraction of methanol and ethanol for i. tinctoria in all concentrations, plant parts, and both types of used fungi. the aqueous extraction has less effect in correlation with ethanol and methanol [table 4]. the inhibition effects of the extracts of i. tinctoria obtained from leaf, stem, and flower parts using different solvents on the two different strains of pathogenic fungi. the results were compared with the each to find the correlation. the aqueous leaf extracts were showed significant results on a. niger atcc 16404, also the flower was showed the significant results by ethanol extraction for c. albicans atcc 10231, although the flower extracts by methanol were showed the significant results for a. niger atcc 16404. other differences are shown in table 4. odabaş-serin, et al.: antifungal activity of isatis spp 87 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 85-89 table 1: fungi test results of i. tinctoria. (control=90 mm) plant plant part strains growth of fungi (mm) water concentration ethanol concentration methanol concentration 25% 50% 75% 25% 50% 75% 25% 50% 75% i. tinctoria flower c. albicans 85.75 (0.96) 82.75 (0.50) 80.25 (0.96) 81.00 (0.82) 79.00 (0.82) 72.50 (1.29) 73.00 (1.15) 71.75 (0.50) 62.00 (1.41) a. niger 84.00 (0.82) 82.50 (1.29) 79.00 (0.82) 81.25 (0.96) 80.00 (0.82) 78.00 (1.41) 69.75 (0.50) 68.00 (0.82) 61.25 (0.50) leaf c. albicans 84.50 (2.38) 80.75 (0.96) 79.50 (1.29) 79.75 (1.26) 78.75 (0.96) 77.75 (0.96) 70.5 (0.58) 69.25 (0.50) 62.00 (1.41) a. niger 84.00 (0.82) 81.00 (0.82) 79.25 (0.96) 80.25 (1.71) 78.75 (0.96) 77.50 (0.58) 70.25 (0.50) 68.50 (1.00) 62.75 (0.50) stem c. albicans 84.75 (0.96) 82.25 (2.06) 80.50 (0.58) 82.00 (0.82) 80.50 (0.58) 73.00 (1.83) 73.50 (1.29) 70.25 (0.50) 63.25 (1.50) a. niger 84.50 (1.29) 82.75 (0.96) 80.00 (0.82) 81.25 (0.50) 79.25 (0.96) 78.50 (0.58) 70.75 (0.96) 69.50 (1.29) 61.75 (0.50) i. tinctoria: isatis tinctoria, c. albicans: candida albicans, a. niger: aspergillus niger table 2: fungi test results of i. busichaina. (control=90 mm) plant plant part strains growth of fungi (mm) water concentration ethanol concentration methanol concentration 25% 50% 75% 25% 50% 75% 25% 50% 75% i. busichaina flower c. albicans 84.75 (0.96) 80.25 (1.26) 79.50 (1.29) 80.00 (0.82) 78.50 (0.58) 76.00 (1.41) 71.00 (0.82) 70.00 (1.63) 63.00 (1.83) a. niger 83.50 (1.29) 79.75 (0.96) 77.75 (2.22) 80.00 (0.82) 78.25 (0.50) 76.00 (0.82) 70.75 (0.96) 68.00 (0.82) 61.5 (1.29) leaf c. albicans 83.25 (0.96) 79.50 (1.29) 77.00 (1.83) 80.00 (0.82) 79.00 (0.82) 75.50 (1.29) 71.00 (1.15) 66.75 (2.06) 61.00 (0.82) a. niger 83.25 (0.96) 80.50 (0.58) 77.25 (1.71) 80.75 (0.96) 78.50 (0.58) 74.25 (0.50) 70.75 (0.50) 66.25 (0.50) 63.00 (1.83) stem c. albicans 84.00 (0.82) 80.75 (1.50) 77.25 (1.50) 81.75 (0.96) 80.00 (0.82) 78.25 (0.96) 72.5 (2.38) 70.50 (0.58) 68.50 (0.58) a. niger 84.75 (1.50) 82.25 (1.26) 79.25 (0.96) 81.75 (0.96) 80.50 (0.58) 79.25 (0.96) 71.75 (0.96) 70.00 (0.82) 69.00 (1.41) i. busichaina: isatis busichaina, c. albicans: candida albicans, a. niger: aspergillus niger table 3: fungi test results of i. lusitanica. (control=90 mm) plant plant part strains growth of fungi (mm) water concentration ethanol concentration methanol concentration 25% 50% 75% 25% 50% 75% 25% 50% 75% i. lusitanica flower c. albicans 83.25 (0.96) 81.50 (1.29) 79.00 (0.82) 80.50 (0.58) 79.50 (1.29) 78.00 (0.82) 70.00 (0.82) 67.75 (0.50) 64.50 (1.29) a. niger 83.25 (0.96) 80.75 (0.96) 79.00 (0.82) 80.50 (0.58) 78.25 (1.71) 76.75 (0.96) 68.50 (1.29) 66.50 (1.00) 63.75 (1.71) leaf c. albicans 82.25 (1.71) 81.75 (1.71) 79.50 (1.29) 80.00 (0.82) 79.00 (0.82) 76.75 (0.96) 69.75 (0.96) 67.50 (0.58) 66.50 (1.00) a. niger 82.00 (0.82) 80.25 (0.96) 79.50 (0.58) 80.25 (0.50) 78.25 (1.71) 77.50 (1.29) 68.25 (0.96) 65.75 (0.96) 65.25 (0.50) stem c. albicans 84.50 (0.58) 82.25 (0.50) 80.50 (1.29) 81.50 (1.29) 80.75 (0.96) 79.00 (0.82) 72.00 (0.82) 70.50 (0.58) 69.00 (0.82) a. niger 83.50 (1.29) 80.75 (0.96) 80.25 (1.26) 81.50 (0.58) 80.00 (0.82) 79.00 (0.82) 72.00 (0.82) 70.25 (0.96) 69.50 (0.58) i. lusitanica: isatis lusitanica, c. albicans: candida albicans, a. niger: aspergillus niger the inhibition effects of extracts of i. busichaina obtained from leaf, stem, and flower parts using different solvents on the two different strains of pathogenic fungi. the results were compared with the each to find the correlation. for c. albicans odabaş-serin, et al.: antifungal activity of isatis spp 88 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 85-89 atcc 10231, each of aqueous leaf extraction, ethanol extraction of flower and leaf, and leaf extraction by methanol was showed the significant result correlations. for a. niger atcc 16404, the aqueous extraction of flower and leaf, the ethanol extraction of flower and leaf, and methanol extraction of flower and leaf too was showed the significant results. the total result statistics shown in table 5. the inhibition effects of the extracts of i. lusitanica obtained from leaf, stem, and flower parts using different solvents on the two different strains of pathogenic fungi. the results were compared with the each to find the correlation. for c. albicans atcc 10231, each of flower and leaf of aqueous, ethanol, and methanol extraction was showed the significant results. for a. niger atcc 16404, the ethanol and methanol extraction of flower and leaf was showed the significant results. the total result statistics is shown in table 6. some factors have to be measured to select a solvent for extracting herbal medicine. about 65–75% methanol extraction at room temperature for 15 h is practical for routine analytical study. although, it is not fully active to extract other plants with same procedure because of dissimilarities in the biological compositions. this method is appropriate for controlling a large number of fixed samples and needs no special equipment.[8] the present study proved that methanol and ethanol showed best results than water extracts. our study was in accordance with several studies.[9] tryptanthrin, indole-3acetonitrile, and p-coumaric acid methylester were identified by physical and spectral data (ms, nmr) and by comparison with authentic synthesized samples of these compounds. tryptanthrin has already been isolated from i. tinctoria and characterized as an antidermatophytic.[10] phenylpropanoids mainly comprised flavonoid and lignan metabolites, have been considered as the primary antiviral constituents, which table 4: fungi arithmetic values and tukey b statistical analysis results of i. tinctoria c. albicans concentration a. niger concentration tukey b plant part 25% 50% 75% tukey b 25% 50% 75% water flower 85.75 (0.96) 82.75 (0.50) 80.25 (0.96) a 84.00 (0.82) 82.50 (1.29) 79.00 (0.82) ab leaf 84.50 (2.38) 80.75 (0.96) 79.50 (1.29) a 84.00 (0.82) 81.00 (0.82) 79.25 (0.96) b stem 84.75 (0.96) 82.25 (2.06) 80.50 (0.58) a 84.50 (1.29) 82.75 (0.96) 80.00 (0.82) a tukey b a b c a b c ethanol flower 81.00 (0.82) 79.00 (0.82) 72.50 (1.29) b 81.25 (0.96) 80.00 (0.82) 78.00 (1.41) a leaf 79.75 (1.26) 78.75 (0.96) 77.75 (0.96) a 80.25 (1.71) 78.75 (0.96) 77.50 (0.58) a stem 82.00 (0.82) 80.50 (0.58) 73.00 (1.83) ab 81.25 (0.50) 79.25 (0.96) 78.50 (0.58) a tukey b a b c a b c methanol flower 73.00 (1.15) 71.75 (0.50) 62.00 (1.41) b 69.75 (0.50) 68.00 (0.82) 61.25 (0.50) b leaf 70.50 (0.58) 69.25 (0.50) 62.00 (1.41) c 70.25 (0.50) 68.50 (1.00) 62.75 (0.50) a stem 73.50 (1.29) 70.25 (0.50) 63.25 (1.50) a 70.75 (0.96) 69.50 (1.29) 61.75 (0.50) a tukey b a b c a b c i. tinctoria: isatis tinctoria, c. albicans: candida albicans, a. niger: aspergillus niger table 5: fungi arithmetic values and tukey b statistical analysis results of i. busichaina plant part c. albicans concentration tukey b a. niger concentration tukey b 25% 50% 75% 25% 50% 75% water flower 84.75 (0.96) 80.25 (1.26) 79.50 (1.29) a 83.50 (1.29) 79.75 (0.96) 77.75 (2.22) b leaf 83.25 (0.96) 79.50 (1.29) 77.00 (1.83) b 83.25 (0.96) 80.50 (0.58) 77.25 (1.71) b stem 84.00 (0.82) 80.75 (1.50) 77.25 (1.50) ab 84.75 (1.50) 82.25 (1.26) 79.25 (0.96) a tukey b a b c a b c ethanol flower 80.00 (0.82) 78.50 (0.58) 76.00 (1.41) b 80.00 (0.82) 78.25 (0.50) 76.00 (0.82) b leaf 80.00 (0.82) 79.00 (0.82) 75.50 (1.29) b 80.75 (0.96) 78.50 (0.58) 74.25 (0.50) b stem 81.75 (0.96) 80.00 (0.82) 78.25 (0.96) a 81.75 (0.96) 80.50 (0.58) 79.25 (0.96) a tukey b a b c a b c methanol flower 71.00 (0.82) 70.00 (1.63) 63.00 (1.83) b 70.75 (0.96) 68.00 (0.82) 61.5 (1.29) b leaf 71.00 (1.15) 66.75 (2.06) 61.00 (0.82) c 70.75 (0.50) 66.25 (0.50) 63.00 (1.83) b stem 72.5 (2.38) 70.50 (0.58) 68.50 (0.58) a 71.75 (0.96) 70.00 (0.82) 69.00 (1.41) a tukey b a b c a b c i. busichaina: isatis busichaina, c. albicans: candida albicans, a. niger: aspergillus niger odabaş-serin, et al.: antifungal activity of isatis spp 89 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2020, 4 (1): 85-89 table 6: fungi arithmetic values and tukey b statistical analysis results of i. lusitanica plant part c. albicans concentration tukey b a. niger concentration tukey b 25% 50% 75% 25% 50% 75% water flower 83.25 (0.96) 81.50 (1.29) 79.00 (0.82) b 83.25 (0.96) 80.75 (0.96) 79.00 (0.82) a leaf 82.25 (1.71) 81.75 (1.71) 79.50 (1.29) b 82.00 (0.82) 80.25 (0.96) 79.50 (0.58) a stem 84.50 (0.58) 82.25 (0.50) 80.50 (1.29) a 83.50 (1.29) 80.75 (0.96) 80.25 (1.26) a tukey b a b c a b c ethanol flower 80.50 (0.58) 79.50 (1.29) 78.00 (0.82) b 80.50 (0.58) 78.25 (1.71) 76.75 (0.96) b leaf 80.00 (0.82) 79.00 (0.82) 76.75 (0.96) b 80.25 (0.50) 78.25 (1.71) 77.50 (1.29) b stem 81.50 (1.29) 80.75 (0.96) 79.00 (0.82) a 81.50 (0.58) 80.00 (0.82) 79.00 (0.82) a tukey b a b c a b c methanol flower 70.00 (0.82) 67.75 (0.50) 64.50 (1.29) b 68.50 (1.29) 66.50 (1.00) 63.75 (1.71) b leaf 69.75 (0.96) 67.50 (0.58) 66.50 (1.00) b 68.25 (0.96) 65.75 (0.96) 65.25 (0.50) b stem 72.00 (0.82) 70.50 (0.58) 69.00 (0.82) a 72.00 (0.82) 70.25 (0.96) 69.50 (0.58) a tukey b a b c a b c i. lusitanica: isatis lusitanica, c. albicans: candida albicans, a. niger: aspergillus niger probably contribute to the outstanding pharmacological activity of radix isatidis.[11,12] it is distinguished that a solvent system for extraction is selected according to the purpose of extraction such as preparation or analysis, the nature of interested components, the physicochemical properties of the matrix, the availability of reagents and equipment, cost, and safety concerns.[13] the solvent systems included complete ethanol at 25°c by different methods, 95% ethanol, ethanol–water for 10 min,[14] methanol–water, methanol–hcl (1.0 n, 85:15, v/v), and h 2 o. in another study that done by in 2012,[15] using the ethanol under pressure, extraction condition may be a well choice for viable productions, meanwhile, ethanol may be reused and fewer solvent left over is generated through production. however, extraction with ethanol may be a blameless means to make effective extracts for analysis and research from a minor number of samples. references 1. h. zwickey and l. lipski. “expanding our view of herbal medicine”. the journal of alternative and complementary medicine, vol. 24, no. 7, pp. 619-20, 2018. 2. c. van wyk, f. s. botha, r. vleggaar and j. n. eloff. “obliquumol, a novel antifungal and a potential scaffold lead compound, isolated from the leaves of ptaeroxylon obliquum (sneezewood) for treatment of candida albicans infections: oorspronklike navorsing”. suid-afrikaanse tydskrif vir natuurwetenskap en tegnologie, vol. 37, no. 1, pp. 1-5, 2018. 3. c. i. montero, y. r. shea, p. a. jones, s. m. harrington, n. e. tooke, f. g. witebsky and p. r. murray. “evaluation of pyrosequencing® technology for the identification of clinically relevant non-dematiaceous yeasts and related species. european journal of clinical microbiology and infectious diseases, vol. 27, no. 9, pp. 821-30, 2008. 4. n. rispail, p. morris and k. j. webb. phenolic compounds: extraction and analysis. in: lotus japonicus handbook. dordrecht: springer, pp. 349-354, 2005. 5. r. m. atlas, a. e. brown and l. c. parks. laboratory manual of experimental microbiology. usa: mosby-year book. inc., 1995. 6. a. ahmet, s. munevver, o. hakan, a. guleray. in vitro antimicrobial and antioxidant activities of methanol and hexane extract of astragalus species growing in the eastern anatolia region of turkey. turkish journal of biology, vol. 33, pp. 65-71, 2009. 7. y. n. al-shekhany. alkaloid and glycoside contents and antioxidant activity of two heliotropium species (boraginaceae) from kurdistan region-northern iraq. garmian university online journal, vol. 2, pp. 963-979, 2012. 8. x. li, s. feng, a. farajtabar, n. zhang, g. chen and h. zhao. solubility modelling, solvent effect and preferential solvation of 6-chloropurine in several aqueous co-solvent mixtures between 283.15 k and 328.15 k. the journal of chemical thermodynamics, vol. 127, pp. 106-116, 2018. 9. p. tane, s. d. tatsimo, g. a. ayimele and j. d. connolly. bioactive metabolites from aframomum species. in: 11th napreca symposium book of proceedings. vol. 214, pp. 214-223, 2005. 10. h. wang, g. gao, l. ke, j. zhou and p. rao. isolation and characterization of a lectin-like protein (sblp) from the dried roots of scutellaria baicalensis (lamiaceae). natural product communications, vol. 13, no. 12, pp. 1409-1414, 2018. 11. m. chen, l. gan, s. lin, x. wang, l. li, y. li, c. zhu, y. wang, b. jiang, j. jiang and y. yang. alkaloids from the root of isatis indigotica. journal of natural products, vol. 75, no. 6, pp. 11671176, 2012. 12. d. zhang, j. li, d. ruan, z. chen, w. zhu, y. shi, k. chen, y. li and r. wang. lignans from isatis indigotica roots and their inhibitory effects on nitric oxide production. fitoterapia, vol. 137, p. 104189, 2019. 13. f. chemat, m. a. vian, a. s. fabiano-tixier, m. nutrizio, a. r. jambrak, p. e. munekata, j. m. lorenzo, f. j. barba, a. binello and g. cravotto. a review of sustainable and intensified techniques for extraction of food and natural products. green chemistry, vol. 22, no. 8, pp. 2325-2353, 2020. 14. k. k. adom and r. h. liu. antioxidant activity of grains. journal of agricultural and food chemistry. vol. 50, no. 21, pp. 61826187, 2002. 15. y. nivoix, a. launoy, p. lutun, j. c. moulin, k. a. p. pang, l. m. fornecker, m. wolf, d. levêque, v. letscher-bru, l. beretz and g. ubeaud-sequier. adherence to recommendations for the use of antifungal agents in a tertiary care hospital. journal of antimicrobial chemotherapy, vol. 67, no. 10, pp. 2506-2513, 2012. tx_1~abs:at/add:tx_2~abs:at 119 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 119-124 research article apply parametric shared frailty models to colorectal cancer patients hevi j. hameed, mohammad m. faqe college of administration and economics, university of sulaimani, kurdistan region, iraq abstract colorectal cancer is a combination of colon and rectal cancer that indicates an abnormal growth of cells in either the colon or rectum and is named according to its original location. after treatment, cancer may return to the primary site of the original tumor or to a different location in the body once or more, which is called recurrent. this paper aimed to model this type of data from 128 colorectal cancer patients collected at hiwa hospital in sulaimani considering the gamma shared and inverse gaussian shared frailty models for analyzing the patient’s survival times with colorectal cancer recurrence and estimate the prognostic factor’s impact on their survival. the results of these frailty models compared to those without frailty models using weibull, log-logistic, and lognormal as a baseline distribution for both models. to identify the best model for the data, the akaike information criterion (aic) and bayesian information criterion (bic) were also used. results showed that the cancer stage was the only significant factor affecting survival in recurrent events, as well as evidence of existing heterogeneity in colorectal patients. according to aic and bic, the weibull as baseline distribution with shared gamma frailty model proved the most efficient model for the colorectal recurrent data. in conclusion, the shared frailty model is better than no frailty when analyzing this type of data. keywords: survival analysis, recurrent events, inverse gaussian shared frailty, failure time model, gamma shared frailty model introduction survival analysis is a branch of statistics that contains survival data analysis methods with the response variable being the time required for occurring an event of interest.[1] the event may be death, the onset of a disease, and the required time the patient takes to therapy response or the interval between response and disease recurrence. the event itself is a state transition, and the time to an event is a variable with a positive real value and a continuous distribution.[2] regression models with the response distribution set to time-to-failure are known as parametric survival models. the capacity to take into consideration the censoring and truncation is another feature that distinguishes survival models from normal regression models. specifically, the response distribution must have positive support, weibull, log-normal, and log-logistic are some examples of such distributions.[3] a frailty model is the random effects time-to-event model, in which the frailty effects the baseline hazard multiplicatively. it can be applied for independent lifetime data (univariate), or for multivariate (dependent) duration times. this multivariate method mostly used for related individuals survival times, such as family members or twins, when the assumption of independence cannot stand, or for recurring occurrences in the same individual, or for times of various events in the same individual, for instance disease start, and relapse recurrence.[4] shared frailty models are the time-to-event data analog to random effects model. it is a model wherever frailties shared by groups of observations or multiple records in the same observation instead of specific observation, thus causing the same group’s observations to be correlated. in these models, the main idea is that each individual has different frailty, and the frailest will experience the event of interest earlier than the less frail subjects in the data.[5] according to the world health organization (who), the second-leading cause of cancer-related mortality globally is colorectal cancer. patients getting curative resection for localized illness had a 5-year survival rate of 70–90%. in contrast, when there are metastases in the regional lymph node, this probability lowers to 40–80%. the most effective cihan university-erbil scientific journal (cuesj) corresponding author: hevi j. hameed, college of administration and economics, university of sulaimani, kurdistan region, iraq. e-mail: hevi.hameed@univsul.edu.iq received: july 18, 2022 accepted: august 11, 2022 published: november 01, 2022 doi: 10.24086/cuesj.v6n2y2022.pp119-124 copyright © 2022 hevi j. hameed, mohammad m. faqe. this is an openaccess article distributed under the creative commons attribution license (cc by-nc-nd 4.0). hameed and faqe: apply shared frailty models to cancer patients 120 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 119-124 technique is a surgical treatment to colorectal cancers’ cure; however, recurrences happen at a steady rate depending on the disease stage.[6] colorectal cancer frequently returns after at least a year. when it is simply an advanced form of the original cancer, or when it occurs within a few months. in this case, the cancer frequently returns, because the first round of treatment did not eliminate all of the cancer cells, the patient may be more tired than usual, and depending on whether and where the cancer has spread, the doctor performs a physical examination and an endoscopy of the colon at regular intervals during these examinations, which usually take place every 3–6 month.[6] ignoring the presence of heterogeneity leads to inaccurate parameter estimates and standard errors in survival analysis. it may also overstate life expectancy. underestimation of covariate effects when heterogeneity is disregarded. discounting frailty causes the estimate of regression coefficients to be biased toward zero by an amount that relies on the distribution and variability of the frailty factors.[7] while there are several studies that have considered frailty and shared frailty models, to the best of our knowledge, none have considered the recurring events of colorectal cancer, and this is the first examination of the factors affecting patient survival without colorectal recurrence using shared frailty model in accelerated failure time (aft) metric. this article demonstrated the application of frailty models to recurrent tumor events in the colorectal cancer dataset. frailty models are the extension of common survival models such as (aft models cox proportional hazard and parametric proportional hazard). this article focused only on aft models and their extension. various extensions from univariate frailty models are possible to multivariate frailty models and one of these is considered in this study, namely, shared frailty models, which will share frailty among groups or clusters, where the cluster can be an individual or a group. here, groups recurring events in the same person in patients with colorectal cancer. materials and methods constructing random effects models can be useful in recurrent occurrences studies over time or other sorts of multivariate survival data; it is a common phenomenon to tackle such problems utilizing frailty models. many statistical strategies for adjusting and evaluating unobserved heterogeneity have been developed. modeling unobserved variability in survival data is a topic that has received a lot of attention in recent decades. unobserved heterogeneity, often known as hidden variability, is a significant source of variation in medical and biological applications. variability is classified into two types: that produced by observable risk variables and that affected by unknown variables that are ideally unpredictable.[8] in the present paper, we considered the no frailty model and the shared frailty model. the no frailty model aft models are discussed without taking frailty into account, while, in shared frailty, model survival models are analyzed using frailty effects. our study interest is about the colorectal cancer recurrent disease. frailty models are best models can be used to describe such unobserved heterogeneity in timeto-event analysis. the effect of heterogeneity, or frailty, has been recognized for a long time, there are several methods to access the frailty effect. we choose gamma distribution and inverse gaussian distribution for evaluating the frailty effect, because they have a simple density function, for which parameters are easily obtained through likelihood estimation. survival analysis functions we start with a time-to-failure random variable (t) and specify the density function for t as f(t), the survivor function s(t) = p(t>t) the cumulative distribution function can be described as f(t) = p(t≤t) = 1−s(t). in this context, one more function that stands out is the hazard function �h t f t s t � � � ( ) ( ) it is the instantaneous failure rate up till time t given survival.[3] cox proportional hazards the cox proportional hazards model is a method of multiple regressions for determining the impact on survival time from several covariates. h t z h t exp zt| ( )� � � � �0 β (1) where h 0 (t) enotes a legitimate hazard function (failure rate) for some unspecified life distribution model, β, is a vector of parameters, and z is a covariate vector.[9] aft model when examining survival time data, an alternative to the proportional hazard (ph) model is the aft model. we investigated the direct effect of explanatory variables on survival time rather than risk using aft models. for the reason that the parameters assess the corresponding covariates effect on survival time, this feature enables the result’s interpretation be more easier than other models.[9] according to the aft model, an individual at time t with covariate z has the survival function as same as an individual at time {t×exp(βtz)} with a baseline survival function, where βt = (β 1 β 2…… β p ) is a regression coefficients vector; also, the aft model can be defined by this relationship: s t z s t e tz | { }� � � � � �0 β for all z (2) the aft model is a log-scale in relation to time that it gives the analogous to the ordinary linear regression method. in this method, we model the log (t) = y, in which it is the survival time’s natural logarithm. this is how positive variables are naturally transformed into observations along the full real line in linear models. for y, a linear model is assumed: log t y zt� � � � � �µ β σε (3) where βt = (β 1 β 2…… β p ) is regression coefficients vector μ = intercept σ = cale parameter the error distribution = ε which assumed to have a specific parametric distribution. when we denote by s 0 the survival function when z = 0. hameed and faqe: apply shared frailty models to cancer patients 121 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 119-124 we adopt the following baseline hazard functions h(t) and survival functions s(t) the parameter λ,p:[9] 1. weibull h t �( ) = λptp−1 (4) s( )t �= e t p−λ (5) 2. lognormal h t logt logt t � � � �� � � � � � � �� � � � � � � � � � � � � � µ σ µ σ σ , 1 (6) s t logt� � � � �� � � � � �1 � µ σ (7) 3. log-logistic h t � � pt t p � p ( ) � � �λ λ 1 1 (8) s( )t tp � � 1 1 λ (9)[9] frailty models a generalization of the survival regression model is a frailty model. in addition to the observed covariates, a frailty model also allows for the latent multiplicative effect to be presence on the risk function. this impact, or frailty, is inferred rather than directly determined from the data. it is supposed having limited variance and its mean equal to one. when the frailty is larger than one, people have a higher risk of failing and are considered to be frailer than others. thus, frailty models can be a valuable alternative to ordinary time –to-event models, while the standard models are unable to fully account for the variation in failure observed times.[3] aft shared frailty model the applicable choice for multivariate time-to-event data is the shared aft frailty model. supposing log t ij is the natural logarithm of the jth recurrent event’s survival time in the ith subject for colorectal cancer patient. where i = 1,2,3,…n i patients number j = 1,2,3,4, recurrent events and z ij be the covariates vector related to each individual. now, the shared frailty model in aft form can be given by, logt z uij ij t i ij� � � �µ β σ (10) where μ is the parameter of intercept β, the unknown regression coefficients vector, the scale parameter is σ the ∈ ij, are the random errors, in which distributed identically independent and the u i are the random effects for specific individual (each patient) which is supposed to be a random variable with density function that is independent identically distributed. f(ui) in this paper, we supposed that the random effect (shared frailty) had inverse gaussian and gamma distribution with variance θ and unit mean, as described in the density functions bellow.[10] gamma distribution f u u exp u u � � � � �( / ) / ( ) ( ) 1 1 11 θ θ θ γ θ θ (11)[10] inverse gaussian distribution f u �� à u u u �� uu � � � � � � � � � � �� �� � � � � � � � � �� 1 2 1 2 0 0 1 2 3 2 2 θ θ θ / / exp , (12) the conditional hazard function and survival function could be written as follows for the jth observation in the ith patient: h t|u t �h uij i ij ij i� � � 1 0σ ( | ) (13) s t|u s uij i ij i� � � 0( | ) (14) where ij ij ij t ilogt z u� � � �µ β σ/ (15) h 0 (.), s 0 (.) are and survival function of ∈ ij espectively, and β refers to the covariate coefficient’s vector associated with the covariates vector z ij measured in the jth event of ith subject.[10] model assessment we employed akaike information criterion (aic) in addition to bayesian information criterion (bic) to compare and select the efficient model among parametric aft models without frailty with three distributions as baseline and those aft models with two different shared frailty model (gamma and inverse gaussian). akaike’s (1974) information criterion defined by akaike is: aic lnl h m� � � � �2 2( ) (16) where h indicates the covariate numbers in the model, for instance m = 2 in weibull, because it has two parameters. the smaller aic value is considered as better model. another fit measure is bayesian information criterion defined by schwarz in (1978), bic lnl ln n� � � �2 2 ( ) (17) the number of data points is represented by n, where n is the sample size. the bic estimate has the main advantage of including the bic penalty for the number of parameters being evaluated. the best model is determined by having the lowest bic values.[11] data description and analysis description of the data in this study, 128 cases of colorectal cancer were recorded at hiwa hospital in sulaimani city. cases were collected over a period of (59) months; beginning from january 7, 2017, to july 20, 2021, for the colorectal cancer patients who were alive at the time that they took part in the study. data from colorectal hameed and faqe: apply shared frailty models to cancer patients 122 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 119-124 cancer patients who were diagnosed at the time the patients entered the study. the event of interest is recurrences of tumors. of those patients, 68 were randomized to the group that had just received chemotherapy, and 60 were randomized to the group that received both chemotherapy and radiation therapy. multiple tumor recurrences occurred in many patients during the study, and at each visit, the new tumors were removed. the dataset contains the first four tumor recurrences of each patient. furthermore, each time of recurrence was measured from the patient’s time of entry into the study. about 28.9% had at least the first recurrence and 11% of the patients had the second recurrence, while 5.5% had the third recurrent event, 2.3% had all four recurrences. the data consist of the following variables as demonstrated in table 1 id, patient identification (this is the sequence number of the subject), follow-up time: the time from the (diagnosis) or the start of the study to the end of it. time 1, time 2, time 3, and time 4, are the times of the four potential recurrences of colorectal cancer in months. a patient with only two recurrences has missing values in time 3 and time 4. four observations are made for each patient, one for each of the four possible tumor recurrences. the treatment type that each patient received is demonstrated as (0 = chemotherapy and 1 = chemotherapy with radiotherapy). initial size and initial tumor size measured in centimeters (cm). stage (the cancer stage at which the patient was first diagnosed), and age (the age of the patient). for the aft without frailty model, the data considered have only the first event (recurrent) or the follow-up time if no relapse is observed. the frequencies and percentages of explanatory variables are given in table 2 that 53.1% of the patients received chemotherapy, while (46.9%) received chemotherapy with radiation therapy. in respect to our data, the patients were table 1: the first and last 8 observations from colorectal cancer patient’s data id follow-up time time 1 time 2 time 3 time 4 treatment initial size stage age 1 31 0 2.6 4 51 2 41 26 36 41 0 4 4 47 3 52 7 12 41 45 0 5 3 42 4 19 0 9 3 62 5 16 0 3.5 2 62 6 24 9 0 4.5 2 60 7 21 0 1.5 2 78 8 39 33 0 4 4 35 121 12 1 5.2 3 79 122 11 1 4.5 2 36 123 10 1 2 1 2.5 3 48 124 9 1 5 3 69 125 11 1 3.5 3 44 126 10 1 4.6 3 66 127 6 1 5 3 66 128 32 8 13 1 5 2 38 table 2: explain frequencies and percentages of explanatory variables variables categories code frequency (n=128), n (%) treatment group chemotherapy (0) 68 (53.1) chemotherapy with radiotherapy (1) 60 (46.9) cancer stage low stage (0) 48 (37.5) high stage (1) 8062.5) initial size ≤2.5 cm (1) 18 (14.06) 2.6–5 cm (2) 52 (40.63) ≥5 (3) 58 (45.31) age <50 (0) 47 (36.7) ≥50 (1) 81 (63.3) response the response is a time to recurrent event or follow-up table 3: the values of bayesian information criterion and akaike information criterion for the parametric accelerated failure time shared frailty models distributions of baseline the distribution of frailty aic bic weibull without frailty gamma inverse gaussian 302.4751 294.9285 296.3048 325.0553 320.7345 322.1108 lognormal without frailty gamma inverse gaussian 305.4182 297.7116 299.0783 327.9984 323.5176 324.8843 log logistic without frailty gamma inverse gaussian 304.4661 297.0295 298.4006 327.0464 322.8355 324.2066 bic: bayesian information criterion, aic: akaike information criterion hameed and faqe: apply shared frailty models to cancer patients 123 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 119-124 diagnosed according to their initial cancer stages with 37.5% which were diagnosed in lower stage (stage one and stage two), while patients of higher stages (stage three and four) made up the top proportion, progressing to (62.5%). regarding patients’ initial tumor sizes (14.06%), their tumors were less than or equal to 2.5 cm, whereas 40.63% their tumor sizes were between 2.6 and 5 cm as well as sizes larger or equal to 5 cm correlated to 45.31% of cases, finally in regards to the patients age groups, 36.7% were less than 50 years, while whose more than or equal to 50 years were (63.3%). the data analyzed with stata 14 program. results according to the results in table 3, the values of aic and bic for several parametric aft models employing inverse gaussian and gamma shared frailty models, as well as models without frailty. the gamma shared frailty model with weibull baseline distribution recorded the lowest value of aic and bic among other models tested, demonstrating the efficiency of this model for describing the colorectal cancer recurrence dataset using some parametric aft models. table 4 demonstrates the results of the weibull aft model (without frailty) and with inverse gaussian as well as gamma shared frailty model using weibull distribution as baseline which was the best model for data from patients with colorectal cancer. the table presented estimated values (β, time ratio (tr), p-value, estimated baseline distribution parameters (p), and frailty variance θ) with their likelihoodratio test. in all three models, the only significant prognostic factor was the patient’s initial cancer stage with time ratio values (0.275, 0.295, and 0.294) for weibull without frailty, weibull with gamma shared model, and weibull with inverse gaussian shared models, respectively, all of them are less than one indicates that patients with a higher initial stage are accelerating the time to relapse for the patient. in addition, the both results of frailty models indicated the significant of heterogeneity (θ) between patients according to the significance-likelihood-ratio test of theta (θ = 0) with values 1.100818 for the gamma shared frailty model and 1.068641 for the shared inverse gaussian model indicating that there is a frailty effect affects the resulting of the data which tell us that the data is not homogeneous, but heterogeneous some patients are more frails than others. although each patient has the same covariate value, the heterogeneity impacts the patient’s recurring event. the estimates from the inverse table 4: weibull accelerated failure time with and without shared frailty model for colorectal cancer patients parameter weibull (without frailty) weibull (gamma frailty) weibull (inverse gaussian frailty) β tr p β tr p β tr p treatment 0.093 1.098 0.705 0.0791 0.804 1.0823 0.0612 1.063 0.846 age 0.2351 1.264 0.356 0.243 0.461 1.275 0.280 1.323 0.388 stage −1.288 0.275 0.001* −1.217 0.005* 0.295 −1.222 0.294 0.005* initial size (cm) ≤2.5 ref 1 ref 1 ref 1 2.6–5 0.489 1.63 0.236 0.359 1.43 0.487 0.384 1.468 0.459 ≥5 0.258 1.29 0.491 0.105 1.11 0.823 0.157 1.17 0.740 intercept 4.549 0.0000 4.5792 0.0000 4.538 0.0000 p 1.078 1.1008 1.097 frailty (θ) --θ=(1.100818) prob ≥ chibar2=0.001* θ=(1.068641) prob ≥ chibar2=0.002* *significance at 99% level or (0.01). ref: reference level which its time ratio always equals to one. tr: time ratio table 5: the survival rate at lower stage (0) and higher stage (1) for weibull with gamma shared id time stage s (t) 1 31 1 0.6443853 2 26 1 0.627423 2 36 1 0.5416252 2 41 1 0.5063942 126 10 1 0.8720858 127 6 1 0.9003416 128 8 0 0.9505355 128 13 0 0.9184979 128 32 0 0.8074583 figure 1: the survival rate at the levels of stage hameed and faqe: apply shared frailty models to cancer patients 124 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 119-124 gaussian shared frailty model are quite close to the results from the gamma shared frailty models, showing the analysis’s robustness with respect to the baseline function choice. from table 5, it is clear that with regular repetition of recurrent events the patient’s survival decreases steadily. figure 1 represents the survival rate at lower stage (0) and higher stage (1) for weibull with gamma shared frailty which was the best fitting model; furthermore, this figure demonstrates the same results as the numerical does that the patients with lower cancer stages at most survive as the higher cancer stage. conclusion and recommendation conclusion from performing the recurrent event or more commonly multivariate survival data, according to the results the paper obtained, the following conclusions were drawn: 1. the shared frailty model, as an extension of the cox model for time-to event data is an excellent choice of recurrent events, particularly when observations of the same subject share an unobservable common frailty. it obviously considers the possibility of dependence between downtimes. 2. based on aic as well as bic to compare the shared frailty aft models, it is concluded that the shared frailty aft model using weibull distribution as the baseline was the most appropriate model to the data set used in this paper. 3. among (treatment group, age, cancer stage, and initial size) as prognostic factors the only variable that affecting survival time to recurrent event was initial cancer stage, it is noted that the higher stage will accelerate the time to event, meaning that the diagnoses with the early stages of cancer reduce the chances of cancer recurrence. 4. ignoring un observed heterogeneity may lead to overestimate or underestimate the estimated coefficients. recommendation 1. performing univariate frailty models and shared frailty models by comparing both results to get more information about the use of frailty models in each circumstance and differences among them. 2. more studies should be done in this field because such studies are important and relate to people’s lives. 3. providing qualified endoscopy machines in several hospitals and care units for the early detection of colon cancer hence lowering the incidence as well as recurring rates of it. 4. data should be registered in health authorities so that researchers can conduct the research in detail and get interesting results. references 1. m. r. karim and m. a. islam. reliability and survival analysis. singapore: springer nature pvt ltd., 2019. 2. d. d. hanagal. modeling survival data using frailty models. boca raton, fl: chapman & hall/crc, 2011. 3. r. g. gutierrez. parametric frailty and shared frailty survival models. the stata journal, vol. 2, no. 1, pp. 22-44, 2002. 4. a. wienke. frailty models in survival analysis. london, united kingdom: chapman and hall/crc, 2010. 5. g. grover and d. seth. application of frailty models on advance liver disease using gamma as frailty distribution. srl, vol. 3, pp. 42-50, 2014. 6. h. kobayashi, h. mochizuki, k. sugihara, t. morita, k. kotake, t. teramoto, s. kameoka, y. saito, k. takahashi, k. hase, m. oya, k. maeda, t. hirai, m. kameyama, k. shirouzu and t. muto. characteristics of recurrence and surveillance tools after curative resection for colorectal cancer: a multicenter study. surgery, vol. 141, no. 1, pp. 67-75, 2007. 7. u. abdulkarimova. frailty models for modelling heterogeneity. canada: mcmasters university, 2013. 8. k. adeleke and g. grover. parametric frailty models for clustered survival data: application to recurrent asthma attack in infants. journal of statistics applications and probability letters, vol. 6, pp. 89-99, 2019. 9. a. gebeyehu. survival analysis of time-to-first birth after marriage among women in ethiopia: application of parametric shared frailty model. jimma: department of statistics, college of natural sciences, jimma university, 2015. 10. k. d. fentaw, s. m. fenta, h. b. biresaw and s. s. mulugeta, time to first antenatal care visit among pregnant women in ethiopia: secondary analysis of edhs 2016; application of aft shared frailty models. archives of public health, vol. 79, no. 1, pp. 1-14, 2021. 11. p. k. swain and g. grover. determination of predictors associated with hiv/aids patients on art using accelerated failure time model for interval censored survival data. american journal of biostatistics, vol. 6, pp. 12-19, 2016. _goback tx_1~abs:at/add:tx_2~abs:at 48 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 48-51 research article effect of hypericum perforatum extract on gastric ulcer development in rat models mohammed m. hussein department of biomedical sciences, cihan university-erbil, kurdistan region, iraq abstract gastric ulcer is a serious condition that results from the tearing of the stomach mucosa. a physiological equilibrium is maintained between mucosal defense and aggressive factors. in this study, the effectiveness of medicinal plant (hypericum perforatum) in the management of gastric ulcers was compared to that of omeprazole, a well-known drug for gastric ulcers. this study involved numerous study groups of male rats comprising the following: the first group (control group exposed to helicobacter pylori and fed with normal animal feed), the second group (rats exposed to h. pylori and treated with two doses (of 250 mg/kg and 500 mg/kg) of aqueous h. perforatum), and the third group (rats exposed to h. pylori and treated with 20  mg/kg of omeprazole). the outcome of the study demonstrated that a high dose of 50.65% h. perforatum prevents the development of stomach ulcers. the group treated with omeprazole revealed a formation of stomach ulcers (24.50%). a high dose of the aqueous plant extracts showed greater effectiveness in preventing gastric ulcer formation compared to the lower dosage. keywords: gastric ulcer, antibacterial activity, helicobacter pylori, gastrointestinal tract, peptic ulcer introduction a highly mobile, spiral-shaped, gram-negative bacteria called helicobacter pylori colonizes the intestines of 50–80% of people around the globe.[1] the bacterium h. pylori damage the gastric mucosa most commonly by causing gastritis, peptic ulcer disease, or even gastric cancer.[2] gastric ulcer is a condition of the stomach lining that has common symptoms such as vomiting, burning, dull abdominal pain, headache, weight loss, low oral resistance, stenosis, perforation, and stomach bleeding.[3] the focus has been on natural plants for the management of many human disease conditions partially because certain pharmaceutical medicines are highly harmful to the patient or cause adverse effects.[4] moreover, in terms of cure and disease prevention, plant products are inexpensive and more affordable contributors to improving human health.[5] hypericum perforatum is a hypericaceae commonly known as st. john’s wort. it has been used as a form of treatment for different diseases in various eastern countries. nevertheless, scholars suggest that it is used to cure mental illnesses caused by demonic forces among the ancient greeks. the plant grows well in temperate open disturbed regions.[6] conventionally, omeprazole is used as a drug for the treatment of conditions where the stomach excretes large quantities of acid. it prevents the activity of a particular enzyme mechanism located within the acid-secreting stomach cells (in the stomach),[7] thus, preventing acid production. omeprazole is used to control heartburn, acute gastritis, duodenitis, esophagitis attributable to recurrence of gastrointestinal material, peptic ulcer, and other secretary diseases.[8] aim of study this research is aimed at comparing the effectiveness of h. perforatum and omeprazole in gastric ulcer treatment using male albino rats as the disease model. materials and methods extraction of the aerial part of h. perforatum fresh aerial plants (stem, leaf, flower, fruit, and seed) of h. perforatum collected from khabat (district of erbil city in the western region of the kurdistan region, iraq) were corresponding author: mohammed m. hussein, department of biomedical sciences, cihan university-erbil, kurdistan region, iraq. e-mail: mohammed.m.hussein@cihanuniversity.edu.iq received: march 10, 2023 accepted: april 21 2023 published: may 20, 2023 doi: 10.24086/cuesj.v7n1y2023.pp48-51 copyright © 2023 mohammed m. hussein. this is an open-access article distributed under the creative commons attribution license (ccby-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) hussein: antibacterial activity of hypericum perforatum 49 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 48-51 extracted for the study. the aerial part of the plant with a high presence of flowers and buds was collected in august 2020 and air-dried in the shade. the extraction process followed a modified method previously used by.[9] the dried and pulverized aerial parts of the flower (25 g) were soaked in 250 ml of water (at 85°c) for high-dose extraction and 25 g in 500 ml for low-dose extraction. the mixture was kept for 24 h before filtering with whatman number 1 filter paper. finally, the filtered extract was kept inside a sterile cup in the refrigerator until used. omeprazole omeprazole is the class of medicines that have been used to treat disorders including peptic ulcers because they act as proton-pump inhibitors. omeprazole inhibits the acid-forming enzymes in the stomach wall. the output of the stomach acid is reduced by blocking these enzymes and thus, the stomach can be healed. omeprazole served as the standard anti-ulcer agent in this study; it was procured from happy pharmacy in erbil city. the medication was administered to the rats by mouth at the dose of 20 mg/kg body weight following suspension in distilled water (5 ml/kg).[10] experimental animals forty-eight male albino rats were used for the experiment; the body weight of the rats was 160 ± 20 g at 7–9 weeks old. the rats were randomly allocated to four groups, each group of 12 rats was purchased from zakho university and housed hygienically in ventilated cages. the rats were fed on a standard diet for 1 week in the animal house unit of the college of education at salahaddin university-erbil. induction of gastric ulcer by h. pylori gastritis was induced by feeding rats with the bacterium h. pylori that were collected from biopsy samples from clinical patients. the rats were fed for 1 week on a basal diet in the animal house unit, college of education at salahaddin university-erbil. in this study, the rats were kept from food in the morning and afternoon, then, inoculated with the plant extracts and immediately inoculated with 1 ml h. pylori suspension (5 × 108–5 × 1010 cfu/ml) using gavage twice daily at 4 h intervals for 3 straight days. the rats were left on normal feeding for 15 days to observe the effect of the plant extracts on h. pylori activity.[11] before inoculating the animals with the plant extract and h. pylori, they were subjected to stool antigen tests to ensure no pre-infection with h. pylori. in the last period of prevention, the rats were anesthetized through intramuscular injection of mixed xylazine–ketamine (1:9) as a single dose in the same syringe. the stomach of each rat from each group was removed and dissected for observation. gross gastric lesions evaluation ulcers located in the gastric mucosa, parallel to the long stomach axis, emerged as elongated bands of hemorrhagic lesions. thus, the damage was studied in each gastric mucosa specimen. in the measurement of the ulcer region (ua), the total of the lesion areas for each stomach was summed, wherein the sum of small squares × 4 × 1.8 = ua mm2 as earlier described by.[12] the inhibition percentage (i %) was estimated using the formula: [13] (i %) = ([ua control – ua treated]/ua control) × 100% measurement and evaluation of acid content of gastric juice (ph) the ph level of the rat stomach was individually recorded for evaluation of the ph level of all the groups of rats; this was done to know the effect and correlation of the plant extract with ph level. microbiological analysis microbiological analysis was directly carried out on the day of sample collection. stool antigen and blood antibody tests were done on all the rat groups using strip h. pylori antigen and antibody tests; this was done to determine the h. pylori infection status of the rats. results and discussion effect of h. perforatum extract and omeprazole on gastric lesion development the anti-ulcer effect of h. perforatum extract against gastric lesion model caused by h. pylori is reported in table 1. the figure 1: the ph level of gastric content of rats treated with hypericum perforatum extract and omeprazole compared to the control group figure 2: effect of omeprazole antibiotic drugs on ph of gastric content hussein: antibacterial activity of hypericum perforatum 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 48-51 table 1: effect of aqueous extract of h. perforatum and omeprazole on gastric ulcer production and gastric ulcer development in albino rats animal group prevention (5 ml/kg) dose ulcer area (mm2) mean ± sem ulcer inhibition % 1 ulcer control group 815.00 0 2 high-dose h. perforatum 402.20 50.65 low-dose h. perforatum 611.45 24.97 3 omeprazole 615.25 24.50 h. perforatum: hypericum perforatum results showed that the ulcer area of the rat’s stomach with a high dose (500 mg/kg) of h. perforatum extracts was inhibited at the rate of 50.65% compared with the ulcer control group. table 1 also showed the anti-ulcer effect of omeprazole in the h. pylori-induced gastric ulcer rat models. the findings demonstrated that the rats’ stomachs were protected by omeprazole while receiving h. pylori solution, thereby showing a decrease in gastric ulcer formation rate by 24.50% compared to the control group. effect of h. perforatum extract and omeprazole on the gastric ph level when compared to the non-treated control group in figure 1, the stomach contents’ acidity in the experimental mice treated with h. perforatum was increased. the experimental rats treated with omeprazole showed an increase (5–5.5) in the gastric contents’ acidity when compared to the control group (3.5). the acidity of the gastric juice of the group treated with h. perforatum extract, however, ranged from 4 to 4.5 see figur e 2. evaluation of stomach color changes in gastric ulcer rat models treated with h. perforatum and omeprazole the color of the stomach is one of the parameters considered in the study; the color of the stomach of h. perforatum-treated rats significantly changed when compared with the control group. a bright white color was found in the stomach of the rats treated with h. perforatum, whereas a pink-to-red color was found in the stomach of omeprazole-treated rats. the control group’s stomach was totally dark red because of the ulcer and inflammation caused by h. pylori, while the color of the protected rat stomachs changed significantly [figure 3]. gastritis is among the most prominent clinical symptoms of h. pylori infection; h. pylori forms widely visible black hemorrhagic injuries on the stomach mucosa. in this study, the plant extract showed a 50.65% inhibition rate of the formation of stomach ulcers compared with the control group at the concentration of 500 mg/kg and 24.97% inhibition at the concentration of 250 mg/kg. however, a previous study[14] also reported the effectiveness of h. perforatum on gastroprotection in rat animal models. as an antibacterial agent, omeprazole, a standard drug used to treat gastric ulcers, plays a major role in preventing gastric ulcer disorders in male albino rat animal models, especially gastric ulcers caused by h. pylori. according to the current study results, the stomach of mice infected with h. pylori and treated with omeprazole showed an observable reduction in gastric ulcer development at the rate of 24.50% compared to the control group. the stomach mucosa of the rats exposed to h. pylori solution showed noticeable black hemorrhagic lesions. the outcome of this study agrees with the report by[15] which showed a healing rate of 61% in a study group that received 20 mg of omeprazole within 4 weeks during a study with 68 patients receiving concurrent nsaids. this is significant because a high percentage of patients in the omeprazole groups were free of symptoms and ulcers. in the 4th week, the healing rate was 61% in the group receiving omeprazole 20 mg, and a significant number of patients in the omeprazole group had no symptoms or ulcers. conclusion the present study demonstrated that the development of gastric ulcers following active h. pylori induction can be prevented significantly using medicinal plants, such as h. perforatum. h. perforatum can serve as a natural alternative to omeprazole (a chemical agent) in the prevention of gastric ulcer formation in people that have been exposed to h. pylori. references 1. p. palamides, t. jolaiya, a. idowu, e. loell, c. onyekwere, r. ugiagbe, i. agbo, o. lesi, d. ndububa, o. adekanle, m. carranza, figure 3: stomach changes in different rat groups: (a) control group, (b) treated with hypericum perforatum extract, (c) treated with omeprazole b ca hussein: antibacterial activity of hypericum perforatum 51 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 48-51 r. ally, h. niom, i. a. adeleye, u. harrison, a. clarke, w. fischer, s. smith and r. haas. helicobacter pylori patient isolates from south africa and nigeria differ in virulence factor pathogenicity profile and associated gastric disease outcome. scientific reports, vol. 10, p. 11409, 2020. 2. o. k. yulizal, a. lelo, s. ilyas and r. l. kusumawati. the effect of channa striata extract and standard eradication regimen on asymmetric dimethylarginine in helicobacter pylori gastritis rat model. veterinary world, vol. 13, pp. 1605-1612, 2020. 3. h. mashayekhi-sardoo, b. m. razavi, m. ekhtiari, n. kheradmand and m. imenshahidi. gastroprotective effects of both aqueous and ethanolic extracts of lemon verbena leaves against indomethacin-induced gastric ulcer in rats. iranian journal of basic medical sciences, vol. 23, pp. 1639-1646, 2020. 4. h. youssef and a. el-mahmoudy. evaluation of the antimicrobial potential of punica granatum leaves hydro-methanolic extract against selected pathogens. american journals of current microbiology, vol. 7, pp. 1-11, 2019. 5. t. belwal, h. p. devkota, m. k. singh, r. sharma, s. upadhayay, c. joshi, k. bisht, j. k. gour, i. d. bhatt, r. s. rawal and v. pande. st. john’s wort (hypericum perforatum). in: nonvitamin and nonmineral nutritional supplements. elsevier inc., amsterdam, p. 415-432, 2019. 6. h. seraji, h. hosseini-vashan, n. afzali, m. h. namayi and a. allahressan. effect of water extract of hypericum perforatum on performance, abdominal fat and blood biochemical parameters of broiler chicken. animal science journal, vol. 28, no. 106, pp. 133-146, 2014. 7. a. m. pedernera, t. guardia, c. g. calderón, a. e. rotelli, n. e. de la rocha, s. d. genaro, l. e. pelzer. anti-ulcerogenic and anti-inflammatory activity of the methanolic extract of larrea divaricata cav. in rat. journal of ethnopharmacology, vol. 105, no. 3, pp. 415-420, 2006. 8. n. m. nasr, m. khider, w. metry and k. atallah. antibacterial activity of lactic acid bacteria against helicobacter pylori evidence by in vivo and in vitro studies. international journal of current microbiology and applied sciences, vol. 6, no. 12, pp. 4235-4247, 2017. 9. g. l. jr. kauffman, and m. i. grossman. prostaglandin and cimetidine inhibit the formation of ulcers produced by parenteral salicylates. gastroenterology, vol. 75, no. 6, pp. 1099-1102, 1978. 10. v. c. njar, j. k. adesanwo and y. raji. methyl angolensate: the antiulcer agent of the stem bark of entandrophragma angolense. planta medica, vol. 61, no. 1, pp. 91-92, 1995. 11. g. zdunić, d. gođevac, m. milenković, d. vučićević, k. šavikin, n. menković and s. petrović. evaluation of hypericum perforatum oil extracts for an antiinflammatory and gastroprotective activity in rats. phytotherapy research, vol. 23, no. 11, pp. 1559-1564, 2009. 12. a. walam, j. p. bader, m. classen, c. b. lamers, d. w. piper, k. rutgersson and s. eriksson. effect of omeprazole and ranitidine on ulcer healing and relapse rates in patients with benign gastric ulcer. the new england journal of medicine, vol. 320, no. 2, pp. 69-75, 1989. 13. n. s. al-wajeeh, m. f. halabi, m. hajrezaie, s. m. dhiyaaldeen, d. a. bardi, s. m. salama, e. rouhollahi, h. karimin, r. abdolmalaki, a. h. s. azizan, h. m. a. s. noor and m. a. abdulla. the gastroprotective effect of vitex pubescens leaf extract against ethanol-provoked gastric mucosal damage in sprague-dawley rats. plos one, vol. 12, no. 5, p. e0179072, 2017. 14. n. seyhan. evaluation of the healing effects of hypericum perforatum and curcumin on burn wounds in rats. evidence based complement alternative medicine, 2020, p. 6462956, 2020. 15. f. seyis, e. yurteri, a. özcan and c. cirak. altitudinal impacts on chemical content and composition of hypericum perforatum, a prominent medicinal herb. south african journal of botany, vol. 135, pp. 391-403, 2020. tx_1~abs:at/add:tx_2~abs:at 141 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 141-146 research article influence of the covid-19 lockdown on the lifestyles and eating behavior of cihan university students salih m.s. zebari1,2, yaseen galali1,3, kazhin younis1, zainab rizgar1, nareen sidiq1, brwa taha1 1department of nutrition and dietetics, cihan university, erbil, kurdistan region, iraq, 2department of animal resources, college of agricultural engineering sciences, salahaddin, university-erbil, iraq, 3department of food technology, college of agricultural engineering sciences, salahaddin university-erbil, erbil, krg, iraq abstract recent studies found that a city confinement as a result to covid-19 outbreak resulted on eating and lifestyle behavior changes. this study investigated the influence of lockdown on eating and lifestyle changes. a  cross-sectional study was done from december 18, 2021, to february 20, 2022. the research composed of three parts. part  i (six questions) contained sociodemographic statues; section two (11 questions) studied dietary behavior through that included a mediterranean diet (md) adherence using 14-scaled items with an md screener tool ranging from 0 to 13; and section three (12 questions) studying lifestyle habit changes such as physical activity, sleeping habits, and smoking habits. the results showed that 17% (63) of individuals that believed their lifestyle had improved, while 38.6% (143) stated that it had worsened. specifically, during lockdown, the physical activity frequency seems reduced from 50% (185) to 33% (127), and sleeping hours for those sleeping more than 9 h were dramatically raised from 17% to 31.4%. in terms of eating habits, 36.8% of participants stated appetite improvement, whereas 22.7% reported a decrease. the appetite change during confinement was not connected with body mass index, although it was significantly associated with weight change (p = 0.0001). similarly, 47% of participants said that they had gained weight. the data also showed that 67% (248) medium adhered to md diet. thus, the covid-19 lockdown has negatively affected both lifestyle and eating behavior of the students. keywords: covid-19, eating behavior, eating pattern, lifestyle, mediterranean diet introduction the covid-19 is a crucial respiratory disease due to the sars coronavirus 2. the virus was discovered in transit from animals to people at the market of seafood hunanchina in december 2019 and very quickly spread globally. the covid-19’s global expansion is a quickly changing condition. the emergency committee of world health organization’s summoned a worldwide health emergency on january 2020, due to increased case reporting rates.[1] the sars-cov-2 virus’s emergence and spread created a global pandemic that led to serious pandemic and a large deaths number. governments around the world responded to the pandemic by establishing a variety of measures to prevent the virus spread including home quarantine and travel restrictions.[2] in kurdistan region of iraq, the confirmed infection with covid-19 was 392,791 from march 1, 2020, to january 18, 2022. therefore, at the beginning to stop the separation of pandemics, the ministry of health introduced a lockdown in march and april of 2020, and a limiting the community activity, parties, gatherings, meetings, and schools.[3] the city lockdown results in rapid and extreme changes in the habits and lives of the people and students, as well as a noticeable reduction in social activities. physical confinement and isolation had an enormous impact on community’s lifestyles, especially their eating patterns and daily activities. home staying works (which includes online schooling, working at home, and limiting outdoor and recreational activity) and saving food (due to shopping restrictions) are the two key impacts. in addition, the changing daily routine work resulted from the lockdown can cause boredom that can be associated with increased food consumption.[4] a person can feel stressed from reading or hearing about the covid-19 in the media on a daily basis, which can contribute corresponding author: yaseen galali, department of nutrition and dietetics, cihan university, erbil, kurdistan region, iraq. e-mail: yaseen.galali@su.edu.krd received: august 8, 2022 accepted: october 18, 2022 published: november 30, 2022 doi: 10.24086/cuesj.v6n2y2022.pp141-146 copyright © 2022 salih m.s. zebari, yaseen galali, kazhin younis, zainab rizgar, nareen sidiq, brwa taha. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) zebari, et al.: eating behavior and life style during covid-19 142 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 141-146 to overeating, particularly “food desiring” comfort foods high in sugar.[5] dietary behavior and changes are possible due to of the covid-19 confinement. on the other hand, a healthy lifestyle and diet are supportive to the body well-being. for that purpose, a number of health organizations including the efad and who and proposed a number of nutritional and lifestyle guidelines to be practiced during confinement.[6] especially, mediterranean diet was recommended as a healthy and immune booster diet style.[7] the main aim of the study included investigating and understanding and the changes of eating behavior including eating pattern, shopping behavior, and lifestyle behavior during the covid-19 lockdown among the cihan university students. materials and methods study design the research used a cross-sectional design to study lifestyle changes and eating behavior in covid19 lockdown (lcebcovid-19) snowball survey sampling among cihan universityerbil students. the study was conducted from december 18, 2021, to february 20, 2022. the study was started by intervening students randomly from different department and sociodemographic backgrounds. there was no inclusion expert for being cihan-university-erbil. sample size a total of 370 undergraduate student of cihan universityerbil (n = 5000) was randomly selected from genders (applied and humanitarian fields). the size of the sample was finalized following the yamane’s formulation considering 50% prevalence at the finite targeted population. n = n/(1 + ne2) = approximately 370 where, n = size of the targeted sample, e = error of margin of 0.05, and n = size of the population that is approximately accounted to 5000. study questionnaire and instruments lceb-covid-19 questionnaire was first created by the previous studies (di renzo et al., 2020) (galali). a total of 43 questions were structured that divided into three parts. • part i: sociodemographic data (five questions) such as age, gender, place of living and weight, and height. • part ii: dietary behavior part (11 questions: including two parts: (a) mediterranean diet (md) adherence utilizing 13-scaled questions and using md screener tool ranging from 0 to13 and (b) set of questions food intake structured style: intake of unhealthy foods; pastry products and packaged sweet, salted snacks, fizzy drinks, and sauces used for dressing on a daily basis. • part iii: lifestyle changes (12 questions): sleeping behavior, smoking habit, physical activity, and shopping. ethical consent ethical approval was not required by the university; however, consent of participating from the participants was taken. statistical analysis statistical analysis was done to analyze data utilizing spss (statistical package for the social sciences v. 21.0, ibm). chisquare test was done to compare variables of different categories. statistically significant level was set at a level of p < 0.05. results demography and lifestyle this questionnaire was launched in 18 des 2021, and data collected were commenced after the confinement was attenuated after 2 months passed. the answer of the participants was analyzed statistically. the total number of participants was 370 as designed after validation of the data, and any missing data were excluded. the number of participants was (78.9%) 292 from erbil city, (8.9%) 33 from sulaimani city, (5.7%) 21 from duhok city, (4.6%) 17 from soran city, and (1.9%) 7 from halabja city [table 1]. data of gender showed that (51.4%) 190 were male and (48.6%) 180 were female according. in relation to participants, body mass index (bmi); normal bmi 54% (198), underweight 8.4% (31), overweight 28% (99), and obese type i was 7% (26), obese type ii was 3.2% (12), and obese type iii was 1% (4). participants aged from 18 to 25 years old represent the majority of the participant with 68.4% (253), following by participants who aged from 26 to 30 years old with 21.4% (79), following by participants who 31–35 years old with 5.9% (22), then participants who more than 35 years old with 4.3% (16). regarding residency, 65.1% (241) stated that they live in cities and 34.9% (129) were from countryside areas. regarding the lifestyle changes, the data showed that the greater number of the students stated that their lifestyle had effected in some way. for instance, 17% (63) of participants reported that their lifestyle was getting better; on the other hand, 38.6% (143) stated that their lifestyle quality had deteriorated [table 2]. the results also showed that the frequency of physical activity seems to be lowering and the students are less active. the percentage of students did not practice exercise before and during the outbreak was 50% and 67%, respectively. furthermore, the weekly sessions of exercise training number were also significantly reduced [table 4]. smoking cigarettes were slightly changing but not significantly different (p > 0.05). only people who smoked 5–10 cigarettes seem to be increasing and people who smoked more than 10 cigarettes decreased. regarding sleeping hours, there are differences before and during covid-19 pandemic lockdown. the percentage of people reporting sleeping <7 h reduced from 36.5% to 30.5%. the percentage of students stated sleeping between 7 and 9 h per night reduced from 46.5% to 38.1% and those reporting more than 9 h sleep increased from 17% to 31.4% [table 6]. the data also revealed the before the covid-19, the most practiced exercise was running (11%), gym (25%), football (9%), basketball (3.5%), swimming (1%), volleyball (0.1%), and tennis (0.8%), respectively. on the other hand, functional training (25%), treadmill running (4.3%), weight lifting (2.5%), and using training tools (2.5%), respectively, were among the most practiced exercise during the pandemic [table 3]. zebari, et al.: eating behavior and life style during covid-19 143 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 141-146 the data also showed that smoking behavior seems to slightly changing (p > 0.05). only people who smoked 5–10 cigarettes seem to be increasing and people who smoked more than 10 cigarettes decreased [table 5]. regarding sleeping hours, there are differences in relation to covid-19 confinement. the students’ percentage reporting sleeping <7 h reduced from 36.5% to 30.5%. furthermore, the students’ percentage confirmed sleeping hours between 7 and 9 h reduced from 46.5% to 38.1% and those exceeding 9 h sleep raised from 17% to 31.4% [table 6]. eating habits eating habits also seems to be effected by the confinement, the students percentage stated an appetite change during covid-19 confinement revealed that 37% (136) of the table 2: perception of participants about lifestyle changes during covid-19 perception frequency % no changes 164 44.3 yes, for worse 143 38.6 yes, for better 63 17.0 total 370 100.0 table 4: physical activity and exercise frequency before and during lockdown frequency before during number % number % nothing 185 50 243 65.6 1–2 times 87 24 90 24.3 3–5 times 52 14 22 5.9 more than 5 46 12 15 4 table 3: exercise types practiced during and before covid-19 confinement types of exercise before types of exercise after no. % no. % no exercise 185 50 no exercise 243 67 running 40 11 functional training 93 25 gym 91 25 treadmill 16 4.3 food boll 33 9 weight lifting 9 2.5 swimming 4 1 using training tools 9 2.5 volley ball 1 0.1 tennis 3 0.8 basketball 13 3.5 table 5: smoking frequency before and after lockdown smoking frequency before during frequency % frequency % no 248 67 248 67.0 <5 cigarettes 40 10.8 38 10.3 5–10 cigarettes 26 7.0 29 7.8 >10 cigarettes 56 15.1 55 14.9 total 370 100 370 100.0 table 1: participants sociodemographic data characteristics total residency erbil sulaimani duhok halabja soran no. (percentage) no. (percentage) no. (percentage) no. (percentage) no. (percentage) no. (percentage) gender male 190 (51.4%) 144 (39%) 20 (5.4%) 13 (3.5%) 4 (1%) 9 (2.5%) female 180 (48.6%) 148 (40%) 13 (3.5%) 8 (2.1%) 3 (0.8%) 8 (2.1%) age 18–25 253 (68.4%) 205 (55.5%) 18 (4.9%) 14 (3.8%) 7 (1.9%) 9 (2.5%) 26–30 79 (21.4%) 62 (16.8%) 9 (2.5%) 6 (1.7%) 0 2 (0.5%) 31–35 22 (5.9%) 15 (4.1%) 3 (0.8%) 1 (0.1%) 0 3 (0.8%) older than 35 16 (4.3%) 10 (2.7%) 3 (0.8%) 0 0 3 (0.8%) bmi underweight 31 (8.4%) 27 (7.3%) 1 (0.1%) 2 (0.5%) 1 (0.1%) 0 normal weight 198 (54%) 153 (41.4%) 23 (6.2%) 10 (2.7%) 2 (1%) 10 (2.7%) overweight 99 (28%) 82 (24%) 5 (1.4%) 8 (2.1%) 1 (0.1%) 3 (0.8%) obesity 1 26 (7%) 17 (5%) 4 (1%) 1 (0.1%) 1 (0.1%) 3 (0.8%) obesity 2 12 (3.2%) 10 (2.7%) 0 0 2 (0.5%) 0 obesity 3 4 (1%) 3 (0.8%) 0 0 0 1 (0.1%) bmi: body mass index zebari, et al.: eating behavior and life style during covid-19 144 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 141-146 students thought appetite improved, however, 23% (84) reported opposite and 40% (150) stated no change appetite [table 7]. according the data [table 8], it can be seen that the percentage of students who put on weight is 36% (133) or gained a lot of weight was 17% (17) whereas 19% (70) of the students thought that they lost weight and 40% (150) was felt no change in weight. the results also showed that the percentage of the students feeling hunger before the main meals was 47.8 % (177), between main meals was 26% (97), and hunger post dinner was 27% (100). food consumption behavior was also researched before and during the pandemic. the data showed that delivered pizza, vegetables, fruit, cereals, homemade pizza, and wine increased. whereas, consumption of hot beverage, delivered pizza, wine and beer, processed meat, snack, and homemade pizza decreased. the data of place of purchase revealed that percentage of students buy food products in supermarkets by 75% (276), grocery 10.2% (38), and local products 12% (44). despite significant decrease in food supply chain of organic and local products in kurdistan, but still 3.2% of participants purchase organic foods from locals, this might be attributed to safety and health reasons [figure 1]. data analysis of the participants showed that here is no relationship between bmi and appetite change (p < 0.05). bmi was also associated with meal number change. for instance, 25.8% of underweight, 19.7% of normal, 12.1% over, and 23.8% of obese participants seem to have more meals during the pandemic. furthermore, the data analysis also showed that there is a significant association between weight change and appetite change (p < 0.0001), but age had no relationship with weight change (p > 0.05). furthermore, 47% of the participants who thought their weight increased it seem that their appetite was also increased. the data analysis also showed that weight change is significantly different among gender groups (p < 0.05). furthermore, 41% of the males and 29% of the females think that their weight has increased [table 8]. md adherence screener tool the md adherence screener tool was used to determine the adherence level to md. according to their adherence level, the participants were categorized into three groups: high, low, and medium adherence. furthermore, it can be seen that the greater number of the participants is medium adhered to 248 (67%) [table 9]. consumption of the foods that have the highest level of adherence included delivery pizza, vegetables, fruit, nuts, cereals, home pizza, wine, dairy products, vegetable juice, and eggs, respectively. according to data, the students seem medium adhered to md fruits and vegetables are particularly highly consumed [figure 1]. discussion this study has been done after the covid-19 lockdown in the cihan university, erbil, iraq, alongside the studies[3] in kurdistan region, iraq, and spain (rodríguez-pérez et al., 2020). the insight of weight increase might be connected with the fact that individuals are less physically active are more sedentary and consume extra frequent meals and/or snacks.[8] many governments implemented safety precautions to flatten positive cases during the early emergency stages of the covid-19 outbreak, including as quarantine and isolation. despite the fact that these protective measures reduced the percentage of cases, along with media coverage of incidence and mortality, as well as employment losses and restrictions on persons’ freedom had a negative impact on psychology, eating behavior, and lifestyle of people and the public. this might be of particular interest when health professionals advise maintaining a positive mental attitude and eating a well-balanced diet to boost the immune system and lower the chance of viral infection, such as covid-19.[9] this is extremely critical when no approved protective or therapeutic drug is registered, and the community should advise caution and maintain a healthy lifestyle and diet. the results revealed that percentage of students who exercised during covid-19 was declining. similarly, due to confinement, the total hours of sleeping are increased. these might address that outbreaks made the students less active particularly and more sedentary outdoors activities. however, there are still students who can create chances to practice exercise at home, especially functional such as treadmill and training. whereas, the percentage who students who smoked between 5 and 10 seem to be increased by 1%. this could explain raising stress and quarantine upset. the data also discovered that almost more than half of students had weight gain perception. this might be linked to an increase in hunger and the introduction of a meal or a snack during confinement, since roughly identical percentages of individuals stated that their appetite and snacks or meals table 6: the sleeping hours before and during lockdown frequency before during frequency percent frequency percent <7 h 135 36.5 113 30.5 7–9 h 172 46.5 141 38.1 more than 9 h 63 17.0 116 31.4 total 370 100.0 370 100.0 table 8: perception of weight changes among the participants during covid-19 lockdown weight changes no. percentage no change 70 19 reduced 84 22.7 increased 133 36 significantly increased 1 17 table 7: perception of appetite changes among the participants during covid-19 lockdown perception appetite changes no. percentage not changed 150 40.5 reduced 84 22.7 increased 136 36.8 zebari, et al.: eating behavior and life style during covid-19 145 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 141-146 number increased during lockdown. a recent study discovered that the impression of weight increase is linked to changes in hunger and the addition of additional meals.[10] people who are more sedentary and less physically active, as well as those who consume more often meals and/or snacks, may perceive weight increase.[8] increasing weight, or the thought of gaining weight, may have a negative impact on an individual’s physiology, psychological well-being, and immune system. obese people are more likely to get covid-19 virus infections, according to recent research and stress and anxiety.[11] as a result, it is worth noting that, in addition to remaining healthful, the psychological dimension of this epidemic is equally vital and useful in better managing it. home-cooked food and only-delivered-pizza consumption appear to be increasing, and individuals are spending more time cooking their own meals, according to the data. furthermore, some people’s unhealthy eating and junk food intake increased significantly after being quarantined. it was to be expected, considering how limited access to fruits and vegetables is under lockdown. kurdistan, in particular, is an area where the vast majority of vegetables and fruits are imported from outside, and the restriction of borders with surrounding countries has made it impossible to obtain. this could have a negative impact on the intake of immune boosting nutrients and general health. however, the mediterranean diet was modestly adhered upon by the majority of the participants as healthy and suggested diet for covid-19. because we have better access to these things before the quarantine, this number may be substantially higher. getting adequate healthy foods improves your health condition and helps you fight infections from table 9: the consumption of md components and adherence to md md components recommended servings (day or week) number percentage vegetables two or more s/day 198 53.5 fruits three or more/day 270 73.0 olive oil main dressing 75 20.3 olive oil four or more s/day 75 20.3 read meat less than 1 s/day 281 75.9 butter less than 1 s/day 83 22.4 legumes three or more/week 212 57.3 fish and seafood three or more s/week 135 36.5 sweet beverage less than 1/day 238 64.3 sweets less than/week 267 72.2 nuts three or more/week 258 69.7 white meat over red weekly 311 84.1 pasta, vegetable, or rice dishes flavored with garlic, tomato, leek, or onion 214 57.8 adherence to md* number percentage high 76 21 medium 248 67 low 46 12 *adherence to md high; of more than 9 components consumed, medium if 5–9 components is consumed, and low if<5 components consumed 67 .2 75 .1 67 41 .3 40 4 2 42 .4 31 31 .3 28 .1 40 .2 50 18 .1 18 28 .1 18 40 4 2 18 .1 4 0 96 19 45 5 1 7 15 2 1. 3 15 .4 12 4 40 .8 12 12 .1 22 22 .4 4 1. 6 11 1 4 2 2 22 39 .1 27 41 12 22 .1 4 1 88 .1 23 58 14 13 f ru it v eg et ab le n ut s h ot b ev er ag es h om em ad e sw ee ts e gg s d ai ry p ro du ct s w hi te m ea t r ed m ea ts s na ck s m ilk a nd y og ur t h om e pi zz a le gu m es f re ss h fis h e ne rg y dr in ks d re ss ig n so uc es s w ee ts v eg et ab le ju ic e c an ne d fis h p ro ce ss ed m ea t d el iv er t p iz za f ro ze n fis h w in e an d be er c er ea ls n on e increased dcereased figure 1: food consumption behavior during covid-19 zebari, et al.: eating behavior and life style during covid-19 146 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 141-146 viruses (silverio et al., 2020). as a result, the enough amount and diet types, such as the md, can give enough immuneboosting micronutrients, such as minerals and vitamins, to reduce the virus pathogenicity. conclusion the aim of this research was to see how covid-19 lockdown affects cihan university students’ food habits and lifestyle modifications. the findings revealed that 38.6% of participants stated that the lockout had a negative influence on their lifestyle. covid-19 has the potential to make the students less active than it was before to the confinement. covid-19’s eating habits appear to have deteriorated as well. extra meals or snacks and appetite were given. as a result, many of the individuals believe that they have gained weight. on the other hand, the participants appear to be moderately committed to the mediterranean diet, with a high consumption of vegetables and fruits. as a result, a wellbalanced diet combined with a more active lifestyle can help to 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activity and dietary behaviours among youth in the compass study. ssm population health, vol. 2, pp. 841-849, 2016. 9. l. zhang and y. liu. potential interventions for novel coronavirus in china: a systematic review. journal of medical virology, vol. 92, no. 5, pp. 479-490, 2020. 10. l. di renzo, p. gualtieri, g. cinelli, g. bigioni, l. soldati, a. attinà, f. f. bianco, g. caparello, v. camodeca, e. carrano, s. ferraro, s. giannattasio, c. leggeri, t. rampello, l. lo presti, m. g. tarsitano and a. de lorenzo. psychological aspects and eating habits during covid-19 home confinement: results of ehlc-covid-19 italian online survey. nutrients, vol. 12, no. 7, pp. 1-14, 2020. 11. s. k. karim, p. h. taha, n. m. m. amin, h. s. ahmed, m. k. yousif and a. m. hallumy. covid-19-related anxiety disorder in iraq during the pandemic: an online cross-sectional study. middle east current psychiatry, vol. 27, no. 1, p. 55, 2020. tx_1~abs:at/add:tx_2~abs:at 133 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 research article the use of tobit and logistic regression models to study factors that affect blood pressure in cardiac patients bekhal s. sedeeq, banaz w. y. meran department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq abstract this research studies the factors that affect blood pressure in cardiac patients using the tobit and logistic regression models. the data have been collected, from 500  patients with heart disease in hospital – heart center – erbil. the two levels of blood pressure, low and high blood pressures, were taken from the patients as dependent variables plus other, independent variables (gender, age, urea, cholesterol, creatinine, and weight). the research shows that the median of blood pressure by means of arterial pressure (map) equation contains each of high and low blood pressures differently. this is due to the threshold value of 99.33, equal to,12/8 mmhg, which represent a normal level of a human blood pressure. the aim of this research is to explain the main concepts and processes of tobit regression analysis (censored and truncated) and logistic regression analysis, which are used for predicting the factors of independent variables that have more effect on the response variables, for example, blood pressureand to compare the outcomes of the two models (tobit regression and logistic regression) in order to determine which of the models best fits our data in which aic and bic are used. the data analysis of this research shows that the logistic regression model best fits our data, as compared with the tobit regression model. the data analysis has been achieved using statistical packages in r programming, matlab and statistical package for the social sciences (spss) v.26. keywords: akaike information criterion, bayesian information criterion, censoring, logistic regression model, tobit regression model, truncation introduction the health sector is one of the most vital sectors that undertake the task of providing health services to all members of society through health institutions to protect and improve society and achieve the well-being of its members. in fact, one of the components of building health in society is to ward off all diseases, and at the lead of these diseases is cardiovascular disease, which is one of the problems that challenge medicine. cardiovascular disease, the leading cause of death worldwide, is greatly exacerbated by high blood pressure. around,54% of strokes-and 47% of coronary heart disease occur worldwide as a result of high blood pressure. high blood pressure is common medical issue that becomes more prevalent as people become older.[1] there have been several statistical studies of individual data that include observations in which a dependent variable is equal to 0, for a digit of observations in the dataset. this behavior is known as censored or truncated data.[2] since james tobin’s work, the tobit regression has received a huge and diverse amount of theoretical interest (1958). its use in practical applications has been developed in fields such as economics, biology, finance, and medicine. tobit regression can be seen as a linear regression model where only the data, on the dependent variable are incompletely, observed.[3] logistic regression is a statistical, method for examining the relationship among a dependent variable of a nominal level and one or other independent variables so that those independent variables are from any type of measurement level.[4] logistic regression is investigation of the best method in the case of the binary dependent variable, according to dayton[5] logistic regression, which is a categorical data statistical modeling method. it is the usages of the same logic as ordinary least squares regression. this study, aims at: 1. utilizing both the tobit and the logistic regression models. corresponding author: banaz w. y. meran, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq. e-mail: banaz.yaqoob@su.edu.krd received: july 18, 2022 accepted: august 12, 2022 published: november 20, 2022 doi: 10.24086/cuesj.v6n2y2022.pp133-140 copyright © 2022 banaz w.y meran, bikhal s. sedeeq. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) meran and sedeeq: tobit and logistic regression models 134 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 2. knowing which factors of independent variables is more effecter on the dependent variable (blood pressure) using both models. 3. comparing the results of the two models to determine which one best fits our data that are using akaike information criterion (aic) and bayesian information criterion (bic). literature reviews shirafkan et al.[6] estimated the potential of tobit regression, as a method to study time to onset of cytomegalovirus in renal recipient transplants. the results of this study revealed that the age of patients was influenced by the time of onset of disease. consequently, the bigger the age, the shorter time required to-start infection. karim et al.,[7] tobit model and the traditional regression model were compared on data taken from patients with kidney disease, and through using some statistical measures, it was concluded that tobit’s model is preferable than the traditional regression model for this type of data. taleb et al.[8] used a binary logistic regression model, estimating the coefficients of this model the least squares method for people with heart disease. the aim of the investigation was to compare the actual reasons of death with the estimated causes of death. the binary logistic regression model concluded that smoking was the leading cause of death. rambeli et al.[9] used the binary logistic model for a study the aim of which was to identify the factors that influence a teacher’s decision to remain in their professional life. the result showed that the income was considered as a key factor in teacher remaining committed to the profession. methodology this research contains three parts: the first part of this research included the introduction, methodology, aim of the study, and review of the literature. the second part of the research included the basic concepts of the two models and the last part contains data applications, implementation of two models, and interpretation of the results. tobit regression model regression analysis is one of the most important ways to find out the significant effect between the dependent variable and explanatory, variables but sometimes, the dependent variable is constrained to the threshold point. in this situation, the use of the traditional regression model is biased. with this kind of data, using the tobit regression model is the best option. logistic-regression, it is used as in this, study. the analysis of the tobit regression means the, statistical method used to examine the relationship between the limited dependent variable and explanatory variables of any type. the analysis in this situation is named a tobit regression. a limited dependent variable is the one whose range of potential values is constrained in some significant way.[10] limited dependent variable models contain: a – censoring, in which some data are lost but other data are present for certain persons in a data collection and b – truncation, in which some individuals are purposefully removed from observation.[11] tobin used a regression model based on household expenditures that particularly took into account the fact that (his regression model’s dependent variable) cannot be negative. tobin coined the phrase “model of constrained dependent variables” to describe his approach, a term invented by goldberger (1964) for the reason that they resemble probit models. these models are frequently denoted to as truncated or censored models. if the observations are lost outside of a carefully defined range and are censored, the model is called a truncated model.[12] the structural tobit model y xi * � �� ei� where, ei ~ n(0, σ 2). y* is a latent variable that is observed y, shown by the following equation y y if y if yi y � � � � � � �� * * ���� * ��� ���� ����� ���� ������ � � � when assume that τ = 0 in the standard tobit model, data are censored next to a value of 0. for use there is:[13] y y if y if y i � � � � � � �� * * * ��� ���� ����� ���� ���� ������ 0 0 0 as it has been shown, the probability function is censored l y µ i n di di � �� � � � � � � � � � � � � �� � � � � � � � � � � �� � � 1 1 1 � � � � � � in the conventional tobit model, one groups τ = 0 and parameterize µ as (x i β). that is tobit model’s likelihood function: l y x x i n i i di i di � �� � � � � � � � � � � � � � � � � � � � � � � � �� � 1 1 1 � � � � � � the tobit model’s log likelihood function is ln ln l d y x d xi n i i i i i � � � �� � � � � � � � � � � � � �� � � � � � �� � 1 1 1 ln ln� � � � � � � �� � � � � � � � � � � � the overall log, likelihood is split into two components. the first component corresponds to a traditional regression for an uncensored observation, while the, second component represents the associated likelihood of censoring the test.[14] standard tobit model censored regression method has been very widespread as a standard tobit model since tobin (1958) first presented it to evaluate the relationship between household expenditure and household income. y i n'* ����� , , .� � � �x ui i� 1 2 y �y ������if��������y � n� �* * y � �if�������y n� �0��������� * where, u i is identical independent distribution (iid). drawings from n(0, σ u ). y i and x i are observed in the sample, but the yi * is unobserved if � *y i2 < 0. the likelihood function for this model is: [12] meran and sedeeq: tobit and logistic regression models 135 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 l x y xi i i� � � � � � � � � � � � � � �� � � � � �� � 0 1 1 1 � � ' '� � � � � censoring a regression, model is said to be-censored when the recorded, data on the dependent variable (the response) cutoff outside a certain-range with multiple observations at the-endpoints of that range. when the data are censored, variation in the observed dependent variable will underestimate the impact of the regression on the actual dependent variable. as a result, coefficient estimates from standard ordinary least squares regression employing censored data are often biased toward zero.[15] truncation truncated data, or missing data, are discovered when an observation is not reported despite of whether it is below or above a specific level that differs. in actuality, these are known as left and right truncation, respectively. the truncation effect can also happen when only a small portion of a larger population is represented in the sample data. in addition, the response variable in the model is truncated if observations are not possible while taking values inside a particular range. consequently, when the dependent variable is within that range, neither the dependent nor the independent variables are observed.[2] logistic regression model what distinguishes a logistic regression model from the, linear regression model, in logistic regression, could be the result variable which is either binary or, dichotomous. this difference among logistic and linear regression is reflected both in the choice of a parametric model and it assumptions, whereas the methods used in a logistic, regression study follow the same fundamental principles, as linear regression.[16,17] logistic regression model circuitously models the response variable created on probabilities linked with the digits of the dependent variable y. we will use p(x) to represent the possibility of a response when y = 1. furthermore, we will define, 1–p(x) which represents the possibility of a response when y = 0. these probabilities are written as follows: p x p y x x xk� � � � �� �1 1 2| , , 1 0 1 2� � � � � �� �p x p y x x xk| , , the logistic regression equation and the regression equation with a straight line, equation ( y x x xk k� � � � ��� � � �1 1 2 2 . ) are related by the formula below. the logistic regression equation form is rewritten as such: logit p x p x p x x x xe k k� log .� � � � � � � �� � � � � � � � � � � � � � �� 1 1 1 2 2�� � � � the logistic regression model can be calculated using the formula below.[17,18] p x x x kxk x x kxk ( ) . � � � � ���� � � � �� �� � e e �� �� � � � � � � 1 1 2 2 1 1 2 21 estimate the parameters of logistic regression model the approach of maximum likelihood estimation will be used. the log likelihood is given as:[19-21] ( )yi 1 yi 1 (  ,   ) [1 ( )] n i l p x p x − = α β = −∏ we’ll use the log likelihood method for estimation: logl logp x p x i ( �, ) ( ) ( ( )� � � � � � � �� � � �� � � � � � � � � � � 1 1 1 n i i n iy n y log �� � � � � � � � � �� � � �� � � � � �u p l p x i ( ) ( , ) / ( ) /( ( � ��� p y n y p x i n i n i 1 1 1 ))) � � � � � � � � cox and snell r2 statistic in the logistic regression model, the determination coefficient r2 used to determine the fittingness of the proposed regression, table 1: five hundred patients sample id y: bp x1: gender x2: age x3: urea x4: cholesterol x5: creatinine x6: weight 1 88 (0) 1 70 68 127 1.26 89 2 76 (0) 2 40 41 117 0.97 96 3 99.33 1 44 34 201 0.59 74 4 85 (0) 2 43 33 221 0.67 107 5 117 1 55 54 226 1.12 88 6 92 (0) 1 67 31 173 0.72 76 7 96.67 1 47 43 165 0.78 65 497 107.67 1 54 31 160 0.87 85 498 93.67 2 63 16 109 0.6 69 499 106.67 1 58 25 100 0.73 67 500 118.33 1 69 38 134 1.11 65 bp: blood pressure meran and sedeeq: tobit and logistic regression models 136 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 models for the study data is changed by the r2 nagelkerke, cox, and snell r2 computation statistics. can be calculated as: � ( / ) r cox�and�snell2 0 1 2 1� � � � � � � � l l n l 0 : maximum likelihood for constant in the model. l 1 : maximum likelihood independent variables in the model n: sample size[22] and, can be calculated as: r r n 2 2 0 21 nagelkerke cox snell l � � � �&� ( / ) the hosmer-lemeshow test the hosmer and lemeshow test is a widely-used test for determining a model’s quality of fit. it accepts any number of explanatory-variables, which can be continuous or categorical. it is used to test the hypothesis: h 0 : the model is adequate for data h 1 : the model is not adequate for data the model will be a useful model if (hosmer and lemeshow) static is >0.5.[22] hl o e nk i n i i i i i � � �� � � �� � �� ��2 2 1 2 1 wald statistic the wald test is used to determine whether or not the effect of the logistic regression coefficient on the independent variables.[23] the wald statistic is calculated according to the following formula: 2 2        . w s eβ  β =      the classification table the use of classification tables is one of the ways to check the quality of matching the model with the data. this method depends on creating tables that put the number of cases that have the desired trait or the cases that do not have the desired trait and that were categorized correctly or incorrectly. the concept behind using the analysis is it leading to better results if the model is data compatible.[24] the aic aic as a model selection criterion for assessing actual data, it has played a key role in solving issues in a wide range of fields, and the model by the lowermost aic is chosen as the best model.[25] aic l k�� �� log *2 2 the bic in statistical model selection, the bic is unique of the best well-known and commonly used tools. bic is calculated for each of the models, and the model with the lower bic value is selected as the best model.[26]  2 log 2 * log *bic l n k=− + data analysis in this study, the two models of tobit and logistic regression models have been applied on a sample taken from 500 patients with heart disease and two levels of blood pressure, high and low blood pressure, in hospital – heart center – erbil. blood pressure is taken from the patients as response variables and some independent-variables (gender, urea, age, cholesterol, creatinine, and weight). the study found that the average of blood pressure by means of arterial pressure (map) equation contains each high and low blood pressure differently because the threshold point was determined to be 99.33. to take the best model for our data in the study, two statistical measures (aic and bic) were used. note: all assumptions and tests related to tobit and logistic regression models have been applied before we started the data analyses in this study. data description for tobit regression analysis in this study, the data are gathered from 500 patients with heart diseases, and the two levels of blood pressure; high and low blood pressure were taken from patients as dependent variables and the variables: gender, age urea, cholesterol, creatinine, and weight as the independent variables (table 1). the researcher set that the medium of blood pressure by map equation contains each highest and lowest blood pressure differently because the threshold point was determined to be 93.33, in regards, the dependent variable in this study has been defined as: y=y* y*>93.33 appropriately, the limitation threshold point y = 93.33 was regarded the limitation threshold point y = 0, in accordance with the stated theoretical presentation, and the model will be referred to as the following model: y=y* y*>93.33 y=0 y*<=93.33 the explanatory variables in this study are the follows: x 1 : gender (male and female) x 2 : age measured by year x 3 : urea (mg/dl) x 4 : cholesterol (mg/dl) x 5 : creatinine (mg/dl) x 6 : weight measured by kg the explanation of the variables is presented in table 2 in which contains independent variables and dependent variable and the table views the maximum, minimum, mean, and standard deviation of data. application of tobit regression model analysis (censored and truncated) because of the researcher initially checked all the necessary assumptions that must be present in the data before starting to analyze the data, and also set a unified standard for the data in this study, it becomes clear for us that there is no problem in terms of our data and we can use the data for analyzing. meran and sedeeq: tobit and logistic regression models 137 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 censored regression model first, let’s start with censored-regression model are: censored (formula = y~ x, -left = 0, right = inf., data= data 1) total (n = 500 observation, left censored = 132 observation, uncensored = 368 observation, left censored (y < 99.3 then y* = 0: observation). table 3 presents the results of the exact regression model. the coefficients of the independent variables sex, age, cholesterol, creatinine, and weight are positive because the variables have a positive relationship with the dependent variable (blood pressure), while the coefficient of the independent variable urea is negative because the variable has a negative relationship with the dependent variable (blood pressure). according to the findings in table 3, the variables age, cholesterol, and creatinine all significantly affect blood pressure. the summarized fit to the censored regression model is log-likelihood = −2125,549, aic = 4265.1, bic = 4294.6. the score for the best model is determined by the lowest value for aic and bic. truncated regression model in this case, the number of observations turns to 368 due to a truncation. table 4 shows the results from truncated regression model. those coefficients of the independent variables such as gender, age, cholesterol, urea, and weight are positive for the reason that the variables take a positive relationship with the dependent variable (blood pressure) whereas the coefficient of the independent variable creatinine is negative because has a negative connection with the dependent variable (blood pressure). it is through the conclusion in table 4 that only the urea variable has the effect on blood pressure. logistic regression analyses in this part, we use binary logistic regression since the dependent variable in this study is blood pressure and the researcher has taken the average of blood pressure by map equation which contains each of the high and low blood pressure differently because of the threshold points which was determined to be 93.33. the patient whose blood pressure is >93.33 is considered to be infected and takes the worth of one while the patient whose blood pressure is ≤93.33 is considered to be uninfected and takes the value of 0, and the other variables are gender, age, cholesterol, urea, creatinine, and weight. y: the dependent variable has a binary response code: y infected uninfected � � � � 1 0 the independent variables are the same as the variables written above. application of binary logistic regression analysis let’s start with the outcome of the classification table starting with the zero stage in which the model is free of independent variables (only the constant). table 5 represents the baseline model, which is a model without our explanatory variables. the overall right percentage was 73.6% which refers the model’s overall explanatory strength . the initial log likelihood function (-2 log likelihood function)= 577.2. omnibus test of logistic model coefficients based on the model coefficients in omnibus tests, we find that the chi-square tests are to illustrate if there is an important variance between the factors of the nil model and the current model. table 2: descriptive statistics of variable variables minimum maximum mean sd bp 70.33 141.00 101.23 10.97 gender 1 2 1.33 0.47 age 17 86 57.73 11.24 urea 11 198 40.11 20.53 cholesterol 63 326 167.36 45.02 creatinine 0.11 10.70 1.58 1.195 weight 48 135 78.83 13.47 bp: blood pressure, sd: standard deviation table 3: censored regression model coefficients estimate se t pr (>t) constant −127.94562 25.77314 −4.964 6.89e-07*** gender 3.57944 5.59007 0.640 0.522 age 1.91136 0.24742 7.725 1.12e-14*** cholesterol 0.25688 0.05688 4.516 6.30e-06*** urea −0.11037 0.12724 −0.867 0.386 creatinine 12.21152 2.09200 5.837 5.31e-09*** weight 0.31290 0.19800 1.580 0.114 *** mean p ≤ 0.001 table 4: truncated regression model coefficients estimate se t pr (>t) constant 98.7995674 4.7143913 20.9570 <2e-16*** gender 0.6703624 0.9215182 0.7275 0.46695 age 0.0645897 0.0448466 1.4402 0.14980 cholesterol 0.0015114 0.0094141 0.1605 0.87245 urea 0.0520044 0.0202811 2.5642 0.01034* creatinine −0.2198352 0.3154896 −0.6968 0.48592 weight 0.0040456 0.0340886 0.1187 0.90553 se: standard error; * mean p ≤ 0.05 and *** mean p ≤ 0.001 table 5: classification table shows that the model has a constant bound zero step observed predicted y percentage correct uninfected infected step 0 y uninfected 0 132 0.0 infected 0 368 100.0 overall percentage 73.6 meran and sedeeq: tobit and logistic regression models 138 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 table 6 displays that the current model is meaningfully more suitable than the null model. model coefficients omnibus test offers significant reduction in the −2 log likelihood value = 426.572= as compared to −2 log likelihood value =577.2 of the null model. this indicates that the chi-square values for step, block, and model are all the same. p < 0.05 illustrating that the model’s accuracy increases when explanatory variables are included. furthermore, the chi-square is significant (χ2 = 150.628, df = 6, p < 0.05). as a result, our new model is bested. hosmer and lemeshow test being a goodness of fit test for logistic regression, it fits the data model. table 7 explains since p = 0.226, it is larger than the level of significance at 5%. we may well conclude that the data are will be suitable to the model. cox and snell r2, nagelkerke’s r2 the cox and snell r2, nagelkerke’s r2 values are utilized to estimate the model’s fit to the data. as the nagelkerke’s r2, cox and snell r2 values given in table 8 are examined, the ratios of interpretation of the independent variables over the dependent variable are shown. the value of nagelkerke r2 is the modified form for the cox and snell r2 coefficient. giving to the outcomes shown in table 8, it is seen that the dependent variables determine 26% of the variance in the independent variables according to the value of cox and snell r2, and 38% according to the value of nagelkerke r2. the amount of the −2 log likelihood statistic is 426.572 in model summary for the whole model. table 9 is corresponding to table 5 but it is based on the model that contains our explanatory variables. the total percentage of correct was 79.2% which replicates the model’s overall explanatory strength. classification table contains the constant term and the rest of the predictors, that is, 91.6% were correct for the infected of blood pressure and correctly classified. this table shows how many events were correctly predicted (59 cases were observed to be uninfected and were correctly predicted to be uninfected; 337 cases were observed to be infected and were correctly predicted to be infected) and how many were not (73 cases are observed to be uninfected but are predicted to be infected; 31 cases are observed to be infected but are predicted to be uninfected). variables in the equation logistic-regression model the variables in the equation logistic regression are the most essential of all the outputs. this table must be studied carefully since it contains the answers to our questions about the common relationship between all variables. table 10 shows the values of wald test that represents the parameter of the test value of the model and it appears that the variables (age, urea cholesterol, and creatinine) represent the significant variables in the research. it is by associating the p-value with the level of significant (0.05) that the p-value represents the significance of the effect of the variable on the patient condition, that is, it is significant when p < 0.05 was considered for the variable under test. it shows that the variables (gender and weight) are not significant variables in the study, and it is through comparing the p-value with the level of significant (0.05) that the p-value represents the non-significance of the effect of the variables on the patient condition, that is, it is no significant when p > 0.05 was considered. as far as table 10 is concerned, the logistic computation coefficients that may be utilized to build a predictive equation could be: y � � � � � � � � �� � � �� � � � ��log p x p x x x xk k1 1 1 2 2 � � � � the effect factors for blood pressure in cardiac patients can be ranked as follows based on the value of the odds ratio. likewise, we can write the logistic regression computation with just significant variables: logistic equation (model): y = −7.689 + 0.080 age 0.016 urea + 0.013 cholesterol + 1.451 creatinine table 10 shows, exp(β) = eb represents the ratio changed in the odds of the event of importance for a one unit variation in the predictor. the value of exp(β) for the variable gender indicates that when the gender changes from the value 0 (female) to the value 1 (male), the probability of disease blood pressure in patients with heart disease increases because the value of exp(β) is >1. such incomes indicate that the blood pressure of males is higher than females. table 6: omnibus test step 1 χ2 df significant step 150.628 6 0.000 block 150.628 6 0.000 model 150.628 6 0.000 table 7: hosmer and lemeshow test step χ2 df significant 1 10.595 8 0.226 table 8: summary of logistic regression step −2 log likelihood cox and snell r2 nagelkerke r2 1 426.572a 0.260 0.380 table 9: classification tables observed predicted y percentage correct uninfected infected step 1 y uninfected 59 73 44.7 infected 31 337 91.6 overall percentage 79.2 meran and sedeeq: tobit and logistic regression models 139 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 the odds ratio for the variable age is >1, exp(β) = 1.083. this means that each additional rise of 1 year in age is in touch with the increase in the odds infected of blood pressure in cardiac patients. the odds ratio for the variable urea is <1. this means that each additional increase of one unit in urea is associated with decrease in the odds infection of blood pressure in cardiac patients with 0.984 times. the odds ratio for the variable cholesterol is >1. this means that each additional increase of one unit in cholesterol is related to the increase in the odds of blood pressure in cardiac patients with 1.013 times. the odds ratio for the variable creatinine is >1. this means that each additional increase of one unit in creatinine is related to the increase in the odds infection of blood pressure in cardiac patients with 4.267 times. the odds ratio for the variable weight is >1. this means that each additional increase of one unit in weight is associated-with the increase in the odds infection of blood pressure in cardiac patients with 1.011 times. discussion there are different techniques for comparing the analysis of two or more models; however, the aic and bic criteria are two that may be worth considering. table 11 shows a comparison between three regression models censored, truncated, and logistic, for choosing the most fit model to our data of blood pressure in cardiac patients, the aic and bic values with the least values are chosen. the results display that the logistic regression model is better and more suitable rather than truncated regression and censored regression for our data, because it’s aic equal to 591.2 and bic = 620.7 are the lowest values contrast with tobit models (censored and truncated). conclusion it has been concluded the following: 1. in the censored, regression model, the explanatory, variables (age, cholesterol, and urea) significantly impacted on blood pressure. 2. the results, from truncated regression model show, that only the urea variable has the effect on blood pressure. 3. according, to the results classification table, logistic model is, correctly classifying the consequences for 79.2% of the cases, compared to 73.6% in the null, model. 4. according to the hosmer-lemeshow test, our data fit the logistic regression based on a χ2 = 10.595 and p-value greater, than a significant level. 5. wald’s test showed that the variables of age, creatinine, cholesterol, and urea, respectively, contributed significantly to the prediction, depending on the p-value (0.000 < 0.005). the variables that do not have a significant, effect are weight and gender. 6. it was concluded, that the logistic regression model for the sample under study or for our data is better, than the censored regression model and the truncated regression model after comparing, their aic and bic values. references 1. c. y. wu, h. y. hu, y. j. chou, n. huang, y. chou and c. li. high blood pressure and all-cause and cardiovascular disease mortalities in community-dwelling older adults. medicine(baltimore), vol. 94, no. 47, p. e2160, 2015. 2. w. h. greene. censored data and truncated distributions, ssrn electron. j, 2005. 3. m. h. odah, b. k. mohammed and a. s. m. bager. tobit regression model to determine the dividend yield in iraq. lumen proceedings, vol. 3, pp. 347-354, 2018. 4. j. s. cramer. the origins of logistic regression: tinbergen institute discussion papers, 2002. 5. c. m. dayton. logistic regression analysis. stat, vol. 474, p. 574, 1992. 6. h. shirafkan, j. yazdani-charati, s. a. mozaffarpur, s. khafri, r. akbari and a. a. pasha. application of tobit model in time until cytomegalovirus infection in kidney transplant recipients. acta medica, vol. 32, p. 1237, 2016. 7. r. m. h. karim and s. m. salh. using tobit model for studying factors affecting blood pressure in patients with renal failure. uhd journal of science and technology, vol. 4, no. 2, pp. 1-9, 2020. 8. h. r. talib and s. a. mazloum. the use of binary logistic table 10: variables in the equation step 1 β se wald df significant exp(β) 95% ci for exp(β) lower upper gender 0.108 0.264 0.168 1 0.682 1.114 0.664 1.868 age 0.080 0.012 41.674 1 0.000*** 1.083 1.057 1.110 urea −0.016 0.007 4.692 1 0.030* 0.984 0.970 0.998 cholesterol 0.013 0.003 21.265 1 0.000*** 1.013 1.007 1.019 creatinine 1.451 0.276 27.595 1 0.000*** 4.267 2.483 7.333 weight 0.011 0.009 1.327 1 0.249 1.011 0.992 1.030 constant −7.689 1.294 35.304 1 0.000 0.000 ci: confidence interval, se: standard error, * mean p ≤ 0.05 and *** mean p ≤ 0.001 table 11: comparison tobit regression model (censored) truncated regression model logistic regression model aic=4265.1 aic=2574.6 aic=591.2 bic=4294.6 bic=2602.0 bic=620.7 aic: akaike information criterion, bic: bayesian information criterion meran and sedeeq: tobit and logistic regression models 140 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 133-140 regression method to analyze the factors affecting heart disease deaths: an applied study on a sample of patients in dhi qar governorate. journal of al-rafidain university college college for sciences, 2020, no. 46, 2020. 9. n. rambeli, e. hashim, f. c. leh, n. s. hudin, m. f. ramli, m. c. mustafa, et al. decision to leave or remain in the career as early childhood educator: a binary logistic regression model. review of international geographical education online, vol. 11, no. 5, pp. 450-456, 2021. 10. g. s. maddala. limited-dependent and qualitative variables in econometrics. new york, ny: cambridge university, 1983. 11. n. m. ahmed. limited dependent variable modelling (truncated and censored regression models) with application. the scientific journal of cihan university sulaimanyia, vol. 2, no. 2, pp. 82-96, 2018. 12. t. amemiya. tobit models: a survey. journal of econometrics, vol. 24, no. (1-2), pp. 3-61, 1984. 13. j. s. long, regression models for categorical and limited dependent variables. thousand oaks, ca: saga publication, 1997. 14. a. flaih, j. guardiola, h. elsalloukh and c. akmyradov. statistical inference on the esep tobit regression model. journal of statistics applications and probability letters, vol. 6, no. 1, pp. 1-9, 2019. 15. k. y. chay and j. l. powell. semiparametric censored regression models. journal of economic perspectives, vol. 15, no. 4, pp. 29-42, 2001. 16. d. hosmer jr., s. lemeshow and r. sturdivant. applied logistic regression. vol. 398. new york: john wiley and sons, 2013. 17. a. m. khudhur and d. h. kadir. an application of logistic regression modeling to predict risk factors for bypass graft diagnosis in erbil. cihan university-erbil scientific journal, vol. 6, no. 1, pp. 57-63, 2022. 18. n. m. m. abd elsalam. binary logistic regression to identify the risk factors of eye glaucoma. international journal of sciences basic and applied research, vol. 23, no. 1, pp. 366-376, 2015. 19. n. s. k. barznj. using logistic regression analysis and linear discriminant analysis to identify the risk factors of diabetes. zanco journal of humanity sciences, vol. 22, no. 6, pp. 248-268, 2018. 20. s. menard. applied logistic regression analysis. vol. 106, new york: sage, 2002. 21. n. h. mahmood, r. o. yahya and s. j. aziz. apply binary logistic regression model to recognize the risk factors of diabetes through measuring glycated hemoglobin levels. cihan university-erbil scientific journal, vol. 6, no. 1, pp. 7-11, 2022. 22. v. bewick, l. cheek and j. ball. statistics review 14: logistic regression. critical care, vol. 9, no. 1, pp.112-118, 2005. 23. d. hosmer and s. lemeshow. applied logistic regression. new york: johnson wiley and sons, 2000. 24. i. r. soderstrom and d. w. leitner. the effects of base rate, selection ratio, sample size, and reliability of predictors on predictive efficiency indices associated with logistic regression models, 1997. 25. s. konishi and g. kitagawa. information criteria and statistical modeling. berlin: springer science and business media, 2008. 26. k. i. mawlood. using logistic regression and cox regression models to studying the most prognostic factors for leukemia patients. qalaai zanist scientific journal, vol. 4, no. 3, pp. 705724, 2019. tx_1~abs:at/add:tx_2~abs:at 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 1-6 research article impact of textiled-poly bags as economic and environmentally friendly recycled fibers on mechanical features of soil heba a. ahmed department of surveying, darbandikhan technical institute, sulaimani polytechnic university, sulaimani, iraq abstract newly, the utilization of polypropene for packaging consumer products was populated significantly, which might make ecological problems. hence, it is substantial to detect environmentally friendly techniques to recycle these waste substances without causing any environmental risks. one such technique could be the utilize poly wastes as stabilizer agents for soils. in the current investigation, textiled-poly bags (tpb) have been recycled and fabricated to be economy and environmentally-friendly fibers. the impact of utilizing different proportions and lengths of these fibers on the mechanical features of clayey soil has been evaluated. the investigation depended on four different fiber-proportions (1, 2, 3, and 4%) of soil weight in two different lengths (1 and 2) cm. geotechnical experimental test consequences demonstrated that the recycled fiber pieces minimize the optimum dry density and the corresponding optimum water content of the treated soil samples. in addition, there was a notable increment in the uniaxial compression test results of the treated soil samples. moreover, the consequences of california bearing ratio tests showed that the utilization of recycled tpb fibers in clayey soil samples enhances the resistance and deformation performance of the soils specially when utilizing 4% of recycled tpb fibers for both lengths (1 and 2) cm. this recycled fiber improves the strength features and dynamic behaviors of clayey soils. therefore, the recycled tpb fibers could be successfully utilized as reinforcement materials for the modification of clayey soils. keywords: environmentally friendly techniques, textiled-poly bags, recycled fibers, geotechnical experimental tests introduction the elimination of waste substances has become prevalent trouble in most of the world’s countries. accumulating this waste in huge amounts causes ecological and monetary troubles.[1] in iraq, the reports revealed that polypropene waste is the second largest solid waste after food waste.[2,3] because most plastic waste is non-biodegradable and can remain for several years, several states have planned to reduce or prevent the impact of such waste by recycling or reutilizing it in efficiently techniques in several fields.[4-7] several civil engineers have performed several investigations to discover successful techniques to minimize the contamination of these substances inclusive of reutilizing and recycling these substances in construction and geotechnical implementations as solutions to preserving the environment from the contamination of polypropene waste substances.[8-16] literature review the utilization of polypropene waste substances as stabilizer agents for soils is one of successful techniques. many studies reported that conventional soil stabilizers such as cement and several pozzolanic substances are vastly utilized for enhancing the geotechnical features of weak bearing soils sherwood,[17] yadav and tiwari,[18] and yadav et al.[19] the efficiency of these stabilizers in enhancing soil’s features has been proved by several geotechnical engineers bell,[20] little,[21] rout et al.,[22] rasul et al.,[23] rasul et al.,[24] rasul et al.,[25] yadav and tiwari.[26] nevertheless, the utilization of these stabilizers in high proportions makes them cost-ineffective, as a result, authors such as obo and ytom) attempted to discover alternate cost-effective stabilizer agents such as polypropene, tires rubber chips, and rice hull.[27] shelema (2020) reported that the utilization of polypropene waste strips as stabilizer substances could enhance the foundation layers of pavements.[28] consequently, this technique could solve the trouble of waste’s cumulative by minimizing its corresponding author: heba a. ahmed, department of surveying, darbandikhan technical institute, sulaimani polytechnic university, sulaimani, iraq. e-mail: hiba.adnan@spu.edu.iq received: november 11, 2022 accepted: june 26, 2023 published: july 10, 2023 doi: 10.24086/cuesj.v7n2y2023.pp1-6 copyright © 2023 heba a. ahmed. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) ahmed: impact of textiled-poly bags on mechanical features of soil 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 1-6 amounts and recycling it for improving the characteristics of soils. one technique of utilizing polypropene for soil stabilization was to utilize the polypropene in the shape of separated fibers since when polypropene substances have been merged with soil’s particles, they conducted identical behavior of fiber-reinforced soils temel and salbas.[29] many geotechnical investigations have been implemented by stacy et al.[30] and (temel and salbas[29] to evaluate the efficiency polypropene waste substances in the shape of separated fibers on soil’s features. they reported that utilizing polypropene waste substances for soil stabilization could enhance the features of weak bearing soil like an enhancement in uniaxial compressive resistance, california bearing ratio, and reduction in soil’s plasticity. singh and hussain (2020) utilized different percentages of waste coir fiber with polypropylene fiber for soil stabilization, they discovered that 75% of the fiber content could be the best ratio to improve the features of the soil.[31] twinkle et al. (2011)[32] have investigated the impact of polypropylene fiber, lime admixture, and period of curing on the geotechnical features of clayey soil. they noted that the increase in fiber content caused an increment in resistance features and shrinkage potential and caused a decrement in the swelling potential. furthermore, the period of curing significantly improved the uniaxial compressive resistance and shear resistance parameters of the treated soil. jadho and nagarnik (2008)[33] have investigated the impact of polypropylene fibers in the geotechnical features of soil treated with fly-ash by utilizing various fiber lengths (6, 12, and 24 mm) with the range of (0–1.5% by dry wt. of the soil). they noted that the optimum improvement in the resistance features could be achieved at a fiber length of (12 mm), and the optimum proportion of fiber content was (1%). according to the literature, the tensile resistance values of strengthened stabilized soil with polypropylene fibers have improved to a great extent. the highest resistance values have been collected with approximately (0.5–1.0%) fiber content for the (12 mm) length. the value of shrinkage limits improved with increasing fiber content and length of the fiber, whereas volume changes and cracks have been minimized.[34-39] however, there are no investigations implemented in stabilizing clayey soil with fabricated textiled-poly bags (tpb) to evaluate the geotechnical features of such stabilized soil. this work is a sustainable and environmentally friendly experimental study focuses on the impact of utilizing different proportions (0, 1, 2, 3, and 4%) and different lengths (1 and 2 cm) of fabricated tpb on the mechanical features of clayey soil. materials and methods soil soil samples which utilized in the current investigation have been collected from raparin area, in the sulaymaniyah governorate of the kurdistan region of iraq, from a depth one meter below the ground level. the collected samples have been immediately preserved in anti-evaporation plastic bags and then transported to the engineering laboratory. the essential characteristics of the collected soils and the corresponding standards which were depended in evaluations are demonstrated in table 1. the collected soils have been categorized as clayey soils based on uscs (astm-d2487-17)[40] and the volume distribution curve was assessed and plotted according to astm-d422-63[41] [figure 1]. the compaction tests curve has been evaluated and plotted according to astmd698-12r2021[42] [figure 2] which gave an optimum water content (owc) value of about 23% with an optimum dry density (odd) of about 1545 kg per cubic meter. recycled fiber the fiber has been obtained from recycling waste tpb. the tpb waste has been cut into strips of two different lengths (1 cm and 2 cm) with widths ranged between 2.5 and 3 mm. these fabricated tpb fibers have a tensile resistance about 125 mpa, with 3000 mpa young’s modulus, and 73% ultimate linear strain. table 1: essential characteristics of the collected soil characteristics values standards g s 2.7 astm-d854-14[43] odd (kg/m3) 1545 astm-d698-12r2021[42] owc (%) 23 astm-d698-12r2021[42] atterberg limits ll (%) 55.8 astm-d4318-17e01[44] pl (%) 29.1 astm-d4318-17e01[44] pi (%) 26.7 astm-d4318-17e01[44] odd: optimum dry density, owc: optimum water content figure 2: compaction curve of collected soil figure 1: volume distribution curve of collected soil ahmed: impact of textiled-poly bags on mechanical features of soil 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 1-6 test methods in current investigation, to evaluate the resistance and geotechnical features of both natural untreated clayey soil and treated soil, several experimental tests have been performed inclusive the evaluation of index characteristics of untreated soil, proctor compaction, uniaxial compressive resistance, and california bearing ratio for both untreated and treated soil. these experimental tests have been performed depending on astm standards. first, the recycled tpb fibers have been gradually added to the air-dried soil and mixed by hand. significant care has been taken to get best homogeneous between soil particles and fiber strips. after that, the desired water has been added to get soil samples ready to test. depending on astm-d698-12r2021,[42] compaction tests have been implemented for evaluating the impact of tpb fibers on owc and odd of treated soil, which have been adopted later for uniaxial compressive resistance test and california bearing ratio test. the astm-d2166-06[45] standard has been depended to implemented the uniaxial compressive resistance test to evaluate the impact of tpb fibers of uniaxial compressive resistance of treated soils. furthermore, astm-d1883-21[46] standard has been depended to evaluate load per cylindrical test sample (150 mm diameter, 175 mm height) demand to penetrate a soil mass with uniform rate of 1 mm/min. this test was depended to evaluate california bearing ratio by dividing the test load by standard load. results and discussion impact of tpb fibers on the parameters of compaction test the results of proctor compaction tests have demonstrated that the utilizing of tpb fibers at different percentages (0, 1,2, 3 and 4%) and different lengths (1 cm and 2 cm) gives dissimilar behavior in terms of omc and odd [figures 3 and 4]. when utilizing tpb fiber at 1-cm length, odd minimized by 2.31% and 1.61% for fiber content 1% and 2%, consecutively, and increased by 1.16% and 1.55% when fiber content was increased to 3% and 4%, consecutively. moreover, when utilizing tpb fiber at a 2-cm length, odd minimized by 0.97%, 0.58%, 0.71%, and 1.23% for fiber content 1%, 2%, 3%, and 4%, consecutively. the relation between odd and length of tpb fiber is demonstrated in figure 5, which could summarize the impact of fiber length and its content on the values of odd. when utilizing tpb fiber at 1-cm length, owc increased with increasing fiber content about 3.18%, 0.45%, and 2.72% for fiber content 2%, 3%, and 4%, consecutively, however at 1% fiber content, the owc minimized by 5.45%. moreover, when utilizing tpb fiber at a 2-cm length, omc minimized by 1.36%, 2.72%, 3.63%, and 1.36% for fiber content 1%, 2%, 3%, and 4%, consecutively. the relation between owc and length of tpb fiber is demonstrated in figure 6, which could summarize the impact of fiber length and its content on the values of omc. impact of tpb fibers on the behavior of uniaxial compressive resistance the results of uniaxial compressive resistance tests [tables 2 and 3] have demonstrated that the utilizing of tpb fibers at different percentages (0, 1, 2, 3, and 4%) and different lengths (1 cm and 2 cm) gives almost similar behavior in terms of improving the resistance. figure 3: compaction curve for tpb fiber at 1-cm length figure 4: compaction curve for tpb fiber at 2-cm length figure 6: the relation between omc and length of tpb fiber figure 5: the relation between odd and length of tpb fiber ahmed: impact of textiled-poly bags on mechanical features of soil 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 1-6 when utilizing tpb fiber at 1-cm length, uniaxial compressive resistance increased by 57.04%, 50.33%, 48.99%, and 48.32% for fiber content 1%, 2%, 3%, and 4%, consecutively. moreover, when utilizing tpb fiber at 2-cm length, the increment in uniaxial compressive resistance reached about 75.16%, 61.74%, 63.75%, and 65.77%, for fiber content 1%, 2%, 3%, and 4%, consecutively. the comparative between tpb fiber at 1-cm length and tpb fiber at 2-cm length in terms of the behavior of uniaxial compressive resistance is demonstrated in figure 7. it could be seen that the utilizing of tpb fiber at 2-cm length gives the highest improvement. impact of tpb fibers on the behavior of california bearing ratio the results of california bearing ratio tests [tables 4 and 5] have demonstrated that the utilizing of tpb fibers at different percentages (0, 1, 2, 3, and 4%) and different lengths (1 cm and 2 cm) gives significant improvement in the values of california bearing ratio. when utilizing tpb fiber at 1-cm length, california bearing ratio increased by 10%, 36.66%, 50%, and 63.33% for fiber content 1%, 2%, 3%, and 4%, consecutively. moreover, when table 2: impact of tpb fibers at 1 (cm) length on the behavior of uniaxial compressive resistance percentage of tpb fiber uniaxial compressive resistance (kpa) increment (%) 0 150 / 1 235 57.04 2 225 50.33 3 223 48.99 4 222 48.32 table 3: impact of tpb fibers at 2 (cm) length on the behavior of uniaxial compressive resistance percentage of tpb fiber uniaxial compressive resistance (kpa) increment (%) 0 150 / 1 262 75.16 2 242 61.74 3 245 63.75 4 248 65.77 table 4: impact of tpb fibers at 1 (cm) length on the behavior of california bearing ratio percentage of tpb fiber california bearing ratio increment (%) 0 4 / 1 4.3 10 2 5.1 36.66 3 5.5 50 4 5.9 63.33 table 5: impact of tpb fibers at 2 (cm) length on the behavior of california bearing ratio percentage of tpb fiber california bearing ratio increment % 0 4 / 1 4.9 30 2 5.2 40 3 5.8 60 4 6.2 73.33 table 6: conclusion of the mechanical behavior percentage of tpb fiber (%) length of tpb fiber (cm) mechanical behavior uniaxial compressive resistance (kpa) california bearing ratio 0 1 150 4 1 235 4.3 2 225 5.1 3 223 5.5 4 222 5.9 1 2 262 4.9 2 242 5.2 3 245 5.8 4 248 6.2 figure 8: comparative in california bearing ratio results figure 7: comparative in uniaxial compressive resistance results ahmed: impact of textiled-poly bags on mechanical features of soil 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 1-6 utilizing tpb fiber at 2-cm length, the increment in california bearing ratio reached about 30%, 40%, 60%, and 73.33%, for fiber content 1%, 2%, 3%, and 4%, consecutively. the comparative between tpb fiber at 1-cm length and tpb fiber at 2-cm length in terms of the behavior of california bearing ratio is demonstrated in figure 8. it could be seen that the utilizing of tpb fiber at 2-cm length gives the highest improvement. conclusion the investigation was implemented to fabricate environmentally friendly fiber by recycled tpb and evaluate its impact on several geotechnical features of clayey soil. the behavior of utilizing this fiber in several content and several lengths has been evaluated and the results could be concluded in the following points: • the utilizing of 3% and 4% of tpb fiber at 1-cm length helps to improve the parameters of compaction test • the utilization of 2-cm tpb fiber instead of 1-cm tpb fiber reduces the parameters of compaction test • tpb fiber improves the uniaxial compressive resistance; this has been noted for all fiber content (1%, 2%, 3%, and 4%) • tpb fiber of 2-cm length gives highest improvement in terms of uniaxial compressive resistance; this has been noted when compared it with tpb fiber of 1-cm length • california bearing ratio values improve when utilizing tpb fiber of 1and 2-cm lengths and this has been noted for all fiber content (1%, 2%, 3%, and 4%) • tpb fiber of 2-cm length gives highest improvement in terms of california bearing ratio; this has been noted when compared it with tpb fiber of 1-cm length. the overall mechanical behavior which gated from the current work could be concluded in table 6 below. references 1. c. g. godswill and c. g. awuchi. impacts of plastic pollution on the sustainability of seafood value chain and human health. international journal of advanced academic research, vol. 5, no. 11, pp. 46-138, 2019. 2. a. f. al-jaf and j. i. al-ameen. modelling of municipal solid waste landfill gas generation. a case study, kirkuk, iraq. journal of materials and environmental sciences, vol. 10, no. 10, pp. 987994, 2019. 3. t. h. hanna and i. h. mosa. effect of using waste polyethelene production on mechanical properties of concrete. pure and 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polyethylene terephthalate in improving poorly graded soil. transportation infrastructure geotechnology, vol. 7, no. 2, pp. 206-223, 2020. 14. r. k. kamil and m. a. aljumaili. strengthening the clayey gravel and sand soils for subbase course highway using different waste materials. iop conference series: materials science and engineering, vol. 888, no. 1, p. 012062, 2020. 15. s. h. fadhil, m. s. al-soud and r. m. kudadad. enhancing the strength of clay-sand mixture by discrete waste plastic strips. journal of applied science and engineering, vol. 24, no. 3, pp. 381391, 2021. 16. n. h. jaber, s. r. mushtaq and a. j. alsaad. ecological applications of polyethylene terephthalate plastic in producing modified subbase soil. iop conference series: materials science and engineering, vol. 1067, no. 1, p. 012006, 2021. 17. p. sherwood. soil stabilization with cement and lime. hmso, london, 1993. 18. j. s. yadav and s. k. tiwari. behaviour of cement stabilized treated coir fibre-reinforced clay-pond ash mixtures. journal of building engineering, vol. 8, pp. 131-140, 2016. 19. j. s. yadav, s. k. tiwari and p. shekhwat. strength behaviour of clayey soil mixed with pond ash, cement and randomly distributed fibres. transportation infrastructure geotechnology, vol. 5, no. 3, pp. 191-209, 2018. 20. f. g. bell. lime stabilization of clay minerals and soils. engineering geology, vol. 42, no. 4, pp. 223-237, 1996. 21. d. n. little. stabilization of pavement subgrades and base courses with lime. kendall/hunt publishing company, united states, 1995. 22. r. k. rout, p. ruttanapormakul, s. valluru and a. j. puppala. resilient moduli behavior of lime-cement treated subgrade soils. in: geocongress 2012: state of the art and practice in geotechnical engineering, pp. 1428-1437, 2012. 23. j. rasul, g. s. ghataora and m. p. n. burrow. permanent deformation of stabilized subgrade soils. in: bituminous mixtures and pavements. vol. 41. taylor and francis group, london, 2015. 24. j. m. rasul, m. p. n. burrow and g. s. ghataora. consideration of the deterioration of stabilised subgrade soils in analytical road pavement design. transportation geotechnics, vol. 9, pp. 96-109, 2016. 25. j. m. rasul, g. s. ghataora and m. burrow. the effect of wetting and drying on the performance of stabilized subgrade soils. transportation geotechnics, vol. 14, pp. 1-7, 2018. 26. j. s. yadav and s. k. tiwari. effect of waste rubber fibres on the geotechnical properties of clay stabilized with cement. applied clay science, vol. 149, pp. 97-110, 2017. 27. c. l. obo and a. b. ytom. study on the use of plastic fibre materials as an alternative solution for soil stabilization. university of ahmed: impact of textiled-poly bags on mechanical features of soil 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 1-6 southeastern philippines bislig campus, bislig, 2014. 28. s. amena. experimental study on the effect of plastic waste strips and waste brick powder on strength parameters of expansive soils. heliyon, vol. 7, no. 11, p. e08278, 2021. 29. t. yetimoglu and o. salbas. a study on shear strength of sands reinforced with randomly distributed discrete fibers. geotextiles and geomembranes, vol. 21, no. 2, pp. 103-110, 2003. 30. s. ziegler, d. leshchinsky, h. i. ling, and e. b. perry. effect of short polymeric fibers on crack development in clays. soils and foundations, vol. 38, no. 1, pp. 247-253, 1998. 31. h. singh, and y. hussain. utilization of waste coir fiber with polypropylene fiber for soil stabilization. journal of green engineering, vol. 10, no. 9, pp. 6076-6089, 2020. 32. s. twinkle, and m. k. sayida. effect of polypropylene fibre and lime admixture on engineering properties of expansive soil. proceedings of indian geotechnical conference, 2011. 33. p. d. jadhao, and p. b. nagarnaik. influence of polypropylene fibers on engineering behavior of soil− fly ash mixtures for road construction. electronic journal of geotechnical engineering, vol. 13, pp. 1-11, 2008. 34. a. modarres and h. hamedi. effect of waste plastic bottles on the stiffness and fatigue properties of modified asphalt mixes. materials and design, vol. 61, pp. 8-15, 2014. 35. f. changizi and a. haddad. strength properties of soft clay treated with mixture of nano-sio2 and recycled polyester fiber. journal of rock mechanics and geotechnical engineering, vol. 7, no. 4, pp. 367-378, 2015. 36. a. fauzi, z. djauhari and u. j. fauzi. soil engineering properties improvement by utilization of cut waste plastic and crushed waste glass as additive. international journal of engineering and technology, vol. 8, no. 1, pp. 15-18, 2016. 37. p. rawat and a. kumar. study of cbr behaviour of soil reinforced with hdpe strips. methodology, vol. 2, pp. 4-10, 2016. 38. s. peddaiah and s. sreedeep. experimental study on effect of waste plastic bottle strips in soil improvement. geotechnical and geological engineering, vol. 36, no. 5, pp. 2907-2920, 2018. 39. salimi, k and m. ghazavi. soil reinforcement and slope stabilisation using recycled waste plastic sheets. geomechanics and geoengineering, vol. 16, no. 6, pp. 497-508, 2021. 40. astm. d2487-17 standard practice for classification of soils for engineering purposes (unified soil classification system). american society for testing of materials, pennsylvania pa, usa, 2015. 41. astm. d422-63 standard test methods for hydrometer analysis of soils. american society for testing of materials, pennsylvania pa, usa, 2015. 42. astm. d698-12r2021 standard test methods for laboratory compaction characteristics of soil using standard effort (12,400 ft-lbf/ft3 (600 kn-m/m3)). american society for testing of materials, pennsylvania pa, usa, 2015. 43. astm. d854-14 standard test methods for specific gravity of soil solids by water pycnometer. american society for testing of materials, pennsylvania pa, usa, 2015. 44. astm. d4318-17e01 standard test methods for liquid limit, plastic limit, and plasticity index of soils. american society for testing of materials pennsylvania pa, usa, 2015. 45. astm, d2166-06 standard test method for unconfined compressive strength of cohesive soil. american society for testing of materials, pennsylvania pa, usa, 2015. 46. astm. d1883-21 standard test method for california bearing ratio (cbr) of laboratory-compacted soils. american society for testing of materials, pennsylvania pa, usa, 2015. tx_1~abs:at/add:tx_2~abs:at 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 7-10 research article clinical prediction of blood parameters associated with breast, colon, thyroid, ovarian, and prostate cancer: diagnostic studies ali j. muhialdin1, zaenah z. alamri2, ali m. hussein3, rahman k. faraj4, zhala b. taha5, mohammed m. hussein3, zeena m. ameen3, naz r. majeed3, maryam a. a. akramc3, mohammed i. jameel6 1department of animal production, college of agricultural engineering sciences, university of garmian, kalar, kurdistan region, iraq, 2department of biology, university of jeddah, college of science, jeddah 21589, saudi arabia, 3department of biomedical sciences, college of science, cihan university-erbil, kurdistan region, iraq, 4department of chemistry, college of science, university of garmian, kalar, kurdistan region, iraq, 5department of forestry, college of agricultural engineering sciences, salahaddin university, erbil, kurdistan region, iraq, 6department of medical microbiology, koya university, kurdistan region, iraq abstract cancer is a cellular change caused by uncontrolled cell growth and division. this disease develops from the growth of abnormal cells that have the potential to invade or spread to other parts of the body. this study’s goal was to determine how blood parameters, such as mean corpuscular volume, mean corpuscular hemoglobin (mch), and mch concentration, relate to one another. white, red blood cells, hemoglobin and other kind of cancer. breast cancer, thyroid gland cancer, ovarian cancer, testicular cancer, brain tumors as well as other types of cancer are based on the cell of the tissue found on or in the body. each of these cancers is accompanied by different health problems that put the patient’s life at risk. to perform a complete blood count (cbc) before and after cancer diagnosis, as abnormalities in the body can cause an increase or decrease in blood components, depending on the cancer type, the patient’s physiological mechanisms, and the structural component. since the cbc test belongs to hematology, drawing a blood sample and putting it into the anticoagulant tube for testing were preferred. in this study, the blood components of almost all patients were normal except for a few of them which may be due to other medical and biological factors. the aim of the study was to see if there is a significant relationship between blood parameters and cancer types. keywords: complete blood count, breast cancer, thyroid gland cancer, testicular cancer, ovarian cancer, blood parameters introduction cancer is a disease that causes deaths worldwide. the increasing risk of cancer disease can be accounted for by elements such as a longer life expectancy and social, economic, and population growth.[1,2] breast cancer originates from the breast tissue, typically from the lining or lobules of milk ducts.[3] thyroid cancer is a type of cancer that affects the endocrine system.[4] there are two different forms of thyroid cancer. papillary thyroid cancer and follicular thyroid cancer are both differentiated thyroid cancer that arise from thyroid follicle cells. medullary thyroid cancer the third form of thyroid cancer, develops from parafollicular c-cells. the last type of thyroid cancer, which is anaplastic thyroid cancer, is a lethal form of the disease that can distinguish over time and occurs from de novo.[5] epithelial ovarian cancer causes gynecological cancer-associated deaths.[6] ovarian carcinoma can develop in one of three ways which include the surface of the ovary, the fallopian tube that connects the uterus to the fallopian, and the peritoneal cavity lined by mesothelium,[7] whereas colon cancer develops when malignant tumors form in the colon tissue. it is one of the most commonly diagnosed cancers and one of the main causes of death.[8] prostate cancer is the second most commonly diagnosed cancer in men after corresponding author: mohammed i. jameel, department of medical microbiology, koya university, kurdistan region, iraq. e-mail: mohammed.isam@koyauniversity.org received: may 19, 2023 accepted: june 26, 2023 published: july 20, 2023 doi: 10.24086/cuesj.v7n2y2023.pp7-10 copyright © 2023 ali j. muhialdin, zaenah z. alamri, ali m. hussein, rahman k. faraj, zhala b. taha, mohammed m. hussein, zeena m. ameen, naz r. majeed, maryam a. a. akramc, mohammed i. jameel. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) muhialdin, et al.: blood parameters associated with breast, colon, thyroid, ovarian, and prostate cancer 8 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 7-10 lung cancer.[9] brain tumor is the most terrible type of cancer. it caused many deaths among children and adults during the past years.[10] the tumor affects and damages the healthy tissues of the brain or intensifies intracranial pressure.[11] testicular cancer is the most common solid tumor in men over the age of 15–40 years. it is extremely aneuploidy, with the iso-chromosome 12p being the most common chromosomal abnormality.[12] ewing’s sarcoma, a small round cell tumor, usually develops in the bones of children and teenagers but may also occur in soft tissue. ewing’s disease develops quickly, and within a few weeks, the skeletal lesion transforms into massive tumors that develop in soft tissue.[14] materials and methods after diagnosing patients with cancer, a complete blood count (cbc) examination is required for further diagnoses of diseases, such as leukemia, anemia, and other hematological diseases.[15] there were two stages of the cbc test such as the phlebotomy technique followed using the blood sample in the cbc instruments. phlebotomy, in a so-called venipuncture, a blood sample is taken by puncturing a vein and collected in edta tube. this was the medical procedure to withdraw the blood sample. this study involved collecting data about a hundred patients diagnosed with cancer. the patients consisted of those with cancer of various types such as breast (30 samples), colon (15 samples), thyroid gland (15 samples), ovarian (20 samples), and prostate cancer (20 samples). they received treatments at kalar hospital in sulaimaniyah, kurdistan region, iraq. data for each patient for this study were collected by an oncologist at the above hospital statistics department. results the variations in blood parameters of patients with cancer of various types are illustrated in the following figures. the highest mean white blood cell (wbc) count was seen in patients with breast cancer 7.3, prostate cancer 7.18, and ovarian cancer 6.6. the mean wbc count was lower (5.4) in patients with colon cancer compared to 5.56 in patients with thyroid gland cancer. however, the standard rate was 4.5–11 (fig. 1). the highest and lowest mean red blood cell (rbc) counts were observed in patients with thyroid gland cancer (5) and prostate cancer (3.5), respectively. the same mean (4.5) rbc count was shown in patients with colon cancer and ovarian cancer. the mean rbc count in patients with breast cancer was 4, which was slightly higher than that of patients with prostate cancer. however, the standard rate was 4–5 (fig. 2). the highest mean hemoglobin (hgb) count (14) was in patients with thyroid gland cancer and ovarian cancer, followed by 13 and 11 in patients with colon cancer and breast cancer, respectively. patients with prostate cancer had the lowest mean hgb count (10). the standard rate was 12–15.5 (fig. 3). patients with ovarian cancer, colon cancer, thyroid gland cancer, and breast cancer presented normal mean corpuscular volume (mcv), mean corpuscular hemoglobin (mch), and mch concentration (mchc) rates, indicating these types of cancer normally do not affect mcv, mch, and mchc (fig. 4) figure 1: the mean white blood cell count in patients with cancer of various types the wbc, rbc, and hgb rates were normal in all patients with ovarian cancer, colon cancer, thyroid gland cancer, and breast cancer. this finding indicates these types of cancer do not normally affect wbc, rbc, and hgb (fig. 5). each of the patients with ovarian cancer, breast cancer, and thyroid cancer had normal levels of platelets. on the other hand, patients with colon cancer had the lowest levels of platelets in comparison to patients with other types of cancer (fig. 6). discussion early prognosis of tumors in the body, especially in relation to early detection of oncological diseases, is a popular issue. figure 2: the mean red blood cell count in patients with cancer of various types figure 3: the mean hemoglobin count in patients with cancer of various types muhialdin, et al.: blood parameters associated with breast, colon, thyroid, ovarian, and prostate cancer 9 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 7-10 the cbc test was administered to patients diagnosed with cancer of various types (e.g., prostate cancer, ovarian cancer, thyroid cancer, colon cancer, and breast cancer). the results of this study displayed average wbc rates in all patients with cancer of various types, with the exception of one breast cancer patient who had a high rate of wbc. these results are consistent with previous study findings.[13,14] other studies showed that, using the cbc profiles to measure a patient’s risk of health outcomes may indicate different infections or leukemia.[16] the rates of rbc were normal in all patients with cancer of various types except one breast cancer patient who had a very low rate of rbc, which indicate some vitamin deficiency or anemia. according to prior studies conducted in 2020, patients are rated based on quantitative changes in blood parameters related to the body’s response to stress caused by malignant tumors.[17] the results of this study also demonstrated that the rates of hgb were almost normal, and there was no association between bleeding and cancer type. these results are in line with the findings of the previous study.[18,19] however, some patients had low rates which may signify anemia. in addition, mchc rates were normal in all patients with cancer of various types.[20,21] conclusion cbc is a well-known laboratory technique that measures the levels of various blood parameters to predict and diagnose a wide range of disorders and infections. cancer is a lethal disease caused by irregular cell division and development. the findings of this study revealed that all patients with ovarian cancer, thyroid cancer, breast cancer, and colon cancer had normal levels of wbcs, rbcs, and hgb. furthermore, the mcv, mch, and mchc rates were normal in all patients with cancer of the same type. the platelet levels were also normal in breast cancer, thyroid gland cancer, and ovarian cancer patients. on the other hand, colon cancer patients exhibited low platelet levels. based on the results of this study, it was suggested that the cbc test does not accurately diagnose cancer. acknowledgment the authors would like to thank cihan university-erbil for granting them permission to use the laboratory and to carry out this research. thanks also to the research center and quality assurance directorate for providing useful guidelines for this study. ethics committee approval and consent for publication the oncologist gathered information from the kalar hospital’s statistics department in sulaimaniyah, iraq’s kurdistan region. medical ethics application was not applicable, as this study did not involve patients. references 1. world health organization. cancer fact sheets. world health organization, geneva, 2020. available from: https://gco.iarc.fr/ today/fact-sheets-cancers [last accessed on 2023 jan 29]. 2. g. salcedo-villanueva, a. a. medina-andrade, d. morenoparamo, m. f. golzarri, e. moreno-paramo, g. y. ortiz-ramirez, u. martinez-aguilar, u. de dios-cuadras, m. jimenez-rodriguez, i. espinosa-soto, x. mira-lorenzo, j. guzman-cerda, a. orozcomoguel, c. becerra-revollo, l. p. orozco-gomez and e. fulda. primary cancer sites and clinical features of choroidal metastasis in mexican patients. clinical ophthalmology, vol. 15, pp. 201209, 2021. 3. g. n. sharma, r. dave, j. sanadya, p. sharma and k. k. sharma. various types and management of breast cancer: an overview. journal of advanced pharmaceutical technology and research, vol. 1, no. 2, pp. 109-126, 2010. 4. m. xing. molecular pathogenesis and mechanisms of thyroid cancer. nature reviews cancer, vol. 13, pp. 184-199, 2013. 5. d. f. schneider and h. chen. new developments in the diagnosis and treatment of thyroid cancer. ca: a cancer journal for clinicians, vol. 63, no. 6, pp. 373-394, 2013. figure 4: the mean corpuscular volume, mean corpuscular hemoglobin (mch), and mch concentration counts in patients with cancer of various types figure 6: the results of the platelet count test for patients with cancer of various types figure 5: the mean white blood cell, red blood cell, and haemoglobin counts in patients with cancer of various types muhialdin, et al.: blood parameters associated with breast, colon, thyroid, ovarian, and prostate cancer 10 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 7-10 6. g. c. jayson, e. c kohn, h. c. kitchener and j. a. ledermann. ovarian cancer. lancet, vol. 384, no. 9951, pp. 1376-1388, 2014. 7. e. lengyel. ovarian cancer development and metastasis. the american journal of pathology, vol. 177, no. 3, pp. 1053-1064, 2010. 8. t. hu, z. li, c. y. gao and c. h. cho. mechanisms of drug resistance in colon cancer and its therapeutic strategies. world journal of gastroenterology, vol. 22, no. 30. pp. 6876-6889, 2016. 9. p. rawla. epidemiology of prostate cancer. world journal of oncology, vol. 10, no. 2. pp. 63-89, 2019. 10. a. rehman, m. a. khan, t. saba, z. mehmood, u. tariq and n. ayesha. microscopic brain tumor detection and classification using 3d cnn and feature selection architecture. microscopy research and technique, vol. 84, no. 1, pp. 133-149, 2020. 11. t. saba, a. s. mohamed, m. el-affendi, j. amin and m. sharif. brain tumor detection using fusion of hand crafted and deep learning features. cognitive systems research, vol. 59, pp. 221230, 2020. 12. g. de vries, x. rosas-plaza, m. a. t. m. van vugt, j. a. gietema and s. de jong. testicular cancer: determinants of cisplatin sensitivity and novel therapeutic opportunities. cancer treatment reviews, vol. 88, p. 102054, 2020. 13. y. iwamoto. diagnosis and treatment of ewing’s sarcoma. japanese journal of clinical oncology, vol. 37, no. 2. pp. 79-89, 2007. 14. j. j. noh, m. c. lim, m. h. kim, y. h. kim, e. s. song, s. j. seong, d. h. suh, j. m. lee, c. lee and c. h. choi. the prognostic model of pre-treatment complete blood count (cbc) for recurrence in early cervical cancer. journal of clinical medicine, vol. 9, no. 9, p. 2960, 2020. 15. n. h. mahmood, p. c. lim, s. m. mazalan and m. a. a. razak. blood cells extraction using color based segmentation technique. international journal of life sciences biotechnology and pharma research, vol. 2, no. 2, pp. 2250-3137, 2013. 16. d. k. kur, d. thøgersen, l. kjeldsen and l. friis-hansen. the hemoscreen hematology pointof-care device is suitable for rapid evaluation of acute leukemia patients. international journal of laboratory hematology, vol. 43, no. 1, pp. 52-60, 2021. 17. m. g. sadovsky, a. a. feller, e. a. martynova, d. v. chernyaev, e. v. semenov, e. v. slepov and r. a zukov. cbc effectively stratifies the patients with different types of malignant tumors. journal of physics: conference series, vol. 1679, p. 042019, 2020. 18. s. yousefian, a. moafi and m. khalilian. the relation between end of induction minimal residual disease and different risk factors in patients with acute lymphoblastic leukemia. iranian journal of pediatric hematology and oncology, vol. 9, no. 4, pp. 211-218, 2019. 19. n. michels, c. van aart, j. morisse, a. mullee and i. huybrechts. chronic inflammation towards cancer incidence: a systematic review and meta-analysis of epidemiological studies. critical reviews in oncology hematology, vol. 157, p. 103177, 2021. 20. a. m. hussein, z. b. taha, a. g. malek, k. a. rasul, d. q. hazim, r. j. ahmed and u. b. mohamed. ddimer and serum ferritin as an independent risk factor for severity in covid-19 patients. materials today: proceedings, 2021. 21. t. s. salih and s. n. r. darogha. incidence of b-cell leukemia among kurdish people in erbil city. cihan university-erbil scientific journal, vol. 2, p. 2, 2017. . 61 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 61-65 research article in vitro screening of seed extracts of medicinal plants for protease inhibitory activity faiyaz khudaboddin shaikh1,2*, sarwan w. bradosty3, saber w. hamad4, ashok a. shinde2 1department of biochemistry, dr. babasaheb ambedkar marathwada university, aurangabad, maharashtra, india, 2department of biotechnology, mgm’s institute of biosciences and technology, aurangabad, maharashtra, india, 3department of biology, cihan university-erbil, kurdistan region, iraq, 4department of field crops production, college of agriculture salahaddin university, erbil, kurdistan region, iraq abstract protease inhibitors (pis) are deployed in the plant kingdom as storage proteins or peptides, regulators of endogenous proteases, and plant protection agents against insect pests and pathogen attack. in humans, they are identified as chemopreventive agents against a range of cancers and have potential as drug to treat an array of disease associated with aberrant activity of proteases. the present investigation reports pis activity data from 30 medicinal plants. the screening for pis activity was done by dot blot assay using x-ray film coated with gelatin. among screened seed extracts, albizia lebbeck, raphanus sativus, mucuna pruriens, achyranthes aspera, and coffea arabica showed high inhibitory activities with trypsin protease. most of seed extracts exhibited moderate activity, whereas ocimum sanctum showed moderate to low activity against trypsin. the presence of varied protein content is reported from all seed extracts with highest in a. lebbeck (50.0 ± 3.4 mg/ml). the data produced in the present investigation could be helpful for further exploration of pis as therapeutic agent. keywords: dot blot, protease inhibitors, proteases, protein, trypsin introduction proteases or peptidases (e.c. 3.4) are one of the largest and the most diverse families of enzymes which selectively catalyze the hydrolysis of peptide bonds in proteins.[1,2] proteases are physiologically vital in all living organisms for cell growth, cell differentiation and death (apoptosis), cell migration, and invasion and are abundant in a wide variety of sources starting from viruses, bacteria, protozoa, metazoan, or fungi ending with plants and animals.[3,4] they perform a variety of functions during the vital processes in organism such as food digestion, blood clotting, embryogenesis, tissue reorganization (e.g., wound healing, regeneration, molting, metamorphosis, etc.), defense mechanisms, and immune responses.[5,6] from the analysis of several genomes, it is estimated and found that about 2–4% of all gene products are proteases with more than 560 members.[7,8] proteases are intricately involved in the protein catabolism and their action can be divided into two different categories: limited proteolysis and unlimited proteolysis. in limited proteolysis, proteases cleave specific peptide bonds in immature proteins or remove the target signals in pre-proteins, whereas the unlimited proteolysis (bulk hydrolysis) of dietary proteins is essential for providing cells with simple metabolites essential for growth and development. on the basis of catalytic types, they can be classified into serine, cysteine, aspartic, threonine, glutamic acid, and metalloproteases.[9] serine proteases are numerous and widespread among the mammals, viruses, insects, bacteria, and eukaryotes, suggesting that they are vital to the organisms. serine proteinases are classically categorized by their substrate specificity, particularly by whether the residue at p1: trypsinlike (lys/arg preferred at p1), chymotrypsin-like (large hydrophobic residues such as phe/tyr/leu at p1), or elastaselike (small hydrophobic residues such as ala/val at p1).[10] they are found to be involved in tissue degradation, blood coagulation, digestion, development, and immune defense.[11] the aberrant activity of these proteases can lead to various diseases and thus has been the focus of intense investigation as potential therapeutic targets. corresponding author: faiyaz khudaboddin shaikh, assistant professor, department of biotechnology, mgm’s institute of biosciences and technology, aurangabad-431003, maharashtra, india. e-mail: faizbiochem@gmail.com received: apr 13, 2019 accepted: apr 23, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp61-65 copyright © 2019 faiyaz khudaboddin shaikh, sarwan w. bradosty, saber w. hamad, ashok a. shinde. thi is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ shaikh, et al.: protease inhibitor activity from medicinal plants 62 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 61-65 protease inhibitors (pis) are molecules that resist the proteolytic actions of proteases and play a key role in regulation of protein catabolism in the body, the aberrant activity of proteases can lead to various diseases and must be regulated.[2] the diseases which could be attributed are metastasis, angiogenesis, alzheimer’s disease, parkinson’s disease, aids, and ischemia.[12,13] in general, regulation could be achieved by differential transcription, post-translational processing, subcellular compartmentalization, and the presence and activity of pis.[14,15] the existence of pis in nature was first reported by fermi and pernossi.[16] they are small proteins which are quite common in nature and also present in all life forms.[17] in plants, pis are generally concentrated (about 10%) in the seeds as seed storage proteins.[18] in general, pis are grouped into five groups as serine pis, cysteine pis, threonine pis, aspartic pis, and metalloproteases.[19] among these, serine pis is predominant followed by cysteine pis and metalloproteases, while aspartic pis and aspartic pis are rare and dispersed in different families. serine pis are widespread, well-characterized, and most-studied class of pis in plants.[20,21] historically, serine pis were first characterized from soybean plant.[22] proteases are proven therapeutic targets and screening of potent pis against aberrant proteolytic activity could lead to the development of the drugs. earlier plant-based pis are found to be effective against cardiovascular diseases, osteoporosis, inflammatory diseases, and neurological disorders. several plant pis are under evaluation in in vitro clinical trials for the treatment of many diseases. the pis have widespread applications in different sectors and are one of the prime candidates in biotechnology and medicine. the present investigation reports the detection and screening of pis from 30 medicinal plants. materials and methods procurement of seeds dry seeds of medicinal plants were purchased from the local market of aurangabad (ms), india. procurement of chemicals trypsin (bovine pancreas, e.c. 3.4.21.4), bovine serum albumin (bsa), hexane, acetone, and polyvinylpyrrolidone (pvp) were obtained from sisco research laboratories, mumbai, india. x-ray films were obtained from fuji film, usa. all other chemicals used in this study were of the highest purity available. extractions of pis from medicinal plants the dried seeds were pulverized to a fine powder in a mixer grinder. the fat was removed from the powder by hexane and acetone washes. defatted powder was suspended in milli-q water (1:6 w/v) containing 1% pvp and incubated overnight at 15°c. the suspension was then centrifuged at ×12,000 g for 20 min at 4°c, and the supernatant was termed as crude pis.[23] detection of pis by dot blot/spot test the dot blot/spot test was carried out to determine the potency of pis against protease trypsin, using x-ray film.[23] three different concentrations of the trypsin and pis were prepared: 1 (1:3), 2 (1:1), and 3 (3:1) v/v. the volume of the reaction mixture was adjusted by 0.1 m tris-hcl (ph 7.8). the total volume was made up to 20 µl using buffer and loaded onto x-ray film. the film with spots was incubated for 20 min at 37°c, then washed with running tap water and dried in air. different ratios of enzyme and inhibitor produced different patterns of gelatin hydrolysis on the x-ray film depending on the efficiency of inhibitor, which may be observed visually and scanned at 300 dpi using an hp digital scanner. protein determinations protein content was estimated by following the method of lowry et al.[24] using bsa as the standard. statistical analysis all experiments were conducted and analyzed in triplicate. the means and standard deviations were calculated and compared using microsoft excel 2010. figure 1: protein content of selected medicinal plants. values are mean±standard deviation for at least three replicates shaikh, et al.: protease inhibitor activity from medicinal plants 63 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 61-65 table 1: classification and protein content of selected medicinal plants botanical name family common name total protein content (mg/ml±sd) hygrophila spinosa acanthaceae talimkhana 12.8±1.4 ocimum basilicum lamiaceae sweet basil 13.9±1.6 raphanus sativus brassicaceae radish 26.8±1.8 daucus carota apiaceae wild carrot 12.1±1.2 mucuna pruriens fabaceae velvet bean 28.3±1.9 putranjiva roxburghii putranjivaceae putranjiva 12.5±1.4 cassia tora leguminosae sickle senna 23.3±1.2 croton tiglium euphorbiaceae jamaal gota 12.0±1.2 albizia lebbeck fabaceae siris tree 50.0±3.4 guizojia abyssynica asteraceae black seed 13.4±1.5 gloriosa superba colchicaceae flame lily 11.3±1.2 cucumis melo cucurbitaceae muskmelon 14.3±2.3 plantago ovata plantaginaceae ispaghul 13.5±1.5 eugenia jambolana myrtaceae jambolan 32.1±1.8 mimosa pudica fabaceae sensitive plant 21.1±1.1 ocimum sanctum lamiaceae tulasi 17.1±1.7 solanum xanthocarpum solanaceae kantakari 26.5±2.7 vitex negundo lamiaceae nirgundi 13.1±1.5 ocimum gratissimum lamiaceae ram tulsi 13.6±2.3 sida cordifolia malvaceae kharinta 19.3±1.3 celastrus paniculata celastraceae malkagani 20.1±1.2 psoralea corylifolia fabaceae bavachi 25.1±2.1 myristica fragrans myristicaceae jatiphala 17.3±1.9 nigella sativa ranunculaceae kalonji 19.1±1.2 gymnema sylvestre apocynaceae gurmar 22.1±2.1 achyranthes aspera amaranthaceae chaff-flower 24.4±2.6 coffea arabica rubiaceae coffee 32.1±2.7 hemidesmus indicus apocynaceae nannari 29.3±1.9 asparagus racemosus asparagaceae shatavari 28.2±2.0 nelumbium speciosum nelumbonaceae lotus/kamal 37.5±3.2 sd: standard deviation results and discussion the past few decades have seen a growing interest in the identification, purification, and characterization of novel pis. they are prime candidates who have numerous applications in agricultural and medicinal biotechnology. plants are good sources of pis which protect them against diseases, insects, pests, and herbivores.[25] they also explored as natural drugs for the treatment of an array of diseases including cancers.[13,26] the current study was undertaken to detect the presence of pis activity in seeds of selected medicinal plants. here, we screened about 30 medicinal plants for pis activity. the initial experiments were designed to explore total protein content from hexane defatted seed powder of these plants. the total protein content was estimated by protein assay developed by lowry et al.[24] in terms of bsa equivalence [figure 1 and table 1]. the protein content was found to be higher in medicinal plants belonging to fabaceae family with highest in albizia lebbeck (50.0 ± 3.4 mg/ml). the higher protein content is attributing to the additional nitrogen that legumes receive through the process of nitrogen fixation.[27] the seed extracts of other medicinal plants were reported to have varied protein content as reported in table 1. the crude seed extracts of these plants were further investigated for pis activity. the presence of pis activity was confirmed by spot test (dot blot) analysis.[23] detection of pis was observed by mixing various concentrations of protease and inhibitor and spotted on the x-ray film, then the clearing zone formed due to gelatin hydrolysis by trypsin and a reduction in the clearing zone by trypsin incubated with an inhibitor was compared. dot blot method was used to screen a large number of pis. the results of detection of pis activity by dot blot are shown in figure 2 and results for screening of seed sample of 30 plants for inhibition of proteases are given in table 2. shaikh, et al.: protease inhibitor activity from medicinal plants 64 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 61-65 among screened samples, a. lebbeck, raphanus sativus, mucuna pruriens, achyranthes aspera, and coffea arabica showed high inhibitory activities against trypsin. at all selected ratio (1:3, 1:1, and 3:1) of trypsin and the inhibitor, all these samples showed inhibition. most of the screened seed samples such as cucumis melo, eugenia jambolana, sida cordifolia, myristica fragrans, nigella sativa, gymnema sylvestre, hemidesmus indicus, asparagus racemosus, and nelumbium speciosum exhibited moderate to high trypsin inhibitory activity, while ocimum sanctum showed moderate to low activity against trypsin. the present study revealed that seed extracts of a. lebbeck, r. sativus, m. pruriens, a. aspera, and c. arabica were effective in inhibiting the trypsin and could be explored as pis source. proteases are enzymes involved in protein digestion. these enzymes are omnipresent in plants, animals, and also most microorganisms. they are importantly involved in health and well-being by performing a pivotal role in survival and maintenance of the organism. they constitute about 2% of the human genome. the conduct of proteases in spite of many advantages has to be closely regulated and controlled to avoid the excess activity of these enzymes, as it may possibly damage its host organism. this task of controlling is carried out by pis. the pis have coevolved with proteases, to control their destructive nature.[28] cancer is a collection of over 100 devastating diseases that share a number of characteristics, a primary hallmark of which is out-of-control growth. there is a positive correlation between the aggressiveness of a tumor and the secretion of various proteases.[29] a number of reviews on various aspects of the use of pis as a mean to combat cancer have been published recently. therefore, the aim of this study was to screen indian medicinal plants for pis and a. lebbeck, r. sativus, m. pruriens, a. aspera, and c. arabica were found to exhibit pis activity. this study could be useful for the identification of pis, designed for therapeutic applications. acknowledgments the financial assistance in the form of the fellowship to faiyaz k. shaikh from the university grant commission and department of ministry of minority affairs, new delhi, india is greatly acknowledged. authors’ contributions faiyaz shaikh designed, performed, and coordinated the main study. sarwan hamad, saber hamad, and ashok shinde helped in the manuscript preparation and editing. table 2: screening of pis from seeds extract of medicinal plants by dot blot method botanical name trypsin 3:1 1:1 1:3 hygrophila spinosa n n n ocimum basilicum n n n raphanus sativus p y y daucus carota n n n mucuna pruriens y y y putranjiva roxburghii n n n cassia tora n n p croton tiglium n n n albizia lebbeck y y y guizojia abyssynica n p p gloriosa superba n n p cucumis melo p y y plantago ovata n n n eugenia jambolana p p y mimosa pudica n n n ocimum sanctum n n p solanum xanthocarpum n n p vitex negundo n n p ocimum gratissimum n n y sida cordifolia n p y celastrus paniculata n n n psoralea corylifolia n n n myristica fragrans p y y nigella sativa p p y gymnema sylvestre p p y achyranthes aspera y y y coffea arabica y y y hemidesmus indicus p p y asparagus racemosus p y y nelumbium speciosum p y y n: no inhibition, p: partial inhibition, y: total inhibition, pis: protease inhibitors table 2: (continued) botanical name trypsin 3:1 1:1 1:3 (contd...) figure 2: detection of protease inhibitor (pi) of trypsin by the dot blot method. three different concentrations of enzymes and inhibitors (i [3:1], ii [1:1], and iii [1:3]) were used for screening of pis. the faint green spot (gelatin) indicates total inhibition of enzymes, while dark blue spot (gelatin hydrolysis) indicates no inhibition. ti profile of hygrophila spinosa (a), ocimum sanctum (b), cucumis melo (c), and albizia lebbeck (d) a b c d shaikh, et al.: protease inhibitor activity from medicinal plants 65 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 61-65 conflicts of interest the authors declare that there are no conflicts of interest regarding the publication of this paper. references 1. a. j. barrett, n. d. rawlings and j. j. f. woessner. “hand book of proteolytic enzymes”. london: acadamy press, 1998. 2. s. g. fan and g. j. wu. “characteristics of plant proteinase inhibitors and their applications in combating phytophagous insects”. botanical bulletin of academia sinica, vol. 46, pp. 273-292, 2005. 3. a. n. flynn and a. g. buret. “proteinase-activated receptor 1 (par-1) and cell apoptosis”. apoptosis, vo. 9, pp. 729-737, 2004. 4. g. a. joanitti, s. m. freitas and l. p. silva. “proteinaceous protease inhibitors: structural features and multiple functional faces”. current enzyme inhibition, vol. 2, no. 3, pp. 199-217, 2006. 5. p. k. lawrence and k. r. koundal. “plant protease inhibitors in control of polyphagous insects”. electronic journal of biotechnology. vol. 1, pp. 93-109, 2002. 6. d. ivanov, c. emonet, f. foata, m. affolter, m. delly, m. fisseha, s. blum-sperisen, s kochhar and f. arigoni. “a serpin from the gut bacterium bifidobacterium longum inhibits eukaryotic elastase like serine proteases”. journal of biological chemistry, vol. 281, no. 25, pp. 17246-17252, 2006. 7. a. j. barrett, n. d. rawlings and e. a. o’brien. “the merops database as a protease information system”. journal of structural biology, vol. 134, pp. 95-102, 2001. 8. x. s. puente, l. m. sanchez, c. m. overall and c. lopezotin. “human and mouse proteases: a comparative genomic approach.” nature reviews genetics, vol. 4, pp. 544-558, 2003. 9. d. n. rawlings, p. d. tol1e and a. j. barrett. “evolutionary families of peptidase inhibitors”. biochemical journal, vol. 378, pp. 705-716, 2004. 10. j. d. tyndall, t. nall and d. p. fairlie. “proteases universally recognize beta strands in their active sites.” chemical reviews, vol. 105, pp. 973-999, 2005. 11. a. j. molehin, g. n. and d. p. gobert. “mcmanus: serine protease inhibitors of parasitic helminthes.” parasitology, vol. 139, no. 6, pp. 681-695, 2012. 12. m. l. j. korsinczky, h. j. schirra and d. j. craik. “sunflower trypsin inhibitor-1”. current protein and peptide science, vol. 5, p. 351, 2004. 13. m. h. lingaraju and l. r. gowda. “a kunitz trypsin inhibitor of entada scandens seeds: another member with single disulfide bridge.” biochimica et biophysica acta, vol. 1784, pp. 850-855, 2008. 14. p. r. shewry. “tuber storage proteins”. annals of botany, vol. 91, pp. 755-769, 2003. 15. n. d. rawlings, a. j, barrett and a. bateman. “merops: the database of proteolytic enzymes, their substrates and inhibitors”. nucleic acids research, vol. 40, pp. 343-350, 2012. 16. c. fermi and l. pernossi. “untersuchungen uber die enzyme, ver-gleichende studie”. zeitschrift für hygiene und infektionskrankheiten, vol. 18, pp. 83-89, 1894. 17. h. fritz. foreword. in: k. von der helm, b. d. korant and j. c. cheronis, editors. “proteases as targets for therapy”. berlin: springer-verlag, pp. 5-6, 2000. 18. f. de leo, m. volpicella, f. licciulli, s. liuni, r. gallerani and l. r. ceci. “plant-pis: a database for plant protease inhibitors and their genes”. nucleic acids research, vol. 30, no. 1, pp. 347-348, 2002. 19. g. fear, s. komarnytsky and i. raskin. “protease inhibitors and their peptidomimetic derivatives as potential drugs.” pharmacology and therapeutics, vol. 113, pp. 354-368, 2007. 20. g. c. mello, m. l. v. oliva, j. t. sumikawa, o. l. t. machado, s. marangoni, j. c. novello and m. l. r. macedo. “purification and characterization of a new trypsin inhibitor from dimorphandra mollis seeds”. journal of protein chemistry, vol. 20, pp. 625-632, 2001. 21. s. haq, s. atif and r. khan. “protein proteinase inhibitor genes in combat against insects, pests, and pathogens: natural and engineered phytoprotection”. archives of biochemistry and biophysics, vol. 431, pp. 145-159, 2004. 22. j. w. read and l. w. haas. “studies on the baking quality of flour as affected by certain enzyme actions. v. further studies concerning potassium bromate and enzyme activity”. cereal chemistry, vol. 45, pp. 59-68, 1938. 23. m. v. padul, r. d. tak and m. s. kachole. “protease inhibitor (pi) mediated defense in leaves and flowers of pigeonpea (protease inhibitor mediated defense in pigeonpea)”. plant physiology and biochemistry, vol. 52, pp. 77-82, 2012. 24. o. h. lowry, n. j. rosebrough, a. l. farr and r. j. randall. “protein measurement with folin phenol reagent”. journal of biological chemistry, vol. 193, pp. 265-275, 1951. 25. c. a. ryan. “protease inhibitors in plants: genes for improving defenses against insects and pathogens”. annual review of phytopathology, vol. 28, pp. 425-449, 1990. 26. p. j. magee, r. owusu-apenten, m. j. mccann, c. i. gill and i. r. rowland. “chickpea (cicer arietinum) and other plantderived protease inhibitor concentrates inhibit breast and prostate cancer cell proliferation in vitro”. nutrition and cancer, vol. 64, pp. 741-748, 2012. 27. j. i. sprent. “legume nodulation: a global perspective”. ames, iowa: wiley-blackwell, pp. 12, 2009. 28. n. vergnolle. “protease inhibition as new therapeutic strategy for gi diseases”. gut, vol. 65 pp. 1215-1224, 2016. 29. c. fields, p. mallee, j. muzard and g. u. lee. “isolation of bowman-birk inhibitor from soybean extracts using novel peptide probes and high gradient magnetic separation”. food chemistry, vol. 134, pp. 1831-1838, 2012. . 80 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 80-84 research article some immunological and hematological parameters among refugees in kawergosk camp – erbil governorate tanya s. salih, safa safin haydar, muhsin h. ubeid, ameena s. m. juma department of biology, cihan university-erbil, kurdistan region, iraq abstract the study included 258 syrian refugees of different ages and sex and another 60 volunteers as control group (c.g). these refugees were in kawergosk camp in erbil governorate. blood was collected from each individual for the estimation of white blood cell (wbc), eosinophil, iron, hemoglobin (hb), and immunoglobulin e (ige) levels. mean serum levels of ige among male and female refugees showed highly significant increasing when compared to c.g. most of the refugees had normal iron levels, where iron concentrations were more than 65 mg/dl among 67 males and more than 50 mg/dl among 104 females and 48 children, while some had iron deficiency in which the majority were female (9 males, 24 females, and 6 children had iron deficiency). in addition, hb concentrations were normal among 65 males (more than 13.0 g/dl), 89 females (more than 11.0 g/dl), and 48 children (more than 12.0 g/dl). however, anemia was found among 8 men, 42 women, and 6 children. it was revealed that there was a highly significant rising in eosinophils in male and female refugees in comparison to c.g. wbc count is non-significantly slightly increased in both male’s and female’s refugees when compared to c.g. keywords: eosinophil count, hemoglobin, immunoglobulin e, iron, refugees, white blood cell count introduction there are about 68.5 million people currently displaced around the world, and 25.4 million of them have crossed international borders seeking shelters and safety. traveling and lodging in different places may cause ill health to many of the refugees, although their health status might have been quite well in their original homes. falling ill may be because of poor living conditions in camps, poor sanitation, and lifestyle change that may include inadequate food and water and stress. all these conditions were well known to lead to depression.[1] erbil city currently has four refugee camps – kawergosk camp, qushtapa camp, darashakran camp, and basirma camp, that host around 50,000 syrian refugees. kawergosk refugee camp was the first camp to be established in erbil in 2013 hosting refugees from the town of qamishli in syria. the overall situation of the camp is fair. however, there are two concerns – health and sanitation due to lack of advanced health services and limited availability of water supply.[2] due to displacement and lack of many health facilities, it is common that the general health of refugees might deteriorate with time. moreover, communicable diseases are common among refugees including tuberculosis, hiv/aids, hepatitis, measles, and rubella.[3] the aim of this study is to investigate the levels of white blood cell (wbc), eosinophil, iron, hemoglobin (hb), and immunoglobulin e (ige) levels among refugees in kawergosk camp – erbil governorate. the change in these parameters may be a predisposing factor to further disease development. materials and methods the study included 258 syrian refugees of different ages and sex and another 60 volunteers as control group (c.g). these refugees were in kawergosk camp in erbil governorate. forms were filled for each individual for information that included age, gender, case history, physical, and laboratory examination reports. four milliliter of blood were withdrawn from each refugee and c.g by sterile disposable syringes. blood samples were divided into two parts – 2 ml were placed into ethylenediaminetetraacetic acid-containing tubes for the performance of hematological tests including wbc, eosinophil corresponding author: tanya s. salih, department of biology, cihan university, erbil, kurdistan region, iraq. e-mail: tanya.salam@cihanuniversity.edu.iq received: apr 23, 2019 accepted: apr 27, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp80-84 copyright © 2019 tanya s. salih, safa safin haydar, muhsin h. ubeid, ameena s. m. juma. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ salih, et al.: some immunological and hematological parameters among refugees 81 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 80-84 count, iron, and hb and 2 ml of blood were placed into plain gel tubes and allowed to clot at room temperature, then centrifuged at 2500 rpm for 15 min for serum collection. the serum was transferred into two labeled eppendorf tubes, 0.5 ml of serum in each and stored at −20°c for later estimation of ige levels. determination of complete blood count was made using the automated hematology analyzer for all samples within 1 hour of sample collection. wbc, eosinophil, iron, and hb were evaluated. the stored serums were used for estimating ige after bringing them to room temperature, then determining ige by an automated cobas e411 immunoassay analyzer and using its ige kit (roche company). statistical analysis data were evaluated statistically, analyzed, and organized in tables. computer program software – statistical package for the social sciences version 18 was used to analyze the data. quantitative variables were compared using the students f-test and independent t-test. results were considered highly significant, if the p < 0.01; significant if the p value was between 0.01 and 0.05; and non-significant if the p value was more than 0.05. results wbc count table 1 represents the mean wbc count. it reveals that wbc count is non-significantly slightly increased in both males and females refugee when compared to c.g (p > 0.05). eosinophil count the mean of eosinophil count of refugees in comparison to c.g. is shown in table 2. it was revealed that there was a highly significant rise in eosinophils in male and female refugees in comparison to c.g. (p < 0.01). iron and hb levels figure 1 shows the level of iron among adult refugees (above 15 years of age) and children. most of the refugees had normal iron levels, where iron concentrations were more than 65 mg/dl among 67 males and more than 50 mg/dl among 104 females and 48 children, while some had iron deficiency in which the majority was females (9 males, 24 females, and 6 children). in addition, hb concentrations were normal among 65 males (more than 13.0 g/dl), 89 females (more than 11.0 g/dl), and 48 children (more than 12.0 g/dl). table 1: mean wbc counts in refugees in kawergosk camp in erbil governorate immunological parameter gender no. mean±se no. mean±s t-test p refugees suffer with asthma (c.g) p value wbc 109/l m 121 7.91±0.57 30 6.74±0.47 0.123 n.s f 137 7.67±0.31 30 6.69±0.44 0.089 n.s p: probability, p≥0.05: non-significant; wbc: white blood cell, se: standard error table 2: mean of eosinophil count in refugees in kawergosk camp in erbil governorate immunological parameter gender no. mean±se no. mean±se t-test p refugees suffer with asthma (c.g) p value eosinophil 109/l m 121 5.60±0.18 30 3.50±0.45 0.001 h.s** f 137 5.81±0.15 30 2.80±0.38 0.000 h.s** p: probability; p≥0.05: non-significant; *p<0.05: significant; **p<0.01: highly significant. se: standard error figure 1: iron and hemoglobin levels in refugees in kawergosk camp in erbil governorate salih, et al.: some immunological and hematological parameters among refugees 82 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 80-84 however, anemia was found among 8 men, 42 women, and 6 children. data were compared between individuals regarding the relation of hb to iron. the first group had normal hb and normal iron; the females are the highest number with 98 females, 65 males, and 37 children. in addition, the second group was classified to have iron-deficiency anemia in which the highest deficient was among females (6 males, 20 females, and 2 children). furthermore, the results have been recorded to have 2 males, 22 females, and 3 children that suffered from anemia, but their blood iron was within normal range. moreover, it has been found that at least one of the males, females, and children had iron deficiency but had no anemia, as shown in figure 2. serum ige mean serum levels of ige among male and female refugees showed highly significant increased levels when compared to c.g. (p < 0.01), as shown in table 3. discussion the general health of most refugees has been found to be bad. there is no provision of basic health care, no clean drinking water, and no primary education for children.[4] although the wbc count was insignificantly increased in the current study, this may have several indications. these cells are very important constituents of blood. they play a vital role in combatting infections and stabilizing health the condition in an individual. a high wbc count may indicate that the immune system is stimulated due to the presence of an infection. physical and emotional stress has also been found to cause an increase in the wbc count. the presence of some cancers in some people cannot be eliminated, which is another cause of increased wbc counts.[5] eosinophils are one of the wbc types that are an important component of the immune system. causes of eosinophilia are many, but among refugees, it may be because of parasitic infections, allergies, or even asthma.[6] in the current study, eosinophilia was prominent among the studied group. the presence of other infections or asthma cannot be excluded in these people, for a lot of them did show signs of asthma. refugees of all ages and ethnicities have been found to have anemia. the prevalence of anemia in newly arrived populations has ranged from 19% among african refugees resettling in australia to 37% among southeast asian refugees resettling in the united states.[7,8] anemia was identified in 12% of 1247 refugee children in massachusetts, with a rate of 29% among children under 2 years.[9] in addition, a study in maine found that 20% of 127 refugee children were anemic at the time of their new arrival medical evaluation.[10] these findings are consistent with the findings of the current study. anemia may result from a wide range of disease processes. common causes of anemia in refugee populations include iron deficiency, inherited hematologic abnormalities (e.g., thalassemias, hemoglobinopathies, and enzyme defects), and infectious diseases (e.g., malaria and intestinal parasitosis). the ultimate cause is often multifactorial.[11] iron-deficiency anemia, probably the most common cause of anemia in immigrant populations, is usually manifested as a microcytic anemia. the groups most commonly affected are children and women;[12] however, refugees of both sexes and all age groups are at risk. although it is frequently figure 2: correlation between iron and hemoglobin levels in refugees in kawergosk camp in erbil governorate table 3: mean serum levels of ige in refugees in kawergosk camp in erbil governorate immunological parameter gender no. mean±se no. mean±se t-test p refuges (c.g) p value ige iu/ml m 121 281.97±44.81 30 34.33±6.48 0.000 h.s** f 137 317.59±47.29 30 33.32±6.38 0.000 h.s** p: probability; p≥0.05: non-significant; *p<0.05: significant; **p<0.01: highly significant. se: standard error salih, et al.: some immunological and hematological parameters among refugees 83 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 80-84 multifactorial, it is primarily caused by deficient dietary iron. chronic blood loss, which frequently adds to iron deficiency, is commonly caused by infection with intestinal parasites, particularly hookworm. helicobacter pylori infections may lead to gastrointestinal blood loss through ulcer formation.[6] ige, named in 1968,[13-15] was the last of the five classes of human antibodies to be discovered, and today is commonly associated with the various manifestations of allergic disease.[16] ige antibodies are well known for their role in mediating allergic reactions, and their powerful effector functions activated through binding to fc receptors fcεri and fcεrii/ cd23.[17] the possible exposure of the refugees to different allergens may be the main reason of the significant increase in ige levels; this could also be the reason for the asthmatic sings in these individuals.[17] one might predict that the high ige levels which present in parasite-endemic regions would be associated with increased rates of atopic disease. however, the data on this point are complicated. a number of studies have pointed to the opposite effect, namely a decrease in allergic conditions in parasitized patients.[18,19] in the setting of chronic filarial infestation, this has been associated mechanistically with an il-10-driven state of t h 1 and t h 2 suppression. in contrast, investigations in ascaris or toxocara species-infected patients point to a parasiteinduced increase in allergen-specific t h 2 responses and allergic symptoms, including wheezing.[20-22] cross-reactivity between parasitic antigens and allergens might play a significant role, as has been suggested by the observation that ige formed in response to the tropomyosin of ascaris lumbricoides recognizes the homologous proteins from the dust mite or cockroach.[23] conclusion mean serum levels of ige among male and female refugees showed highly significant increasing. most of the refugees had normal iron levels, while some had iron deficiency in which the majority were female. in addition, hb concentrations were normal among most of them. however, anemia was found. it was revealed that there was a highly significant rising in eosinophils in male and female. wbc count is non-significantly slightly increased in both male’s and female’s refugees. recommendations follow-up tests or refugees are recommended to avoid any health complications and to ensure the treatment for them. references 1. world health organization. “10 things to know about the health of refugees and migrants”. world health organization, 2019. available from: https://www.who.int/news-room/featurestories/detail/10-things-to-know-about-the-health-of-refugeesand-migrants. [last accessed on 2019 april 10]. 2. “kurdistan regional government, syrian refugees in erbil governorate”, 2019. available from: http://www.cabinet.gov.krd/p/ page.aspx?l=12&s=000000&r=401&p=483&h=1&t=407. [last accessed on 2019 april 10]. 3. “copenhagen: who regional office for europe; migration and health: key issues”. in: health topics, 2015. available from: http://www.euro.who.int/en/health-topics/health-determinants/ migrationand-health/migrant-health-in-the-european-region/ migration-and-health-key-issues. [last accessed on 2018 apr 12]. 4. a. abdi. “a hard life for kurds in the makhmour refugee camp”, 2015. available from: https://www.thekurdishproject.org/ahard-life-for-kurds-in-makhmour-refugee-camp. [last accessed on 2019 april 10]. 5. l. smith. “what to know about high white blood cell count, medical news today”, 2018. available from: https://www. medicalnewstoday.com/articles/315133.php. [last accessed on 2019 april 10]. 6. cdc. “refugee health guidelines, complete blood count with red blood cell indices, white blood cell differential, and platelet count”, 2019. available from: https://www.cdc. gov/immigrantrefugeehealth/guidelines/domestic/general/ discussion/complete-blood-count.html. [last accessed on 2019 april 10]. 7. a. c. tiong, m. s. patel, j. gardiner, r. ryan, k. s. linton, k. a. walker, j. scopel and b. a. biggs. “health issues in newly arrived african refugees attending general practice clinics in melbourne”. medical journal of australia, vol. 185, pp. 602-606, 2006. 8. a. catanzaro and r. j. moser. “health status of refugees from vietnam, laos, and cambodia”. jama, vol. 247, pp. 1303-1308, 1982. 9. p. l. geltman, m. radin, z. zhang, j. cochran and a. f. meyers. “growth status and related medical conditions among refugee children in massachusetts, 1995-1998”. american journal of public health, vol. 91, pp. 1800-1805, 2001. 10. e. b. hayes, s. b. talbot, e. s. matheson, h. m. pressler, a. b. hanna and c. a. mccarthy. “health status of pediatric refugees in portland me”. archives of pediatric adolescent medicine, vol. 152, no. 6, pp. 564-568, 1998. 11. w. m. stauffer, d. kamat and p. f. walker. “screening of international immigrants, refugees, and adoptees”. primary care, vol. 29, no. 4. pp. 879-905, 2002. 12. k. pottie, p. topp and f. kilbertus. “case report: profound anemia. chronic disease detection and global health disparities”. canadian family physician, vol. 52, pp. 335-336, 2006. 13. t. a. platts-mills, p. w. heymann, s. p. commins and j. a. woodfolk. “the discovery of ige 50 years later”. annals of allergy, asthma and immunology, vol. 116, pp. 179-182, 2016. 14. h. h. bennich, k. ishizaka, s. g. o. johansson, d. s. rowe, d. r. stanworth and w. d. terry. “immunoglobulin e, a new class of human immunoglobulin”. bulletin of the world health organization, vol. 38, pp. 151-152, 1968. 15. k. ishizaka, t. ishizaka and m. m. hornbrook. “physicochemical properties of reaginic antibody. v. correlation of reaginic activity with γe globulin antibody”. journal of immunology, vol. 97, pp. 840-853, 1966. 16. h. j. gould and b. j. sutton. “ige in allergy and asthma today”. nature reviews immunology, vol. 8, pp. 205-217, 2008. 17. b. j. sutton, a. m. davies, h. j. bax and s. n. karagiannis. “ige antibodies: from structure to function and clinical translation”. antibodies, vol. 8, no. 1, p. 19, 2019. 18. a. h. van den biggelaar, r. van ree, l. c. rodrigues, b. lell, a. m. deelder,p. g. kremsner and m. yazdanbakhsh. “decreased atopy in children infected with schistosoma haematobium: a role for parasite-induced interleukin-10”. lancet, vol. 356, no. 9243, pp. 1723-1727, 2000. 19. p. j. cooper, m. e. chico, l. c. rodrigues, m. ordonez, d. strachan, g. e. griffin and t. b. nutman. “reduced risk of atopy among school-age children infected with geohelminth parasites in a rural area of the tropics”. the journal of allergy and clinical immunology, vol. 111, no. 5, pp. 995-1000, 2003. 20. g. m. hunninghake, m. e. soto-quiros, l. avila, n. p. ly, c. liang, j. s. sylvia, b. w. hollis and s. t. weiss. “sensitization to ascaris lumbricoides and severity of childhood asthma in costa rica”. salih, et al.: some immunological and hematological parameters among refugees 84 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 80-84 the journal of allergy and clinical immunology, vol. 119, no. 3, pp. 654-661, 2007. 21. s. dold, j. heinrich, h. e. wichmann and m. wjst. “ascarisspecific ige and allergic sensitization in a cohort of school children in the former east germany. the journal of allergy and clinical immunology, vol. 102, no. 3, pp. 414-420, 1998. 22. m. m. alshishtawy, a. m. abdella, l. e. gelber and m. d. chapman. “asthma in tanta, egypt: serologic analysis of total and specific ige antibody levels and their relationship to parasite infection”. international archives of allergy and immunology, vol. 96, no. 4, pp. 348-354, 1991. 23. m. j. sereda, s. hartmann and r. lucius. “helminths and allergy: the example of tropomyosin”. trends in parasitology, vol. 24, no. 6, pp. 272-278, 2008. tx_1~abs:at/add:tx_2~abs:at 68 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 68-75 research article green synthesis of gold nanoparticles using zingiber officinale rhizomes aqueous extract and its gastroprotective activity against ethanol-produced stomach ulcer in rats suhayla h. shareef1,2, ismail n. mahdi3, mustafa a. badeea3, mahmood a. abdulla3 1department of biology, college of education, salahaddin university, erbil 44001, kurdistan region, iraq, 2department of medical biochemical analysis, college of health technology, cihan university, erbil, kurdistan region, iraq, 3department of medical microbiology, college of science, cihan university, erbil, kurdistan region, iraq abstract synthesis of gold nanoparticles exhibition wide range of bioactivities and was conventionally used for the remedy of numerous disorders. our study aims to measure the gastroprotective effect of synthesized gold nanoparticles on absolute ethanol-induced stomach ulceration in rats. the rats were noticeably administered with two doses of gold nanoparticles (175 ppm and 350 ppm) with absolute alcohol and showed a significant improvement in the adverse effect of ethanol-produced stomach damage as confirmed by decreasing ulcer index and raising % of ulcer prevention. increased mucus ph of stomach content reduced ulcerated portion, condensed or nonappearance of edema, and white blood cells permeation subcutaneous layer. in gastric tissue homogenate, synthesized gold nanoparticles presented a substantial increase in superoxide dismutase (sod), catalase (cat) actions, and a substantial lessening of malondialdehyde (mda) amount. besides, gold nanoparticle increases the intensity of periodic acid schiff stained (pas) of the gastric epithelium of the stomach. gold nanoparticles confirmed gastro-protective effects which might be qualified to its antioxidant, increase mucus secretion, increase endogenous enzymes (sod and cat), and reduce mda quantity. keywords: synthesized zingiber officinale, gold nanoparticle, gastric ulcer, histology, pas introduction a peptic ulcer is the most communal gastrointestinal tract ailment that is moving a huge number of individuals universally and has therapeutic and worldwide financial attention. several factors are included in the progress of stomach ulceration, for instance, stress, smoking, malnutrition, administration of nonsteroidal antiinflammatory medications, genetic tendency, and contagion by h. pylori.[1] even though there are, countless medications presently used in the scientific arena to achieve stomach ulcer illness, the mainstream of them displays numerous adversative responses. therefore, there is necessary to notice additional active and fewer poisonous antiulcer mediators.[2] the great attention to herbal medications and the pharmacologic actions of their bioactive composites has given rise to the detection of abundant usual medicines or herbal excerpts.[3] in specific, a huge quantity of therapeutic plants established anti-ulcer possessions,[4] with fairly a smaller amount cost and an extensive variety of protection boundaries.[5] zingiberaceae plant family furthermost plentiful in southeast asia, extensively utilized outdated remedies.[6] zingiber officinale is one of the zingiberaceae species, categorized by its important financial consequence; meanwhile, it is used as a flavor and as a traditional medication.[7] the rhizomes are prevalent in numerous humid nations. the rhizomes have been exhaustively deliberate and found to display an extensive variety of pharmacologic actions for instance febrifuge, painkilling belongings, anti-inflammatory, chemo-preventive actions,[8] antinociceptive, antioxidant, antineoplastic, antimicrobic, antihyperglycemic, antiallergic, and antiplatelet actions.[9] the purpose of the current study is to make stable gold nanoparticles and to observe their gastroprotective significance in ethanol-produced stomach ulceration in rats. corresponding author: suhayla h. shareef, department of medical biochemical analysis, college of health technology, cihan universityerbil, kurdistan region, iraq. e-mail: suhayla.shareef@cihanuniversity.edu.iq received: june 19, 2022 accepted: july 30, 2022 published: august 22, 2022 doi: 10.24086/cuesj.v6n2y2022.pp68-75 copyright © 2022 suhayla h. shareef, ismail n. mahdi, mustafa a. badeea, mahmood a. abdulla. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) shareef, et al.: anti-ulcer effect of gold nanoparticles in ethanol-induced gastric ulcer in rats 69 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 68-75 materials and methods uv-vis’s fascination ranges were established on uv-vis spectroscopic analysis, whereas the size and shape of elements were quantified on scanning electron microscope (sem0, and atomic force microscopic (afm). reliability elements recognized by zeta analyzer. crystallography construction decided samples regulate utilizing high-resolution x-ray diffractometric system jdx-3532 utilizing one-color cu-k𝛼 radiation wavelength 1.5418 ˚a. chemicals and reagents reagents and chemicals were of the analytical score. gold nitrate crystal was purchased from merck in germany; zingiber officinale rhizome and arabic gum were obtained from the local market. omeprazole was purchased from a dispensary dissolved in 10% tween 20 forced feeding by mouth to rats at a dosage of 20 mg/kg (5 ml/kg), as recommended by several investigators’.[10-12] omeprazole was purchased from the pharmacy, ketamine, and xylazine and was purchased from a veterinary clinic. preparation of rhizome extract fresh zingiber officinale rhizomes were gathered and documented by the herbarium of biology department, university of salahaddin-erbil (no erb 8513. the rhizomes were cleaned carefully with distilled water many times to remove the dirt, cut into small pieces, and desiccated under shadow at room temperature for 2 weeks.[13] dry rhizomes make well as ash using an electrical mixer. the ash saturated distilled water (200 g/1000 ml) for 5 days and mixed every 3 h and was filtered using sieve paper and rotating evaporator. finally, prepared aqueous extract solution stored at -20°c. arabic gum (0.15 g) additional to 50 ml purified water 70oc in 100 ml pyrex mug. this solution supplementary aqueous excerpt zingiber officinale rhizomes through rousing by a magnetic stirrer. synthesis of gold nanoparticles a mix of zingiber officinale extracts besides arabic gum is located in a 250 ml bottle fiery 60–70°c. ph mixture accustomed 7.5. after that, 50 ml of 1mm gold nitrate solution additional slowly with rousing. the color of the solution changed from a straight line to violet suggesting the progress of gold nanoparticles. the response was appropriated for 20 min, later, the hotplate detached solution persevered rousing till grasped 25oc. color violet no alteration later four months. ethics statement this training was accessible official ethical assessment panel for endorsement, an agreement approved (ethical number bio/25/03/2021/maa). entirely in vivo investigational techniques were achieved in the sterilized state in the tentative animal unit succeeding their strategies. altogether rats acknowledged humane attention, conferring to the principles delineated “director care and utilize workroom animals” ready nationwide conservatory disciplines available countrywide organization healthiness. initiation of stomach ulceration rats reserved individually distinct pens widespread net wire bottom evade coprophagia. rodents fed normal pellets food permit allowed water admission. investigational animals separate randomly into four collections of six animals correspondingly. rats reserved in animal house for 2 weeks for variation. immediate animals nourished as follow: gathering 1 fed orally 0.5% cmc (w/v) (5 ml/kg) collection 2 gavage omeprazole 20 mg/kg in cmc (5 ml/kg) as the reference group.[14] assemblies 3 and 4 were nourished with gold nanoparticles (175 and 350 ppm/p.o), correspondingly.[15] after 1 h, the entire collection is gavage by mouth absolute alcohol. sixty minutes future, altogether rats were sacrificed under overdose ketamine and xylazine gastric eliminate promptly.[16,17] gross evaluations of stomachs stomach respectively trial rat opened lengthwise greater curvature gastric rinsed cold buffered. stomach epithelium was examined for any injury using a dissecting microscope. stomach ulceration on the epithelium appears as long bands of red hemorrhagic damages. ulcers typically seemed analogous to the long alliance of the stomach. the coldness (mm) and thickness (mm) of every crew were restrained by utilizing an integrator [(10 mm × 10 mm = ulcer portion) under strew microscopy (1.8×)]. ulcer zone of every laceration designed through count numeral small squares, 2 mm × 2 mm, cover dimension thickness of every hemorrhagic ulcerated band.[18] the summary of the parts of complete abrasions for respective gastric practical cunning ulcerated part (ua) the totality of small squares × 4 × 1.8 = ua mm2. inhibition % (i %) was calculated by the following preparation inhibition percentage (i%) = (ua control– ua treated/ ua control) × 100[19] measurement of stomach ph stomach content composed spined estimate ph of stomach’s fluid supernatant through ph metrical titration through 0.1 n naoh solutions utilizing numerical ph pattern and acidity restrained in meq/l.[13,20] quantity of stomach mucus content stomachs rinsed with cold phosphate buffer saline. the stomach mucosa of respectively rat’s stomach was slightly scrubbed off using a clean slid and mass of secretion quantity utilizing correct electric equilibrium.[16] formulation of gastric tissue homogenate small piece glandular gastric wall from the respective rat washed away prudently cold pbs. utilizing homogenizer. a minor shareef, et al.: anti-ulcer effect of gold nanoparticles in ethanol-induced gastric ulcer in rats 70 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 68-75 portion of every gastric wall standardized cold pbs including a mammal protease inhibitor mixture. gastric homogenates centrifuged 1000 g 10 min 4°c. clean supernatant utilized measure amounts of superoxide dismutase (sod), catalase (cat), and malondialdehyde (mda). these assessments were completed based on the consistent manufactural method (cayman, usa).[21] measurements of endogenous antioxidant enzymes sod and cat activities stomach homogenate measured utilizing lucrative usual kits. the manufacturer’s trials were used to measure their amounts in homogenate supernatant.[22] measurements of lipid peroxidation (mda) level of stomach homogenate mda level glandular stomach homogenate was quantified using merchantable kits consulting to manufactural technique (cayman chemical co).[23] histological evaluation minor sections (1–2 cm), respectively, stomach was fixed instantly in 10% formalin overnight followed by tissue dehydrated with ethanol, permission with xylene, and permeation with paraffin using tissue processing apparatus. respectively stomach tissue was entrenched in wax sections 5 𝜇m (leica rotation microtome, germany).[24] hematoxylin and eosin stain slides-stained monotonous hematoxylin and eosin (h & e stain) for histopathology examination using a light microscope.[25] periodic acid schiff (pas) stains to measure mucus excretion of stomach’s glandular portion, slices of 5 𝜇m width stains with pas to imagine gastric mucus and differences in both acid and base glycoproteins following the construction exercise method.[26] statistical data investigation the statistical investigation was achieved using a one-way analysis of variance (anova) method shadowed through a post hoc trial with turkey’s post hoc test. normality testing was performed using the kolmogorov–smirnov test. altogether morals are designed by way of mean ± sem. value of p < 0.5 measured significant. results influence of zingiber officinale gold nanoparticle on gross assessment of stomach rats fed with zingiber officinale gold nanoparticles showed meaningful reductions in the ulceration portion of gastric comparison to the ulcerated gathering [figure 1]. flatten stomach epithelial surface decrease of mucosa damage also noticed macroscopical estimation of investigational rat’s stomach nourished with zingiber officinale gold nanoparticle in comparison with ulcered control assemblage [figure 1]. influence of zingiber officinale gold nanoparticle on gastric mucus content experimental rats fed with zingiber officinale gold nanoparticles presented a considerable upsurge in mucus excretion stomach comparison ulcerated cluster [table 1]. effect of zingiber officinale gold nanoparticle on ph of the stomach investigational rats nourished with zingiber officinale gold nanoparticle preceding given of absolute alcohol suggestively augmented stomach ph likened to ulcered control cluster [table 1]. influence of zingiber officinale gold nanoparticle on the histological estimation of absolute ethanol-induced gastric ulceration in rats h & e stain ulcer control collection obtainable widespread damage to stomach mucosa, and lacerations infiltrated intensely into the gastric epithelium with edema and white blood cell infiltration of subcutaneous coating. rats in trial assemblages nourished with zingiber officinale gold nanoparticle exhibited relatively enhanced defense stomach epithelium for instance showed through the lessening of ulcerated portion, lessening or nonappearance edema, provocative cell penetration submucosal coating [figure 1]. pas stain trial rodents nourished with zingiber officinale gold nanoparticles showed a moderately increased pas staining intensity of glycoprotein of stomach glandular epithelial, in comparison with ulcered control collection [figure 1]. table 1: effect of the gold nanoparticle on mucus weight, ph of the stomach, ulcer area, and inhibition percent of ulcer area in rat’s stomachs animal groups pre-feeding (5ml/kg) mucus weight (g) ph ulcer area (mm)2 inhibition (%) g1 ulcer control 10% tween 20 0.676 2.88±0.04 779.3±3.50 g2 omeprazole 20 mg/kg omeprazole 1.907 5.81±0.02 100.2±1.93 100.2 g3 low dose 175 ppm gold nanoparticle 1.565 5.16±0.02 153±2.08 52.6 g4 high dose 350 ppm gold nanoparticle 1.773 5.35±0.02 136.8±0.79 36.5 shareef, et al.: anti-ulcer effect of gold nanoparticles in ethanol-induced gastric ulcer in rats 71 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 68-75 figure 1: plate 1 (a): influence of gold nanoparticles on the macroscopical appearance of absolute ethanol-produced gastric damage in rats. (a) ulcer control cluster showed severe hemorrhagic lacerations of the stomach epithelium. (b) omeprazole collection presented slight injuries to stomach mucosa. (c and d) investigational assemblies showing a clear reduction in gastric mucosa. plate 2 (b): influence of gold nanoparticle on histology of gastric mucosa against ethanol-induced mucosal damages in rats. a. ulcered control cluster viewing severe mucosal injury together with necrosis, edema, and leukocytes penetration of sub-mucosal coating. b. omeprazole assemblage observing small mucosal impairment. (c and d) experimental rats exhibition reduction mucosal epithelium (h&e stain). plate 3 (c): influence of gold nanoparticle on periodic acid schiff (pas) stains of stomach glycoproteins secretion on ethanol-produced stomach damages in rats. a. ulcer control cluster revealed lessening or nonappearance of pas stains of gastric epithelium with extensive mucosal injuries. (b) omeprazole set demonstration strong pas stain. (c and d) trial groups exhibited dense up-take of pas stains b c d a a b c shareef, et al.: anti-ulcer effect of gold nanoparticles in ethanol-induced gastric ulcer in rats 72 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 68-75 effects of zingiber officinale gold nanoparticle on endogenous antioxidant enzymes of stomach tissue homogenate rats in ulcered control assortment demonstrated expressively lesser sod and cat activities compared to the normal collection. animals in experimental gatherings nourished zingiber officinale gold nanoparticle offered meaningfully returned the reduction of sod and cat to normal range [figure 2]. influences of zingiber officinale gold nanoparticle on malondialdehyde (mda) in gastric epithelium homogenate animals in ulcered control collection displayed a considerable increase in mda level of stomach tissue homogenate compared to normal assemblage [figure 2]. mda of gastric homogenate dramatically condensed in rodents nourished with zingiber officinale gold nanoparticles. mda was used as an indicator for lipid peroxidation. discussion absolute alcohol necrotizing agent brings abundant adverse possessions on gastric tissue, for example, damages causing lessening plentiful protein concentration.[7,9,26,27] the mucous membrane is recognized chief line of defense organization in the stomach; consequently, absolute alcohol eliminates mucus, and changeable permanent damage predictable.[3,28] the stomach mucosa performance barrier does not allow disgrace enzymes, for instance, pepsin effusive stomach wall.[24,29,30] chiefly, mucus owns real slight permeability large elements, perhaps pepsin, nonetheless closely subjects, alike tension, absolute alcohol increases permeability reason liberation of vasoactive crops, and finally vascular damage. vascular damaging enhanced alcohol success in gastric tissue and consequence disproportions removal of bicarbonate and building mucus.[2,23,31] likewise, proposed construction reactive oxygen species (ros) influenced complete absolute alcohol significant percentage ulceration development.[27,32-34] it is fine recognized that omeprazole drips additional majority acid intended gastric, omeprazole is effective in acid self-sufficient imitations, analogous ethanol ulceration, employment mucosal protection.[35,36] consistently, expected antioxidants, include the ability of gastroprotection.[3,29,32,37] in the current study, rats pre-nourished with gold nanoparticle enhanced construction of stomach mucosa increase wide improvement protein concentration in gastric secretions.[38,39] significances of the present exercise were similar with other amendments quantity gastroprotective and antiulcer achievement gold nanoparticle and herbal composites.[4,40,41] pre-feeding with the gold nanoparticle, challengingly impassable gastric acid likewise takeover stomach injury in rats forced feeding ethanol, characteristic gastro-protective effect gold nanoparticle easy relatively protection of gastric wall.[42,43] designated acute inflammation is attracted through ethanol neutrophils penetration the stomach partition.[1,44] the current values recognized submucosal infusion proficiently congested through pre-feeding rodents by the gold nanoparticle. an extensive release of reactive oxygen species (ros) and free radicals’ reason metabolic lessening everlasting damage.[45] pre-feeding rats with the gold nanoparticle, expressively increased the stomach mucus structure weakened acidity of gastric contented.[5,43] gastric mucosal protection, reduction of hemorrhagic mucosal area, decrease or nonappearance of edema, and provocative responses of sub-mucosal coating rats nourished gold nanoparticle compared to ulcerated control collection long-established by the valuation of the gastric histology.[8,46,47] periodic acid-schiff (pas) staining results displayed increased mucus creation in gastric of rodents pre-nourished gold nanoparticle indorses gastro-protective achievement of gold nanoparticle intensity since of defense of mucus release. these significances consistent abundant other exercises that obtainable substantial upsurge of stomach mucus concentration (pas) stains nourished several compounds against ethanol produced gastric damages.[48-50] pas stains figure 2: effect of the gold nanoparticle using aqueous rhizomes extract of zingiber officinale on antioxidant enzyme activities (sod and cat) and mda level in the liver. statistically significant differences were examined using one-way anova and tukey’s post hoc multiple comparisons test. ****p < 0.0001, ***p < 0.001, **p < 0.005, *p < 0.05 above columns specify significant differences from the normal control group and above lines for the differences between treated groups. ns p > 0.05 denotes to non-significant differences shareef, et al.: anti-ulcer effect of gold nanoparticles in ethanol-induced gastric ulcer in rats 73 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 68-75 decrease in ethanol contrary augmented through gold nanoparticle feeding likewise formed increased glycoprotein content.[5,49] superoxide radical anion is formed through polymorph nuclear neutrophils subsequent in reply by cellular lipids formation lipid peroxides. supplementary actual indicators oxidative compression motives mucosal injury could be mda principal metabolite of fat peroxidation.[1,51] conferring to results of existing exercise, oral forced feeding of gold nanoparticles protects against gastric ulcer through upsurge sod and cat actions, and lessening mda quantity consistent with consequences specified through plentiful investigators concerning sod and cat actions enhanced gold nanoparticle nourished clusters in comparison to absolute alcohol gavage rodents.[43,52] furthermore, reduction in mda quantities were specified rats forced feeding with many herbal and artificial composites.[50,53] ros, for example, hydroxyl radicals, superoxide anions, and lipid peroxides damaging programs approved to worsen gastric ulcer development.[54] antioxidants hunt free-radical formation exhibition principal portion in the protection of cellular damage. rats fed gold nanoparticle enhanced antioxidant enzymes activities in contrast to ulcer collection evading free radical construction might happen through ulcered development. comparable outcomes have been described through numerous academics.[28,38] conclusions according to the significance of the existing study, gold nanoparticles revealed significantly gastroprotective results in inhibition of ethanol-induced gastric ulcers in rats as accepted by macroscopic appearance, histology, and pas stain. gold nanoparticle intensely increases the serum concentration of cat and sod, while significant reduction of hepatic mda. the gastroprotective significance of gold nanoparticles in ethanol-induced ulceration could be due to its measurements to discount oxidative stress, lipid peroxidation, and its antioxidant and free-radical scavenger possessions. statement of rival interest the authors announce that they have no acknowledged rival financial welfares or individual relations that could have seemed to affect the work stated in this manuscript. writer aids altogether novelists paid to train beginning and enterprise. s.h. s and m.a.a: leading the trials, information gathering, and examination, document research. students: analysis and clarification of information, studied breezes of the script. students subsidized reagents materials, understanding of statistics, and studied breezes of text. references 1. f. a. armah, i. t. henneh, j. alake, w. ahlidja, b. amoani, e. g. ofori, b. asante-kyei, g. i. temitayo, c. k. adokoh. allanblackia floribunda seed extract attenuates the ethanol-induced gastric ulcer in rats via the inhibition of tnf-α and inf-γ levels and modulation in the expression of ki67 protein. biomedical research 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gastric ulcer in mice. evidence-based complementary and alternative medicine, vol. 2021, pp. 9998982, 2021. 53. s. lu, s. kong, y. wang, z. hu, l. zhang and m. liao. gastric acidresponse chitosan/alginate/tilapia collagen peptide composite hydrogel: protection effects on alcohol-induced gastric mucosal injury. carbohydrate polymers, vol. 277, pp. 118816, 2022. 54. j. lawrence, g. swerhone, j. dynes, a. hitchcock and d. korber. complex organic corona formation on carbon nanotubes reduces microbial toxicity by suppressing reactive oxygen species production. environmental science nano, vol. 3, no. 1, pp. 181189, 2016. tx_1~abs:at/add:tx_2~abs:at 153 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 research article pre and early post-operative iron assessment in obese patients underwent laparoscopic sleeve gastrectomy salih m. s. zebari1, nashwan q. mahgoob2, maryam n. qahtan1, noorhan m. jassim1, hafsa o. adil1, amenah q. hashim1 1department of nutrition and dietetic, cihan university-erbil, kurdistan region, iraq, 2advance laparoscopy and obesity center, soran private hospital, erbil, kurdistan region, iraq abstract obesity is a chronic disease associated with mortality and morbidity. bariatric surgery as a long-term weight reduction treatment has been increased. this study investigated the early state of iron storage and deficiency in patients underwent laparoscopic sleeve gastrectomy (lsg). thirty patients were selected as a prospective study underwent lsg according to specialized inclusion and exclusion criteria at soran private hospital in erbil, from january 01, 2021, to september 01, 2021, after signing a consent form. all selected patients underwent preoperative and 3 months’ post-operative hemoglobin (hb), mean corpuscular volume (mcv), and serum ferritin (sf) assessments. they were instructed to follow specialized diet regimen for 3 months’ post-operative and not to take any drugs and supplements. results showed a strong positive correlation between pre-operative and post-operative assessment for both hb and mcv respectively (r = 0.72; p < 0.001), (r = 0.76; p < 0.001), and a moderate correlation for sf (r = 0.41; p < 0.024). significant difference (p  <  0.05) was found between pre-operative and post-operative for the three markers (hb, mcv, and sf). their values were decreased from pre-operative to post-operative for the majority of the cases, but they were still within the normal range. the finding demonstrated that after 3 months from lsg, no patients showed iron deficiency anemia. however, there was decrease in sf level which indicated that the storage capacity of iron was decreasing. this suggests that the iron supplement has to be started since the food regime followed in by the patients was not sufficient to maintain normal iron level. keywords: severe obesity, sleeve gastrectomy, iron deficiency, regime follow-up, weight reduction introduction obesity as a chronic disease is considered as a major threaten to the public health,[1] reaching a pandemic state.[2] factors leading to obesity consist of association of genetic, metabolic, food habits, behavioral, social, cultural, and other aspects.[3] recent data by the world health organization (who) demonstrated that worldwide obesity has been increased.[4] over six-hundred million adults globally are suffering from obesity, and more than 1.9 billion adults were overweight.[5] the association among obesity and accelerated mortality is a fact.[6] for example, an average increase in mortality by 30% is resulted by each 5 kg/m2 increase in body mass index (bmi) above 25 kg/m2. this increasing rate of mortality is mainly due to vascular diseases and diabetes.[7] furthermore, the existence expectancy is decreased around 5–20 years in obese patient with a bmi higher than 35.[8] bariatric surgery (bs) reduces weight for obese patients and keeps their weight loss by limiting the consumption and/or induce mal-absorption of food. it is often used when other treatments such diet, exercise and drugs have failed.[9] decreasing the size of the stomach is due to the restraining strategies, and hence restrict the energetic intake and initiate satiety.[10] laparoscopic sleeve gastrectomy (lsg) method is rapidly gaining popularity inside academic medical centers of the usa and worldwide.[11,12] despite the benefit of bs, many complications and the nutritional deficiencies are a worrying trend that need attention.[13] the deficiencies develop because of the change inside the physiology and anatomy of the gastrointestinal corresponding author: salih m. s. zebari, department of nutrition and dietetic, cihan university-erbil, kurdistan region, iraq. e-mail: salih.zebary@cihanuniversity.edu.iq received: august 20, 2022 accepted: november 23, 2022 published: december 30, 2022 doi: 10.24086/cuesj.v6n2y2022.pp153-160 copyright © 2022 salih m. s. zebari, nashwan q. mahgoob, maryam n. qahtan, noorhan m. jassim, hafsa o. adil, amenah q. hashim. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 154 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 tract.[14] the most obvious nutrients deficiency is the risk of iron deficiency anemia (ida). restricted dietary intake, reduced appetite, increased satiety, inadequate intake of iron, and reduced secretion of hydrochloric acid are substantially caused to the development of iron deficiency after lsg.[15,16] in general, different factors contributed to iron deficiency such as poor adherence to dietary guidelines, recommended supplementation, and insufficient professional guidance.[17] therefore, it is advocated that patients underwent lsg ought to acquire postoperative follow-up according to the accreditation and standards from the “american society for metabolic and bs (asmbs)” and the “international federation for the surgery of obesity and metabolic disorders.”[18] however, it has been revealed that 47–90% of patients do not acquire dietary intervention following surgery treatment.[19] iron deficiency prevalence ranged between 1% and 54.1% for the duration of a follow-up of maximal 5 years following lsg and the prevalence of anemia varies between 3.6% and 52.7%.[20] affected patients rate by iron deficiency rely on the suggested iron status markers and their standard ranges. none of the iron marker is an ideal determinant. consequently, the desired screening technique is the combination of markers. therefore, hemoglobin (hb), mean corpuscular volume (mcv), and serum ferritin (sf) levels are the markers that should be included for investigation of iron deficiency.[21,22] this study aimed to evaluate the early state of iron storage and iron deficiency in patients who underwent lsg for weight reduction at preand post-operative using three markers (hb, mcv, and sf levels), and to find out the effectiveness of the diet follow-up plan. patients and methods this research is a prospective study that included 30 patients, selected from 55 patients undergoing lsg for managing their weight loss that held at soran private hospital in erbil, kurdistan region of iraq from january 01, 2021, to september 01, 2021, using the most updated surgical technique. all surgeries were performed by single laparoscopic and bariatric after taking and signing patients written consent form. clinical reassessment was carried out each month postoperatively during the period of the study. all patients under went preoperative and 3 months post-operative hb, mcv, and sf assessments at the same laboratory. a deficiency was described as a serum plasma level determined under the regular reference range. while, excess level was defined as a serum plasma level over the reference range for normal standard. the values of the tests were measured to compare with the described values from the references records as follows: the normal value of hb in men is 13.8–18.0 g/dl and in women is 12.1–15.1 g/dl. the normal reference range of mcv is 80–100 fl. while, the value of sf ranged between 30 and 300 ng/ml for males, and 30–160 ng/ml for females, and a value <50 ng/ml is considered as iron deficiency.[23] all selected patients were instructed to follow the specialized diet regimen for 3 months’ post-operative and not to take any drug or herbal supplements during the study time. the inclusion standard for the patients in this study include 1. age between 18 and 40 years 2. both male and female were included 3. bmi from 35 to 45 kg/m2 4. normal preoperative blood test value (hb, mcv, and sf) 5. no comorbid conditions (ht, dm, and blood disease) 6. accept the procedure, follow-up, and signed the consent form. the exclusion criteria for the patients in this study were as follows 1. patient with abnormal pre-operative blood tests 2. patients who developed intraor post-operative complications (bleeding, leak, and conversion of surgery) 3. failure to follow the regimen given. the food regime that followed by the patients for postoperative composed of 2 stages, stage one for the 1st month and stage two for the next 2 months: [24,25] stage one week 1: clear liquids to give stomach time to heal and help avoid post-operative implications. a clear liquid diet is often the preliminary regimen. the patients start with sips of clear liquid. the goal is to drink a minimum of 2.5 l each day. the clear liquids include: water, tea including herbal or de-caffeinated coffee, fruit, clear chicken, beef, vegetable broth, watery fruit juice such as apple, grape or cranberry, diluted free of sugar squash, and flavored water. liquids to be avoided: caffeine, sugary drinks, and carbonated drinks. week 2: full liquids: the patients should keep drinking more of clear fluids as well as incorporating into fuller liquids that are loaded in protein. full liquid choices include: low fat milk, homemade smoothies, yogurt drinks, smooth soups including cream of tomato, chicken, meat, ice cream without sugar, and sorbet. foods to be avoided as follows: high-fat foods (wholemilk yogurt), sweets foods, and food that has any lumps, caffeine, sugary drinks, and carbonated drinks. week 3: pureed foods: the patients were guided to eat 60 g of protein daily. in addition, they are recommended to add milk to mixture foods, ensuring that it is free of lumps. patients are suggested to eat small portions, and introduce new foods slowly including mashed fruit like bananas, avocados, and canned fruit, blended steamed fish in a low-fat sauce, tuna and salmon, mashed potato, lowfat cheese and soft cheeses, liquidized meat, chicken or fish stew, steamed or boiled vegetables, thick blended soups, and scrambled eggs. foods not to eat include: meals rich in starch (pasta, rice, and bread), hard uncooked vegetables, skin and seeds from vegetables, fatty foods (oils and butter), and spicy food. week 4: smooth meals: the patients can add soft foods and lumps to diet. well-cooked chicken and turkey, soft fish, lean mince meats, vegetables cooked well, cheese with low-fat, zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 155 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 eggs, soups with lumps, soft fruit, sweet potatoes, rice, and cereal with low-sugar. foods not to consume: difficult digested foods including steak, tough vegetables, nuts, white potatoes, and meals with excessive fat. stage two on the week 5 the patients can consume solid food smoothly. after the 1st month following lsg surgery diet instructions for patient is to eat a healthy diet, high protein, and low fat. it is necessary to have three small meals a day, have protein in each meal. plan the meals, drink lots of water, if feel hungry between meals try taking a drink first. it is advised to wait for at least 30 min from the time of eating to drinking. drinking fluids during meals may cause to an overfull stomach and vomiting. stop eating when feel satisfied, overeating will stretch stomach pouch and can cause vomiting. foods not to eat: food with empty calories, alcohol as it has high calories and absorption will steadily increase after surgery. some drink cause bloating and can increase stomach size such as fizzy drinks. high fat foods may cause nausea and will not help to lose weight. tough meats are difficult to chew and hard to digest. the clinical follows-up assessment was done each month through the period of study (3 times evaluation). it included general examination, assessments of vital signs and searching for symptoms and signs of anemia concentrating on the following: headaches, fatigue, weakness, shortness of breath, dizziness, pale skin, irregular heartbeats, cold hands and feet, hair loss, brittle nail, and glossitis. table 1: the preand post-operative values of hb, mcv, and sf patient code age/year sex preoperative tests 3 months post-operative tests hb/g/dl mcv/fl sf/ng/ml hb/g/dl mcv/fl sf/ng/ml 1 21 f 12.5 80 85 11 75 45 2 33 f 14 90 92 12 90 56 3 25 m 16 80 120 14 80 45 4 40 f 14.5 85 43 14 80 51 5 22 f 12 82 60 11 75 60 6 45 f 15 86 50 12 85 66 7 30 f 14 83 45 13 78 45 8 18 f 12.5 87 100 12 80 80 9 30 f 12 80 66 11.5 75 54 10 22 m 12 84 65 11 82 50 11 40 f 14 90 80 13 85 76 12 18 f 13 86 55 12 80 50 13 22 f 15 82 100 14 75 65 14 40 m 14.5 88 96 14 82 58 15 45 f 12.5 80 100 13 80 84 16 33 f 12.5 87 130 12.5 80 55 17 20 f 14 92 105 12 85 67 18 25 m 15 90 110 14 82 55 19 30 f 14 89 120 14 80 64 20 32 f 13 90 94 12 85 80 21 35 f 13 86 90 11.5 82 43 22 18 f 13.5 88 86 13 85 60 23 40 f 12 81 65 12 75 55 24 25 f 15 85 140 13 80 93 25 30 m 14 87 97 12 83 55 26 28 m 12.5 86 100 12 78 80 27 40 f 13.5 90 152 11.5 80 77 28 45 f 14 92 140 13 85 60 29 32 m 12.5 80 105 12 78 58 30 40 f 14 90 80 12.5 85 60 hb: hemoglobin, mcv: mean corpuscular volume, sf: serum ferritin zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 156 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 statistical analysis statistical package for the social sciences (ibm spss version 24) was used. a statistically significant was considered with p ≤ 0.05. measurements are provided as mean ± standard deviation. a number of descriptive methods were used such as frequency distributive and descriptive statistical for demographic variables. a few statistical tests such as t-test paired samples and t-test two independent samples were used to examine the differences in the average measurement before and after the operation for some studied variables. pearson’s test was used to find out the relationships between some relevant variables. results and discussion this study aimed to assess the pre and early post-operative iron assessment in obese patients underwent lsg. this is due to the fact that lsg is one of the most widely used techniques and is account for about 37% of all range of bariatric methods achieved across the world in 2013. however, to ensure that the procedure is success, patients should adhere to comprehensive lifestyle changes. therefore, the follow-ups practice is very important. for this purpose, a multidisciplinary team is necessary that should involve a surgeon, dietitian, psychologist, and family physician or endocrinologist.[25] dietitians in particular have a vital role in this group before and after bs.[26] for instance, loss of medical and nutritional follow-up’s patients did not achieve considerable weight reduction and they were at higher risk of developing nutritional deficiency according to several studies.[27,28] the present study was a prospective, follow up study that included 30 patients after selection to eliminate bias, there were 23 female (76.5%) and 7 male (23.5%) patients. the median age was 30 and their age ranged from 18 to 40 years old [table 1]. their bmi ranged from 35 to 45 kg/m2. all patients underwent smooth recovery without any considerable post-operative complications. table 1 presented the results of preand post-operative tests of the three markers (hb, mcv and sf) for all patients, and the mean, median and mode of the results are presented in table 2. the significant levels between pre-operative and post-operative for the three markers were statistically analyzed [tables 3-5], and the pearson correlation coefficient between pre-operative and post-operative of the three markers were calculated [table 4]. in our study, hb level was not significantly changed after 3 months’ post-operative which means that no patient has anemia, the mcv showed insignificant decrease, while sf showed significant decrease which means that mild deficiency has been occurred. the normal ranges of hb according to the who guidelines should not be <13 g/dl in men and not <12 g/dl in women.[29] a positive correlation (r = 0.76; p < 0.05) was found between hb level at pre and post-operative assessment tests. although these values of hb were dropped from preoperative (mean value 13.5) to postoperative (12.4) as shown on tables 1-4, the values were still within the normal range. this decrease may be due to the implication of the bs that often decrease the level of hb. for example, according to the study conducted by santos et al., the hb levels were slightly decreased from pre-operative to post-operative after 3 months of surgery and were significantly decreased after 1 year of surgery.[30,31] similarly, the mean value of mcv showed a significant (p < 0.05) decrease from 85.5 at pre-operative tests to 80.8 at post-operative tests, while the normal value ranged from 80 to 100 fl. furthermore, the mean value of sf showed a significant table 2: the mean, median, and mode of pre and post-operative results iron markers pre-operative post-operative mean median mode mean median mode hb 13.5 13.75 14 12.4 12 12 mcv 85.8 86 90 80.8 80 80 sf 92.3 95 100 61.5 59 60 hb: hemoglobin, mcv: mean corpuscular volume, sf: serum ferritin table 3: t-test of two paired samples for three variables hb, mcv, and sf paired samples descriptive statistics mean sd std. error mean pair 1 pre hb 13.533 1.0981 0.2005 post hb 12.483 0.9692 0.1770 pair 2 pre mcv 85.87 3.893 0.711 post mcv 80.83 3.788 0.692 pair 3 pre sf 92.37 28.440 5.192 post sf 61.57 12.955 2.365 hb: hemoglobin, mcv: mean corpuscular volume, sf: serum ferritin table 4: paired samples correlations pearson’s correlation n correlation sig. pair 1 pre hb and post hb 30 0.721 0.000 pair 2 pre mcv and post mcv 30 0.765 0.000 pair 3 pre sf and post sf 30 0.411 0.024 hb: hemoglobin, mcv: mean corpuscular volume, sf: serum ferritin table 5: t-test for independent two samples. descriptive statistics of hb descriptive statistics of hb sex n mean sd std. error mean pre hb male 7 13.786 1.4960 0.5654 female 23 13.457 0.9760 0.2035 post hb male 7 12.714 1.2536 0.4738 female 23 12.413 0.8874 0.1850 hb: hemoglobin zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 157 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 (p < 0.05) decrease from 92.3 at pre-operative tests to 61.5 in post-operative tests (normal value 30–300 ng/ml for males, and 30–160 ng/ml for females), the statistical analyses are shown in tables 3-7. recommendation of iron intake and other nutrients are available within the “dietary reference intakes (dris)” set by the “food and nutrition board” at the “institute of medicine of the national academies”. dri is the general term for a set of reference values used for planning and assessing nutrient intakes of healthy people.[31] according to the practice guidelines of the asmbs, iron status have to be assessed at every follow-up visit following bs. in case of id, a daily iron dosage of 150–200 mg should be provided via supplements.[24] this because of the fact that patients with bs are at high risk of suffering from id before and after surgery.[32] a study revealed an estimation of 15% of the prevalence of iron deficiency 1 year after sleeve gastrectomy.[33] therefore, preoperative education is essential for better understanding the complication of lsg. in both pre-operative and postoperative, the coordination between expertise in medicine, surgery, psychology, and nutrition is required.[10] although the impact of variable sex on the three markers at pre-and post-operative assessment were not effective [tables 6-10], hb value at pre-operative for all female cases was at normal range. whereas for seven male cases, only three cases had lower value at pre-operative assessment compared to normal range. in addition, there was a significant decrease on post-operative of the most patients. there were no significant differences in the mean value of hb between male and female neither at pre nor at post-operative assessment. this refers that sex variable has no effect on the hb vale. similarly, the results of mcv of preoperative in all case were within the normal range. however, there was a significant decrease on mcv at post-operative compared to preoperative. at preoperative assessment test, only two female cases had a value <50 ng/ml which is considered as iron deficiency. while, at postoperative assessment 4 patients had lower value than normal range. in addition, there was a significant decrease on the value of sf for most cases from pre-operative to post-operative assessment. several studies have documented that iron intake after lsg is reduced below the minimal daily requirement (8.7 mg a day for men over 18. 14.8 mg a day for women).[34] in addition, some others factors make contribution to the high occurrence of iron deficiency after lsg such as failure to provide supplements consistently,[34] lack of patient adherence to supplementation,[35] the link of iron supplement with troublesome gastrointestinal side effects,[24] large gaps in dietary follow-up, and the absence of specialized nutritional care after surgery. hence, led to iron deficiency progresses from mild iron deficiency, to iron-deficiency erythropoiesis (erythrocyte production), and finally to ida.[36,37] for iron supplementation following lsg, there are two phenomena of thought. few bariatric surgeons choose to monitor laboratory levels of iron and then have patients start iron once their iron levels begin to decrease. on the other hand, others bariatric surgeons prefer to have patients start a daily table 6: t-test for two independent samples with respect to sex variable and hb t-test for sex and hb variables pre hb pos hb equal variances assumed equal variance not assumed equal variances assumed equal variance not assumed levene’s test for equality of variances f 3.364 3.474 sig. 0.077 0.073 t-test for equality of means t 0.688 0.548 0.714 0.592 df 28 7.620 28 7.920 p-value 0.497 0.600 0.481 0.570 mean difference 0.3292 0.3292 0.3012 0.3012 se difference 0.4784 0.6010 0.4220 0.5087 95% confidence interval of the difference lower −0.6507 −1.0687 −0.5631 −0.8738 upper 1.3091 1.7271 1.1656 1.4763 hb: hemoglobin table 7: t-test for independent two samples. descriptive statistics of mcv descriptive statistics of mcv sex n mean sd se mean pre mcv male 7 85.00 3.873 1.464 female 23 86.13 3.946 0.823 post mcv male 7 80.71 2.059 0.778 female 23 80.87 4.214 0.879 hb: hemoglobin, mcv: mean corpuscular volume zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 158 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 dose of iron soon after their operation to be able to prevent levels from declining. the most important thing is to continue to get blood work done as advised so iron supplementation can be adjusted as needed and can keep normal iron levels. however, sometimes patients’ loss to follow-up. if there is a more effective and evidence-based guideline for repletion of iron deficiency, patients are more likely to be participated. therefore, it is preferable to give patients who are not serious about follow-up for any reason iron treatment, since our study supported a decrease in iron stores in patients after the 3rd month of the operation.[38] conclusion the finding data of patients participated in this study showed decrease in their sf after 3 months from operation even with adequate nutritional support but not to a level of causing ida. therefore, patients undertaken lsg should start iron supplements at 3 months or earlier as the patient can tolerate the supplements. follow-up is mandatory to detect early iron deficiency. further assessments are needed on iron state after 6 months and 1 year and then annually to have more information about the quality of nutrition state of patients after sleeve gastrectomy. it is suggested that iron supplements should not be used until patients have been evaluated by their primary healthcare provider such as nutritionists, dietitian or their surgical team. this is to ensure that supplementations were taken appropriate and under their supervision. when patients are counseled about iron supplementation, it is important to remind them that iron supplements may be associated with gi irritation, nausea, constipation, dark stools, and abdominal pain. to prevent iron toxicities, patients using iron supplements should be reminded not to take multivitamin supplements that contain iron. patients should be encouraged to contact their primary health-care provider if they experience any adverse effects or their symptoms worsen. to monitor their iron level, patients should be advised to maintain routine checkups. conflicts of interest the authors confirmed that they have no conflicts of interest of the research submitted. acknowledgment the authors thank soran private hospital team, noor laboratory for make us share and use the patients’ information table 8: t-test for two independent samples with respect to sex variable and mcv t-test for sex and mcv variables t df sig. (2-tailed) 95% confidence interval of the difference lower upper pre mcv equal variances assumed −0.666 28 0.511 −4.606 2.345 equal variances not assumed −0.673 10.115 0.516 −4.866 2.605 pos mcv equal variances assumed −0.093 28 0.926 −3.564 3.253 equal variances not assumed −0.132 21.515 0.896 −2.592 2.282 mcv: mean corpuscular volume table 9: t-test for independent two samples. descriptive statistics of sf descriptive statistics of sf sex n mean sd std. error mean pre sf male 7 99.00 17.166 6.488 female 23 90.35 31.106 6.486 post sf male 7 57.29 11.041 4.173 female 23 62.87 13.431 2.801 table 10: t-test for two independent samples w. r. to sex variable and sf t-test for sex and sf variables t df sig. (2-tailed) 95% confidence interval of the difference lower upper pre mcv equal variances assumed 0.699 28 0.491 −16.721 34.025 equal variances not assumed 0.943 18.851 0.358 −10.560 27.864 pos mcv equal variances assumed −0.998 28 0.327 −17.040 5.872 equal variances not assumed −1.111 11.960 0.288 −16.538 5.370 zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 159 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 and tests as well as the support from the department of nutrition and dietetics, cihan universityerbil. references 1. e. m. h. mathus-vliegen, d. nikkel and h. s. brand. oral aspects of obesity. international dental journal, vol. 57, no. 4, pp. 249256, 2007. 2. d. ryan, s. barquera, o. b. cavalcanti and j. ralston. the global pandemic of overweight and obesity: addressing a twenty-first century multifactorial disease. in: handbook of global health. springer international publishing, champaign, pp. 739-773, 2021. 3. j. m. 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csendes, k. papapietro, f. pizarro, m. valencia, zebari, et al.: iron assessment after laparoscopic sleeve gastrectomy 160 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 153-160 j. l. westcott, k. m. hambidge and n. f. krebs. heme-and nonheme-iron absorption and iron status 12 mo after sleeve gastrectomy and roux-en-y gastric bypass in morbidly obese women. the american journal of clinical nutrition, vol. 96, no. 4, pp. 810-817, 2012. 33. m. coupaye, k. puchaux, c. bogard, s. msika, p. jouet, c. clerici, e. larger and s. ledoux. nutritional consequences of adjustable gastric banding and gastric bypass: a 1-year prospective study. obesity surgery. vol. 19, no. 1, pp. 56-65, 2009. 34. k. k. mahawar, w. r. j. carr, n. jennings, s. balupuri and p. k. small. simultaneous sleeve gastrectomy and hiatus hernia repair: a systematic review. obesity surgery, vol. 25, no. 1, pp. 159-166. 2015. 35. o. jamil, r. gonzalez-heredia, p. quadri, c. hassan, m. masrur, r. berger, k. bernstein and l. sanchez-johnsen. micronutrient deficiencies in laparoscopic sleeve gastrectomy. nutrients, vol. 12, no. 9, pp. 2896, 2020. 36. t. diamantis, k. g. apostolou, a. alexandrou, j. griniatsos, e. felekouras and c. tsigris. review of long-term weight loss results after laparoscopic sleeve gastrectomy. surgery for obesity and related diseases, vol. 10, no. 1, pp. 177-183, 2014. 37. b. lowry, k. hardy and a. vergis. iron deficiency in bariatric surgery patients: a single-center experience over 5 years. canadian journal of surgery, vol. 63, no. 4, pp. e365-e369, 2020. 38. j. l. roberts and a. d. stein. the impact of nutritional interventions beyond the first 2 years of life on linear growth: a systematic review and meta-analysis. advances in nutrition, vol. 8, no. 2, pp. 323-336, 2017. tx_1~abs:at/add:tx_2~abs:at 11 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 11-16 research article the opinion and awareness of citizens on the commitment of bakers, butchers, and food sellers to health instructions and their application during covid-19 outbreak kareem f. aziz1, hazhar t.a. blbas2, yaseen galali3,4, ahmed aj. jabbar5 1department of nursing, college of nursing, hawler medical university, 44001, erbil, iraq, 2department of statistics, college of administration and economics, salahaddin university-erbil, iraq, 3department of food technology, college of agricultural engineering sciences, salahaddin university-erbil, kurdistan region, iraq, 4department of nutrition and dietetics, college of health technology, cihan university, erbil, iraq, 5department of medical laboratory technology, erbil technical health and medical college, erbil polytechnic university, erbil, 44001, iraq abstract many countries follow the recommendation of the world health organization by practicing physical gathering avoidance as one health measure to decrease covid-19 the spread. the current study identifies the opinions and awareness of citizens about the commitments of workers (bakers, butchers, and food sellers) to health instructions and prevention. this descriptive cross-sectional study was done in different premises in kurdistan region-iraq from september 20  2021 to april 20  2022 using online platforms and printed forms. the study included socio-demographic characteristics, citizens’ opinions, and commitment-related questions. a thousand copies were collected and analyzed. the results showed that the majority of citizens have positive opinions and awareness and agreed that workers (bakers, butchers, and food sellers) are committed to covid-19 regulations. there was a significant correlation between the variables of opinions and awareness of the commitments of workers to the prevention of covid-19. all professional workers have a good obligation to prevention from covid-19 so that the population in erbil city has accepted those workers to deal with food and meat and with complete prevention from covid-19. these perfect commitments of workers to prevention may help to break down the chain of covid-19 infection which consequently reduces mortality and morbidity rate. keywords: bakers, covid-19, food sellers butchers, preventive measurement commitment introduction china announced a number of unknown cases in december 2019 of what is later known as covid-19. later, the origin area of the virus causing covid19, sars-cov-2 was allegedly belonged to the wuhan city of china. covid-19 has been deemed as a novel zoonotic disease that is transmitted to humans from animals.[1] no other previous communicable disease including, the spanish flu, had influenced the market as powerfully as this outbreak. the intake of meals rich in nutrients and purposeful meals can improve the immune system to combat viral diseases.[2,3] besides, supple-mentation vitamins d and e can improve the body’s immune against covid-19.[4] an evaluation of the 2019 historical proof concerning sars influenza and h1n1 prevention inside the excessive-danger populace suggests that chinese natural formulation ought to offer a suitable method for the reduction of covid-19 spread.[5] the recommendations and the natural formulation used for the supportive care of sufferers with covid-19 were revised by cai et al.[6] herbal academic researchers and meal zone specialists will encounter many tremendous obstacles, for example, making sure that meals are safe.[7] tremendous studies have shown that food and nutrients can heavenly impact the immune system and the way that the body reacts toward foreign bodies.[8-12] food quality and shelf life; a crowded situation for employees in meat and rooster processing centers ought to bring about the excessive danger of sars-cov-2 spread.[13] cases of corresponding author: yaseen galali, department of food technology, college of agricultural engineering sciences, salahaddin university-erbil, kurdistan region, iraq, e-mail: yaseen.galali@su.edu.krd received: february 02, 2023 accepted: june 23, 2023 published: july 30, 2023 doi: 10.24086/cuesj.v7n2y2023.pp11-16 copyright © 2023 yaseen galali, kareem f. aziz, hazhar t.a. blbas, ahmed aj. jabbar. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) galali, et al.: covid-19 and food retailer’s commitments 12 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 11-16 covid-19 were located in different congregate settings, such as acute care hospitals, long-time period care centers, shelters for the homeless and correctional centers.[14] as a part of the countrywide covid-19 response, the diagnosed danger to meet and rooster operational facility calls for a spark-off motion to lower dangers to workers, keep the facility function, and hold the meals supply. the world health organization (who) formally called the disease “covid-19”.[13,15] the world faced an unimaginable threat due to covid-19. numerous countries follow who recommendations regarding introducing physical distance measures to reduce the diseases transmission. maintaining all food production and supply chain workers in the safe and healthy environment is crucial to survive during the pandemic. keeping the food chain is an important role in which all stakeholders along with the food chain need to involve. it is also necessary to sustain and consumer confidence and trust in food safety and availability.[16] the new covid-19 is an easily communicable and transmissible disease among the community through sneeze, cough, and respiratory droplets. the signs and symptoms of covid-19 are noticed after a few from the infection which include flu (e.g., coughing and headache) and others such as muscle aches and sore throat[17] and loss of taste as well as smell. covid-19 has close clinical characteristics and symptoms to two other familiar lower respiratory tract diseases; severe acute respiratory syndrome: “sars-cov” “mers”.[18] the outbreak of covid-19, in relation to the onset of suddenness, the disease communicability, and its straight pressure on the market and infrastructure of health, is the main community health issue.[19] slaughterhouses as well as meatpacking factories are the main risk factors for covid19 transmission during the pandemic. they influence the entire population, have far-reaching implications, and need significant interventions by public.[20] among the initial symptoms that are known about coronavirus by countries in the west were quick stockpile of different consumer stuff: masks, sanitizer, and microbial wipes went first. canadian fruit and vegetable markets were significantly impacted by the spread of the novel coronavirus sars-cov-2 (and covid-19 disease), beginning in march 2020.[21] indonesia has been more impacted the outbreak of the virus, with the mortality rate case being 8.9% by the end of march 2020.[22] the closure of the restaurants, cafes, and others imposed further pressure on food supply and retailing and made people work at home. it is believed that just more than 30% of canada’s expenditures of food typically occur in restaurants.[23] this makes necessary to create novel policy to improve education and train supervisors and employees in relation to health and safety information.[24] the food sector and stakeholders are also at the target since food is crucial for the survival of the community and impossible to avoid. health authorities have to take quick actions and pinpoint the most dangerous threats to the food chain during viral outbreaks to proceed preventive measures.[7] in the kurdistan region of iraq, bakers, butchers, and food sellers must commit to health instructions and precautionary prevention in time of covid-19. the pandemic curve in iraq and kurdistan region increased on 1st june, and the mortality and morbidity rates increased. the current study aims to identify citizens’ perception and awareness about the baker, butchers, and food sellers’ commitments to health instructions and prevention from covid-19, and to identify associations between sociodemographic information of citizens and their opinions of commitments during covid-19. materials and methods study design the descriptive cross-sectional research was conducted in different cities in erbil city/iraq via an online application form, beginning from september 20 2021 to april 20 2022. the sample size included 1000 participants in different settings through an online and interview form. the inclusion criteria were anyone who can use computers and the internet and can fill out the online survey and work in the food supply chain. tools and data collection methods data were collected based on the probability sampling techniques, and the questionnaire format through the online application. the tool has three main parts. part one was about the socio-demographic characteristic of the volunteers including (age, gender, family status, education level, family number, address, and occupation). the second part consisted of items related to citizens’ opinions and perceptions regarding commitments to health guidelines for prevention from covid19 depending on the scoring system (1 for agree, 2 for neutral, and 3 for disagree). part three included items related to citizens’ awareness and commitments about preventive measures (for agree was 1, neutral was 2, and disagree was 3). a panel of experts before commencement assessed the validation of questions. ethical approval the research was ethically by the college of nursing-ethical and the scientific committee. data analysis row data were analyzed statistically utilizing the spss package program version 23. (statistical package for social science), descriptive analysis, frequency, and chi-square tests were employed. the problems of this study included opinions and awareness of citizens about the commitments of workers to health guidelines for of (covid-19) prevention. results the characteristics of the socio-demographic of the sample are shown in table 1. the data revealed that the majority of them were females representing 51% whose most of them were above 55 years old. 40%, of the participants, were graduated from secondary school representing 39.8%, and married representing 65.4%. the majority of them were galali, et al.: covid-19 and food retailer’s commitments 13 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 11-16 sellers about covid-19 preventive measures, represented 40.5% and 32.5% of them, have neutral opinions about that, while 27% of citizens were not happy about commitments of butchers, bakers, and food sellers for covid-19 preventive instructions [table 2]. data outcomes indicated an association between citizens’ perceptions of the commitments of butchers, bakers, and food sellers about covid-19 preventive measures. the results indicated the presence statistically significant (p = 0.001). the correlation between participants’ socio-demographic characteristics to the perceptions of the covid-19 preventive measure commitment of butchers, bakers, and food sellers is presented in table 3. the results also indicated that the majority of the citizen’s awareness agreed with the commitments of butchers, bakers, and food sellers about covid-19 prevention, which represented 47.7%, and 25.6 % of them have neutral awareness of that, while 26.7% of citizens have not agreed with awareness about commitments of butchers, bakers, and food sellers for covid19 prevention [table 4]. the data analysis showed a significant association between citizens’ awareness for commitments of butchers, bakers, and food sellers about covid-19 and their nuclear families with 65.4%, most of them from the urban area 69%, while most of them were employed by 41.3% [table 1]. the current study shows that the majority of participants agreed with the commitments of butchers, bakers, and food table 1: socio-demographic characteristics of the participants variables frequency percent sex male 490 49.0 female 510 51.0 total 1000 100.0 age group 18–27 201 20.1 28–38 216 21.6 39–49 180 18.0 55 and above 403 40.3 total 1000 100.0 educational level secondary 398 39.8 institute 290 29.0 college 235 23.5 msc 51 5.1 phd 21 2.1 total 1000 100.0 family status marriage 654 65.4 single 307 30.7 divorce 21 2.1 widow 18 1.8 total 1000 100.0 type of family nuclear 654 65.4 extended 346 34.6 total 300 100.0 residency urban 690 69.0 suburban 295 29.5 rural 12 1.2 refugee 3 0.3 total 1000 100.0 occupation employee 413 41.3 private 277 27.7 retired 40 4.0 student 270 27.0 total 1000 100.0 table 2: participants (butchers, bakers, and food sellers) opinion in relation to commitments to covid-19 preventive measures items frequency percentage agree 405 40.5 neutral 325 32.5 disagree 270 27.0 total 1000 100.0 table 4: levels of participants’ awareness with covid-19 prevention measures item frequency percentage agree 477 47.7 neutral 256 25.6 disagree 267 26.7 total 1000 100.0 table 3: the association between participants (butchers, bakers, and food sellers) commitments about covid-19 and their variables variable value df p-value sex 715.579a 2 0.001 age 1559.088a 8 0.001 education level 660.797a 8 0.001 family status 581.426a 6 0.001 type of family 742.679a 2 0.001 address 837.792a 6 0.001 occupation 1905.360a 6 0.001 galali, et al.: covid-19 and food retailer’s commitments 14 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 11-16 table 6: the percentage of opinion and commitment response to health guidelines preventive measures covid-2019 no. criteria satisfy neutral no satisfied opinion 1 i feel that bakers were committed to health authority guidelines and precautionary guidance 40.5% 40.6% 18.9% opinion 2 in my opinion bakers have good knowledge about health instruction 47.1% 31.1% 21.8% opinion 3 i agree that bakers depending on personal sanitary guidelines to protect themselves and others 41.7% 39.4% 18.9% opinion 4 i am satisfied with butchers’ behaviors to deal with complete personal and environmental sanitation 51.0% 30.4% 18.6% opinion 5 when i go to buy meat, i am satisfied with clean and personal sanitation of them 44.9% 28.8% 26.3% opinion 6 i am sure butchers are dealing with animals properly with complete prevention 52.8% 27.4% 19.8% opinion 7 in my opinion food sellers in mall are committed with the local guideline for prevention 44.5% 32.0% 23.5% opinion 8 i am satisfied with food sellers in my location and have clean behaviors 49.2% 28.7% 22.1% opinion 9 food sellers have good information of covid-19 so that they do their best 44.9% 32.5% 22.6% opinion 10 i am satisfied with the cleanness of the food sellers in my location 52.5% 28.0% 19.5% table 5: the association of participants’ awareness with covid-19 commitments variable value df p-value sex 884.970a 2 0.001 age 1300.208a 8 0.001 education level 292.358a 8 0.001 family status 567.969a 6 0.001 type of family 618.272a 2 0.001 address 617.024a 6 0.001 occupation 1244.119a 6 0.001 values with the same superscript with the same column refer to statistically non-significant at p<0.001. to discontinue the covid-19 infection chain and hence positively affect mortality and morbidity among the community in the kurdistan region of iraq. the current results are in agreement with the previous study stating that prevention is not just a concern about the screening, but it will also consider monitoring the prevention.[18] previous studies indicated that most of the kurdish population has a good awareness of the commitments of butchers, bakers, and food sellers during the pandemic of covid-19, which was very decisive for preventing and encouraging those workers to apply precautionary measures properly including online shopping. these findings were similar to studies done in canada, so most people in this country also applied online shopping to decrease the outbreak of covid19.[21] the study’s other findings indicated a significant association between the citizen’s variables and their opinion and awareness of these workers (bakers, butchers, and food sellers). this was that all of these workers have to apply all health instructions and guidelines of the profession during their daily work to limit the outbreak of covid-19 among the kurdish population in the kurdistan region of iraq. the local government in this region has an active role in educating citizens about precautionary prevention, so we have seen the outcome of that, so the number of death is less than in other countries near iraq as iran, jordan, and turkey. these findings agreed with a study done in indonesia that recommended the prevention of covid-19 and early detection.[22] it is necessary for all workers dealing with food and meat, they have to apply personal and public prevention by social distancing and wearing personal protective equipment (ppe) as mentioned in the study by jonathan et al.[24] workers in markets must have sufficient knowledge about prevention from covid-19 in the workplace to break the chain of infection as mentioned by usman et al.[25] accordingly, several studies have shown that long-term infrastructure commitments of population to the covid-19 regulations are considered as one of the variables. these commitments were significant for the prevention and breakdown of the chain of infection for covid-19 [table 5]. the present data analyzed that the percentage of satisfaction from workers committed among participants was higher than that of non-satisfied groups [table 6]. the current study found that the percentage of satisfaction from workers’ awareness among participants was higher than that of non-satisfied groups [table 7]. discussion the outcome of the work unrevealed that the majority of citizens’ opinions agreed with the commitments of butchers, bakers, and food sellers about covid-19 prevention, representing 40.5%, and 32.5% of them have neutral opinions. while nearly 27% of participants have not agreed with opinions about the commitments of butchers, bakers, and food sellers for covid-19 prevention. their commitments to covid-19 prevention were significant galali, et al.: covid-19 and food retailer’s commitments 15 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 11-16 table 7: percentage of awareness and commitment response to health guidelines preventive measures covid 2019. no. criteria satisfy neutral not satisfy awareness 1 butchers usually apply personal protective equipment during working 49.6% 31.5% 18.9% awareness 2 i see butchers’ commitments about precautionary guidance in their shop 52.2% 25.7% 22.1% awareness 3 when i buy meat from butchers i satisfy with personal and environmental sanitation 50.4% 31.0% 18.6% awareness 4 i see always bakers wearing protective uniform to prevent themselves and others from covid-19 48.6% 32.1% 19.3% awareness 5 always bakers use clean technic in preparing bread every day 51.9% 27.6% 20.5% awareness 6 always bakers order clients to be a distance between two people more than 2 meters for prevention 47.7% 31.2% 21.1% awareness 7 i satisfy with food sellers in mall or shops at my location so they deal properly with prevention measures 54.6% 31.3% 14.1% awareness 8 usually, food sellers clean items and surfaces in the mall 47.7% 35.5% 16.8% awareness 9 i satisfy with food sellers because they bring excellent items to their mall or shops 53.1% 29.5% 17.4% awareness 10 as general i satisfy with bakers, butchers, and food sellers in my location because they are commitments with all health recommendation and other precautionary measures 50.4% 31.6% 18.0% valuable on-going and maintained actions to back up the resilience of physicians and healthcare against the ongoing pandemic.[25-28] conclusion the opinions and awareness of citizens were satisfied and agreed with workers’ commitments as (butchers, bakers, and food sellers) about covid-19 prevention. there was a significant correlation between their variables, opinions, and awareness about workers’ commitments to prevention from covid-19. the proper commitment of workers to prevention may help to break down the chain of covid-19 infection which consequently reduces mortality and morbidity rate. the limitation of this study includes moving limitations due to the confinement. therefore, the authors suggest future studies in that regard including more participants from different countries around the globe and direct interviews. acknowledgments the authors would like to thank all participants for their contribution. authors’ contributions all the authors contributed to data analysis, explanation, writing original draft, and final 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drivers affecting adherence to who guidelines against covid-19 amongst rural ugandan market vendors. frontiers in public health, vol. 8, p. 380, 2020. 26. c. meyer, f. becot, r. burke and b. weichelt. rural telehealth use during the covid-19 pandemic: how long-term infrastructure commitment may support rural health care systems resilience. journal of agromedicine, vol. 25, no. 4, pp. 362-366, 2020. 27. a. a. mohammed, a. ferraris and c. troise. csr practices and creativity during covid-19 pandemic in the emerging market: investigating the mediating effect of affective commitment. management decision, vol. 60, p. 2669-2693, 2021. 28. v. filimonau, b. derqui, and j. matute. the covid-19 pandemic and organisational commitment of senior hotel managers. international journal of hospitality management, vol. 91, p. 102659, 2020. . 71 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 71-74 prospective and semiexprimental the piriformis syndrome: evaluation of seven cases farhad mustafa mousa*, zainab aziz bakr department of physiotherapy, cihan university-erbil, kurdistan region, iraq abstract low back pain is one of the causes of piriformis syndrome. low back pain is the second most common medical complaints encountered by physicians. more than 50% of the population will complain of low back pain at the same time. piriformis syndrome may constitute up to 5% of cases of low back pain, buttock pain, and lower limb pain. this is a prospective and case–control study. in this study, we study 268 cases with low back pain; in 268 cases, seven cases had piriformis syndrome. we studied pain, severity pain, radicular pain, limping, and painful sitting. evaluation of musculoskeletal, bmi, and physical examination of piriformis sign, freiberg sign, and pace sign. neurological evaluation was taken by paraclinical examination elevated through x-ray, computed tomography scan, magnetic resonance imaging, and electromyography. data show as mean±sd and we used ssps software for analysis. in 268 cases with conservative method, 100% of cases were cure treatment after 4 months, in contrast to surgery methods after 1 month. the result illustrated the significantly improved between two methods of treatment (p < 0.0025). in seven cases, four cases had a history of blunt trauma and one case had a history of surgery that after 6 months had complication of surgery and scare tissue that with the second surgery release of scare was done with 100% cure. anomaly of piriformis muscle was seen in one case that surgery was done. tumor was seen in one case that had pressure effect on sciatic nerve that surgery was done. heterotopic ossification was seen in two cases. bmi of all cases was in normal range. all cases with diagnosis of disc herniation were excluded from the study. they were need to surgical operation for herniated disc. piriformis syndrome is differentiated primary from secondary type, in which it is done according to history and physical examination. according to the etiology of piriformis syndrome, the treatments are different. if you see mass that compressed the sciatic nerve, you must remove it. if the patient had sacroiliitis, it must be treated in early phase. surgery of piriformis is not accepted by all surgeons. it is limited to release piriformis tendon and insertion to femur. keywords: piriformis syndrome, low back pain, buttock pain introduction low back pain is the second most common medical complaints encountered by physicians. more than 50% of the population will complain of low back pain at same time. most of us, at 1 time or more, have experienced of low back pain. one of the most differential diagnoses of back pain is sciatica pain and piriformis syndrome is one causes of differential diagnosis to sciatica pain. the piriformis muscle is a pear-shaped muscle in the buttocks.[1-3] the piriformis muscle has origins from several anatomical locations including the anterior surface of the lateral process of the sacrum, the spinal region of the gluteal muscles, and the superior/gluteal surface of the ilium near the margin of the greater sciatic notch. the muscle passes through the greater sciatic notch and inserts on the greater trochanter of the femur. it is one of the six muscles in the hip short external rotator group, coursing parallel to the posterior margin of the gluteus medius.[4,5] piriformis syndrome is an uncommon cause of buttock and leg pain. piriformis syndrome may constitute up to 5% of cases of low back, buttock, and leg pain. piriformis syndrome is an often misdiagnosed cause of sciatica, leg or buttock pain, and disability. dysfunction of the piriformis muscle can cause signs and symptoms of pain in the sciatic nerve distribution, that is, in the gluteal area, posterior thigh, posterior leg, and lateral aspect of the foot. the sciatic nerve may be compressed within the buttock by the piriformis muscle, with pain increased by muscular contraction, palpation, or prolonged sitting.[6-8] it is often not included in the differential diagnosis of back, buttock, and leg pain. in addition, it has received minimal recognition because it is often seen as a diagnosis of exclusion. pain is increased in prolonged sitting; specific physical findings are tenderness in the sciatic notch and buttock pain in flexion, corresponding author: farhad mustafa mousa, department of physiotherapy, cihan university, erbil, kurdistan region, iraq. e-mail: farhadm.atrushi@gmail.com received: apr 20, 2019 accepted: apr 23, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp71-74 copyright © 2019 farhad mustafa mousa, zainab aziz bakr. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ mousa and bakr: piriformis syndrome 72 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 71-74 adduction, and internal rotation (fadir) of the hip. an initial description was given by yeoman, in 1928. in 1934, freiberg recognized the signs specific to this syndrome. however, it was only in 1947 that robinson called this clinical entity the “piriformis” or pyramidal syndrome. the piriformis muscle has been said to receive innervation from l5 to s3 ventral rami with most sources using s1 and s2 ventral rami as the most common innervation this muscle. the most common nerve branches to the piriformis are from the superior gluteal nerve and the ventral rami of s1 and s2. more recently, many etiologies of sciatic nerve entrapment around the gluteal region or the non-discogenic area have been identified, resulting in the use of a new term “the deep gluteal syndrome.”[9-11] causes • an injury, such as fall, a blow to area or car accident • overuse, such as from frequent running, excessive exercise, or overstretching, trauma to the buttocks or gluteal region is the most common cause of piriformis syndrome • a sedentary lifestyle, especially with long periods of sitting • changing from a sedentary lifestyle to more frequent exercise • buttock muscles wasting away • muscle tension and excess weight due to pregnancy • skiers, truck drivers, tennis players, and long-distance bikers are at high risk • in morton’s foot, the prominent head of the second metatarsal causes foot instability and a reactive contraction of the external rotators of the hip during gait • spinal stenosis can lead to bilateral piriformis tenderness • anatomic variations of the divisions of the sciatic nerve above, below, and through the belly of the piriformis muscle may be causative factors.[12] piriformis syndrome is a diagnosis of exclusion. diagnosis of piriformis syndrome is based on a review of the patient’s medical history, a physical examination, and possibly diagnostic tests. piriformis syndrome is often a diagnosis made through a process of ruling out other possible conditions that may be causing the patient’s symptoms such as a lumbar disc herniation or sacroiliac joint dysfunction. the history and physical findings are key elements in differentiating the more figure 3: pace test: pain and weakness are present on resisted abduction-external rotation of the thigh aq1 figure 4: it is combination of abduction and external rotation test, it is positive in all cases aq1 figure 1: piriformis sign: more external rotation in the same side of pain aq1 figure 2: freiberg sign: pain occurs with passive internal rotation of the extended thigh when the patient is supine aq1 mousa and bakr: piriformis syndrome 73 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 71-74 common forms of lbp and piriformis syndrome. many patients who underwent unsuccessful surgery in the lumbosacral region were later found to have piriformis syndrome. the female-tomale incidence ratio of piriformis syndrome is 6:1. in one study at a regional hospital, 45 of 750 patients with lbp were found to have piriformis syndrome. another author estimated that the incidence of piriformis syndrome in patients with sciatica is 6%. the function of the piriformis muscle is to externally rotate and abduct the thigh. in piriformis syndrome, the only true-positive sign is tenderness over the gluteal region.[13] the pain can be reproduced with maximum elongation of the piriformis muscle in fadir of the hip. weakness can be observed with resisted external rotation and abduction of the hip. for diagnosing piriformis syndrome, several authors describe the use of the following signs: 1. lasegue sign: pain is present in the vicinity of the greater sciatic notch during extension of the knee with the hip flexed to 90°, tenderness to palpation of the greater sciatic notch is noted 2. freiberg sign: pain occurs with passive internal rotation of the extended thigh when the patient is supine 3. pace sign: pain and weakness are present on resisted abduction-external rotation of the thigh. robinson, who first described the syndrome, stated that piriformis syndrome had six cardinal features:[14] 1. sausage-shaped mass over the piriformis muscle 2. positive lasegue sign 3. gluteal atrophy in chronic cases 4. trauma to the region 5. pain exacerbated by lifting and relieved by traction on the affected extremity 6. pain in the sacroiliac joint region, gluteal muscles, or greater sciatic notch. beatty explained two maneuvers to reproduced the pain of piriformis syndrome in the following way:[10] 1. the patient lies with the painful side up and the involved leg flexed 2. the knee of the affected side rests on the table. pain in the buttocks is reproduced when the patient lifts the leg and knee slightly above the table. diagnostic considerations • bursitis or weaver’s bottom (i.e., ischiogluteal bursitis) • sciatica caused by compression secondary to a fibrotic band, hematoma, or aneurysm of the inferior or superior gluteal artery • pelvic mass, tumor, or endometriosis (in women) • gout • referred pain to the buttock caused by pronator forces of the foot • spinal canal stenosis. differential diagnoses • hamstring muscle injury • lumbosacral disc herniation injuries • sacroiliac joint injury • lumbosacral discogenic pain syndrome • lumbosacral facet syndrome • lumbosacral spine sprain-strain injuries • lumbosacral spondylolysis • lumbosacral radiculopathy • lumbosacral spondylolisthesis. imaging evaluation computed tomography (ct) scan and magnetic resonance imaging (mri) • ct scans can be used to identify spinal stenosis or arthritic changes • mri findings can rule out other causes of back pain, such as a herniated disc, spinal tumor, or abscess • mri may demonstrate a large mass anterior to the piriformis muscle · the piriformis muscle also can appear enlarged on t1or t2-weighted images.[4] electromyography (emg) • in piriformis syndrome, abnormal patterns are found in the gluteus maximus and piriformis muscles • in piriformis syndrome, emg findings in the gluteus minimum, gluteus medius, and tensor fasciae late muscles are normal • emg can also be performed to determine neurologic or muscular changes • in the case of a herniated disc, all muscles that are posteriorly and anteriorly innervated should be affected. with a herniated disc, a significant delay in the hoffman reflex or h-reflex (motor nerve fiber response after activation of afferent sensory spindle fibers) in the affected limbs is often present during emg.[12] methods this is prospective and case–control study. in this study, we evaluated patients with low back pain or lower limb pain (268 patients). all cases with diagnosis of disc herniation were excluding. in 268 cases, seven cases had piriformis syndrome; these seven cases had low back pain and radicular pain with mean average of 42 years old (12–67 years old). in the first physical examination was done, we studied pain, severity pain, radicular pain, limping, and painful sitting which were taken. for evaluation of musculoskeletal we used bmi and physical examination of piriformis sign, freiberg sign, pace sign. neurological evaluation was taken. and after that, paraclinical examination elevated x-ray, ct-scan, mri, and emg. data show as mean±sd and we used ssps software for analysis. results in table 1, specifications of all seven cases, with conservative method 100% of cases, were cure treatment after 4 months, in which in surgery methods after 1 month that is significant between two methods of treatment (p < 0.00025). in seven cases, four cases had a history of blunt trauma. one case had a history of surgery that after 6 months had complication of surgery and scare tissue that with the second surgery release of scare was done with 100% cure. anomaly of piriformis muscle was seen in one case that surgery was done. heterotopic mousa and bakr: piriformis syndrome 74 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 71-74 ossification was seen in two cases. bmi of all cases was in normal range. discussion in this study, we evaluated patients with low back pain or lower back pain (268 patients). all cases with diagnosis of disc herniation were excluded from the study. they are need surgical operation for disc herniation. the best and most point in piriformis syndrome are differentiating primary from secondary. it is done according to history and physical examination.[18] according to the etiology of piriformis syndrome, the treatments are different. you see mass that compressed the sciatic nerve you must remove it. if the patient had sacroiliitis, it must be treated in early phase. the most differential diagnosis of secondary piriformis syndrome is sacroiliitis in patients with buttock pain. in general, condition sacroiliitis are without sciatica pain.[7,8] however, it may be secondary to tension of piriformis muscle (hiltz – 1976) about 8.22% of sacroiliitis was seen in all low back pain patients. according to this for differentiated, we need to injection to this joint.[10] surgery of piriformis is not accepted in all surgeons. it is limited to release piriformis tendon and insertion to femur.[18] surgery itself has complication and morbidity, scar tissue, and compressed the sciatic nerve. surgery must be limited to only remove tissue around sciatic notch and inferior border of piriformis muscle that limited space around the nerve. the secondary type of piriformis syndrome was seen more in inflammatory process.[15-18] surgery and release of piriformis muscle decrease and removed pain in sacroiliitis indirectly. however, direct treatment of sacroiliitis joint must be done with sacroiliitis girder, manipulation, injection physical therapy and at the end, surgery of joint and fusion the joint. references 1. k. hopayian and a. danielyan. “four symptoms define the piriformis syndrome: an updated systematic review of its clinical features”. european journal of orthopaedic surgery and traumatology, vol. 28, no. 2, pp. 155-164, 2018. 2. l. m. fishman and m. p. schaefer. “the piriformis syndrome is underdiagnosed”. muscle and nerve, vol. 28, no. 5, pp. 646-649, 2003. 3. k. meknas, a. christensen and o. johansen. “the internal obturator muscle may cause sciatic pain”. pain, vol. 104, no 1-2, pp. 375-80, 2003. 4. a. g. filler, j. haynes, s. e. jordan and j. prager. “sciatica of nondisc origin and piriformis syndrome: diagnosis by magnetic resonance neurography and interventional magnetic resonance imaging with outcome study of resulting treatment”. journal of neurosurgery spine, vol. 2, no. 2, pp. 99-115, 2005. 5. m. r. foster. “piriformis syndrome”. orthopedics, vol. 25, no. 8, pp. 821-825, 2002. 6. l. m. fishman, g. w. dombi and c. michaelsen. “piriformis syndrome: diagnosis, treatment, and outcome-a 10-year study”. archives of physical medicine and rehabilitation, vol. 83, no. 3, pp. 295-301, 2002. 7. h. t. benzon, j. a. katz, h. a. benzon and m. s. iqbal. “piriformis syndrome: anatomic considerations, a new injection technique, and a review of the literature”. anesthesiology, vol. 98, no. 6, pp. 1442-1448, 2003. 8. s. k. han, y. s. kim, t. h. kim and s. h. kang. “surgical treatment of piriformis syndrome”. clinics in orthopedic surgery, vol. 9, no. 2, pp. 136-144, 2017. 9. p. m. barton. “piriformis syndrome: a rational approach to management”. pain, vol. 47, no. 3, pp. 345-352, 1991. 10. r. a. beatty. “the piriformis muscle syndrome: a simple diagnostic maneuver”. neurosurgery, vol. 34, pp. 512-514, 1994. 11. r. p. beauchesne and s. f. schutzer. “myositis ossificans of the piriformis muscle: an unusual cause of piriformis syndrome. a case report”. journal of bone and joint surgery american, vol. 79, no. 6, pp. 906-910, 1997. 12. j. a. brown, m. a. braun and t. c. namey. “pyriformis syndrome in a 10-year-old boy as a complication of operation with the patient in the sitting position”. neurosurgery, vol. 23, no. 1, pp. 117-119, 1998. 13. z. durrani and a. p. winnie. “piriformis muscle syndrome: an underdiagnosed cause of sciatica”. journal of pain and symptom management, vol. 6, no. 6, pp. 374-379, 1991. 14. a. h. freidberg. “sciatic pain and its relief by operation on muscle and fascia”. archives of surgery, vol. 34, pp. 337-349, 1937. 15. j. w. frymoyer. “back pain and sciatica”. the new england journal of medicine, vol. 318, no. 5, pp. 291-300, 1988. 16. j. j. jankiewicz, w. l. hennrikus and j. a. houkom. “the appearance of the piriformis muscle syndrome in computed tomography and magnetic resonance imaging. a case report and review of the literature”. clinical orthopaedics, vol. 262, pp. 205-209, 1991. 17. s. p. cass. “piriformis syndrome: a cause of nondiscogenic sciatica”. current sports medicine reports, vol. 14, no. 1, pp. 41-44, 2015. 18. c. steiner, c. staubs, m. ganon and c. buhlinger. “piriformis syndrome: pathogenesis, diagnosis, and treatment”. the journal of the american osteopathic association, vol. 87, no. 4, pp. 318323, 1978. table 1: specifications, age, sex, symptom, causes, treatment, and results of seven cases of piriformis syndrome age sex symptom cause treatment results 61 female hip pain trauma conservative after 6 months cure 45 male lower limb pain trauma heterotopic ossification surgery after 1 month cure 41 male hip and lower limb pain scare of surgery surgery after 3 months cure 67 female numbness and pain in lower limb trauma heterotopic ossification conservative after 4 months cure 42 male low back pain and hip pain anomaly of piriformis muscle surgery after 2 months cure 52 male sciatica blunt trauma conservative after 3 months cure 12 female sciatica tumor of gluteal area surgery after 1 month cure author query??? aq1:kindly cite figures 1-4 in the text part . 75 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 75-79 research article exploration of amylases producing competency of helicoverpa armigera gut bacterial strain, bacillus subtilis rtsba6 6.00 ashok a. shinde1*, faiyaz k. shaikh1,2, sarwan w. bradosty3, manvendra s. kachole1 1department of biotechnology, mgm’s institute of biosciences and technology, aurangabad, maharashtra, india, 2department of biochemistry, dr. babasaheb ambedkar marathwada university, aurangabad, maharashtra, india, 3department of biology, cihan university-erbil, kurdistan region, iraq abstract the helicoverpa armigera (hubner) (lepidoptera: noctuidae) is a polyphagous insect pest of agriculturally important crops. the alkaline gut of this insect pest possesses diverse bacterial communities which may assist in digestive physiology. as part our investigations of understanding the role of gut bacterial communities in insect gut, here amylase producing competency of earlier identified h. armigera gut bacterial strain, i.e., bacillus subtilis rtsba6 6.00 is reported. initial screening for amylase activity was assessed by starch agar plate. upon 7% sodium dodecyl sulfate polyacrylamide gel electrophoresis amylase zymography, bacterial culture supernatant produced seven amylase bands on the gel. the observed molecular weights of amylases were 191.2  kda, 158.0  kda, 131.7  kda, 54.0  kda, 31.3  kda, 67.2 kda, and 44.6 kda, respectively. considerable amylase activity was observed in neutral to alkaline ph with optimum at ph 6.8. the optimal activity temperature of amylases was found to be 50°c, and the activity decreased dramatically at temperatures above 75°c. keywords: amylase, bacillus subtilis rtsba6 6.00, helicoverpa armigera, zymography introduction the symbiotic associations of gut bacterial communities with the host insects are imperative for the host fitness leading to nutrition, development, resistance to pathogens, and reproduction.[1,2] this class of microorganisms is not only contributing in metabolic activities but also behavior of insect hosts.[3,4] improved digestion efficiency of host insects is attributed to digestive enzymes expressed by gut bacterial communities.[3] the secretion of cellulases and lignases enzymes by gut bacterial communities provides glucose and fatty acid to host insect.[5] the bacterial communities associated with lepidopteran insects recently gained attention from scientific community. some of these insects are considered the most damaging agricultural and forest pests worldwide. helicoverpa armigera is a devastating insect pest of agricultural crops such as cotton, chickpea, pigeon pea, tomato, sunflower, okra, and corn.[6] the use of modern chemical and biological insecticides has limited success in management due to adaptation acquired by this insect pest.[7,8] the detailed exploration of gut bacterial community and their contribution in host digestive physiology and adaptation to insecticides is necessary to cope of these insect pests. the multitrophic association of gut bacterial communities with h. armigera explored.[9-11] the bacterial strain bacillus subtilis rtsba6 6.00 isolated from gut of h. armigera is reported to secrete proteases enzymes.[10] this proteolytic arsenal may help h. armigera to contribute in digestion and adaptation. earlier, gut bacterial strain with serine protease activity from velvetbean caterpillars was reported.[12] in present communication, the h. armigera gut isolated bacterial strain b. subtilis rtsba6 6.00 explored further for its ability to produce amylase enzymes. amylases are digestive enzymes that act in the first step of maltopolysaccharide digestion. they cleave α 1–4 glycosidic linkage exists in starch. insects feed on plants for carbohydrates (starch and cellulose) during different stages of their lives and dependent on starch hydrolyzing enzymes for its survival. the amylase action is necessary to improve digestive performance of insects leading to survival within different living conditions and increase their biological fitness.[13] furthermore, the bacterial communities expressing amylases is receiving more significance due to their potential role in food, pharmaceutical, and fine chemical cihan university-erbil scientific journal (cuesj) corresponding author: ashok a. shinde, department of biotechnology, mgm’s institute of biosciences and technology, aurangabad, maharashtra, india. e-mail: ashokshinde05@gmail.com received: apr 13, 2019 accepted: apr 23, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp75-79 copyright © 2019 ashok a. shinde, faiyaz k. shaikh, sarwan w. bradosty, manvendra s. kachole. this is an open-access article distributed under the creative commons attribution license. https://creativecommons.org/licenses/by-nc-nd/4.0/ shinde, et al.: amylase producing bacteria from h. armigera 76 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 75-79 industries. we identified and characterized several isoform of amylases produced by b. subtilis rtsba6 6.00. initial detection of amylase activity was carried on starch agar plate. the optimum temperature and ph were estimated by solution assays. amylase isoforms analysis was done by zymography technique. materials and methods isolation, cultivation, and identification of bacteria earlier isolated h. armigera gut bacterial strain, identified, and characterized by fatty acid methyl ester-based analysis[10,14] were used in the present investigation. detection of amylase by agar-plate method primary screening of bacterial strain b. subtilis rtsba6 6.00 for amylase activity was done by starch hydrolysis on the solid media containing 1% starch, 0.3% beef extract, and 1.2% agar. strain was inoculated on the media plate and incubated for 24 h at 37°c.[15] plate was stained with starch specific lugol’s solution (0.1% iodine and 1% ki), and zone of clearance (α-amylase activity) was observed visually. amylase production the strain of b. subtilis rtsba6 6.00 was cultured in starch broth. the starch broth was prepared by mixing all ingredients except starch together in milli-q water. the ph of broth was adjusted to alkaline condition before sterilization at 121°c for 15 min. starch solution was separately sterilized at 121°c for 15 min. these solutions were mixed aseptically before use. the culture was incubated at room temperature on shaker for 48 h. the cells were harvested by centrifugation at 6000 rpm, 4°c for 20 min. the supernatant was used as crude enzyme source for further studies. the protein concentration was estimated by folin–phenol reagent[16] using bovine serum albumin as standard. starch zymography for starch zymography, 7% sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds-page) was carried out according to the method of laemmli.[17] 0.1% soluble starch was incorporated in the gel. about 100 µg of crude protein was loaded on gel with standard molecular weight markers (genei). after electrophoresis, gel was washed twice at room temperature in a solution of 2.5% triton-x 100 for 30 min. after gentle washing with milli-q water, the gel was incubated in 0.1 m tris–hcl (ph 8) at 37°c for overnight. the gel was stained with lugol’s solution, and the zones of starch clearance were detected. molecular weight determination the molecular weight of α-amylase isoforms was determined by comparing its electrophoretic mobility with that of standard proteins having known molecular weights. sds-page was carried out using 7% resolving gel.[17] enzyme activity assay amylase activity was determined by spectrophotometric method as described by fisher and stein.[18] according to the procedure, 1.0 ml of crude enzyme into a test tube and 1 ml of 1% soluble starch in 0.1 m tris–hcl (ph 8) were added in test tube. the test tubes were covered and incubate at 35°c for 15 min in water bath. then, 2.0 ml dnsa (1% 3, 5-dinitrosalicylic acid, 30% sodium potassium tartrate, and 0.2 m naoh) reagent was added in each tube to stop the reaction and kept in boiling water bath for 5 min. after cooling at room temperature, the absorbance was read at 540 nm by spectrophotometer. effect of ph on amylase activity the effect of different ph on the activity of amylase was examined using standard dnsa assay as described previously. amylolytic pattern of amylase at different ph was observed on 7% starch zymography. after electrophoresis, the gel was cut and the stripes of gel were washed twice at room temperature in a solution of 2.5% triton-x 100 for 30 min. each strip of gel was incubated in 0.1 m acetate buffer for ph 2.8, 3.8; 0.1 m phosphate buffer for ph 5.8 and 6.8;0.1 m tris–hcl ph 7.8, 8.8 and 9.8. 0.1 m tris–naoh for ph 11.8, respectively at 37°c for overnight and the gel was stained with lugol’s solution and the zones of starch clearance were detected. effect of temperature on enzyme activity the influence of heating on the catalytic activity of amylases was determined by pre-incubation for 10 min at different temperatures ranging from 35, 40, 45, 50, 55, 60, and 65°c, followed by cooling and measuring the remaining activity. the stability of amylase at different temperature was observed by loading pre-incubated sample on 10% starch zymography as described previously. results and discussion this study deals with deciphering amylase producing competency of bacterial strain which was isolated and identified as b. subtilis rtsba6 6.00 from gut of fourth instar larvae of h. armigera.[10] the strain was gram-positive spore forming. this strain was screened for amylase activity using starch agar plate method. the starch agar is a differential medium that assesses the ability of an organism to produce starch hydrolyzing enzymes, namely amylase. in principle, the inoculated microorganisms hydrolyze starch present in medium upon incubation at an appropriate temperature. starch can be stained with lugol’s solution which turns blue-black. absence of the blue-black color indicates starch hydrolysis. here upon initial detection, the presence of clear zone of starch hydrolysis was observed around the growth of b. subtilis rtsba6 6.00 on the plate [figure 1]. amylase cleaves the starch into diand oligosaccharides which are transported into the cell to be catabolized. in general, all bacillus species have potential to produce the amylase.[19] the pure culture of b. subtilis rtsba6 6.00 was cultured in starch broth for amylase production with optimum parameters. the supernatant recovered after centrifugation was used for further characterizations. the amylase in shinde, et al.: amylase producing bacteria from h. armigera 77 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 75-79 the supernatant was separated into seven bands on the non-reducing 7% sds-page amylase zymography [figure 2]. this is an electrophoretic technique used to study amylase by means of thin gels containing copolymerized starch, under non-reducing conditions. the formation of clear zones against a dark background in the gel stained with lugol’s solution indicated the presence of amylolytic activity.[20] the zone of starch clearing corresponded to a migration distance of amylase by comparison to the molecular weight standards used in the study. these amylases showed differential mobility on the gel resulting different molecular masses. the major amylase bands were observed with molecular weights 191.2 kda, 158.0 kda, 131.7 kda, 54.0 kda, and 31.3 kda, respectively. two moderate intensity bands were also observed with molecular weights 67.2 kda and 44.6 kda. in our study, we found that, amylase isolated from b. subtilis rtsba6 6.00 showed optimum activity in slightly neutral ph (6.8) as shown in figures 3 and 4. this parameter assessed under standard assay conditions and zymographically. these amylases had a relatively low activity in acidic ph range, i.e., ph 2.8. the amylases were found to be most active at the neutral to alkaline ph range. the optimal ph for amylases of most of the insect species was found to be neutral to alkaline ph range, which is near to our finding for b. subtilis rtsba6 6.00 isolated from gut of h. armigera.[21] the stability of these amylases in the alkaline region indicates that they may be potentially used in digestive physiology of insect. beside ph, temperature is crucial factors that affect enzymatic activity. bacterial amylases were optimally active at temperatures ranging from 35 to 55°c, with a decline in activity above 75°c [figures 5 and 6]. the optimum temperature was found to be 50°c. beyond temperature 75°c decline in activity might be attributed to denaturation of amylases. earlier amylase with optimum activity in slightly higher temperature figure 1: screening of bacillus subtilis rtsba6 6.00 for amylase activity. strain was streaked on agar plate containing 1% starch, incubated for 24 h at 37°c. the clear zone indicated the hydrolysis of starch figure 2: non-reducing 7% sodium dodecyl sulfate polyacrylamide gel electrophoresis amylase zymography of proteases produced by bacillus subtilis rtsba6 6.00 figure 3: non-denaturing 7% sodium dodecyl sulfate polyacrylamide gel electrophoresis zymography showing the influence of ph on the amylase activity of bacillus subtilis rtsba6 6.00 figure 4: effect of different ph values on amylolytic activity of bacillus subtilis rtsba6 6.00. samples were assayed by standard procedure using starch as substrate shinde, et al.: amylase producing bacteria from h. armigera 78 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 75-79 range than the physiological temperature was reported form bacillus species.[22] the gut of h. armigera has an alkaline ph, and b. subtilis rtsba6 6.00 was found to grow and degrade starch at neutral to alkaline ph. the presence of bacteria producing several amylases with optimal activity in alkaline region could be advantageous to h. armigera. here, it might be possible that these bacterial amylases contribute to amylolytic arsenal of h. armigera, which feeds on carbohydrate rich part of plants. optimal activity at near alkaline ph in lepidopterans may be an adaptive response for the digestion of their diets.[23] the characterization of microbial amylase with thermo stability and activity in wide ph range may also found industrial applications, and hence, our future investigation will focus on the detailed investigations of these microbial amylases. references 1. n. a. moran, p. h. degnan, s. r. santos, h. e. dunbar and h. ochman. “the players in a mutualistic symbiosis: insects, bacteria, viruses, and virulence genes.” proceedings of the national academy of sciences of the united states of america, vol. 102, pp. 16919-16926, 2005. 2. j. c. clemente, l. k. ursell, l. w. parfrey and r. knight. “the impact of the gut microbiota on human health: an integrative view”. cell, vol. 148, pp. 1258-1270, 2012. 3. r. dillon and v. dillon. “the gut bacteria of insects: nonpathogenic interactions”. annual review of entomology, vol. 49, pp. 71-92, 2004. 4. f. m. pilon, l. e. visotto, r.n.c. guedes and m. g. a. oliveira. “proteolytic activity of gut bacteria isolated from the velvet bean caterpillar anticarsia gemmatalis”. journal of 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parasitoid campoletis chlorideae”. entomologia experimentalis et applicata, vol. 126, pp. 1-8, 2008. 9. p. k. mishra and s. m. tandon. “gut bacterial flora of helicoverpa armigera (lepidoptera, noctuidae)”. indian journal of microbiology, vol. 43, no. 1, pp. 55-56, 2003. 10. a. a. shinde, f. k. shaikh, m. v. padul and m. s. kachole. “bacillus subtillis rtsba6 6.00, a new strain isolated from gut of helicoverpa armigera (lepidoptera: noctuidae) produces chymotrypsin-like proteases”. saudi journal of biological sciences, vol. 19, pp. 317-323, 2012. 11. p. n. gayatri, a. ojha, m. k. kajla, a. raj and r. rajagopal. “host plant induced variation in gut bacteria of helicoverpa armigera”. plos one, vol. 7, pp. e30768, 2012. 12. l. e. visoˆtto, m. g. a. oliveira, a. o. b. ribon, t. r. maresguia and r. n. c. guedes. “characterization and identification of proteolytic bacteria from the gut of the velvetbean caterpillar (lepidoptera: noctuidae)”. environmental entomology, vol. 38, no. 4, pp. 1078-1085, 2009. 13. r. kaur, n. kaur and a. k. gupta. “structural features, substrate specificity, kinetic properties of insect α-amylase and specificity of plant α-amylase inhibitors”. pesticide biochemistry and physiology, vol. 116, pp. 83-93, 2014. 14. j. s. buyer. “improved fast gas chromatography for fame analysis of bacteria”. journal of microbiological methods, vol. 54, pp. 117120, 2003. 15. s. sharmin, m. d. towhid and m. n. anwar. “isolation and characterization of a protease producing bacteria bacillus amoviv-orus and optimization of some factors of culture conditions for protease production”. journal of biological sciences, vol. 5, no. 3, pp. 358-362, 2005. 16. o. h. lowry, n. j. rosebrough, a. l. farr and r. j. randall. “protein measurement with the folin phenol reagent”. journal of biological chemistry, vol. 193, pp. 265, 1951. 17. u. k. laemmli. “cleavage of structural proteins during the assembly of the head of bacteriophage t4”. nature, vol. 227, pp. 680-685, 1970. 18. e. h. fisher and e. a. stein. “alpha amylase from human saliva”. biochemical preparations, vol. 8, pp. 27-32, 1961. 19. g. g. priest. “extracellular enzyme synthesis in the genus bacillus”. figure 5: non-denaturing 7% starch sodium dodecyl sulfate polyacrylamide gel electrophoresis showing the influence of temperature on the amylase activity of bacillus subtilis rtsba6 6.00 figure 6: effect of different temperatures on amylolytic activity of bacillus subtilis rtsba6 6.00. samples were assayed by standard procedure using starch as substrate mentioned in material and methods shinde, et al.: amylase producing bacteria from h. armigera 79 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 75-79 bacteriological reviews, vol. 41, no. 3, pp. 711-753, 1977. 20. a. andrades and l. m. contreras. amylase zymography”. methods in molecular biology, vol. 1626, pp. 301-308, 2017. 21. f. zeng and a. c. cohen.“partial characterization of a-amylase in the salivary glands of lygus hesperus and lygus lineolaris”. comparative biochemistry and physiology part b, vol. 126, pp. 9-16, 2000. 22. p. c. aryati, a. pangastuti and s. l. a. sari. “amylase production potentials of bacterial isolates obtained from the gut of oryctes rhinoceros larvae”. iop conference series: materials science and engineering, 2017. 23. t. m. clark. “evolution and adaptive significance of larval midgut alkalinization in the insect superorder mecopterida”. journal of chemical ecology, vol 25, pp. 1945-1960, 1999. . 50 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 50-55 research article antibiotic susceptibility pattern of enterococcus spp. isolated from poultry feces raid d. thanoon1, essam a. makky2,3*, yashab kumar4 1department of biology, cihan university-erbil, kurdistan region, iraq, 2department of industrial biotechnology, faculty of industrial sciences and technology, university malaysia pahang, 26300 gambang, kuantan, pahang, malaysia, 3center of excellence for advanced research in fluid flow, university malaysia pahang, 26300 gambang, kuantan, pahang, malaysia, 4department of microbiology and fermentation technology, sam higginbottom university of agriculture technology and sciences, allahabad, uttar pradesh, india abstract enterococci, especially enterococcus faecalis and faecium, have emerged as an important nosocomial pathogen and represent a serious threat to patients with impaired host defenses. e. faecalis and faecium are part of the normal intestinal microbial flora of poultry and man under most conditions, they are considered as an opportunistic pathogen. in the current study, an investigation of enterococcus spp. isolated from poultry feces and their antibiotic susceptibility pattern was studied, due to the worldwide attachment with poultry by human being. samples were collected from different sites of allahabad, india, 80 samples collected screened for the presence of e. faecalis and e. faecium and identified based on cultural and biochemical characteristics. thirty-five isolates were identified as e. faecalis (57.37%), while 26 were e. faecium (42.62%). the pathogens isolated were tested for their susceptibility toward 10 different commonly prescribed antibiotics. most of the isolates showed resistance toward antibiotics under study. e. faecalis strain suggested a higher percentage of possibility of infection estimated by 15% in comparison with e. faecium as it was found to be less in a screening. the high resistance rate also indicates the negative impact of the antibiotic therapy. to evaluate the extent of transmission and impact of such transmission on the effectiveness of the antibacterial use in human medicine, further study is imperative. periodic monitoring of antibiotic resistance pattern to detect any change in it would be necessary for the effective treatment against these pathogens. enterococci revealed an alarming rate of resistance to the standard antimicrobial agents used for therapy and raised mic values to vancomycin. the importance and infection control were stressed. keywords: enterococcus faecalis, enterococcus faecium, poultry feces, vancomycin resistance introduction enterococci are members of the normal intestinal microflora in humans and animals, and they are common in environments affected by animal and human fecal material. these organisms are not considered primary pathogens, but due to their ability to acquire high-level resistance to antimicrobial agents, enterococci have emerged as nosocomial pathogens worldwide.[1] concern has especially been focused on enterococci that show high-level resistance to the glycopeptide antibiotic vancomycin (vancomycin-resistant enterococci) (vre), which recently has been the drug of the last resort against multiresistant enterococci and methicillinresistant staphylococcus aureus.[2] genetical similarities between animal and human originated enterococci have been reported and role of natural transmission of enterococci from food animals and contaminated foods to human tract cannot be ruled out.[3] enterococci cause food intoxication through the production of biogenic amines and worrisome opportunistic infections due to the virulence traits.[4] some strains are resistant to many antibiotics, but antibiotic resistance alone cannot explain the virulence of enterococci.[3] the differentiation of apparently safe and non-safe enterococci strains is not simple, especially due to effective horizontal gene transfer mechanisms.[5] enterococcus faecalis and enterococcus faecium are the most relevant species of enterococcus genus with regard to clinical aspects.[6] corresponding author: essam a. makky, faculty of industrial sciences and technology, universiti malaysia pahang, 26300 gambang, kuantan, pahang, malaysia. phone: +6013-9553169. fax: +609-5492766. e-mail: essam22001@gmail.com received: apr 10, 2019 accepted: apr 23, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp50-55 copyright © 2019 raid d. thanoon, essam a. makky, yashab kumar. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ thanoon, et al.: antibiotic susceptibility pattern of enterococcus spp. 51 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 50-55 vancomycin as a definition is an antibiotic at the end of the last resort for the treatment of gram-positive microorganisms such as mrsa and the multidrug-resistant staphylococcus epidermidis [figure 1].[7-10] in addition to the emergence of vre,[11] clinical isolates of mrsa strains with decreased susceptibility to vancomycin[12] (vancomycin intermediate-resistant s. aureus) and more recently with high-level vancomycin resistance [figure 1].[13] avoparcin is a glycopeptide antibiotic that has been used as a growth-promoting agent for food animals in many countries, except the united states and canada. in human medicine, glycopeptide-resistant enterococci (gre) have become an increasingly serious problem in the treatment of nosocomial infections.[14] in europe, gre have been isolated from animals, meat, environment, and healthy humans outside hospitals.[15-17] the widespread use of glycopeptides in hospitals has led to the emergence of vre which is a major concern for healthcare professionals. vre is frequently reported from hospitals in the usa and europe.[19] there is a paucity of information on vancomycin resistance in enterococci from our country.[20] since chicken meat and products are highly consumed by human beings and influx of virulence genes from enterococci of chicken origin to human intestinal tract is a possible route; therefore, it is desirable to study the antibiotic susceptibility pattern of enterococcus spp. to ensure the safe consumption of chicken and their products.[21] in this study, the incidence of enterococcus spp. has been isolated, identified, and tested for their antibiotic susceptibility pattern. materials and methods sample collection fresh poultry fecal samples were collected from (mahewa purab patti, naini, atala, and civil lines) allahabad, india, and used in the present study. a total of 80 fecal samples (20 from each site) were collected in pre-autoclaved glass sample bottles and immediately transported to the laboratory for further studies. isolation of enterococcus spp. about 0.5 g of each fecal sample was diluted in 4.5 ml of sterile physiological water and roughly homogenized. serial dilutions ranking from 10−3 to 10−7 prepared. afterward, 100 µl of each dilution has been plated in duplicate into a bile esculin azide agar (bea) medium and incubated for 24–48 h at 30°c, ph 7.1 ± 0.2. the isolates identified using standard morphological, cultural, and biochemical tests.[22] after incubation time, samples were inspected, and those containing from 50 to 100 colonies were selected for preliminary identification based on morphological and physical characteristics like gram staining and production of catalase, oxidase, and acid from glucose fermentation. morphological characteristics various cultural and morphological characteristics of the isolates have been examined, namely, color, elevation, odor, and surface of the colonies on bea agar plates. biochemical tests slide catalase test a glass rod has been used to pick up a colony from a culture plate and has been placed in a drop of 3% hydrogen peroxide on a glass slide. a positive catalase reaction would show gas bubbles. nitrate reduction test this test has been used to identify whether an organism that was able to reduce nitrate into nitrite and further into ammonia or molecular nitrogen. the isolates have been inoculated into nitrate broth media and incubated at 37 ± 2°c for 24 h. nitrate reagent (solution a) sulfanilic acid + acetic acid and solution b – α-naphthylamine + acetic acid added to observe the color change into red indicate positive result. sugar fermentation test this test has been performed to determine the ability of enterococcus spp. to ferment various carbohydrates under anaerobic condition in fermentation tube. for this purpose, basal media have been prepared and 4.5 ml of this media have been transferred into test tube along with inverted durham’s tubes and autoclaved at 121°c for 15–20 min. different carbohydrates, namely, glucose, lactose, galactose, mannitol, sorbitol, sucrose, maltose, xylose, and dextrose have been prepared in 1% concentration in distilled water and autoclaved at 10 lbs/inch² separately. then, 0.5 ml aliquot from sterile figure 1: (a) structure of vancomycin 1 and a binding scheme between vancomycin and a cell wall precursor analog, dada. (b) a cartoon illustration of the type of click vancomycin dimers[18] a b thanoon, et al.: antibiotic susceptibility pattern of enterococcus spp. 52 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 50-55 stock solution of each sugar added in 4.5 ml of basal medium. one loopful of isolated strains inoculated in each sugar solution and incubated at 37°c for 24–48 h. after incubation, the tubes have been observed for color change and gas production. uninoculated tube period has been taken as control. starch hydrolysis test starch agar media were prepared and autoclaved at 121°c for 15–20 min. then, the plates have been streaked with the bacterial isolates and incubated at 37°c for 24 h. after incubation period, the plates flooded with iodine solution (1 m) and observed for clear zone around the colonies. formation of clear zone around the colonies indicates positive starch hydrolysis. motility test this test has been done to check the motility of the bacteria. tube containing motility agar has been stab inoculated and incubated at 37°c for 24 h. growth around the stab line indicates a positive test. no growth around the stab line indicates a negative test. indole production test test tubes containing peptone broth will be inoculated with test organism and incubated at 37°c + 1°c for 24–48 h. after incubation, the 10 drops of kovac’s reagent will be added to it. a red color ring on the top of the peptone water indicated positive result. growth at various temperatures the isolates have been streaked on the bea agar plates and incubated at three different incubation temperatures, i.e., 10, 45, and 50°c for 24 h. after incubation period, the plates have been observed for microbial growth. study of antibiotic susceptibility pattern of identified bacterial isolates the antibiotic susceptibility of the isolated and identified enterococcus species has been assessed by the disc diffusion method.[23] for this test, melted and cooled muller-hinton agar has been poured in sterilized petri dishes and swabbed with overnight culture of enterococcus isolates. under aseptic conditions, antibiotics placed onto the surface of inoculated plates. following overnight incubation at 37°c, zone of inhibition for each antibiotic measured (in mm). to determine the susceptibility pattern of isolated enterococcus spp., the following antibiotics were used which are as follows: chloramphenicol (30 µg), penicillin g (10 µg), erythromycin (15 µg), vancomycin (30 µg), teicoplanin (30 µ), ciprofloxacin (5 µg), gentamycin (10 µg), streptomycin (10 µg), ampicillin (10 µg), and tetracycline (30 µg). results and discussion incidence of enterococcus spp. isolated from different sites in allahabad city in the present study, among 80 samples collected from poultry feces of allahabad region, 61 bacterial isolates were obtained, of which 35 were identified as e. faecalis (57.38%) and 26 were e. faecium (42.62%) [figure 2]. the percentage incidence of enterococcus spp. was, however, found to be observed that there is a significant difference in primary screening of e. faecalis strain, suggesting a higher percentage of possibility of infection estimated by 15% in comparison with e. faecium as it was found to be less in a screening, isolation, and most importantly the pathogenicity of it at their level of opportunistic pattern in contrast with e. faecalis. among both e. faecium and e. faecalis, some strains produced β-hemolysin with the incidence of this trait being higher for e. faecalis strains (21.3%) than for e. faecium strains (8.3%). hemolysin plays an important role in enterococcal virulence, as it may increase the severity of the infection [table 1].[24] the identification of enterococci isolated from the commercial poultry production environment did not reveal any unusual species, although eight isolates require more discriminate analysis before definitive identification. while multiple isolates were occasionally recovered from the same sample, the elimination of isolates with indistinguishable antibiograms from the same farm provided a collection that was conservative in its estimation of diversity but did not substantively affect the relative proportions of species isolated. antibiotic susceptibility pattern the antimicrobial susceptibility test was performed by disc diffusion method or kirby–bauer technique.[23] e. faecalis was tested for their sensitivity against various antibiotics. most of the antibiotics were found to be sensitive, vancomycin (30 µg) was resistance (10%), and some of them were found to be intermediately resistance against e. faecalis [table 2]. e. faecalis was found to be sensitive to five antibiotics, namely, penicillin g (10 µg), erythromycin (15 µg), ciprofloxacin (5 µg), streptomycin (10 µg), and tetracycline (30 µg), (50%). it was found to be resistant to vancomycin (30 µg) (10%). it was found to be intermediately resistant to gentamycin (10 µg), ampicillin (30 µg), teicoplanin (10 µg), and chloramphenicol (30 µg) (40%) [figure 3]. among the antibiotics, vancomycin, an inhibitor of cell wall synthesis is of major concern as it is one of the last antibiotics broadly effective against clinical infections caused by multidrug resistance pathogens.[25] e. faecium was found to be sensitive to penicillin g (10 µg), ciprofloxacin (5 µg), and tetracycline (30 µg), and it was resistant vancomycin (30 µg) and streptomycin (10 µg). furthermore, it was found to be intermediately resistant to chloramphenicol 61 35 26 total isolates e. faecalis (57.37%) e. faecium (42.62) figure 2: incidence of enterococcus spp. percentages isolated from different sites of allahabad city thanoon, et al.: antibiotic susceptibility pattern of enterococcus spp. 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 50-55 table 1: morphological characteristics and biochemical tests of enterococcus spp. parameter enterococcus spp. e. faecalis e. faecium cultural characteristics entire, flat, non-mucoid, white transparent, elevated, smooth, and entire margin gram reaction +ve +ve shape cocci cocci arrangement monococci monococci growth at 10°c 40°c 50°c +ve +ve growth at ph 9.6 +ve +ve biochemical tests nitrate residuals +ve +ve catalase +ve +ve starch hydrolysis −ve −ve indole test −ve −ve motility −ve −ve esculin hydrolysis +ve −ve sugar fermentation glucose a+g− a−g− lactose a−g− a−g− maltose a−g− a−g− sucrose a−g− a−g− mannitol a+g− a−g− galactose a−g− a−g− dextrose a+g− a+g− sorbitol a−g− a−g− xylose a+g− a+g− a+: acid positive, a−: acid negative, g+: gas positive, g−: gas negative, e. faecalis: enterococcus faecalis, e. faecium: enterococcus faecium table 2: antibiotic susceptibility pattern antibiotics enterococcus spp. clear zone (mm) e. faecalis e. faecium chloramphenicol (30 µg) 15 ++ 12 ++ penicillin g (10 µg) 26 +++ 19 +++ erythromycin (15 µg) 24 +++ 17 ++ vancomycin (30 µg) 10 + 10 + teicoplanin (30 µg) 12 ++ 17 ++ ciprofloxacin (5 µg) 21 +++ 21 +++ gentamycin (10 µg) 12 ++ 12 ++ streptomycin (10 µg) 18 +++ 10 + ampicillin (10 µg) 15 ++ 15 ++ tetracycline (30 µg) 23 +++ 24 +++ +++: sensitive, ++: intermediate, +: resistant. degree of inhibition = +: moderate inhibition zone (6–9 mm), ++: strong inhibition zone (10–14 mm) +++: very strong inhibition zone (15–18 mm or above), –: no inhibition zone, e. faecalis: enterococcus faecalis, e. faecium: enterococcus faecium (30 µg), ampicillin (30 µg), teicoplanin (10 µg), gentamycin (10 µg), and erythromycin (15 µg) [table 2]. conjugal transfer of vancomycin resistance genes from enterococci to other grampositive bacteria has been accomplished in vitro [figure 4]. the gram-positive organisms include group a and viridans group streptococci, listeria monocytogenes, and staphylococcus aureus.[26] this gives rise to concern that such transfer in humans under natural conditions might be feasible. ten (20.8%) e. faecium strains and 6 (12.8%) e. faecalis strains were susceptible to all antibiotics tested. all e. faecalis strains and all but one e. faecium strain were susceptible to vancomycin. e. faecium strains were mostly resistant to vancomycin (30 µg) (2.1%) and streptomycin (4.2%). while ciprofloxacin (56.3%), followed by penicillin (45.8%), erythromycin (27.1%), chloramphenicol (10.4%), tetracycline (6.3%), and gentamicin (2.1%) showed both intermediate and sensitive activity and none were resistant against ampicillin [table 2]. in contrast, e. faecalis strains were mostly resistant to figure 3: antibiotic susceptibility pattern of enterococcus faecalis isolates figure 4: antibiotic susceptibility pattern of enterococcus faecium isolates thanoon, et al.: antibiotic susceptibility pattern of enterococcus spp. 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 50-55 vancomycin (30 µg) (2.1%) while chloramphenicol (63.8%), followed by streptomycin (46.8%), tetracycline (44.7%), erythromycin (31.9%), ciprofloxacin (27.7%), gentamicin (25.5%), penicillin (12.8%), and ampicillin (2.1%) were found to be intermediate and sensitive. the present results of thesis study showed that a larger number of e. faecium strains than e. faecalis strains were resistant to vancomycin, which may be explained by e. faecium being generally more resistant to vancomycin than e. faecalis. the incidence of streptomycin and vancomycin resistance for both e. faecium and e. faecalis was low, indicating that most of the strains tested did not acquire resistance determinants for these antibiotics. in a study of european cheeses, teuber (1999)[27] also reported a low (4%) incidence of vre. while the incidence of aminoglycoside-resistant enterococci was low among e. faecalis isolates, it was considerably higher for e. faecium. these results indicate that especially e. faecium strains with acquired, high-level aminoglycoside resistance can occur in traditional cheeses.[28] frank (1996)[29] reported that isolates were found in 36 poultry flocks and 10 pig herds and among two of the human isolates, whereas no vancomycin-resistant e. faecium isolates were found among the isolates from calves. the pathogens isolated were tested for their susceptibility toward commonly prescribed antibiotics, namely, chloramphenicol, penicillin g, erythromycin, vancomycin, teicoplanin, ciprofloxacin, gentamycin, streptomycin, ampicillin, and tetracycline. from the present study, the following observations were made in incidence of e. faecalis in poultry feces samples which were maximum (57.37%) and minimum in e. faecium (42.62%). most of the isolates showed resistance toward a few of antibiotics, suggesting that vancomycin (30 µg) (2.1%) and streptomycin (4.2%) cannot be used against e. faecium and e. faecalis. the high resistance rate also indicates the negative impact of the antibiotic therapy. to evaluate the extent of transmission and impact of such transmission on the effectiveness of the antibacterial use in human medicine, further study is required. periodic monitoring of antibiotic resistance pattern to detect any change in it would be necessary for the effective treatment against these pathogens. the presence of high-level resistance to vancomycin eliminates a valuable therapeutic option in the management of serious enterococcal infections. resistance to glycopeptides in these organisms is caused by synthesis of modified bacterial cell wall precursors that demonstrate decreased affinity for vancomycin and teicoplanin.[30] enterococci revealed an alarming rate of resistance to the standard antimicrobial agents used for therapy and raised mic values to vancomycin. the importance and infection control are stressed. conclusion the total isolates of e. faecium and e. faecalis were found to be resistant to vancomycin e. faecium. vancomycin-resistant e. faecalis isolates were detected only among isolates from four poultry flocks. the results of this study illustrate that enterococcus spp. from poultry production and processing operations are frequently resistant to multiple antimicrobials and that some of these patterns may very well reflect the use of approved antimicrobials in poultry. this work also establishes a baseline of resistance among enterococcus spp. that will be useful in monitoring the dynamics of resistance longitudinally. considering some of the current estimates of the extent of antimicrobial use in the poultry production industry for growth enhancement, the increasing potential of such an intensive agricultural operation to affect antimicrobial resistance must be weighed against the reasonable risk that the treatment of human bacterial infections may be compromised. acknowledgment this work was carried out by the first author. the author also acknowledges university malaysia pahang and for providing guidance to fulfill this work. references 1. p. k. linden and c. b. miller. “vancomycin-resistant enterococci: the clinical effect of a common nosocomial pathogen”. diagnostic 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systematic and applied microbiology, vol. 23, pp. 510-522, 2000. 24. b. d. jett, m. m. huycke and m. s. gilmore. “virulence of enterococci”. clinical microbiology reviews. vol. 7, pp. 462-478, 1994. 25. a. p. johnson, a. h. c. uttley, n. woodford and r. c. george. “resistance to vancomycin and teicoplanin: an emerging clinical problem”. clinical microbiology reviews, vol. 3, pp. 280-291, 1990. 26. p. courvalin. “resistance of enterococci to glycopeptides”. antimicrobial agents chemotherapy, vol. 34, pp. 2291-2296, 1990. 27. m. teuber, l. meile and f. schwarz. “acquired antibiotic resistance in lactic acid bacteria from food”. antonie van leeuwenhoek, vol. 76, pp. 115-137, 1999. 28. b. e. murray. “diversity among multidrug-resistant enterococci in poultry feces”. emergency infectious diseases, vol. 4, pp. 37-47, 1990. 29. f. m. aarestrup, p. ahrens, m. madsen, l. v. pallesen, r. l. poulsen and h. westh. “glycopeptide susceptibility among danish enterococcus faecium and enterococcus faecalis isolates of animal and human origin and pcr identification of genes within the vana cluster”. antimicrobial agents and chemotherapy, vol. 40, no. 8, pp. 1938-1940, 1996. 30. r. leclercq and p. courvalin. “resistance to glycopeptides in enterococci”. clinical infectious diseases, vol. 24, pp. 545-546, 1997. tx_1~abs:at/add:tx_2~abs:at 67 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 67-73 research article psychological stress of the covid-19 pandemic on the mental health of healthcare workers in the kurdistan region delan j. qadir1, jamal k. shakor2, trifa m. mohammed3, raza m. abdulla4 1department of adult, college of nursing, university of sulamani, sulaimani, iraq, 2department of nursing, sulaimani polytechnic university, darbandikhan technical institute sulaimani, iraq, 3department of psychiatric nursing and mental health, college of nursing, university of sulamani, sulaimani, iraq, 4department of statistic and computer, college of commerce, university of sulamani, sulaimani, iraq abstract coronavirus disease-19 (covid-19) pandemic can lead to terrific condition among healthcare workers (hcws) and severe stress reactions can raise the risk of secondary trauma. the aim of this study was determine the psychological burden of the covid-19 outbreak on hcws. the cross-sectional quantitative survey was conducted online from september 1 to september 18, 2020. online questionnaires employing scales including the generalized anxiety disorder (gad-7), patient health questionnaire (phq-9), and perceived stress scale (pss-10) were used to investigate anxiety, depression, and stress. this study has shown that percentage of severe anxiety, stress, and depression were (22%), (3.9%), and (11%) respectively among hcws. there was a significant association of phq score with age, gender, marital status, number of children, kind of employment, and work experiences. gad score was found to have a significant relationship (p = 0.05) with gender, job title, and healthcare experiences in this study. the pss score of women who worked in hospitals was found to be considerably higher. health-care professionals in kurdistan experienced high anxiety and stress during the covid-19 epidemic. keywords: anxiety, coronavirus disease-19, depression, health care workers, kurdistan introduction the coronavirus disease infection (covid-19) first appeared in china in december 2019 and quickly spread to nearly every country on the planet. the world health organization declared the virus a global pandemic in march 2020.[1] the kurdistan region of iraq (kri) alone saw 10,595 illnesses and 402 fatalities on march 30, 2020, according to statements made by the kurdistan regional government (krg).[2] to address this urgent matter, the government mandated a public holiday from february 26 to march 10, 2020, for all public and private institutions, including kindergartens. public and private universities were likewise closed from february 29 to march 10. they will remain closed for spring break from march 10 through march 23 and reopen on march 24 if conditions improve. the krg has also made the decision to declare a halt to all religious rituals, activities, and events until further notice. this includes friday sermons in mosques, churches, and temples throughout the kri. even though the lockdown’s time limit was ultimately extended, the health-care professionals continued to work to address the situation. due to their direct and indirect interactions with covid-19 patients as well as their susceptibility to infection,[3] they are at risk of developing mental health difficulties.[4] in facilities that have been trained and specialized for covid-19. iraq had reported approximately 319,035 cases of infection and 8555 deaths as of september 30, 2020, with the majority of sickness and deaths coming from western europe and north america.[5] covid-19,[6] the immediate response from departments of respiratory medicine and intensive care units (icus),[7] or the stress on mental health faced by critical care practitioners were all factors that contributed to the onset of covid-19,[6] the immediate response from departments of respiratory medicine and intensive care units (icus),[7] or the place stress on mental health faced by practitioners in critical care.[8] health-care professionals (hcps) who care for covid19 patients were said to have a negative impact on them. corresponding author: jamal k. shakor, department of nursing, sulaimani polytechnic university, darbandikhan technical institute sulaimani, iraq. e-mail: jamal.shakor@spu.edu.iq received: august 22, 2022 accepted: may 03, 2023 published: june 20, 2023 doi: 10.24086/cuesj.v7n1y2023.pp67-73 copyright © 2023 jamal k. shakor, delan j. qadir, trifa m. mohammed, raza m. abdulla. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) cihan university-erbil scientific journal (cuesj) qadir, et al.: covid-19 mental health related among healthcare workers 68 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 67-73 physical exhaustion and sleep troubles have been recorded in hcps, as well as worry and fear of infection.[9] as well as, the spread of the virus among relatives, as a family was hampered by severe preventative measures.[10] evaluation support and mental healthcare are important aspects of the response to the covid-19 outbreak,[11] and these mental health issues have been addressed in both international and local legislation. hcps dealing with the covid-19 epidemic have noticed several unusual symptoms.[12] mers cov has a low overall human-to-human transmission capability; nonetheless, 38% of all confirmed illnesses are occasionally aggravated in the healthcare setting.[13] mers cov infections caused by healthcare workers occur for 1–27% of all mers cov cases.[14] healthcare worker (hcws) are said to be more insecure, and the severity of the psychological effect was found to be connected to the length of the quarantine period.[15] it is feared that such a negative impact would persist and have long-term consequences. hcws are the backbone of any country’s health-care system, and they may suffer from a variety of mental health issues as a result of their work during the covid-19 epidemic. as a result, the goal of this study was to determine the levels of depression, anxiety, and stress among a group of health-care employees who answered our online survey. review of literature the research studies from china, india, italy, spain, iran, and kurdistan/iraq were among those in the review. all the others have a cross-sectional layout. the increase in mental health issues has been noted in editorials, scientific words, viewpoints, and commentary in scientific literature as well as reporting in print and visual media. experts asked for mental health support as they were concerned over the rising number of mental health issues.[11] another a significant global public health concern during this epidemic is the rise in mental health issues in every community and age group across all countries.[16-23] to relieve psychological anguish and its impacts, experts have recommended appropriate and affordable solutions.[24] this new predicament with mental health issues has received a lot of attention. however, there is still a lack of quantifiable data regarding the rise in mental health issues brought on by the pandemic. before making the proper arrangements for tackling this issue of growing mental health difficulties, authorities need to be aware of the scope of the issue. this scoping review was carried out to estimate the number of different mental health issues brought through covid-19. methodology design of the study this cross-sectional quantitative survey was done online from september 1–18, 2020, following an 8-month lockdown in the kurdistan region due to a coronavirus pandemic. administrative arrangement the proposal for the study was accepted by the council of nursing college and approved by the scientific committee of nursing college/university of sulaimani. participant this cross-sectional design quantitative survey was done online from september 1–18, 2020, following an 8-month lockdown in the kurdistan region due to a coronavirus pandemic. the online poll drew the participation of 334 hcws. after reading the study purpose, the survey was generated in google survey (www.google.com) and the link was provided through email with invitations to possible participants who were eligible for the written consent section in the first section of the online survey. if they agreed, they could then proceed to fill out the questionnaire. tools and measurement age, sex, marital status, degree of education, work, year of experience, economic situation, possible direct interaction with covid-19 patients, and disease severity were some of the sociodemographic issues mentioned. simply checking the answer checkbox takes an average of 5 min to complete the inquiry. the kurdish translation of the perceived stress scale (pss-10),[25] patient health questionnaire (phq-9),[26] and generalized anxiety disorder (gad-7) was a significant aspect of the questionnaire for recording the degree of stress, grief, and anxiety.[27] pss10 scores vary from 0 to 13 (low), 14 to 26 (moderate), and 27 to 40 (high) (extreme stress perceived). the phq-9 scale has scores of 5–9, 10–14, 15–19, and 20–27 for mild, moderate, moderate, intense, and severe depression, respectively. the 7-item gad scale was used to assess anxiety. the gad-7 scale is a self-reporting scale with great reliability and validity. if the score is <5, there is no anxiety, whereas the cutoffs for moderate, mild, and severe anxiety are 5, 10, and 15 respectively. in this study, a cutoff of five on the phq-9 and gad-7 was used to identify persons who had or did not have any symptoms of depression or anxiety. statistical analysis after data were automatically registered in the survey’s excel file and imported into spss, statistical analysis was performed using spss version 22. for descriptive analysis, the frequency with percentage and mean with standard deviation was used. as inferential analysis, the t-test and anova were utilized to evaluate for significance. informed consent the study protocol was accepted by sulaimani university. results table 1 shows that the participants were largely from sulaimani (79.9%), that they were mostly men (52.1%), that they were mostly married (71.3%), and that they had 1–3 children (56.6%). a responder was mostly a nurse (74.9%), worked in a critical department of a hospital (46.4%), had 1–9 years of experience (44.0%), and was employed permanently (79.9%) with monthly 100–120 h of work in hospitals (45.2%). table 2 illustrated that most responders had direct contact with covid-19 infected patient, and (31.7%) was infected qadir, et al.: covid-19 mental health related among healthcare workers 69 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 67-73 table 2: covid-19 suffers and contacts among participants place of resident frequency percentage from the beginning of the covid-19 outbreak, did you have contact with infected peoples yes 243 72.8 no 91 27.2 total 334 100.0 due to contact, did you have infected by coved-19 yes 106 31.7 no 162 48.5 i don’t know 66 19.8 total 334 100.0 if you are infected, how do you rate your symptoms? severe 27 8.1 mild 99 29.6 no symptoms 35 10.5 no infected 173 51.8 total 334 100.0 from the beginning of the covid-19 outbreak, did you have used personal protective measures? yes 279 83.5 no 55 16.5 total 334 100.0 by covid-19 due to their work with mostly mild symptoms (29.6%). meanwhile, nearly (84%) have used personal protective measures since the covid-19 outbreak. table 1: sociodemographic and work status of the participants variables frequency percentage place of resident sulaimani 267 79.9 hawler 37 11.1 kirkuk 13 3.9 halabja 17 5.1 total 334 100.0 age groups 20–29 118 35.3 30–39 119 35.6 40–49 67 20.1 50–59 30 9.0 total 334 100.0 gender male 174 52.1 female 160 47.9 total 334 100.0 marital status single 97 28.7 married 238 71.3 total 334 100.0 number of children no child 116 34.7 1–3 children 189 56.6 4 children and more 29 8.7 total 334 100.0 job title nurse 250 74.9 physician 49 14.7 lab technicians 7 2.1 others 25 7.5 total 331 99.1 the hospital departments where the staff work at them word 87 26.0 critical department (icu, emergency) 155 46.4 lab 13 3.9 administration 32 9.6 health centre 47 14.1 total 334 100.0 health service experience <1 year 24 7.2 1–9 years 147 44.0 10-19 years 93 27.8 20 and more years 70 21.0 total 334 100.0 (contd...) variables frequency percentage type of employments public employee 267 79.9 bond (temporary employment) 34 10.2 volunteer 33 9.9 total 334 100.0 how long do you work in the month in hospitals 100–120 151 45.2 121–144 80 24.0 145–168 51 15.3 169–192 52 15.6 total 334 100.0 economic status sufficient 69 20.7 barley sufficient 239 71.6 insufficient 26 7.8 total 334 100.0 table 1: (continued) qadir, et al.: covid-19 mental health related among healthcare workers 70 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 67-73 table 3 demonstrates the psychiatric condition of health staff. this study has shown that percentages of severe anxiety, stress, and depression were (22%), (3.9%), and (11%), respectively. table 4 has shown the significant association of phq score (p < 0.05) with age, gender, marital status, number of children, kind of employment, and work experiences. mean of phq score was significantly higher in the age group 20–29 years (10.36 ± 4.22), female (9.71 ± 4.20), single marital status (10.23 ± 4.28), not have children (9.92 ± 4.21), volunteered employed (11.91 ± 4.16), and <1 years have experienced (10.58 ± 4.31). this study showed the significant association of gad score (p < 0.05) with gender, job title, and health service experiences. the mean gad score was significantly higher in females (7.41 ± 3.65), administrative staff (7.72 ± 4.17), and <1 year have experienced (8.08 ± 3.53). similarly, the significant association of pss score (p < 0.05) was observed with gender, kind of employment, and work hours in hospitals. the mean pss score was significantly higher in females (18.07 ± 5.91), voluntary employment (18.76 ± 6.35), and have 121–144 h of work in the hospitals (18.44 ± 5.74). table 5 explained the relationship between the psychiatric health condition and experience with covid-19 among health staff. there was a significantly high phq score among those who had covid-19 contact (9.43 ± 4.16), those who were infected by covid-19 (10.18 ± 4.18), and had severe symptoms (10.30 ± 4.54). similarly, there was a significant high gad score among those who were infected by covid-19 (7.69 ± 3.70) and had severe symptoms (7.85 ± 3.91). discussion covid-19 has caused some psychiatric problems in hcws because of their sensitivity to infection or other diseases.[19] table 3: the level of the psychiatric condition among health staff psychiatric conditions frequency percentage anxiety levels minimal (normal) 90 26.9 mild anxiety 170 50.9 moderate anxiety 65 19.5 severe anxiety 9 2.7 total 334 100.0 stress levels low perceived stress 84 25.1 moderate perceived stress 237 71.0 severe perceived stress 13 3.9 total 334 100.0 depression levels mild perceived depression 194 58.1 moderate perceived depression 101 30.2 moderate severe depression 34 10.2 severe perceived depression 5 1.5 total 334 100.0 covid-19 anxiety and stress were found to be higher in hcws than mers-cov and seasonal influenza.[20] hcws are concerned about infecting their family members and coworkers, therefore table 4: the association of sociodemographic and work experience with psychiatric health conditions variables mean±sd gad score phq score pss score place of resident sulaimani 6.87±3.56 9.24±4.08 17.09±5.77 hawler 6.84±4.27 8.97±3.73 17.49±6.32 kirkuk 5.69±2.50 6.77±4.32 14.23±6.93 halabja 6.71±4.54 9.88±5.45 17.06±7.28 total 6.81±3.65 9.15±4.14 17.02±5.96 p value 0.730 0.170 0.376 age groups 20–29 7.21±3.73 10.36±4.22 17.63±6.26 30–39 7.00±3.55 8.56±3.92 16.86± 40–49 5.79±3.61 8.75±4.21 16.24±5.82 50–59 6.77±3.61 7.57±3.46 17.07±5.72 p value 0.072 0.000 0.483 gender male 6.26±3.58 8.63±4.03 16.06±5.87 female 7.41±3.65 9.71±4.20 18.07±5.91 p value 0.004 0.016 0.002 marital status single 7.08±3.61 10.23±4.28 17.34±6.36 married 6.70±3.68 8.71±4.02 16.90±5.82 other 8.00± 11.00± 17.00± p value 0.649 0.009 0.834 economic status sufficient 6.10±2.82 8.19±3.63 15.71±5.67 barley sufficient 6.88±3.79 9.38±4.28 17.35±5.97 insufficient 8.08±4.04 9.58±3.84 17.54±6.35 p value 0.054 0.095 0.119 number of children no child 7.04±3.71 9.92±4.21 17.22±6.46 1–3 children 6.85±3.57 8.83±3.98 17.13±5.73 four children and more 5.66±3.88 8.14±4.56 15.59±5.33 p value 0.184 0.031 0.395 type of employments public employee 6.64±3.63 8.70±3.95 17.13±5.70 bond (temporary employment) 7.12±4.22 10.00±4.51 14.50±6.89 volunteer 7.85±3.12 11.91±4.16 18.76±6.35 p value 0.178 0.000 0.011 job title nurse 6.80±3.55 9.27±4.23 16.91±5.99 (contd...) qadir, et al.: covid-19 mental health related among healthcare workers 71 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 67-73 table 5: the association of psychiatric health conditions with covid-19 variables mean±sd gad score phq score pss score from the beginning of the covid-19 outbreak, did you have contact with infected peoples yes 6.99±3.66 9.43±4.16 17.01±5.88 no 6.33±3.60 8.40±4.03 17.05±6.22 p value 0.140 0.042 0.954 due to contact, did you have infected by coved-19 yes 7.69±3.70 10.18±4.18 17.42±5.71 no 6.22±3.70 8.30±0.00 16.78±5.75 i don’t know 6.86±3.19 9.56±4.39 16.97±6.87 p value 0.005 0.001 0.690 if you are infected, how your symptoms are? severe 7.85±3.91 10.30±4.54 17.30±5.82 mild 7.48±4.05 10.13±4.39 17.32±6.09 no symptoms 6.49±3.15 8.86±3.40 17.51±5.57 no infected 6.33±3.40 8.46±3.95 16.71±6.02 p value 0.031 0.005 0.798 from the beginning of the covid-19 outbreak, did you have used personal protective measures? yes 6.73±3.61 8.97±4.15 16.77±6.02 no 7.24±3.89 10.05±4.03 18.29±5.54 p value 0.346 0.075 0.085 gad: generalized anxiety disorder, phq: patient health questionnaire, pss: perceived stress scale, sd: standard deviation variables mean±sd gad score phq score pss score physician 7.04±3.74 8.86±3.62 18.00±4.69 lab technicians 2.86±3.24 6.00±3.74 12.71±9.32 others 7.72±4.17 9.40±4.28 17.40±6.89 p value 0.019 0.207 0.163 hospital department of work of the participant word 6.99±3.42 9.70±4.10 17.05±5.85 critical department (icu, emergency) 7.19±3.66 9.42±4.18 17.70±5.51 lab 4.85±4.10 8.38±4.25 16.00±7.74 administration 5.87±3.43 7.72±3.72 15.25±5.83 health center 6.40±3.88 8.40±4.14 16.23±6.97 p value 0.079 0.091 0.195 how long do you work in the month in hospitals 100–120 h 6.46±3.74 8.92±4.27 17.03±5.91 121–144 h 7.58±3.34 9.44±3.79 18.44±5.74 169–192 h 7.17±4.00 9.77±4.50 15.42±6.86 p value 0.091 0.469 0.032 health service experience <1 year 8.08±3.53 10.58±4.31 15.13±8.20 1–9 years 7.23±3.89 9.82±4.23 17.96±5.65 10–19 years 6.29±3.16 8.59±4.07 16.52±5.86 20 and more years 6.19±3.63 7.99±3.64 16.39±5.66 p value 0.031 0.003 0.056 gad: generalized anxiety disorder, phq: patient health questionnaire, pss: perceived stress scale, icu: intension care unit table 4: (continued) they wear protective clothing.[21] psychiatric conditions of hcws were altered during the covid-19 epidemic in the present study, with the percentages of severe anxiety, stress, and depression being (22%), (3.9%), and (11%) accordingly. the anxiety rate in this study was higher than that of hws in suadi, at 11%,[20] and depression was also higher than that of hcws in china, at 6.2% (mean phq-9: 15.1).[22] these findings are in line with a significant amount of research that has been published[28,29] and show that hcws experience anxiety at this time. in the present study, there were considerably more people who reported having moderate or severe anxiety (22.5%) than there were in the general population (5%), as had been observed prior to the pandemic.[30] meanwhile, anxiety and depression prevalence rates among hcws during covid-19 were found to be lower in the present study than in the systematic review (232 and 228%, respectively).[29] the difference in psychiatric levels is primarily due to the use of various techniques and metrics. the prevalence of moderate-to-severe psychiatric illnesses was rated in the current investigation. the majority of study participants had direct contact with covid-19-infected patients and worked more than 120 h each month, and one-third of them was infected with covid-19. since the covid-19 outbreak, roughly four-fifths had adopted personal preventive measures. according to a study, the high workload causes a shortage of medical protective supplies as well as staff behavioral issues.[31] in the present study, demographic status and job conditions were the most important factors of hws’ psychological problems. phq score was significantly higher (p = 0.05) in the age group 20–29 years, female, single marital status, no children, volunteers employed, and had <1 year of experience. phq score was shown to be considerably higher among females, most outbreaks placed, nurses, frontline job, non-volunteer employed, self-infected, or colleague infected in several wuhan research.[2,4] in an indian study, however, gender had no bearing on the severity of psychiatric problems.[32] gad score, p = 0.05 was found to have a significant relationship with female gender, administrative staff, and experience of <1 year in this study. this finding was in line with a wuhan study that found that the critical department qadir, et al.: covid-19 mental health related among healthcare workers 72 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 67-73 of the hospital, gender, and marital status were all linked to high gad and phq[33] levels.[4] nurses were shown to be more anxious than other medical care professionals in earlier studies in china.[32] the majority of the study’s participants were nurses who worked in the hospital’s critical care units, although a high psychiatric disorder score had no bearing on nursing professionalism. other italian research has found no link between phq and gad scores and nursing professionalism, icu work, or frontline work; nonetheless, nursing and frontline work had high phq and gad scores.[34] phq and gad scores were shown to be considerably higher in those who had covid-19 contact, were infected with covid-19, and had severe symptoms in the current investigation. the leading determinants of stress among hcws were found to be demographics and working conditions. according to an italian study, stress is caused by youthful age, female gender, front-line employment, and nursing professionalism.[35] meanwhile, there was a significant relationship between pss score (p = 0.05) and female gender, voluntary employment, and having 121–144 h of work in hospitals in the present study. a chinese study also discovered a link between pss score and age, working years, and current job title.[33] younger employees are almost inexperienced, and they would be unable to handle stressful circumstances such as working on the front lines for more than 120 h. the female gender is likewise more susceptible to psychological problems. the association of covid-19-related stress with female gender was confirmed among physicians in another study in kurdistan, moderate/ high stress was high among female physicians.[36] conclusion during the covid-19 outbreak, healthcare workers in kurdistan experienced severe anxiety and stress due to unpleasant psychiatric effects. screening for negative psychological outcomes 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(preprint article) 29. s. pappa, v. ntella, t. giannakas, v. g. giannakoulis, e. papoutsi and p. katsaounou. prevalence of depression, anxiety, and insomnia among healthcare workers during the covid19 pandemic: a systematic review and meta-analysis. brain, behavior, and immunity, vol. 92, p. 247, 2021. 30. a. shechter, f. diaz, n. moise, d. e. anstey, s. ye, s. agarwal, j. l. birk, d. brodie, d. e. cannone, b. chang and j. claassen. psychological distress, coping behaviors, and preferences for support among new york healthcare workers during the covid-19 pandemic. general hospital psychiatry, vol. 66, pp. 1-8, 2020. 31. q. liu, d. luo, j. e. haase, q. guo, x. q. wang, s. liu, l. xia, z. liu, j. yang and b. x. yang. the experiences of health-care providers during the covid-19 crisis in china: a qualitative study. the lancet global health, vol. 8, no. 6, pp. e790-e798, 2020. 32. m. y. si, x. y. su, y. jiang, w. j. wang, x. f. gu, l. ma, j. li, s. k. zhang, z. f. ren, r. ren, y. l. liu and y. l. qiao. psychological impact of covid-19 on medical care workers in china. infectious diseases of poverty, vol. 9, no. 1, p. 113, 2020. 33. l. q. wang, m. zhang, g. m. liu, s. y. nan, t. li, l. xu, y. xue, m. zhang, l. wang, y. d. qu and f. liu. psychological impact of coronavirus disease (2019) (covid-19) epidemic on medical staff in different posts in china: a multicenter study. journal of psychiatric research, vol. 129, pp. 198-205, 2020. 34. r. buselli, m. corsi, s. baldanzi, m. chiumiento, e. del lupo, v. dell’oste, c. a. bertelloni, g. massimetti, l. dell’osso, a. cristaudo and c. carmassi. professional quality of life and mental health outcomes among health care workers exposed to sars-cov-2 (covid-19). international journal of environmental research and public health, vol. 17, no. 17, p. 6180, 2020. 35. r. rossi, v. socci, f. pacitti, g. di lorenzo, a. di marco, a. siracusano and a. rossi. mental health outcomes among frontline and second-line health care workers during the coronavirus disease 2019 (covid-19) pandemic in italy. jama network open, vol. 3, no. 5, p. e2010185, 2020. 36. b. a. saeed, n. p. shabila and a. j. aziz. stress and anxiety among physicians during the covid-19 outbreak in the iraqi kurdistan region: an online survey. plos one, vol. 16, no. 6, p. e0253903, 2021. tx_1~abs:at/add:tx_2~abs:at 1 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 research article age of women as a parameter affecting ca-125, tsh, and cbc: a correlation study muhsin h. ubeid1, ameena s. m. juma1, tanya s. salih2, shilan s. kamel1, shakir m. al-awqati3 1department of medical microbiology, college of science, cihan university-erbil, kurdistan region, iraq, 2department of physiotherapy, cihan university-erbil, kurdistan region, iraq, 3department of nutrition and dietetics, cihan university-erbil, kurdistan region, iraq abstract one of the cancer’s antigens is the ca125, which is a protein antigen, found at excessively high levels in females characterized with ovarian cancer. it is suggested for ovarian cancer screening of women that are at higher risk to get the disease, despite its limited sensitivity and specificity. this study is conducted to identify possible correlations between ca125 and thyroid stimulating hormone (tsh) levels with parameters of hematological origin among women of different ages. this study included 44 healthy looking females, and 20 individuals as control group most but not all patients were from erbil. the study revealed that, ca125 and tsh levels were not associated. furthermore, the elevation of the ca125 levels in the females could be related to other non-cancerous factors such as menstrual cycle, or postmenopausal. furthermore, wbc parameters and hb levels were not affected by ca125. the only parameter that was affected by ca125 elevation was rbcs. the research recorded a significant difference of rbc between patient group and control group. also recorded, no difference to be considered as an indication related to the difference in women age. keywords: cancer antigen 125, thyroid stimulating hormone, white blood cells, red blood cells, pregnant women introduction identified women with ovarian cancer are likely to have the cancer protein antigen ca125 at excessively high levels their blood of yurkovetsky et al.[1] sikaris[2] stated that the serosal epithelial cells produce ca125 naturally. and this product can be found in both malignant and benign serosal fluids. meanwhile, bast et al.[3] advocated in a small segment of diseased patients that the ascending of the ca125 levels facilitated the triggering of transvaginal sonography. further, yurkovetsky et al.[1] stated that the blood serum of a large segment of healthful females contained levels lower than 35 units/µl of ca125. besides, there were indicators of noncancerous origin contributed to the elevation of ca125 levels. other scholars stated, when thyroid hormons extensively exposed they have the tendency to influence ovarian tumors development.[4] in addition,[5] in their vitro research illustrated that at the physiological levels both hormones 3,5,3’-triiodo-l-thyronine and l-thyroxin, (t3) and (t4), respectively, were capable of the expansion and improvement of the ovarian cancer cells. accordingly, these cells were grown much slower when exposed to (t3) and (t4) below normal serum concentrations levels. hence, thyroid functionality evaluation would be important to adjudge whether it is an elevation in the tumor markers or high-protein ascites.[6] a positive correlation was found between ca125 and the total white, neutrophil, monocyte and neutrophil-tolymphocyte ratio (nlr) counts. but conversely, a negative correlation recorded with lymphocytes.[7] the current study by ceran et al.[8] demonstrated that the rise in neutrophil and platelet counts, nlr, along with platelet-to-lymphocyte ratio (plr) correlated to an adverse clinic pathological results subsequent to various cancer types which comprised the epithelial ovarian cancer (eoc). while a previous study mentioned, that van derr zee et al. shown whether the ovarian tumors were malignant or benign the elevation in interleukin-6 level in cystic fluids is related to a decline corresponding author: muhsin h. ubeid, department of medical microbiology, college of science, cihan university-erbil, kurdistan region, iraq. e-mail: muhsin.hmeadi@cihanuniversity.edu.iq received: october 16, 2022 accepted: december 2, 2022 published: january 30, 2023 doi: 10.24086/cuesj.v7n1y2023.pp1-7 copyright © 2023 muhsin h. ubeid, ameena s. m. juma, tanya s. salih, shilan s. kamel, shakir m. al-awqati. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) cihan university-erbil scientific journal (cuesj) ubeid, et al.: parameter affecting ca-125, tsh, and cbc 2 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 in the hemoglobin level and an inclination in platelet count.[9,10] based on lengye study, the ovary cell transformation occur due to ovarian carcinoma that is consequent to genetic and epigenetic alterations.[11] ovarian cancer etiology is still not verified. however, the incessant ovulation theory is one of the models to explain the issue. it states that women have a continual ovulation regardless to age.[12] another proponents stated that high contributors to ovary carcinoma were, that is, infertility alongside to menopause age. yet, determinants particularly hysterectomy, pregnancy, and tubal ligation showed to diminish such risk.[13,14] abdominal size, bloating, urinary symptoms, and pelvic pains are the symptoms that were frequently informed by women prior diagnosing them with ovarian cancer. such symptoms are likely indicators causing cancer more than benign. yet, distinguishing cancer symptoms from those associating with benign is difficult. consequently, results, regardless how sensitive and specific pelvic examinations that were carried out to detect asymptomatic ovarian cancer recorded poor also, not supporting the physical examination in same way as the screening method.[15] according to national academies of sciences and medicine women and their designated health care providers repeatedly look upon as such symptoms (ovarian cancers) as non-specific and not referring to critical illness until they are dramatically worsen or developed.[16] which concurrent with rauh-hain et al.[17] who suggested that 70% of women with ovarian cancer were diagnosed at late stages of the disease hence, these women have a high fatality rate and have a 5 year survival rates with advanced illness ranging between 20% and 30%. in contrast, early diagnosed women as the disease is encompassed within the ovary the cure percentage were 70–90%. more into this, due to an association between the disease stage and survival, early detection provides higher hope to reduce mortality besides long-term disease control. globally, the ovarian cancer is ranked the 7th disease that is commonly diagnosed cancer while, it is classified the 8th among cancer diseases causing death. 239,000 women were diagnosed with this illness in 2012, 152,000 did not survive.[18] yet, the disease mortality rates and its frequencies vary from one region to another. and the variance is due to genetic factors and environmental influences.[19] moradi et al. stated that iran is witnessing an inclination in ovarian cancer cases, specifically in the central and northwestern regions. they suggested that this increment is stemming from changes in the patterns of risk factors regarding this disease, that is, changes in the population’s lifestyle that caused this escalation.[20] meanwhile, it was illustrated that, the risk of confronting severe and death from this disease in the developed countries are higher and nearly the double from those of less developed countries. yet, the latter countries recorded higher ovarian cancer cases because of the large populations. comparing the diagnosed cases number per year china recorded 34,575, india 26,834 and the usa 20,874.[21] after more than two decades from the discovering ca125, its antigen was recommended for screening women with high risk with ovarian cancer at clinical level regardless to its sensitivity and specificity limitation.[22] current results advise that ca125 possibly be a foretelling indicator of perinvasive ovarian cancer. the ca125 measuring approaches had progressed toward sensitive and reliable double-determinant elisa assays.[23] while others findings that, the escalated levels of ca125 serum are not correlated to the size of the tumor of both age groups.[24] as a glycosylated membrane protein and a high molecular weight, the ca125 can be identified in the serum which is levitated in over 80% of ovarian cancer patients. developing ca125 from the coelomic epithelium facilitated the marking of the epithelial. it had marked 90% of advanced and 50% of early ovarian cancer cases but, 20% were marked as low or had no expression. the ca125 is likely to arise its level if any disruptions occurred to the peritoneum epithelial lining. similarly, pregnancy, leiomyomata, ovarian cysts, endometriosis, appendicitis, and diverticulitis which are all benign conditions the ca125 level were high. equally, lung, pancreas, colon and uterine cancers showed and escalation in the ca125 levels. therefore, it is lacking the sensitivity and specificity and thus, it will not be adequate to identify the ovarian cancer at early stages.[25,26] gyftaki et al. stated that, the functionality of a normal thyroid is regulated by the thyroid stimulating hormone (tsh) by attaching to the thyroid receptor tsh receptor that is located at its cells surface. they further demonstrated that among other tissues, other than the thyroid, specifically the surface of a normal ovarian epithelium is amply expressed on.[27] the encoding thyroid hormone with estrogen receptors (tr) and (er) were among various nuclear hormone receptor genes that are distributed in ovarian surface epithelial cells (ose). equally important, from the epidemiology perspective, evidently correlate the ovarian surface inflammation of the ovary and hyperthyroidism with the ovarian cancer (hormonedependent).[28] hyperthyroidism is where hormones of the thyroid t3 and t4 excessive concentrations travel through blood stream through the human body reaching the ovarian tissue and consequently causing its inflammation. the receptors in the ose cells are responsive to thyroid and estrogen hormones. however, direct effects of ovarian epithelial cells inflammation are compelled by the t3. once the latter hormone attaches to its corresponding ose receptor the erα and mrna expressions are increased then the estrogen receptor action is mirrored resulting in encoding the er isoforms.[29] they further suggested that, ovarian cancer is highly correlated with isoforms and likely to be linked with hyperthyroidism. another research expressed that in vitro the thyroid hormones existing in ovaries cells infected with cancer changed both the genes expression that regulates cell cycle and the apoptosis of cancer cell. this action can be demonstrated by the inclination in the genes expression of which are responsible for the tumor’s protein suppression p12 and p16, besides, its significant role in regulating cell cycle.[30] anomalous hematological incidences recorded with ovarian cancer patients when compared with similar benign cases such as: first, hemoglobin hb and hematocrit hct levels were substantially low, second, high platelet count, and thirdly lymphocytes values were low.[31-33] ubeid, et al.: parameter affecting ca-125, tsh, and cbc 3 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 van der zee et al. illustrated that within malignant disease the presence of a conceivable relation between hemoglobin il-6 levels and platelets. they also stated that, as such patients show negative predicting factors when there is an elevated pre-treatment platelet counts in addition to, low levels of pretreatment hemoglobin. they additionally spotted an association between il-6 cystic fluid levels with the level of hemoglobin and platelet counts.[9] other scholars suggested similar association in those patients with progressive illness where anemia is probably related to il-6 levels.[10] which agrees with nomelini et al.[34] identification of the prognostic factors, which are resulting from a probable role of il-6 tumor derived alongside with irregular rates of hemoglobin and platelet levels. a late research weighed other ovarian cancer predictive factors for survival. it comprises the ratios of nlr along with plr. it also added that these ratios highlight the possibility to differentiate whether the ovarian masses are malignant or benign. yet, because of their values it does not have a total consensus as an ovarian cancer screening tool. moreover, the research illustrated another survival predictive factor concerning eoc patients whose illness is in early and advanced stages. this factor signifies the absolute neutrophil count to absolute monocyte count ratio (anc) and (amc), respectively.[15] aim of the study many women can survive from ovarian cancer if early diagnosis of the disease occurs. ca125 protein test may help to early diagnosis and help others whose life are at risk because of the disease, alongside with high tests’ accuracy. many noncancerous factors can elevate ca125 levels. in this study, we attempted to evaluate how tsh levels affect the ca125 levels among women of different ages, and how does ca125 affect the levels of some hematological parameters. materials and methods all samples 64, 20 control group and 44 non-cancerous patients were collected at “alfa lab for medical analysis” in erbil city. about 6 ml of blood were collected from each, 2 ml were used for hematological performance routine tests including blood film, wbc, hb., plt., and dlc. the remaining 4 ml samples were put aside for clotting in gel tubes and centrifuged for 15 min at 2500 rpm. then, the serum was used for conducting the test of electrochemiluminescence immunoassay by adding 0.5 ml of serum in a special tube for measuring serum ca-125 and an automated medonic coulter was used to determine the complete blood count (cbc) to all collected samples. statistical analysis since the study in hand is quantitative the augmented data were evaluated, and analyzed statistically, which was organized in tables. afterward data were analyzed with the version 18 of the statistical package for the social sciences software. then the appointed variables were compared following students f-test and t-test. later, the p value indicates whether a certain variable is significant, if it was either <0.01 or 0.05. results this study included 44 non-cancerous female patients. their ages ranged from (15 to 59) years, with a mean age of (37.8) year. in addition to, 20 healthy individuals were included as control group. table 1 shows and compare the ratio of tsh and ca125 in both control groups and patient group while [table 2] demonstrates the results of cbc parameters, alongside with (7) figures were created accordingly, to compare between mean of ca125 with mean of each (cbc) parameter separately. in addition to, a figure illustrating the ratio of ca125 level as the age rises. table 1 shows no significant variance in tsh levels between members of both study groups; however, it identified a high substantial variation between ca125 of both groups. while [figure 1] highlights the correlation between ca125 and tsh ratio in patient and control groups. [table 2] shows different cbc parameters, the wbc shown significant variance in patient and control groups. meanwhile, there was no substantial variation in lymphocyte, monocyte, granulocyte, and hemoglobin, but it identified a high significant difference between rbcs of patient group and control group. in [figure 2], there is a difference between wbc ratio in patient group and control group as the ca125 ratio rises. in [figure 3], lymphocyte ratio results show no variance in patient group and control group as the ca125 rises. figure 4 shows the similarly, monocyte ratio shows no difference in patient group and control group as the ca125 ratio rises. in [figure 5], there is difference between granulocyte ratio in patient group and control group as the ca125 ratio rises. in [figure 6], there is difference between rbc ratio in patient group and control group as ca125 ratio rises. [figure 7] shows that there is non-significant difference between hb ratio in patient group and control group as ca125 ratio rises. figure 8 shows, ca125 ratio as the age rises, where there is no difference to be considered as indication related to the difference in the age of women, despite the high peaks in age (34–37) years and (50) years. thus, the age of women does not give any sign to be mentioned. discussion this study is aiming to discover possible correlations linking tsh level and ca125 with hematological parameters among women of different ages. in the present study, no association was found between tsh and ca125 levels. as the ca125 levels elevated no significant change were recognized in tsh levels. nine patients with high ca125 levels above ubeid, et al.: parameter affecting ca-125, tsh, and cbc 4 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 the normal were found, but their tsh levels were still at normal range as other (35) patients. meanwhile, the failure of ovulation is considered owing to severe hypothyroidism, whereas, menstrual anomalies stem from hypothyroidism and hyperthyroidism.[35] rae et al. suggested, ovaries sensitivity to estrogen and its inflammatory state be affected due to the status of the thyroid hormone, which consequently raise a gynecological health and disease impact.[28] according to kang et al. advocated the absence of possible relationship in the history of hyperthyroidism and hypothyroidism with ovarian cancer.[36] however, current hyperthyroidism history was linked with the increment of 5-year mortality among ovarian cancer patients. yet, a mildly decreased mortality risk was associated with hypothyroidism.[4] while other scholars stated that, although, hyperthyroidism, and oc association apparently persist only at an epidemiological level. yet, the demonstration of the association mechanism has not been prevailed.[37] table 1: tsh and ca125 ratio of patient group and control group parameters no. all patients no. c.g t-test p value p mean±se mean±se tsh (miu/ml) 44 2.20±0.17 20 2.00±0.25 0.526 n.s ca-125 (u/ml) 44 27.68±4.79 20 14.43±0.84 0.009 h.s** p: probability, *p<0.05: significant, p≥0.05: non-significant, **p<0.01: highly significant table 2: cbc parameters of patient group and control group hematological parameters no. all patients no. c.g t-test p value p mean±se mean±se wbc (×109/l) 44 8.19±0.33 20 6.79±0.39 0.010 s* lymphocyte (×109/l) 44 2.55±0.13 20 2.95±0.23 0.158 n.s monocyte (×109/l) 44 0.46±0.02 20 0.45±0.05 0.884 n.s granulocyte (×109/l) 44 5.16±0.22 20 4.63±0.34 0.213 n.s hb (×109/l) 44 12.06±0.26 20 12.59±0.30 0.203 n.s rbc (×109/l) 44 4.48±0.06 20 5.03±0.14 0.002 h.s** p: probability, *p<0.05: significant, p≥0.05: non-significant, **p<0.01: highly significant figure 1: correlation between ca125 and tsh ratio in patient group and control group figure 2: correlation between ca125 and wbc ratio in patient group and control group figure 3: correlation between ca125 and lymphocyte ratio in patient group and control group figure 4: correlation between ca125 and monocyte ratio in patient group and control group ubeid, et al.: parameter affecting ca-125, tsh, and cbc 5 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 bishara et al. reported that even when a fractional count of lymphocyte to wbcs total is significantly associated with mortality. while an elevated count of monocyte showed a substantial recurrence association. however, a raised count of eosinophil was a mortality and recurrence predictor among the ovarian cancer women. equally, bekmezci and gunenc[38] reported that, in older women both preoperative nlr alongside to ca125 are to be a conceivable prognostic malignant adnexal mass indicator.[39] yildirim et al. demonstrated that in patients of ovarian cancer, nlr, and plr values are significantly high when compared to patients of benign ovarian tumor.[40] furthermore, no change was observed in granulocyte, monocyte, and lymphocyte values as the ca125 was elevated.[41] obermair et al. illustrated that the inversed hb concentrations were associated to the disease stage along with the chronic inflammatory status severity. while, förhécz et al.[42] suggested that red cell distribution width (rdw) was linked with the stage of the ovarian cancer, if the stage is advanced or high, equally the rdw is high.[43] similarly, it was positively correlated with ca125 levels. where, an elevated rdw in peripheral blood smears probably to correlate with iron, vitamin b12, or folic acid, and hb deficiencies; hemolysis or the transfusion of the blood is a reason of elevating the rdw.[44] in the present work, there was no significant change of hb levels, as the ca25 levels elevated among non-cancerous women. but a lower rbc count of patients was recognized. lehtovirta et al. and juma and ubeid demonstrated that, the ca125 test should not be conducted directly, before, or during menstrual cycle, as the ca125 physiological elevation levels possibly to provide false and positive results. in the present study, patients with high ca125 were found, with normal levels of each parameter wbc, hb, and tsh, but without respect to their menstrual cycle. as it was revealed by the previous studies, that hyperthyroidism and mortality rate are correlated to patients of ovarian cancer. in addition, high rdw signifies the disease advanced stage. whereas, ca125 was inadequate biomarker for diagnosing early ovarian cancer.[45,46] as the age rises, many factors will have effect on human health. in this regard many determinants impact the ca125 level especially in females who passed menopause and ovarian cancer disease free. these factors are: age, hysterectomy, obesity, race/ethnicity, and the smoking background.[47] the serum of carbohydrate antigen ca125 serves an indicator to the inclination risk of ovarian cancer. it is also obesity indicator which is considered a significant element to the metabolic syndrome.[48] it is worthy to mention that, the level of serum ca125 showed a significant inclination in patients of chronic heart failure.[49,50] conclusion the study revealed that there was no association regarding ca125 serum and the levels of tsh. however, the elevated ca125 levels in healthy females could be related to other noncancerous factors such as menstrual cycle, or postmenopausal. figure 5: correlation between ca125 and granulocyte ratio in patient group and control group figure 6: correlation between ca125 and rbc ratio in patient group and control group figure 7: correlation between ca125 and hb ratio in patient group and control group figure 8: correlation between ca125 and age ubeid, et al.: parameter affecting ca-125, tsh, and cbc 6 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 furthermore, wbc parameters and hb levels were not affected by ca125. the only parameters that were affected with ca125 elevation were rbc. the study results show that there was no association between ca125 and women age. references 1. z. r. yurkovetsky, f. y. linkov, d. e malehorn and a. e. lokshin. multiple biomarker panels for early detection of ovarian cancer. future oncology, vol. 2, no. 6, pp. 733-741, 2006. 2. k. a. sikaris. ca125-a test with a change of heart. heart, lung and circulation, vol. 20, no. 10, pp. 634-640, 2011. 3. r. c. bast, d. badgwell, z. lu, r. marquez, d. rosen, j. liu, k. a. baggerly, e. n. atkinson, s. skates, z. zhang, a. lokshin, u. menon, i. jacobs and k. lu. new tumor markers: ca125 and beyond. international journal of gynecologic cancer, vol. 15, pp. 274-281, 2005. 4. a. n. minlikeeva, j. l. freudenheim, r. a. cannioto, k. h. eng, j. b. szender, p. mayor, j. l etter, d. w. cramer, b. diergaarde, j. a. doherty, t. dörk, r. edwards, a. defazio, g. friel, m. t goodman, p. hillemanns, e. høgdall, a. jensen, s. j. jordan, b. y. karlan, s. k. kjær, r. klapdor, k. matsuo, m. mizuno, c. m. nagle, k. odunsi, l. paddock, m. a. rossing, j. m. schildkraut, b. schmalfeldt, b. h. segal, k. starbuck, k. l. terry, p. m. webb, e. zsiros, r. b. ness, f. modugno, e. v. bandera, j. chang-claude and k. b. moysich. history of thyroid disease and survival of ovarian cancer patients: results from the ovarian cancer association consortium, a brief report. british journal of cancer, vol. 117, no. 7, pp. 1063-1069, 2017. 5. e. shinderman-maman, k. cohen, c. weingarten, d. nabriski, o. twito, l. baraf, a. hercbergs, p. j. davis, h. werner, m. ellis and o. ashur-fabian. the thyroid hormone-αvβ3 integrin axis in ovarian cancer: regulation of gene transcription and mapkdependent proliferation. oncogene, vol. 35, no. 15, pp. 19771987, 2016. 6. r. kimura, k. imaeda, t. mizuno, k. wakami, k. yamada, n. okayama, y. kamiya and t. joh. severe ascites with hypothyroidism and elevated ca125 concentration: a case report. endocrine journal, vol. 54, no. 5, pp. 751-755, 2007. 7. k. a. williams, s. i. labidi-galy, k. l. terry, a. f. vitonis, w. r. welch, a. goodman and d. w. cramer. prognostic significance and predictors of the neutrophil-to-lymphocyte ratio in ovarian cancer. gynecologic oncology, vol. 132, no. 3, pp. 542-550, 2014. 8. m. u. ceran, u. tasdemir, e. colak and t. güngör. can complete blood count inflammatory parameters in epithelial ovarian cancer contribute to prognosis?-a survival analysis. journal of ovarian research, vol. 12, no. 1, p. 16, 2019. 9. a. g. van der zee, h. w. de bruijn, m. krans, e. m. de cuyper, h. hollema, p. c. limburg, h. w de bruijn, h. hollema, j. bijzet, m. krans and e. g. de vries. higher levels of interleukin‐6 in cystic fluids from patients with malignant versus benign ovarian tumors correlate with decreased hemoglobin levels and increased platelet counts. cancer, vol. 75, no. 4, pp. 1004-1009, 1995. 10. a. maccio, c. madeddu, d. massa, m. c. mudu, m. r. lusso, g. gramignano, r. serpe, g. b. melis and g. mantovani. hemoglobin levels 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j. park, h. y. kim, h. b. kim, s. b. koh and j. namkung. diagnostic accuracy of inflammatory markers for distinguishing malignant and benign ovarian masses. journal of cancer, vol. 9, pp. 1165-1172, 2018. 16. committee on the state of the science in ovarian cancer research; board on health care services; institute of medicine; national academies of sciences, engineering, and medicine. ovarian cancers: evolving paradigms in research and care. national academies press, washington, dc, 2016. 17. j. a. rauh-hain, t. c. krivak, m. g. del carmen and a. b. olawaiye. ovarian cancer screening and early detection in the general population. reviews in obstetrics and gynecology, vol. 4, no. 1, pp. 15-21, 2011. 18. m. thun, m. s. linet, j. r. cerhan, c. a. haiman and d. schottenfeld. cancer epidemiology and prevention: oxford university press, oxford, 2017. 19. n. mahdavifar, r. pakzad, m. ghoncheh, h. s. gandomani and h. salehiniya. epidemiology, incidence, and mortality of gallbladder cancer and its relation with development in the world. annals of tropical medicine and public health, vol. 10, pp. 563-517, 2017. 20. y. moradi, m. jafari, s. chaichian, s. khateri, a. akbarian, b. moazzami, k. mansori, y. mahmodi and s. samie. trends in ovarian cancer incidence in iran. international journal of cancer management, vol. 9, p. e5452, 2016. 21. n. bhatla and a. jones. the world ovarian cancer coalition atlas. global trends in incidence, mortality and survival. the world ovarian cancer coalition, toronto, on. 2018. 22. r. bast, m. feeney, h. lazarus, l. m. nadler, r. b. colvin and r. c. knapp. reactivity of a monoclonal antibody with human ovarian carcinoma. the journal of clinical investigation, vol. 68, no. 5, pp. 1331-1337, 1981. 23. n. scholler and n. urban. ca125 in ovarian cancer. biomarkers in medicine, vol. 1, no. 4, pp. 513-523, 2007. 24. h. yesilyurt, b. seckin, a. aktulay and s. ozyer. age-stratified analysis of tumor markers and tumor characteristics in adolescents and young women with mature cystic teratoma. journal of the chinese medical association, vol. 81, pp. 499-504, 2018. 25. k. a. habib, m. g. jumaa and m. j. hussein. determination of serum ca125 and evaluate its efficiency as screening tool for early detection of ovarian tumors. baghdad science journal, vol. 12, pp. 55-62, 2015. 26. m. h. ubeid. prevalence of leukocytes in type 2 diabetic patients in erbil city. medical journal of babylon, vol. 17, p. 19, 2020. 27. c. l. gyftaki, e. politi, m. liontos, k. saltiki, n. theocharis papageorgiou, d. h. thomakos, d. haidopoulos, a. rodolakis, m. alevizaki, a. bamias and a. dimopoulos. differential transcriptional and protein expression of thyroidstimulating hormone receptor in ovarian carcinomas. international journal of gynecological cancer, vol. 24, no. 5, pp. 851-856, 2014. 28. m. rae, o. gubbay, a. kostogiannou, d. price, h. critchley and s. hillier. thyroid hormone signaling in human ovarian surface epithelial cells. the journal of clinical endocrinology and metabolism, vol. 92, pp. 322-327, 2007. 29. m. rasool, m. i. naseer, k. zaigham, a. malik, n. riaz, r. alam, a. manan, i. a. sheikh, m. asif. comparative study of alterations in tri-iodothyronine (t3) and thyroxine (t4) hormone levels in breast and ovarian cancer. pakistan journal of medical sciences, vol. 30, p. 1356, 2014. 30. k. żak, s. niedobylski, m. leziak, k. maliszewska, m. czapska and k. skórzyńska-dziduszko. have thyroid hormones an impact ubeid, et al.: parameter affecting ca-125, tsh, and cbc 7 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 1-7 author queries??? aq1: kindly provide running title aq2: kindly check the author name does not match with reference list aq3: kindly provide complete reference aq4: kindly provide foot note for significant symbol of “**” aq5: kindly provide foot note for significant symbol of “*” on ovarian cancer? journal of education, health and sport, vol. 8, pp. 803-809, 2018. 31. m. den ouden, j. ubachs, j. stoot and j. van wersch. whole blood cell counts and leucocyte differentials in patients with benign or malignant ovarian tumours. european journal of obstetrics and gynecology and reproductive biology, vol. 72, pp. 73-77, 1997. 32. a. s. juma, m. h. ubeid and t. s. salih. anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers of erbil governorate. cihan university-erbil scientific journal, vol. 3, pp. 85-89, 2019. 33. a. s. juma, m. h. ubeid, s. s. haydar and t. s. salih. some immunological and hematological parameters among refugees in kawergosk camp-erbil governorate. cihan university-erbil scientific journal, vol. 3, pp. 80-84, 2019. 34. r. s. nomelini, t. m. da silva, b. m. t. murta and e. f. c. murta. parameters of blood count and tumor markers in patients with borderline ovarian tumors: a retrospective analysis and relation to staging. international scholarly research notices, vol. 2012, p. 947831, 2012. 35. g. e. krassas. thyroid disease and female reproduction. fertility and sterility, vol. 74, pp. 1063-1070, 2000. 36. j. h. kang, a. s. kueck, r. stevens, g. curhan, i. de vivo, b. rosner, erik alexander, s. s. tworoger. a large cohort study of hypothyroidism and hyperthyroidism in relation to gynecologic cancers. obstetrics and gynecology international, vol. 2013, p. 743721, 2013. 37. t. m. markman, m. markman and b. w. clark. case of ovarian cancer in a woman with undiagnosed graves’ disease: a case report and review of the literature. case reports in oncology, vol. 10, pp. 452-454, 2017. 38. m. bekmezci and o. gunenc. the role of neutrophil/lymphocyte ratio (nlr) with ca 125 in preoperative predictıon of malignancy in adnexial mass. medicine, vol. 7, pp. 857-60, 2018. 39. s. bishara, m. griffin, a. cargill, a. bali, m. gore, s. kaye, j. h. shepherd, p. o. van trappen. pre-treatment white blood cell subtypes as prognostic indicators in ovarian cancer. european journal of obstetrics and gynecology and reproductive biology, vol. 138, pp. 71-75, 2008. 40. m. yildirim, b. d. cendek and a. f. avsar. differentiation between benign and malignant ovarian masses in the preoperative period using neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios. molecular and clinical oncology, vol. 3, pp. 317-321, 2015. 41. t. s. salih, a. s. juma and m. h. ubeid. occupational exposures and associated health effects among dumpsite workers. cihan university-erbil scientific journal, vol. 5, pp. 1-5, 2021. 42. y. qin, p. wang, z. huang, g. huang, j. tang, y. guo, p. huang, z. lai and f. lin. the value of red cell distribution width in patients with ovarian cancer. medicine, vol. 96, p. e6752, 2017. 43. a. obermair, a. handisurya, a. kaider, p. sevelda, h. kölbl and g. gitsch. the relationship of pretreatment serum hemoglobin level to the survival of epithelial ovarian carcinoma patients: a prospective review. cancer, vol. 83, pp. 726-731, 1998. 44. z. förhécz, t. gombos, g. borgulya, z. pozsonyi, z. prohászka and l. jánoskuti. red cell distribution width in heart failure: prediction of clinical events and relationship with markers of ineffective erythropoiesis, inflammation, renal function, and nutritional state. american heart journal, vol. 158, pp. 659666, 2009. 45. p. lehtovirta, d. apter and u. h. stenman. serum ca 125 levels during the menstrual cycle. british journal of obstetrics and gynaecology, vol. 97, pp. 930-933, 1990. 46. a. s. juma and m. h. ubeid. a microbiological survey of ear infections in inhabitants of erbil city. cihan university-erbil scientific journal, vol. 6, pp. 62-67, 2022. 47. c. c. johnson, b. kessel, t. l. riley, l. r. ragard, c. r. williams, j. l. xu, s. s. buys and prostate, lung, colorectal and ovarian cancer project team. the epidemiology of ca-125 in women without evidence of ovarian cancer in the prostate, lung, colorectal and ovarian cancer (plco) screening trial. gynecologic oncology, vol. 110, pp. 383-389, 2008. 48. n. s. joo, k. n. kim and k. s. kim. serum ca125 concentration has inverse correlation with metabolic syndrome. journal of korean medical science, vol. 26, pp. 1328-1332, 2011. 49. n. t. kouris, i. d. zacharos, d. d. kontogianni, g. s. goranitou, m. d. sifaki, h. e. grassos, e. m. kalkandi and d. k. babalis.the significance of ca125 levels in patients with chronic congestive heart failure. correlation with clinical and echocardiographic parameters. european journal of heart failure, vol. 7, pp. 199-203, 2005. 50. h. m. turk, h. pekdemir, s. buyukberber, a. sevinc, c. camci, r. kocabas, tarakcioglu and n. m. buyukberbe. serum ca 125 levels in patients with chronic heart failure and accompanying pleural fluid. tumor biology, vol. 24, pp. 172-175, 2003. . 85 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 85-89 research article anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers of erbil governorate ameena s. m. juma, muhsin h. ubeid, tanya s. salih* department of biology, cihan university-erbil, kurdistan region, iraq abstract the present study aimed to detect the presence of anti-toxoplasma, anti-rubella, and anti-cytomegalovirus (cmv) antibodies in the sera of dumpsite workers of erbil governorate. eighty nine male dumpsite (kany qrzhala, erbil governorate) workers were included in this study. serum was obtained for the detection of anti-toxoplasma, anti-rubella, and anti-cmv antibodies using an automated cobas e411 immunoassay analyzer. no anti-toxoplasma igm antibodies were detected in any of the workers’ sera, while (25.84%) showed a positive result for anti-toxoplasma igg antibodies. all workers’ sera had no anti-rubella igm and igg2 antibodies, while (62.92%) of them revealed the presence of igg1 in their sera. anti-cmv igm was found in (2.25%) of the sera, while (13.50%) of the sera revealed the presence of anti-cmv igg antibodies. keywords: cytomegalovirus, dumpsite workers, municipal solid waste, rubella, solid waste disposal, toxoplasma gondii introduction workers handling wastes are at high risk to be exposed to hazardous biological and chemical factors including different vapors, smoke, fumes, and dust in addition to being exposed to chemical substances and infectious pathogens, including viruses, bacteria, fungi, and even parasites. according to some studies on workers in this sector, they have been found to experience more work-related symptoms and illnesses than other occupational groups.[1,2] waste materials provide a favorable environment in which microorganisms can thrive because of the presence of nutrients and moisture. pathogenic microorganisms are well known to be present in medical and clinical waste or the waste derived from animal and human origins, posing a risk of infection to those handling these materials.[3] irritation of the mucous membrane due to exposure to toxic materials or allergenic effects experienced after inhalation of a large number of microorganisms and their fragments is usually the greatest work-related hazard.[4] waste may include medical and clinical laboratory waste that may include extremely infectious agents, including a wide range of viruses. parasitic diseases can also be acquired by these workers through vectors such as insects or domestic or wild animals such as rodents or cats and dogs that are harboring disease-causing microorganisms or acting as mechanical vectors. these kinds of vector-borne diseases include toxoplasmosis.[5] toxoplasmosis is caused by the intracellular parasite toxoplasma gondii that infects up to a third of the world’s population. infection is mainly acquired by ingestion of food or water that is contaminated with oocysts shed by cats or by consuming contaminated meat containing tissue cysts. in addition, the infection may be acquired by coming in contact with cat feces containing oocysts.[6,7] primary infection is usually subclinical, but in some patients, cervical lymphadenopathy or ocular disease can be present. in immunocompromised patients, reactivation of latent disease can cause life-threatening encephalitis.[8] rubella (german measles) is caused by an rna virus of the togavirus group. it is a contagious disease. most people who get rubella usually have a mild illness, with symptoms that can include a low-grade fever, sore throat, and a rash that starts on the face and spreads to the rest of the body, in addition to the drastic effect of the virus on the fetus in pregnant women.[9] cytomegalovirus (cmv) is an infection caused by a dna virus of the herpes group. the common modes of infections corresponding author: tanya s. salih, department of biology, cihan university, erbil, kurdistan region, iraq. e-mail: tanya.salam@cihanuniversity.edu.iq received: apr 14, 2019 accepted: apr 26, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp85-89 copyright © 2019 ameena s. m. juma, muhsin h. ubeid, tanya s. salih. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ juma, et al.: anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers 86 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 85-89 are through saliva, urine, stool, breast milk, and unscreened blood transfusion.[10] the diagnosis of acute toxoplasma, rubella, and cmv infection can be established by the demonstration of seroconversion in paired sera or by the demonstration of specific igm antibodies.[11] this study was conducted to detect the presence of antitoxoplasma, anti-rubella, and anti-cmv antibodies in the sera of dumpsite workers of erbil governorate. this may reflect the health status of those workers and their exposure to infectious agents. materials and methods subject selection eighty-nine male dumpsite (kany qrzhala, erbil governorate) workers were included in this study. a questionnaire sheet [figure 1] was filled out for each worker included in the study. sample collection five milliliter of venous blood was collected by disinfecting the antecubital fossa with 70% ethanol and using a disposable syringe questionnaire sheet name: no. gender: status married single age years weight kg education primary secondary college non residence rural urban smoker yes no diabetic yes no hypertensive yes no hypersensitivity allergy asthma eczema other diseases any drug intake other notes figure 1: questionnaire sheet for each worker included in a study concerning the detection of different antibodies in the sera of kany qrzhala dumpsite workers, erbil governorate juma, et al.: anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers 87 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 85-89 with a 23-gauge needle after applying a tourniquet. the blood was placed in a plain tube and left to the stand for 1 h at room temperature for clot formation. the tube was centrifuged for 10 min at 4°c at 450×g for serum collection. the serum was then aspirated by using a pasteur pipette and was dispensed into sterile glass tubes (0.5 ml in each) and was stored at −20°c until used. detection of anti-toxoplasma, anti-rubella, and anti-cmv antibodies the stored sera were used for the detection of anti-toxoplasma igg and igm antibodies; anti-rubella igg1, igg2, and igm antibodies; and anti-cmv igg and igm antibodies, using automated cobas e411 immunoassay analyzer, following the instructions provided with the kits. all sera and test reagents were brought to room temperature. results workers’ information this study involved 89 men with ages ranging from 12 years to 65 years. 92.13% of the workers lived in rural areas, while 7.87% lived in urban areas. 58.43% were smokers and 41.57% were nonsmokers. 38.20% were single, while 61.80% were married. 37.08% were illiterate, 40.45% had primary school education, 17.97% had secondary school education, 2.25% had a diploma, and 2.25% had a b.sc. degree [table 1]. frequency of anti-toxoplasma antibodies none of the workers’ sera revealed the presence of antitoxoplasma igm antibodies, while 25.84% were found to be positive for anti-toxoplasma igg antibodies [table 2]. frequency of anti-rubella antibodies none of the workers’ sera revealed the presence of anti-rubella igm and igg2 antibodies, while 62.92% were found to be positive for anti-rubella igg1 antibodies [table 3]. frequency of anti-cmv antibodies igm antibodies against cmv were detected in two workers (2.25%). 13.5% of the workers revealed the presence of anticmv antibodies in their sera [table 4]. a summary of the results is summarized the figure 2. discussion about 40% of the world’s waste ends up in large rubbish dumps that are usually located close to urban populations in underdeveloped and developing countries. this poses a serious threat to the environment, in addition to human health.[12] increase of solid waste is mainly due to urbanization and population growth and that in turn creates problems for municipal corporation to properly manage that kind and amount of waste. it has been shown that excreta and other liquid and solid waste from household and the community represent a health hazard and are the causes of many serious infection spread.[13] improper handling of wastes represents a serious risk to human health, especially those working in the dumpsites; table 1: demographic description of kany qrzhala dumpsite workers, erbil governorate parameter n (%) live in rural areas 82 (92.13) live in urban areas 7 (7.87) smoker 52 (58.43) non-smoker 37 (41.57) single 34 (38.20) married 55 (61.80) illiterate 33 (37.08) primary school education 36 (40.45) secondary school education 16 (17.97) diploma degree 2 (2.25) b.sc. degree 2 (2.25) table 2: anti-toxoplasma igm and igg antibodies in the sera of kany qrzhala dumpsite workers, erbil governorate parameter ig number of workers number of positive cases % number of negative cases % toxoplasma igm 89 0 0 89 100 igg 89 23 25.84 66 74.16 table 3: anti-rubella igm, igg1 and igg2 antibodies in the sera of kany qrzhala dumpsite workers, erbil governorate parameter ig number of workers number of positive cases % number of negative cases % rubella igm 89 0 0 89 100 igg1 89 56 62.92 33 37.08 igg2 89 0 0 89 100 table 4: anti-cytomegalovirus igm and igg antibodies in the sera of kany qrzhala dumpsite workers, erbil governorate parameter ig number of workers number of positive cases % number of negative cases % cytomegalovirus igm 89 2 2.25 87 97.75 igg 89 12 13.50 77 86.50 juma, et al.: anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers 88 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 85-89 protection of these people from these health threats should be a major concern. these wastes may also include hospitals and clinics wastes that pose a major health issue, in addition to municipal wastes. moreover, it is well known that insect and rodent vectors are attracted to waste sites and can spread diseases such as cholera and dengue fever. public health service in the u.s.a. identified 22 human diseases that are linked to improper management of solid wastes. waste worker and pickers in developing countries are seldom protected from direct contact and injury.[14-16] despite the fact that the results of the current study showed low number of individuals profiling antibodies against the three microorganisms included; however, their presence does implicate that such workers are at risk of acquiring serious infections. dumpsite workers are continuously exposed to dust that contains many harmful substances including bioaerosols, which are airborne materials and are associated with microorganisms. this will impose a biological hazard due to direct contact, inhalation, or ingestion of these particles by the workers, leading to diseases.[17] t. gondii is an obligate intracellular protozoan parasite that is capable of infecting a variety of intermediate hosts including humans. infected definitive hosts (cats) shed oocysts in feces that rapidly mature in the soil and become infectious. toxoplasmosis is acquired by humans through ingestion of food or water contaminated with cat feces or through eating undercooked meat containing viable oocysts. vertical transmission of the parasite through the placenta can also occur, leading to congenital toxoplasmosis. following the primary infection, t. gondii can remain latent for the life of the host; the risk for reactivation is highest among immunosuppressed individuals.[18] a positive toxoplasma igg result is indicative of the late current or past infection with t. gondii. a single positive toxoplasma igg result should not be used to diagnose recent infection. individuals with negative toxoplasma igg results are presumed to not have had previous exposure to t. gondii. however, the negative results may be seen in cases of remote exposure with subsequent loss of detectable antibody. seroconversion from negative to positive igg is indicative of t. gondii infection subsequent to the first negative specimen.[19] 25.84% of the individuals included in this current study showed a positive result for anti-toxoplasma igg antibodies. this may indicate a previous exposure to the parasite, which could have been acquired during their work in the dumpsite. rubella (german or 3-day measles) is a member of the togavirus family, and humans remain the only natural host for this virus. transmission is through inhalation of infectious aerosolized respiratory droplets.[20] the presence of igg antibodies is an indicator of previous infection, which is the case in the workers included in the current study. a negative cmv igm result suggests that the patient is not experiencing acute or active infection. however, a negative result does not rule out primary cmv infection. positive cmv igm results indicate a recent infection (primary, reactivation, or reinfection). igm antibody responses in secondary (reactivation) cmv infections have been demonstrated in some cmv mononucleosis patients, in a few pregnant women, and in renal and cardiac transplant patients. positive cmv igg results indicate the past or recent cmv infection. these individuals may transmit cmv to susceptible individuals through blood and tissue products.[21] 2.25% of the workers demonstrated a possible active infection, while 13.50% demonstrated a possible previous infection. those with active infections pose a hazard on the community and could pass on the infection to susceptible individuals. periodic surveys and treatment of these workers are essential to keep them healthy and prevent spread of diseases through them. workers should also be trained on the safe and proper techniques of dealing with wastes and how to protect themselves from injury and infection. in addition, protective clothing is advised to be provided to these workers to prevent any harm caused by handling waste. acknowledgments special thanks are due to kany qrzhala dumpsite workers of erbil governorate for providing the samples. figure 2: anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in the sera of kany qrzhala dumpsite workers of erbil governorate juma, et al.: anti-toxoplasma, anti-rubella, and anti-cytomegalovirus antibodies in dumpsite workers 89 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 85-89 references 1. j. bünger, b. schapper-scheele, r. hilgers and e. hallier. “a 5-year follow-up study on respiratory disorders and lung function in workers exposed to organic dust from composting plants”. international archives of occupational and environmental health, vol. 80, pp. 306-312, 2007. 2. i. m. wouters, s. spaan, j. douwes, g. doekes and d. heederik. “overview of personal occupational exposure levels to inhalable dust, 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p. a. jenum, w. foulon, a. e. semprini and d. t. dunn. “sources of toxoplasma infection in pregnant women: european multicentre case-control study. european research network on congenital toxoplasmosis”. bmj, vol. 321, pp. 142-147, 2000. 7. a. b. ismael, d. sekkai, c. collin, d. bout and m. n. meralece. “the mic 3 gene of toxoplasma gondii is a novel vaccine candidate against toxoplasmosis”. infection and immunity, vol. 71, pp. 6222-6228, 2003. 8. j. g. montoya and o. liesenfeld. “toxoplasmosis”. lancet, vol. 363, pp. 1965-1976, 2004. 9. c. coulter, r. wood and j. robson. “rubella infection in pregnancy”. communicable diseases intelligence, vol. 23, pp. 93-96, 1999. 10. s. stagno, m. e. dworsky, r. e. henderson, e. g. moore, p. d. walton and c. a. alford. “congenital cytomegalovirus infection: the relative importance of primary and recurrent maternal infection”. the new england journal of medicine, vol. 306, no. 16, pp. 945-949, 1982. 11. c. shashi, u. arora and a. aggarwal. “prevalence of igm antibodies to toxoplasma, rubella and cytomegalovirus infections during pregnancy”. jk science, vol. 6, no. 4, 190-192, 2004. 12. j. vidal. “smelly, contaminated, full of disease: the world’s open dumps are growing”. global development, supported by bill and melinda gates foundation, 2017. 13. p. singh. “impact of solid waste on human health: a case study of varanasi city”. international journal of scientific and engineering research, vol. 4, no. 11, pp. 1840-1842, 2013. 14. s. rathi. “alternative approaches for better municipal solid waste management in mumbai, india”. waste management, vol. 26, no. 10, pp. 1192-1200, 2006. 15. m. sharholy, k. ahmad, r. c. vaishya and r. d. gupta. “municipal solid waste characteristics and management in allahabad, india”. waste management, vol. 27, pp. 490-496, 2007. 16. n. gupta, k. k. yadav and v. kumar. “a review on current status of municipal solid waste management in india”. journal of environmental sciences, vol. 37, pp. 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[last accessed on 2019 apr 01]. 18. a. m. tenter, a. r. heckeroth and l. m. weiss. toxoplasma gondii: from animals to humans. international journal for parasitology, vol. 30, no. 12-13, p. 1217, 2000. 19. test id: toxgp: toxoplasma gondii antibody, igg. mayo clinic laboratories. available from: https://www.mayocliniclabs.com/ test-catalog/clinical+and+interpretive/34972. [last accessed on 2019 apr 01]. 20. american academy of pediatrics. rubella. in red book. 2012 report of the committee on infectious diseases. edited by lk pickering, elk grove village, il, 2012. 21. cytomegalovirus (cmv) antibodies, igm and igg, serum. centra care laboratory servicespowered by mayo clinic laboratories. available from: https://www.centracarelaboratoryservices.testcatalog.org/ show/0205369-1; https://www.centracarelaboratoryservices.test catalog.org/show/0205369-1. [last accessed on 2019 apr 01]. . 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 56-60 research article immunological aspects of trichomonas vaginalis infection in women attending maternity teaching hospital and some public health centers in erbil governorate, northern iraq avreen salahaddin nouraddin1, hadi mahdi alsakee2* 1department of serology and virology, public health laboratory, erbil health directorate, erbil, iraq, 2department of biology, cihan university-erbil kurdistan region, iraq abstract trichomonas vaginalis infection is one of the sexually transmitted diseases. it is a health problem all over the world, including iraq. it has also been associated with adverse outcomes of pregnancy and increased risk of hiv. trichomoniasis typically elicits aggressive local cellular immune responses with inflammation of the vaginal epithelium and exocervix in women and urethra in men. the present study was aimed to assess serum level of interferon gamma (ifn-γ), interleukin 10 (il-10), c-reactive protein, antiphospholipid, anticardiolipin antibodies with eosinophil count as well. four hundred and forty women with ages ranging between 16 and 60 years old (average of 34.2 years) who attended maternity teaching hospital and a number of public health centers in erbil governorate were screened for trichomoniasis by direct wet mount preparation and culture technique. serum ifn-γ, il-10, antiphospholipid, and anticardiolipin were assessed by enzyme-linked immunosorbent assay. c-reactive protein (crp) and eosinophil count were assessed by i croma and coulter, respectively. the results revealed that il-10 level (96.46 ± 1.97 pg/ml vs. 91.86 ± 1.48 pg/ml), eosinophil count (0.1 ± 0.07 × 10³/μl vs. 0.1 ± 0.01 × 10³/μl), and crp concentration (2.55 ± 0.74 mg/l vs. 2.27 ± 0.37 mg/l) were non-significantly (p > 0.05) changed in infected women in comparison with negative control group. however, serum ifn-γ level (484.83 ± 38.35 pg/ml vs. 372.15 ± 9.49 pg/ml) was significantly (p < 0.05) elevated in infected women in comparison to the control group. the results also revealed significant increase (p < 0.05) of antiphospholipid immunoglobulin g (igg) antibodies (4.75 ± 0.3 u/ml vs. 3.79 ± 0.11 u/ml), anticardiolipin igg, and anticardiolipin igm antibodies in the sera of infected women, but antiphospholipid immunoglobulin m (igm), were non significantly (p > 0.05) altered in response to t. vaginalis infection. keywords: anticardiolipin antibodies, antiphospholipid antibodies, c-reactive protein, cytokines, il-10, interferon gamma, trichomonas vaginalis infection, trichomoniasis introduction trichomoniasis is the most prevalent non-viral sexually transmitted infection worldwide, caused by an anaerobic, parasitic, flagellated protozoan, and trichomonas vaginalis. this infection is not a potentially dangerous disease that can go undiagnosed for years and is often passed on by an asymptomatic carrier.[1] humans are the only known host, the trophozoites transmitted principally through vaginal sexual intercourse and rarely through fomites. it is very successful as a pathogen causing roughly the same number of sexually transmitted diseases as chlamydia trachomatis, the most prevalent sexually transmitted bacterial pathogen.[2] worldwide, the prevalence of t. vaginalis varies from 2% to >50% depending on region, country, gender, and demographics of the population specifically evaluated.[3] an estimated 180 million infection acquired annually worldwide.[4] it is emerging as a serious reproductive tract pathogen, mainly affecting minorities and people living in poor or disadvantaged communities. t. vaginalis is listed as one of the five neglected parasitic infections in the united states.[5] nearly half of all women with t. vaginalis are asymptomatic. signs of infection in symptomatic women include vaginal discharge, odor, edema or erythema, and colpitis macularis (strawberry cervix) which is characterized by punctate hemorrhagic lesions, vulval irritation, and inflammation. other complaints include dysuria, a yellowish-green frothy discharge, pruritis, dyspareunia, and lower abdominal pain.[2,6] the infected males are usually asymptomatic and infection is usually self-limiting, and thus, diagnosis is often difficult. corresponding author: hadi mahdi alsakee, department of biology, cihan university, erbil, kurdistan region, iraq. e-mail: hadialsakee@gmail.com received: nov 13, 2018 accepted: apr 17, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp56-60 copyright © 2019 avreen salahaddin nouraddin, hadi mahdi alsakee. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ nouraddin and alsakee: immunological aspects of trichomonas vaginalis infection in women 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 56-60 the general infection areas include urethra, external genitalia, prostate, and epididymis.[7] it is also a known leading cause of prostatitis and urethritis.[8] furthermore, serious complications may be associated with trichomoniasis such as premature rupture of the placental membranes, premature labor, and low birth weight in pregnant women infection, infertility, cervical cancer, pelvic inflammatory disease, birth outcomes, and increase in human predisposition to immunodeficiency virus (hiv), its transmission, and acquisition.[9,10] infection with t. vaginalis is associated with higher genital hiv-1 levels. the treatment of women infected with t. vaginalis results in a 4.2-fold reduction in the quantity of hiv-1 in vaginal secretions.[9] some recent studies have shown that trichomoniasis may be considered as one of the predisposing factors for some human disease such as prostate and cervical cancer.[11] the outcome of infection with trichomonas may be due to genetic variability of the isolates and the host immune response.[12] in iraq, several studies have been conducted in respect to t. vaginalis epidemiology and diagnosis.[13-15] however, little is known concerning the immunological aspects of trichomoniasis among women in erbil. the present study was aimed to assess some immunological parameters including interferon gamma (ifn-γ), interleukin 10 (il-10), c-reactive protein, antiphospholipid and anticardiolipin antibodies, and eosinophil count as well. materials and methods patients and study setting t. vaginalis infection was screened in 440 women with ages ranging between 16 and 60 years old (average of 34.2 years) who attended maternity teaching hospital and a number of public health centers in erbil governorate, northern iraq, over a period of 10 months from september 2012 to july 2013. as a control group, 32, age matched, women who were apparently healthy with no history or clinical evidence of other diseases and who revealed negative results for trichomoniasis by both direct wet mount and culture technique, were enrolled as negative control group. both patients and healthy women were interviewed personally before collection of blood sample and vaginal swab. for each subject, a well-structured questionnaire was filled carefully and included patient name, mobile number, age, address, residency, occupation, marital status, educational level, gestation, and symptoms or history of symptoms (vaginal discharge, fever, and cervix), menstrual cycle, history of allergy or allergic condition, and history of other diseases. specimen collection high vaginal swabs for each subject, two high vaginal swabs were collected under aseptic condition using a sterile cotton swab. one kept in 2 ml saline solution and used for direct microscopic examination, while the second swab was kept in 2 ml of culture medium which was used for the recovery of t. vaginalis. blood sample collection and processing blood samples were collected from the patients for the assessment of immunological parameters (il-10, ifn-γ, anticardiolipin antibodies, antiphospholipid antibodies, c-reactive protein (crp), and eosinophil count). an amount of 5 ml of peripheral blood was collected from each subject by venipuncture. half of the blood was centrifuged for serum separation, aliquot and kept at −40°c, and the rest was used as whole blood for quantitative determination of eosinophils by fully automated hematology coulter. detection of t. vaginalis t. vaginalis infection was detected initially by direct wet mount preparation, in which one of the duplicate vaginal swabs was used to make a direct smear on a glass slide with a drop of normal saline, the slide was initially scanned at ×100, looking for motile trichomonads and then at ×400 to confirm the motility, flagella movement, and morphologic features of the organism. the high vaginal swabs were also cultivated on trichomonas medium (oxoid, uk) supplemented with 8% heat-inactivated horse serum (oxoid, uk) following the manufacturer’s instructions. to suppress bacterial and fungal growth, 500 μg/ml of streptomycin (cox pharmaceutical ltd, uk) and 100 μg/ml of chloramphenicol (oxoid, uk) were added to the complete medium. the ph of complete working medium was adjusted using 1 n hcl and the working medium was sterilized using millipore filter unit (0.2 μm). inoculated cultures were incubated at 37°c and followed up, microscopically for the presence of motile trophozoites at 24, 48, and 72 h of incubation.[14] quantitative estimation of human il-10 by enzyme-linked immunosorbent assay (elisa) the omnikine™ human il-10 and ifn-γ elisa kits (assay biotech, usa) were used for quantitative estimation of il-10 and ifn-γ in the sera of the subjects in accordance with the leaflet provided by the manufacturer. quantitative estimation of high sensitive crp (hs-crp) in the sera of the studied groups hs-crp was estimated in the sera of the studied groups using i-chroma™ hs-crp kit (boditech med inc., uk). the procedure was done according to the instructions provided by the manufacturer. quantitative estimation of cardiolipin antibody immunoglobulin g (igg) and immunoglobulin m (igm) anticardiolipin antibodies (igg and igm) and antiphospholipid antibodies (igg and igm) were quantitatively estimated using elisa kits (aesku diagnostics, germany) and following the instructions provided by the manufacturer. ethical consideration this study was approved by medical ethics committee in the college of medicine, hawler medical university, erbil, and an informed consent was taken from each recruited woman. statistical analysis the association of two categorical variables or difference between two proportions was assessed by chi-square test. nouraddin and alsakee: immunological aspects of trichomonas vaginalis infection in women 58 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 56-60 analysis of quantitative data was estimated using student’s t-test and anova. calculation of means and standard deviation was made for different parameters. p ≤ 0.05 was considered statistically significant. results out of 440 vaginal discharge samples, 14 (3.18%) revealed positive culture for t. vaginalis; however, the number of positive cases was only 12 (2.73%) when direct wet mount technique was used [table 1]. thus, considering culture technique as a gold standard for the diagnosis, the sensitivity (78.5%) of direct wet mount technique was lower than that revealed by cultivation. the 14 positive samples were subjected to immunological parameters. table 2 illustrates the mean concentration of il-10, ifn-γ, and crp in addition to eosinophil count in women with trichomoniasis and women of healthy control group. ifn-γ level (484.83 ± 38.35) was significantly elevated in the sera of women with trichomoniasis when compared with those of healthy control group (372.15 ± 9.49). however, il-10 level, crp concentration, and eosinophil count were non-significantly (p > 0.05) altered in response to t. vaginalis infection. the mean concentration of serum antiphospholipid antibodies and anticardiolipin antibodies for both infected women and control group revealed a significant increase (p < 0.05) in antiphospholipid igg, anticardiolipin igg, and anticardiolipin igm. however, the level of antiphospholipid igm was non-significantly altered (p > 0.05) [table 3]. discussion crp is a known marker of systemic inflammation.[16] in the current study, the level of crp was non-significantly altered in infected women when compared with the control. this finding was inconstant with that obtained by shaker and hussein[17] who revealed that the level of serum crp was significantly elevated in women with trichomoniasis. vaginal bacterial, fungal, and parasitic coinfection significantly change the serum levels of many immunological parameters including crp and cytokines.[18] the results revealed non-significant changes in the eosinophil count in both infected and non-infected control groups. this result was in agreement with that obtained by al-gazali et al.[19] eosinophilia is a common feature of allergic conditions and parasitic infections, in particular, helminthic infection. allergens and also excretory-secretory products of helminths activate th2 arm of the immune system through which il-5 is expressed and this later is an eosinophil growth factor.[20] in trichomoniasis, t helper type 1 (th1) is more likely activated since it could be explained by a significant level of ifn-γ obtained in this study in women with trichomoniasis. ifn-γ level was significantly increased in response to t. vaginalis infection, but no significant change was observed in il-10 level. controversial studies have been published concerning cytokines profile that may be induced in response to t. vaginalis infection. some previous studies in vitro revealed that exposure to t. vaginalis stimulated il-8 secretion in human monocytes,[21] neutrophils,[22] and table 1: trichomonas vaginalis infection among women using direct wet mount and culture techniques diagnostic method number of examined number of positive samples (%) number of negative samples (%) direct wet mount preparation 440 11 (2.5) 429 (97.5) culture 440 14 (3.2) 426 (96.8) χ2=0.371 df=1 p=0.686 table 2: immunological parameter in women with trichomoniasis and control groups immunological parameter infected women (n=12) (mean±se) control group (n=30) (mean±se) t-test p value il-10 (pg/ml) 96.46±1.97 91.86±1.48 1.86 >0.05 ifn-γ (pg/ml) 484.83±38.35 372.15±9.49 2.85 <0.05 crp (mg/l) 2.55±0.74 2.27±0.37 0.33 >0.05 eosinophil (103/μl) 0.1±0.07 0.1±0.01 0.49 >0.05 crp: c-reactive protein, ifn: interferon gamma, il-10: interleukin 10 table 3: serum level of antiphospholipid and anticardiolipin antibodies in women with trichomoniasis and control group autoantibody infected women (n=12) (mean±se) control group (n=30) (mean±se) t-test p value antiphospholipid antibody (u/ml) igg 4.75±0.3 3.79±0.11 2.91 <0.05 igm 3.004±0.45 2.61±0.11 0.82 >0.05 anticardiolipin antibody (gpl/ml) igg 3.32±0.31 2.42±0.07 2.75 <0.05 igm 2.89±0.46 1.85±0.07 2.2 <0.05 igg: immunoglobulin g, igm: immunoglobulin m nouraddin and alsakee: immunological aspects of trichomonas vaginalis infection in women 59 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 56-60 vaginal epithelial cells.[23] it was further demonstrated that the cytokines il-2 and ifn-γ were secreted by murine lymphocytes infected with t. vaginalis. furthermore, t. vaginalis was also stimulated the secretion of gm-csf but not il-6 and il-1β in pregnant women.[24] however, han et al.[25] reported that tnf-α, il-1β, and il-6 were significantly expressed by human macrophages on exposure to t. vaginalis in vitro. the parasite strain may also be concerned in the immunological studies of trichomoniasis.[25] malla et al.[26] found that il-2 and ifn-γ were significantly higher in mice infected with t. vaginalis isolated from asymptomatic women as compared to isolates from symptomatic women group and in control uninfected animals. in the present study, polymorphonuclear cells that are associated with the infection may be implicated in the induction of significance levels of ifn-γ in t.vaginalis infected women in comparison to non-infected control group. recruitment of polymorphonuclear cells and other inflammatory cells to vaginal milieu in response to t. vaginalis infection is induced by a parasite surface adhesion molecules. fichorova et al.[27] have found that t. vaginalis lipophosphoglycan (lpg), but not lpg from tritrichomonas foetus, the causative agent of bovine trichomoniasis induced a selective upregulation of chemotactic cytokines by human endocervical, ectocervical, and vaginal epithelial cells, which do not express toll-like receptor 4/ md2.[27] t. vaginalis lpg triggered interleukin 8 (il-8), which promotes the adhesion and transmigration of neutrophils across the endothelium, and macrophage inflammatory protein 3α, which is a chemoattractant for immune cells and is essential for dendritic cell maturation. these effects were dose dependent that relived in the absence of cytotoxicity and il-1β release utilizing, in part, a signaling pathway independent from the toll-like/il-1 receptor adaptor protein myd88. this study may also support our finding that crp was non-significantly changed in response to t. vaginalis infection. despite of the presence of inflammatory cells in the vaginal milieu that is later released as a symptomatic vaginal discharge, crp concentration was non-significantly changed that may lead to a suggestion that parasite lpg induced inflammatory response in the cervical-vaginal epithelial cells through induction of il-8 and macrophage inflammatory protein 3α but not tnf-α and il-1β, two pro-inflammatory cytokines needed for the synthesis of systemic crp by hepatocytes.[27] in the present study, there was non-significant difference in il-10 level between t. vaginalis infected women and those non-infected. il-10 contributes to inhibition of th1 immunity and favors a t helper type 2 response.[18,24] in respect to autoantibodies, the levels of anticardiolipin (igg and igm) antibodies and antiphospholipid igg were significantly elevated in women with trichomoniasis. however, antiphospholipid igm level was non-significantly elevated in women with trichomoniasis in comparison to uninfected control women. such elevated levels of these autoantibodies could be explained by the antigenic mimicry between the parasite antigens and host proteins, as cardiolipin and phospholipids are abundant in most cells of multicellular organisms, the former is an important component of the inner mitochondrial membrane, where it constitutes about 20% of the total lipid composition,[27,28] while phospholipids are a class of lipids and are a major component of all cell membranes as they can form lipid bilayers.[29] in respect to our results, the detection of significant titers of anticardiolipin igg, igm, and antiphospholipid igg in the sera of trichomoniasis cases may be explained by the fact that all the infected women were chronic cases as they have been infected weeks before seeking medical advice. that is why igg antibodies against both cardiolipin and phospholipids were detected in their sera. however, the detected levels are still within the normal range that is referenced by the manufacturer. t. vaginalis lack mitochondria instead; it has double membrane-bound organelles, called hydrogenosomes, produced molecular hydrogen. phylogenetic and biochemical analyses of hydrogenosomes indicate a common origin with mitochondria; however, the identification of hydrogenosomal proteins and studies on its metabolism and chemical composition has been limited.[7] however, the presence of anticardiolipin antibodies in the sera of the patients may reflect the presence of cardiolipin in the membrane of hydrogenosome as it’s, phylogenetically, belong to the same origin with the mitochondria. the presence of antiphospholipid antibodies in the sera of the women with trichomoniasis, even low levels, must not be neglected as it can be resulted in undesired, serious consequence in pregnant women. antiphospholipid syndrome is a leading cause of miscarriage and maternal and fetal morbidity. this condition is characterized by thrombosis and pregnancy loss that occur in the presence of antiphospholipid antibodies.[30] conclusion there were no significant differences in crp level and eosinophil count between infected women and control group. ifn-γ level was significantly elevated in the sera of women with trichomotiasis. the inhibitory cytokine, il-10 level was non-significantly changed in response to t. vaginalis infection. trichomoniasis induced significant titers of anti-cardiolipin antibodies (igg and igm) and anti-phosplolipid igg antibodies. references 1. f. xueqiang, z. yingzhi, y. yanfang, d. yutao and l. huiqing.“prevalence 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mirgorodskaya, r. t. trifonova, r. r. sassi and c. e. costello. “structural details and composition of trichomonas vaginalis lipophosphoglycan in relevance to the epithelial immune function,” glycoconjugate journal, vol. 26, no. 1, pp. 3-17, 2009. 24. r. c. vilela and m. benchimol. “il-10 release by bovine epithelial cells cultured with trichomonas vaginalis and tritrichomonas foetus”. memórias do instituto oswaldo cruz, vol. 108, no. 1, pp. 110-112, 2013. 25. i. han, s. y. goo, s. park, s. hwang, y. kim, m. s. yang, m. ahn and j. ryu. “proinflammatory cytokine and nitric oxide production by human macrophages stimulated with trichomonas vaginalis”. korean journal of parasitology, vol. 47, no. 3, pp. 205-212, 2009. 26. n. malla, m. yadav and i. gupta. “kinetics of serum and local cytokine profile in experimental intravaginal trichomoniasis induced with trichomonas vaginalis isolates from symptomatic and asymptomatic women”. parasite immunology, vol. 29, no. 2, pp. 101-105, 2007. 27. r. n. fichorova, r. t. trifonova, r. o. gilbert, c. e. costello, g. r. hayes, j. j. lucas and b. n. singh. “trichomonas vaginalis lipophosphoglycan triggers a selective upregulation of cytokines by human female reproductive tract epithelial cells. parasite immunology, vol. 74, no. 10, pp. 5773-5779, 2006. 28. h. m. a. al-sakee. “autoantibodies in human cystic echinococcosis.” tikrit medical journal, vol. 17, no. 1, pp. 53-63, 2011. 29. w. dowhan. “understanding phospholipid function: why are there so many lipids”? journal of biological chemistry, vol. 292, no. 26, pp. 10755-10766, 2017. 30. f. a. f. di prima, o. valenti, e. hyseni, e. giorgio, m. faraci, e. renda, r. de domenico and s. monte. “antiphospholipid syndrome during pregnancy: the state of the art”. journal of prenatal medicine, vol. 5, no. 2, pp. 41-53, 2011. . 66 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 66-70 research article prevalence of anti-helicobacter pylori antibodies among students of cihan university, erbil hadi mahdi alsakee*, sarah hoshyar maroof, gulistan hussein muhammad, asia sherzad muhammad, ilaf abdulrazaq rashid, naza khan tofiq sdiq department of biology, cihan university-erbil, kurdistan region, iraq abstract helicobacter pylori is resident in human stomach and causes chronic disease (peptic ulcer and gastritis). the mouth and colon were both known to host a large number of microbes. this study was carried out to investigate the seroprevalence of h. pylori infection among cihan university students. a total of 197 blood samples were collected from the students (53 females and 144 males) from 13 departments of cihan university, erbil, and tested for anti-h. pylori antibodies, using rapid immunochromatography assay. among 197 students tested, 44 (22.3%) showed positive reaction for h. pylori, 32 males and 12 females. it was non-significantly higher among students with ages ranged between 29 and 32 years old. twenty-one of infected students were using tap water for drinking. twenty-six (59.1%) of positive students experienced no symptoms and 18 (40.9%) were symptomatic (13.63% epigastric pain and 27.2% abdominal pain). keywords: anti-h. pylori antibodies, helicobacter pylori, immunochromatography assay, prevalence introduction helicobacter pylori was originally known as campylobacter pylori, is a gram-negative, curved rods, and inhabits the human stomach. it was discovered in 1982 by two australian scientists, barry marshall and robin warren, who detected it in a person with chronic gastritis and gastric ulcers, a condition not previously believed to have a microbial etiology. it is also linked to the development of duodenal ulcers and stomach cancer.[1] however, over 80% of the cases are asymptomatic, and it may play an important role in the natural stomach ecology.[2] more than 50% of the world’s population have h. pylori in their upper gastrointestinal tract.[1] the infection rates are higher among people in developing countries compared to other western countries.[3] despite high infection rates that reported in certain areas of the world, the incidence of h. pylori infection is declining.[4] however, antibiotic resistance is appearing in h. pylori, many metronidazoleand clarithromycin-resistant strains are found in most parts of the world.[5] the way of transmission of h. pylori is unclear.[6] recent studies showed transmission through either fecal-oral or oral-oral route. it may be directly related to the source of drinking water.[1] infection with h. pylori is related to many diseases, for example, iron deficiency anemia, migraine, and coronary heart disease.[7] epidemiological studies demonstrated that h. pylori infection increases with age. it is higher in developing countries and among population with low socioeconomic status.[8] epidemiological data suggest strong associations of h. pylori infection and carcinoma of stomach and that significantly associated with mucosa-associated lymphoid tissues lymphomas (malt).[9] the infection is classified as a human carcinogen by the world health organization. the risk of cancer is highest among patients, in whom the infection induces chronic gastric inflammation. treatment of h. pylori infection significantly reduced the incidence of carcinoma of stomach and malt in endemic areas.[10] since little information concerning h. pylori have been published in erbil, in particular, among university students, the present study was aimed to investigate the prevalence of anti-h. pylori antibodies among university students in erbil as well as studying the risk factors and possible routes of transmission of this bacterium with its relation to the biomass index (body mass index [bmi]). corresponding author: hadi mahdi alsakee, department of biology, cihan university, erbil, kurdistan region, iraq. e-mail: hadialsakee@gmail.com received: apr 16, 2019 accepted: apr 20, 2019 published: jun 30, 2019 doi: 10.24086/cuesj.v3n1y2019.pp66-70 copyright © 2019 hadi mahdi alsakee, sarah hoshyar maroof, gulistan hussein muhammad, asia sherzad muhammad, ilaf abdulrazaq rashid, naza khan tofiq sdiq. this is an open-access article distributed under the creative commons attribution license. cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ alsakee, et al.: prevalence of anti-helicobacter pylori antibodies among 67 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 66-70 materials and methods study design and setting the present study was carried out in cihan university, erbil in erbil governorate, northern iraq. the total number of students in cihan university, erbil, is 5226 students of both genders divided on 20 departments. the period of the study was 7 months from november 2017 to may 2018. random samples of 197 students of both genders (male and female), who are currently studying in cihan university, erbil, from different departments were invited to participate in the study. the aims of the study and its significance, for both, the university community and public health, were explained and formal consents were obtained from the participants. design of questioner and data collection a closed-ended questionnaire was designed including information on the participants; age, weight, length, home address, current residence, socioeconomic level, marital status, animal handling (history of animal contact), sources of drinking water, type of meals (homemade and fast food), number of fast-food meals per week, history of other diseases (asthma and allergic manifestation), and history of clinical manifestations (abdominal pain, diarrhea, epigastric pain, vomiting, nausea, and anorexia). the data were collected through direct interview with the students who agreed to participate in the study. collection of blood specimens and serum extraction blood specimens were collected from the participants, in clean, sterile, test tubes (without anticoagulants). a specific numeric code was used for each sample. the blood specimens were left to clot at room temperature for an hour, the clotted blood then centrifuged at ×2000 g for 5 min. after centrifugation, the clean supernatant was collected by micropipette and dispensed in a clean sterile 1.5 ml capacity eppendorf tubes and stored at –50°c until used for the detection of anti-h. pylori antibodies. detection of anti-h. pylori antibodies h. pylori antibodies rapid immunochromatography test (plasmatec, united kingdom) was used for qualitative detection of anti-h. pylori antibodies in the sera of the participants. the test is one-step rapid chromatography immunoassay. statistical analysis the collected data were interred into the microsoft excel sheets and then translated into codes using a specific designed coding sheet. an expert statistical analysis was done using the statistical package for the social sciences version 17.0 pc software. the association between two or more categorical variables was analyzed and assessed by chi-square test. count and percentages were also used to describe the frequency of different variables between all possible paired combinations of study groups. p ≤ 0.05 was considered statistically significant. results a total of 197 students from 13 departments of cihan university, erbil, were involved in the study (53 biology, 10 computer sciences, 24 law, 5 banking and finance, 14 business administration, 34 international relations, 2 civil engineering, 4 architecture engineering, 12 health administration, 19 accounting, 6 media, 8 english, and 6 translation). of those students, 44 (22.3%) were revealed positive reactions for anti-h. pylori antibodies [table 1] with no significant (p > 0.05) differences between males (22.2%) and females (22.6%) [table 2]. eight (18.18%) of the positively reacted tests were of students who are always have house-made meals, whereas 15 (34.11%) of the positively reacted tests were of students who have always had restaurant’s fast-food meals. the students who had mixed restaurants and house-made meals constituted 21 (47.72%) of the positive cases. the infection rate was significantly (p = 0.047) higher among student who had their meals in the restaurants [table 3]. eighteen (40.9%) of the positive cases were with episodes of epigastric pain (13.63%) and abdominal pain (27.27%); however, 59.1% of the students who revealed positive reaction to anti-h. pylori antibodies experienced no symptoms. the statistical analysis revealed no significant (p > 0.05) differences in respect to clinical manifestations among h. pylori-infected students [table 4]. in regard to bmi, 27 (61.36%) of the students who revealed positive reactions to anti-h. pylori antibodies were with normal bmi. the statistical analysis revealed no significant (p > 0.05) association of h. pylori infection and bmi [table 5]. table 1: prevalence of anti-helicobacter pylori antibodies among cihan university students according to the academic departments department number of samples number of positive (%) number of negative (%) biology 53 11 (20.75) 42 (79.25) computer sciences 10 0 (0) 10 (100) law 24 4 (16.7) 20 (83.3) banking 5 2 (40) 3 (60) business 14 5 (35.7) 9 (64.3) international relations 34 9 (26.5) 25 (73.5) civil engineering 2 0 (0) 2 (100) architecture engineering 4 0 (0) 4 (100) health administration 12 0 (0) 12 (100) accounting 19 5 (26.3) 14 (73.7) media 6 5 (83.3) 1 (16.7) english 8 3 (37.5) 5 (62.5) translation 6 0 (0) 6 (100) total 197 44 (22.3) 153 (77.7) p=0.009 alsakee, et al.: prevalence of anti-helicobacter pylori antibodies among 68 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 66-70 of the 44 h. pylori-infected students, 5 (11.36%) had a history of animal handling, whereas 39 (88.64%) of them did not have a history of animal contact. no significant association (p > 0.05) of h. pylori infection and a history of animal contact [table 6] was detected. h. pylori infection rate was higher among students within age groups 29–32 years old (36.84%) followed by the students with age groups ranged between 25–28 years (30.55%) and 21–24 years old (22.99%). however, statistical analysis revealed no significant (p > 0.05) differences among different age groups studied in this research, in respect to having h. pylori infection [table 7]. in respect to the sources of drinking water, 21 (47.73%) of h. pylori-positive students were routinely using governmental drinking water supply (tap water) comparing with 14 (31.82%) and 9 (20.45%) of the positive students who are always using bottled water and who are using both (tap and bottled) for drinking and cooking purposes, respectively. however, no significant (p > 0.05) association of h. pylori infection and source of drinking water was noticed [table 8]. discussion h. pylori infection infects more than half of the world’s population. the current medical literature is described h. pylori as group 1 carcinogen. the transmission of this infection is rapidly decreasing in developed countries due to improvement in the sanitation.[11,12] in the present study, the prevalence of anti-h. pylori antibodies among university students was 22.3%, similar results were obtained by al-madi et al.[13] among saudi medical students. the prevalence of anti-h. pylori according to academic departments was significant (p = 0.009) which might be due to the lack of information about h. pylori disease which was higher than howler medical university students[14] due to the medical background and different lifestyle from rest of the society. according to the gender distribution, male students were more infected than female students, and this might table 2: gender distribution of anti-helicobacter pylori antibodies among cihan university, erbil, students department male female total number of samples number of negative (%) number of positive (%) number of samples number of negative (%) number of positive (%) biology 27 21 (77.8) 6 (22.2) 26 21 (88.76) 5 (19.24) 53 computer sciences 9 9 (100) 0 (0) 1 1 (100) 0 (0) 10 law 19 16 (84.2) 3 (15.8) 5 4 (80) 1 (20) 24 banking 3 2 (66.67) 1 (33.33) 2 1 (50) 1 (50) 5 business 12 8 (66.67) 4 (33.33) 2 1 (50) 1 (50) 14 international relations 26 19 (73.1) 7 (26.9) 8 6 (75) 2 (25) 34 civil engineering 1 1 (100) 0 (0) 1 1 (100) 0 (0) 2 architecture engineering 3 3 (100) 0 (0) 1 1 (100) 0 (0) 4 health administration 9 9 (100) 0 (0) 3 3 (100) 0 (0) 12 accounting 17 12 (70.5) 5 (29.5) 2 2 (100) 0 (0) 19 media 4 1 (25) 3 (75) 2 0 (0) 2 (100) 6 english 8 5 (62.5) 3 (37.5) 0 0 (0) 0 (0) 8 translation 6 6 (100) 0 (0) 0 0 (0) 0 (0) 6 total 144 112 (77.8) 32 (22.2) 53 41 (77.4) 12 (22.6) 197 p=0.350 table 4: clinical manifestations experienced by positively reacted students to anti-helicobacter pylori test clinical manifestations number of positive male (%) number of positive female (%) total (%) epigastric pain 3 (9.4) 3 (25) 6 (13.63) abdominal pain 8 (25) 4 (33.33) 12 (27.27) diarrhea 0 (0) 0 (0) 0 (0) asymptomatic 21 (65.6) 5 (41.67) 26 (59.1) total 32 12 44 p=0.468 table 3: prevalence of anti-helicobacter pylori antibodies among cihan university, erbil, students according to meal type feeding habits number of positive male (%) number of positive female (%) total (%) homemade 3 (37.5) 5 (62.5) 8 (18.18) restaurant 12 (80) 3 (20) 15 (34.1) both 17 (80.9) 4 (19.1) 21 (47.72) total 32 12 44 p=0.047 alsakee, et al.: prevalence of anti-helicobacter pylori antibodies among 69 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 66-70 table 5: association of helicobacter pylori infection and bmi bmi number of positive students (%) number of negative students (%) total <16 (severe thinness) 1 (2.27) 0 (0) 1 16–17 (moderate thinness) 0 (0) 4 (2.61) 4 17–18.5 (mild thinness) 0 (0) 2 (1.3) 2 18.5–25 (normal) 27 (61.36) 95 (62.1) 122 25–30 (overweight) 14 (31.8) 46 (30) 60 30–35 (obese class i) 2 (4.55) 6 (3.9) 8 35–40 (obese class ii) 0 (0) 0 (0) 0 >40 (obese class iii) 0 (0) 0 (0) 0 total 44 153 197 p=0.189 table 6: association of h. pylori infection and history of animal contact h.phylori-positive students history of animal contact total yes (%) no (%) male 4 (12.5) 28 (87.5) 32 female 1 (8.33) 11 (91.67) 12 total 5 (11.36) 39 (88.64) 44 p=0.468. h. pylori: helicobacter pylori table 7: age distribution of helicobacter pylori infection among cihan university students age groups total number of samples number of positive number of negative total number of positive (%) female (%) male (%) female (%) male (%) 17–20 40 2 (13.33) 3 (12.0) 13 (86.67) 22 (88.0) 5 (12.5) 21–24 87 8 (27.58) 12 (20.69) 21 (7.42) 46 (79.31) 20 (22.99) 25–28 36 1 (25.0) 10 (31.25) 3 (75.0) 22 (68.75) 11 (30.55) 29–32 19 1 (25.0) 6 (40.0) 3 (75.0) 9 (60.0) 7 (36.84) 33–36 3 0 (0) 0 (0) 0 (0) 3 (100) 0 (0) 37–40 10 0 (0) 1 (11.11) 1 (100) 8 (88.89) 1 (10) 41–44 1 0 (0) 0 (0) 0 (0) 1 (100) 0 (0) 45–48 1 0 (0) 0 (0) 0 (0) 1 (0) 0 (0) total 12 32 41 112 197 p = 0.601 be due to meals habit as the males have more fast-food meals in the restaurants than females. however, this finding was inconsistent with that reported by hooi et al.,[15] as in nasarawa state university, females were more infected due to accumulating risk factors and other social activities such as bed-sharing.[16] the prevalence of anti-h. pylori antibodies according to feeding habit was significant (p = 0.047) which might be due to contamination of food materials with intestinal pathogens including h. pylori bacteria.[17] in the current study, clinical manifestations that experienced by the infected students were non-significantly (p > 0.05) associated with h. pylori infection and might be due to the existence of more than one strain of the organism that responsible for this illness in erbil community that needs more investigations to be proven.[18,19] association of anti-h. pylori antibodies and bmi was non-significant (p > 0.05) and this could be explained by the fact that most of the positively reacted students experienced no symptoms. h. pylori infection impairs secretion balance of pro-inflammatory cytokines and c-reactive proteins, angiotensinogen, free fatty acids, and leptin hormone, and thus, reactive oxygen species begin to accumulate.[12] subclinical chronic inflammation induced by h. pylori infection occurs through impaired cytokine balance and stimulated macrophages.[20] thus, bmi was not influenced by the infection as most of the cases were subclinical. association of h. pylori infection and animal contact was non-significant (p > 0.05). this finding might be explained by the fact that human is the natural reservoir of h. pylori and this bacterium inhabits the gastric mucosa of human.[16] according to the age distribution of h. pylori infection, students with ages between 29 and 32 years old were more susceptible to be infected. however, this increased frequency was non-significant (p > 0.05) comparing with other age groups that studied in this research. dore et al.[21] were found that h. pylori infection is increasing with age due to the increased risk and opportunity of acquiring the infection.[21] h. pylori infection was non-significantly associated with drinking water sources; however, the infection rate was more detected among the students who use tap water for drinking and cooking purposes. this may be due to recent improvement in water basic system and service. alsakee, et al.: prevalence of anti-helicobacter pylori antibodies among 70 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2019, 3 (1): 66-70 conclusion the prevalence of anti-h. pylori antibodies is relatively high among students of cihan university, erbil, with no significant differences between male and female in respect to having h. pylori infection. the infection rate was significantly higher among students from literate and administrative departments than scientific departments. h. pylori infection was significantly associated with daily feeding habits of the students. bmi, clinical manifestations, age groups, history of animal contact, and source of drinking water are non-significantly associated with h. pylori infection. both university and public communities should be aware of infections caused by the pathogen such as gastritis, peptic ulcer, and about its potential association with malignant transformation. h. pylori should be listed as a public health challenge in erbil community by local health authorities and doing further studies to investigate the pathogenesis, strain variations, and drug resistance of the local strains of this bacterium. references 1. m. amieva and r. m. jr. peek. “pathobiology of helicobacter pylori induced gastric cancer manuel”. gastroenterology, vol. 150. no. 1, pp. 64-78, 2017. 2. m. j. blaser. “who are we? indigenous microbes and the ecology of human diseases”. embo reports, vol. 7, no. 10, pp. 956-960, 2006. 3. l. fuccio, r. m. zagari, l. h. eusebi, l. laterza, v. cennamo, l. ceroni, d. grilli and f. bazzoli. “meta-analysis: can helicobacter pylori eradication treatment reduce the risk for gastric cancer?” annals of internal medicine, vol. 151, no. 2, pp. 121-128, 2009. 4. h. m. malaty. “epidemiology of helicobacter pylori infection”. best practice & research. clinical gastroenterology, vol. 21, no. 2, pp. 205-214, 2007. 5. f. mégraud. “h pylori antibiotic resistance: prevalence, importance, and advances in testing”. gut, vol. 53, no. 9, pp. 1374-1384, 2004. 6. p. malfertheiner, f. megraud, c. o’morain, f. bazzoli, e. el-omar, d. graham, r. hunt, t. rokkas, n. vakil and e. j. kuipers. “current concepts in the management of helicobacter pylori infection: the maastricht iii consensus report”. gut, vol. 56, no. 6, pp. 772-781, 2007. 7. l. m. brown. “helicobacter pylori: epidemiology and routes of transmission”. epidemiologic reviews, vol. 22, no. 2, pp. 283-297, 2000. 8. a. tonkic, m. tonkic, p. lehours and f. megraud. “epidemiology and diagnosis of helicobacter pylori infection”. helicobacter, vol. 17, no. 1, pp. 1-8, 2012. 9. m. i. pereira and j. a. medeiros. “role of helicobacter pylori in gastric mucosa-associated lymphoid tissue lymphomas”. world journal of gastroenterology, vol. 20, no. 3, pp. 684-698, 2014. 10. k. e. l. mccoll. “helicobacter pylori infection”. the new england journal of medicine, vol. 362, no. 17, pp. 1597-604, 2004. 11. t. yucel, d. aygin, s. sen and o. yucel. “the prevalence of helicobacter pylori and related factors among university students in turkey”. japanese journal of infectious diseases, vol. 61, no. 3, pp. 179-183, 2008. 12. a. a. hassan, g. m. elnemr, m. a. almourgi, a. k. alzahrani, n. lilian and o. m. mehanna. “study of the effect of hypoxia on the prevalence of helicobacter pylori infection among saudi students at taif university”. research journal of microbiology, vol. 4, pp. 18-24, 2017. 13. m. a. al-madi, a. aljebreen, f. tounesi and a. abdo. “helicobacter pylori prevalence among medical students in a high endemic area”. saudi medical journal, vol. 28, no. 6, pp. 896-898, 2007. 14. b. hussen, s. s. qader, h. f. ahmad and s. h. ahmed. “the prevalence of helicobacter pylori among university students in iraq”. indian journal of science and technology, vol. 6, pp. 4-8, 2013. 15. j. k. y. hooi, w. y. lai, w. k. ng, m. m. y. suen, f. e. underwood, d. tanyingoh, p. malfertheiner, d. y. graham, v. w. s. wong, j. c. y. wu, f. k. l. chan, j. j. y. sung, g. g. kaplan and s. c. ng. “global prevalence of helicobacter pylori infection: systematic review and meta-analysis”. gastroenterology, vol. 153, no. 2, pp. 420-429, 2017. 16. s. agah, h. khedmat, m. e. ghamar-chehred, r. hadi and a. aghaei. “female gender and helicobacter pylori infection, the most important predisposition factors in a cohort of gastric cancer: a longitudinal study”. caspian journal of internal medicine, vol. 7, no. 2, pp. 136-141, 2016. 17. r. m. nyarango, p. a. aloo, e. w. kabiru and b. o. nyanchongi. “the risk of pathogenic intestinal parasite infections in kisii municipality, kenya”. bmc public health, vol. 8, pp. 237, 2008. 18. j. g. kusters, a. h. m. van vliet and e. j. kuipers. “pathogenesis of helicobacter pylori infection”. clinical microbiology reviews, vol. 19, no. 3, pp. 449-490, 2006. 19. m. j. blaser, y. chen and j. reibmann. “does helicobacter pylori protect against asthma and allergy?” gut, vol. 57, no. 8, pp. 1178-1179, 2008. 20. j. michalkiewicz, a. helmin-basa, r. grzywa, m. czerwionkaszaflarska, a. szaflarska-poplawska, g. mierzwa, a. marszalek, m. bodnar, m. nowak and k. dzierzanowska-fangrat. “innate immunity components and cytokines in gastric mucosa in children with helicobacter pylori infection”. mediators of inflammation, vol. 10, pp. 1-7, 2015. 21. m. p. dore, h. m. malaty, d. y. graham, g. fanciulli, g. delitala and g. realdi. “risk factors associated with helicobacter pylori infection among children in a defined geographic area”. clinical infectious diseases, vol. 35, no. 3, pp. 240-245, 2002. table 8: association of drinking water sources and helicobacter pylori infection source of drinking water number of positive male (%) number of positive female (%) total tap water 17 (53.13) 4 (33.33) 21 (47.73) bottle water 8 (25.0) 6 (50.0) 14 (31.82) both 7 (21.88) 2 (16.67) 9 (20.45) total 32 12 44 p=0.601 tx_1~abs:at/add:tx_2~abs:at 125 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 research article the preventive effects of urtica dioica extract and nanoparticle on oxidative stress and lipid profile in hyperlipidemic male rats banw a. othman1, nadir m. nanakali2 1department of pharmacognosy, college of pharmacy, hawler medical university, erbil, iraq, 2department of biology, college of education, salahaddin university-erbil, kurdistan region, erbil, iraq abstract cardiovascular diseases (cvds) are the major factor in human premature death and disability and they are becoming. oxidative stress is thought to be a major factor in the development of atherosclerosis. one of the cvds risk factors are elevated lipid profile. significant gaps in the treatment of cvds still exist, despite making healthy lifestyle and using lipid-lowering medications as part of primary cvds prevention. natural extracts are a rich source of biomolecules that can be used as reagents for the biosynthesis of silver nanoparticles (agnps). the present study estimates the antioxidant and protective activity of urtica dioica (ud) in induced hyperlipidemia rats divided into six groups; group 1 fed on normal diet, group 2 fed a high-fat diet, 200 mg/dl water extract, and ethanol extract, respectively, for groups 3 and 4, group 5 receives 15 mg/dl agnps, and group 6 receives 20 mg/dl atorvastatin all fed on a high-fat diet and then evaluates lipid profile and malonaldehyde, superoxide dismutase and reduced glutathione of the serum after 40 days. the results of characterization test demonstrated the successful synthesis of agnps, also shows that the ud extract and agnps have a favorable impact on lipid profile significantly reduced the level of total cholesterol, low-density lipoprotein (ldl), and markedly decrease liver enzymes also can function as a good antioxidant that can be employed as a protective agent against rising lipids and oxidative stress. however, further research studies need to investigate the safety and activity of different doses of agnps of ud. keywords: hyperlipidemia, antioxidant, urtica dioica, silver nanoparticle, cardiovascular disease introduction hyperlipidemia is thought to be a very harmful factor contributing to cardiovascular and cerebrovascular diseases, in particular, atherosclerosis.[1] this happens as a result of a malfunction in the body’s lipid metabolism, raising the serum lipid concentration over the usual range. the occurrence of hyperlipidemia has increased dramatically over time and is now one of the most prevalent pathological disorders in humans as a result of inheritance, diet, nutrition, medicine, and other variables.[2] the development of numerous diseases, including cancer, agerelated disorders, neurodegenerative diseases, autoimmune disorders, and cardiovascular disease (cvd), is thought to be significantly influenced by oxidative stress. oxidative stress can have an impact on cvd development in a variety of ways. according to the research on atherosclerosis, the formation of atherosclerotic plaques is said to begin with the oxidation of low dispersed lipid (ldl) particles in the arterial endothelium.[3] pharmaceutically dynamic bioactive substances found in medicinal plants have additive and synergistic therapeutic actions that are helpful in the treatment of metabolic diseases. utilizing the knowledge of local populations, the majority of pharmaceutical medications are made by isolating the primary active ingredients from medicinal plants.[4] a significant advancement in modern medicine has resulted from the systematic research of medicinal plants and the investigation of their biologically active phytochemical components in the treatment of metabolic diseases.[5] contrary to current medical systems, polyherbal formulations have drawn increased interest because of their ability to treat several conditions while having fewer negative effects.[6] corresponding author: banw a. othman, department of pharmacognosy, college of pharmacy, hawler medical university, kurdistan region, erbil, iraq. e-mail: banw.othman@hmu.edu.krd received: august 07, 2022 accepted: august 19, 2022 published: november 01, 2022 doi: 10.24086/cuesj.v6n2y2022.pp125-132 copyright © 2022 banw a. othman, nadir mustafa nanakali. this is an openaccess article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) othman and nanakali: the preventive effects of urtica dioica extract and nanoparticle on oxidative stress 126 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 urtica dioica l. (ud) in north europe and parts of asia, the stinging nettle family, or urticaceae, is widely distributed and is typically found in rural areas.[7] traditional medicine has long used this herb to treat a variety of ailments, including dyslipidemia. in vivo studies have shown that ud has a beneficial effect against dyslipidemia.[8] more recently, it was demonstrated that a combination of aqueous extracts from peganum harmala, rhus coriaria, and ud improved metabolic and histological parameters in diabetic rats.[9] another study demonstrates that methanol extracts from various common herbs, including ud, can be helpful in the formation of silver nanoparticles (agnps) verify their free radical scavenging activity and reducing power.[6] this inspired us to examine the potential preventive benefits of ud ethanol, water, and nps extracts on hyperlipidemia in a high-fat diet fed animal model with hyperlipidemia. methodology and experimental design preparation of the plant extracts the dried leaves of the ud were gained from local herbal shop in erbil city, which has been dried in shade at room temperature. the plant was identified by professor abdulla shakur, salahaddin university-erbil, college of education, department of biology. a 25 g of the powdered plant material was mixed with 250 ml of 70% ethanol 1 sample:10 solvent ratio(w/v) and sonicated 60 min power sonic 405/lab tech kept at 40°c. following that, whatman no. 1 filter paper with buchner funnel was used to filter the obtained extract. the filtrate was concentrated in a rotary evaporator under a controlled vacuum. after air drying, the concentrated extract was preserved at 4°c in an airtight container.[10] furthermore, another 25 g of the powdered plant material was mixed with 250 ml distilled water 1 sample:10 solvent sample ratio (w/v) then using the same producer as in ethanol extract. agnps synthesis for the preparation of the nanoparticle, 10 g of dried plant powder was mixed with 300 ml of boiled distilled water then boiled for 10 min with a magnetic stirrer and filtered by whatman no. 1 filter paper then stored at 4°c, 100 ml of prepared extract was added to 1000 ml of 1 mm of aqueous agno 3 and put on a hot plate for 60 min at 40°c with magnetic stirrer up till the color turned dark brown and adjusting the ph to 11 by naoh. uv–vis spectra were used to establish whether agnps were formed following the centrifuging of the wholly reduced solution at 5000 rpm for 30 min. the obtained pellet was dissolved in deionized water after the supernatant was discarded. to get rid of any materials that were adsorbed on the surface of the agnps, the same washing procedure was carried out 2 or 3 times by deionized water and ethanol.[11] characterization of agnps the absorbency of all samples was measured using uv–vis spectra for confirming the formation of agnps, fouriertransform infrared (ftir) spectroscopy was performed on shimadzu ftir spectrometer to detect the possible functional groups in biomolecules present in the plant extract. the x-ray diffraction (xrd) measurement was carried out on x-ray diffractometer (panalytical x’pert-pro pw3050/60) operated at 45 kv and 40 ma and the spectrum was recorded by cu radiation with a wavelength of 1.54060 å in the 2q range of 20e80. a scanning electron microscope (sem) was used to investigate the agnps’ size and surface shape. experimental animals the male albino rats were kept in a room with a temperature of 22 ± 2°c and a 12:12 h light/dark cycle. each rat weighed between 250 and 300 g, the experiment on the animal was conducted in the animal house of pharmacy college/ hawler medical university, the rats were kept for 1 week for acclimatization. induction of hyperlipidemia for inducing hyperlipidemia, the standard pellet of the rat is mixed with 5% cholesterol and 0.5% cholic acid according to a method described by mojarradgandoukmolla et al.[12] for determining whether the rats were induced for hyperlipidemia, a pretest was done after 30 days of feeding them a cholesterolenriched diet, the rats were starved for 24 h before measuring their cholesterol serum level by enzymatically diagnostic kits. doses and toxicity according to a study done by kedi et al.,[13] he concluded that silver nps have advantage to treat inflammation in low doses (100, 200, and 400 mg/kg). research on acute toxicity was conducted by jacob et al.[14] on healthy female albino rats with one dose of 2000 mg/kg reduced by plant extract. the study revealed that the agnps synthesized by ficus carica were relatively non-toxic. another study uses 250 and 500 mg/kg of ud phenolic extract and 5–10 mg/kg agnp of the same plant.[15] experimental procedure the animals were divided randomly into six groups of six each made up of six rats (n = 6). the experiment lasted for 40 total days. • group 1 rats were served a standard pellet diet and they were considered a negative control group • group 2 rats were served with cholesterol-enriched diet and they were considered as a positive control • group 3 rats were served with 200 mg/kg (ud) water extract with cholesterol-enriched diet • group 4 rats were served with 200 mg/kg (ud) ethanol extract with cholesterol-enriched diet • group 5 rats were served with 15 mg/kg (ud) agnps with cholesterol-enriched diet • group 6 rats were served with 10 mg/kg of atorvastatin with cholesterol-enriched diet. body weight rats were weighed once a week to observe changes in body weight throughout the duration of the 40-day study. when the experimental period ends the rats fasted overnight, the othman and nanakali: the preventive effects of urtica dioica extract and nanoparticle on oxidative stress 127 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 next day anesthetizing of the rats was done by giving them a combination of ketamine 35 mg/kg with xylazine 5 mg/kg.[16] then, the blood samples were drawn by puncture of the heart from each rat in each group and gathered in serum separating tubes, the blood was centrifuged at 3000 rpm for 15 min to prepare serum and determination of biochemical and liver function assay for antioxidant test the serum stored in −20°c. determination of serum lipid profiles and liver function test using a method where the color intensity of the dye formed is directly proportional to the concentration of the tested parameters, the measurement of serum total cholesterol (tc), serum ldl-c, serum triglyceride, and serum hdl cholesterol was performed using a med lab reagent kit/cobas, usa.[17] plasma concentrations of vldl were determined by the method of friedewald and levy.[18] liver function assay involving alanine aminotransferase (alt) and aspartate aminotransferase (ast) with gamma-glutamyl transferase (ggt) was determined by colorimetric method using med lab reagent kit/cobas/usa.[19] oxidative stress evaluation for determining the level of malondialdehyde (mda), superoxide dismutase (sod), and glutathione (gsh) in the serum a solarbio kit/china was used. mda content is calculated by the difference between the absorbance at 532 nm, 450 nm, and 600 nm.[17] sod can remove o⁻⁻ and inhibits the formation of methionine, sod activity decreases with increasing darkness of the reaction solution’s blue color, and the absorbency is detected at 560 nm.[20] gsh, 5,5-dithiobis-(2-nitrobenzoic acid), and gsh will react to form 2-nitro-5-mercaptobenzoic acid and gsh disulfide (gssg). the yellow product 2-nitro-5-mercaptobenzoic acid has a maximum absorbance of 412 nm.[20,21] all absorbencies are detected by a microplate reader. statistical analysis the mean and standard error were used to express the study’s data.[13] analyzing of results performed by one-way analysis of variance to test the difference between the groups, tukey multiple comparisons test in graphpad prism 8 version 8.0.1 for windows 10 64x used. the analysis was conducted at a 95% confidence level and a significance level of 5%. whenever p = 0.05 or less was regarded as statistically significant (p ≤ 0.05). results the formation of agnps the formation of agnps was established by the color change observed visibly from bright green to a deep brownish color, further confirmation done by uv–vis spectra, as shown in figure 1. the result of absorbance of agnps from 200 to 800 nm gives the maximum absorbance between 440 and 450 nm and confirmed the presence of agnps. sem was used to investigate the shape and size of agnps which was spherical in shape [figure 2]. the result of xrd shows diffraction peaks at 37.6024°, 43.8755°, 64.0193°, and 76.9627° [figure 3]. the figure 1: the result of uv–vis spectra of urtica dioica agnps figure 2: the result of sem of urtica dioica agnps figure 3: the result of xrd of urtica dioica agnps othman and nanakali: the preventive effects of urtica dioica extract and nanoparticle on oxidative stress 128 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 biosynthesis of agnps was investigated by ftir spectroscopy and the result of the functional group is shown in figure 4. lipid profile the total serum cholesterol in the positive control group was significantly (p < 0.01) increased (105 ± 6.776 mg/dl) when compared with the negative control group (65.90 ± 2.446 mg/dl), but in the hyperlipidemia rats treated with ud water extract (88.29 ± 1.523 mg/dl), ethanol extract (79.65 ± 2.677 mg/dl), nanoparticles nps (83.82 ± 4.275 mg/dl), and atorvastatin (65.47 ± 2.827 mg/dl), the total serum cholesterol was decreased, these changes were significant (p < 0.01) as compared with positive group. the serum level of triglyceride in the positive control rats (116.2 ± 3.629 mg/dl) elevated significantly (p < 0.01) when compared with the negative control (71.67 ± 2.109 mg/dl), while this elevation was decreased significantly in the treated groups with ud water extract (80.4 ± 4.581mg/dl), ethanol extract (87.23 ± 2.639 mg/dl), nps (77.73 ± 4.987 mg/dl), and atorvastatin (86.50 ± 3.221 mg/dl) when compared with positive control rats. the negative control (48.1 ± 2.65 mg/dl) and treated groups with ud water extract (40 ± 2.984 mg/dl) and ethanol extract (38.38 ± 1.257 mg/dl) and nps (37.07 ± 1.711 mg/dl) are significantly (p < 0.01) improved in comparison with the positive control (23.23 ± 0.6712 mg/dl), while the atorvastatin group is non-significant with the positive control group. there is a significant difference in the level of ldl in the negative control and ethanol extract and nps and atorvastatin group in comparison with the positive control, however, the result shows no significant change in the comparison between water extract and positive control rats. the obtained result shows a significant decrease in the level of serum vldl in groups treated with ud water extract (16.08 ± 0.9163 mg/dl) and nps (15.55 ± 0.9979 mg/dl) and atorvastatin (17.30 ± 0.6441 mg/dl) and negative control comparing with the positive control, also, comparing ethanol extract with the positive control, we found significant difference as shown in table 1. liver function table 2 shows that a significant difference in serum alt was observed between the positive control group (54.1 ± 2.652 iu/l) and ethanol extract (42.22 ± 1.756 iu/l) and nps group (41.1 ± 2.473 iu/l). whereas, the rats treated with water extract (46.25 ± 0.6994 iu/l) and atorvastatin (46.32 ± 2.42 iu/l) were not significant difference between positive control as well as the negative control (44.77 ± 2.683 iu/l). the serum level of ast in the negative control (119.3 ± 3.981 iu/l) was significantly different in comparison with the positive control (145.6 ± 7.471 iu/l), as well as the rats that were treated with water extract (117.9 ± 4.357 iu/l), ethanol extract (119.6 ± 6.491 iu/l), and nps (122.5 ± 2.064 iu/l), but table 1: effect of urtica dioica water and ethanol extract and its nanoparticle and atorvastatin on serum lipid profile in hyperlipidemic rats group/parameters cholesterol (mg/dl) triglyceride (mg/dl) hdl_c (mg/dl) ldl_c (mg/dl) vldl_c (mg/dl) negative control 65.90±2.446**** 71.67±2.109**** 48.1±2.65**** 26.85±1.805**** 14.33±0.421**** positive control 105±6.776 116.2±3.629 23.23±0.6712 57.97±5.682 23.25±0.725 water extract 88.29±1.523* 80.4±4.581**** 40±2.984**** 45.85±3.460 16.08±0.916**** ethanol extract 79.65±2.677*** 87.23±2.639**** 38.38±1.257**** 37.43±3.681** 17.44±25.93**** nps 83.82±4.275** 77.73±4.987**** 37.07±1.711*** 42.25±2.819* 15.55±0.997*** atorvastatin 65.47±2.827**** 86.50±3.221**** 27.53±1.491 38.85±2.560** 17.30±0.644**** significant differences between treatment groups were denoted by superscript asterisks symbols * significant at p<0.05, **p>0.01, ***p>0.001, and ****p>0.0001 figure 4: the result of fourier-transform infrared spectroscopy othman and nanakali: the preventive effects of urtica dioica extract and nanoparticle on oxidative stress 129 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 table 2: effect of urtica dioica water and ethanol extract and its nps and atorvastatin on liver function tests in hyperlipidemic rats group/parameters alt (iu/l) ast (iu/l) ggt (u/l) negative control 44.77±2.683 119.3±3.981* 0.81±0.005**** positive control 54.1±2.652 145.6±7.471 3.833±0.600 water extract 46.25±0.699 117.9±4.357* 1.122±0.158**** ethanol extract 42.22±1.756** 119.6±6.491* 1.178±0.171**** nps 41.1±2.473** 122.5±2.064* 0.9483±0.017**** atorvastatin 46.32±2.42 144.3±5.673 1.263±0.217**** significant differences between treatment groups were denoted by superscript asterisks symbols *significant at p<0.05, **p>0.01, ***p>0.001, and ****p>0.0001 table 3: effect of urtica dioica water and ethanol extract and its nanoparticle and atorvastatin on blood glucose, total protein, amylase, and blood urea in hyperlipidemic rats group/parameters glucose (mg/dl) total protein (g/dl) amylase (u/dl) blood urea (mg/dl) negative control 220.3±11.72**** 6.46±0.149 21.95±0.956 32.42±1.593**** positive control 320.4±15.23 6.175±0.091 24.26±0.619 56.53±2.122 water extract 146.7±5.735**** 6.803±0.096* 23.72±1.144 43.63±1.907** ethanol extract 312.9±9.531 6.937±0.176** 25.55±0.979 44.7±3.344** nps 133±11.94**** 6.623±0.127 23.87±1.517 36.43±1.819**** atorvastatin 230.8±15.58*** 6.407±0.1177 23.63±0.785 54.33±1.745 significant differences between treatment groups were denoted by superscript asterisks symbols *significant at p<0.05, **p>0.01, ***p>0.001, and ****p>0.0001 the rats that treated with atorvastatin were non-significant different with the level of serum ast in positive control. decreased levels of serum ggt were observed in the negative control, water extract, ethanol extract, nps, and atorvastatin (0.81 ± 0.005164 u/l), (1.122 ± 0.1584 u/l), (1.178 ± 0.1712 u/l), (0.9483 ± 0.01721 u/l), and (1.263 ± 0.2177 u/l), respectively, and they are significantly different from positive control rats. in the present study, rats in the negative control show a reduction in the level of blood glucose (220.3 ± 11.72 mg/dl) compared with the positive control as in the treated groups also observe a significant difference in the level of blood glucose for water extract (146.7 ± 5.735 mg/dl), nanoparticle (133 ± 11.94 mg/dl), and atorvastatin (230.8 ± 15.58 mg/dl) in comparison with the positive control (320.4 ± 15.23 mg/dl), but in rats that were treated with ethanol extract, we observe no significant difference. results show that serum levels of total protein in treated groups with water extract (6.803 ± 0.09646 g/dl) and ethanol extract (6.937 ± 0.1768 g/dl) increased significantly when compared with positive control rats (6.175 ± 0.09139 g/dl). furthermore, the treatments effect of hyperlipidemic rats with nanoparticles (6.623 ± 0.1273 g/dl) and atorvastatin (6.407 ± 0.1177 g/dl) was not significant when compared with untreated hyperlipidemic rats, also the rats in the negative control (6.46 ± 0.1497 g/dl) have no significant difference with the positive control (6.175 ± 0.09139 g/dl). as shown in table 3, no significant variations existed between untreated and treated animals in the level of serum amylase. while the level of blood urea in negative control and water extract and ethanol extract and nanoparticles decreased which is significantly different from the level of blood urea in the positive control, but when we compare the level of blood urea of atorvastatin (54.33 ± 1.745 mg/dl) with the positive control (56.53 ± 2.122 mg/dl), there was a non-significant difference. there was an insignificant (p > 0.05) decrease in the mda in the negative control group (1.135 ± 0.1453 nmol/ml), water extract (1.115 ± 0.3712 nmol/ml), ethanol extract (1.076 ± 0.3645 nmol/ml), and nps (1.024 ± 0.1353 nmol/ml) compared with the positive control group (2.722 ± 0.2852 nmol/ml), and also, the mda was not changed significantly in atorvastatin group when compared with the rats of the control positive. significant difference in sod was observed between rats in the water extract group (54.57 ± 1.390 u/ml), ethanol extract group (53.23 ± 0.768 u/ml), and nps group (54.09 ± 1.629 u/ml) with the positive control (61.48 ± 0.8206 u/ml). furthermore, the hyperlipidemic rats treated with atorvastatin (60.96 ± 1.601 u/ml) did not change sod significantly when compared with the positive control rats as well as the result of the negative control was non-significant. regarding the gsh, results of the ethanol extract (36.83 ± 2.216 mg/ml) and nps treated rats (42.59 ± 6.71 mg/ml) showed a significant rise (p < 0.01) in the gsh as compared to the positive control rats (21.06 ± 0.7582 mg/ml). however, a non-significant variation (p > 0.05) was detected among the water extract (34.24 ± 1.191 mg/ml), atorvastatin (30.69 ± 4.475 mg/ml) group, and the positive control group in gsh [figure 5]. discussion if the serum levels of tc, ldl, and tg in the hyperlipidemic rats are taken into consideration, our results revealed that the othman and nanakali: the preventive effects of urtica dioica extract and nanoparticle on oxidative stress 130 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 water, ethanol, and nps extracts of ud hold a favorable effect in controlling these parameters and ultimately preventing the risk of atherosclerosis and related cvds. stinging nettles have been shown to be richer in individual polyphenols than other wild plants.[22] many studied found that the phenolic compound has the ability to normalize hyperlipidemic disturbance.[23,24] another study showed that the ud extract lowers the level of lipid profile due to a significant decrease of apolipoprotein b (apob) levels irrespective of the fat in the diet.[25] apob is a surface marker of vldv and ldl and tc.[26] the findings of this research indicated that the effect of ethanol extract is greater than the water extract which is similar to the studies done by avci et al.,[8] nassiri-asl et al.[27] and also greater than the nps. the nettle leaves contain sterols: campesterol and sitosterol.[28] plant sterols resemble cholesterol in structure, they may replace cholesterol from mixed micelles because compared to cholesterol, they are more hydrophobic, this replacement causes a decrease in micellar cholesterol concentrations and consequently lowers cholesterol absorption.[8] in research by rau et al. to determine which of two ethanolic herbal extracts activated the peroxisome proliferator-activated receptors, which play a crucial role in lipid homeostasis, the ethanolic extract of stinging nettle was one of the most active.[29] the results of this study showed a significant effect of nps on hyperlipidemic rat’s lipid profile, which also may be due to the presence of phenolic acids in the plant which have a high reducing power for nps synthesis.[6] hdl also increased significantly as shown in the result of this study and has in agreement with the result of nassiri-asl et al.[27] it was hypothesized that the extract may directly affect the production and metabolism of lipoproteins. several serum parameters were examined in the current investigation, other than lipid profile to evaluate the inhibitory effect of the ud on hepatic marker enzymes such as alt, ast, and ggt which give a significant decrease except those in atorvastatin, and elevation of ast, alt, and ggt observed in hyperlipidemic rats which have an agreement with the study done for the effect of some plants; one of them was ud as an antioxidant and they concluded that the ethanolic extract of ud has antihepatotoxic potential and hypercholesterolemia causes fatty liver and eventually elevation of the liver enzyme.[30] furthermore, in two other studies, their results demonstrate the hepatoprotective effect of stinging nettle.[31,32] the majority of prior investigations demonstrated that harmful chemicals adsorbed on the surface of chemically produced agnps at higher dosages may induce liver or kidney damage.[33,34] some investigations showed that modest concentrations of agnps had no harmful impact on hepatic and renal functioning.[33,35] similarly, the levels of liver enzymes were not affected by the injection of lesser doses of 13–40 nm sized agnps (5 or 10 mg/kg), according to research by dhermendra et al.,[25] while intact rats exposed to greater dosages of agnps (13–40 nm, 20–40 mg/kg) had impaired liver function.[36] however, the results of a different study showed that, although both chemically and eco-friendly agnps at a dose of 2.5 mg/kg were able to lower blood sugar levels in diabetic rats, green agnps were more protective than chemically made agnps at the same dose in terms of regulating and enhancing liver function.[7] the results of the present study showed that a 15 mg/kg dose of green agnps synthesized using ud for about 6 weeks significantly reduced the serum concentration of liver enzyme levels including alt, ast, and ggt compared with the hyperlipidemic rats. to learn more about the potential processes behind the beneficial benefits of green produced agnps utilizing ud extract on liver function in hyperlipidemic rats, more research is required. the glucose level in the hyperlipidemia rats in this study was elevated which is agree with the previous study done by mojarradgandoukmolla et al.[12] the result shows a significant decrease in the level of treated rats with ud. the findings demonstrate that the ud extract’s ability to reduce blood glucose was caused by langerhans islets’ increased insulin production.[37] coumarins derivative has also been identified in nettle roots.[38] a review of several studies on the effect of coumarins on blood glucose concluded that numerous in vivo and in vitro studies show that coumarins help reduce blood sugar levels in people with diabetes by inhibiting proteintyrosine phosphatase 1b (ptp1b), reducing inflammation and oxidative stress, enhancing pancreatic function, and correcting abnormal insulin signaling. as a result, they are useful and need more study and development. in addition, several coumarins, including osthole and psoralen, have the ability to activate adenosine monophosphate-activated protein kinase (ampk), enhancing the translocation of glucose transporter type 4 (glut4) to the plasma membrane, and facilitating glucose uptake. an important justification for creating insulin mimics figure 5: effect of urtica dioica extracts and agnps on mda, sod, and gsh biomarkers othman and nanakali: the preventive effects of urtica dioica extract and nanoparticle on oxidative stress 131 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2022, 6 (2): 125-132 is the fact that ampk may increase glut4 translocation in the absence of insulin.[39] it has been hypothesized that hypercholesterolemia raises the amount of lipid peroxidation in the serum of affected guinea pigs and rabbits.[40] hence, we evaluate the level of mda, sod, and gsh, our result shows the increasing level of mda, sod and decreasing gsh in hyperlipidemic rats which agrees with das et al.[40] but in the treated rats, this level was significantly different from the hyperlipidemic rats, and according to the findings of a study, there is a positive linear association between total phenolic and total flavonoid content and antioxidant activity. according to stanojević et al., nettle leaf extracts in aqueous methanol might serve as an alternative to synthetic additions as a source of natural antioxidants.[41] our results also have an agreement with two other studies; the hydroalcoholic extract of ud plant has shown significant results for antioxidant activity with half inhibitory concentration (ic50) value of 88.33 ± 2.88 µg/ml[42] and the aqueous extract has significant reducing power, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities.[43] in another study, it has been shown that the ethyl acetate fraction of ud has potent antioxidant activities as compared to other fractions.[44] as a result, the antioxidant capabilities of ud may have been impacted by the extraction technique or the tissue or location examined. the activity of the ud ethanol extract increased noticeably.[8] this proves the result of gsh level of ethanol extract and nps which was significant while the rats in water extract are not significant. due to the protective capping of agnps by polyphenols and the corresponding size of the agnps, biosynthesized agnps utilizing pongamia pinnata leaf extract have been demonstrated to have a better antioxidant radical scavenging capability than chemically synthesized agnps.[7] conclusion our results show that ethanol, water extracts, and agnps of ud reduced tc, ldl, and tg levels while raising hdl levels. further research should be done on this plant to use as an active medicinal agent that bears fewer side effects on drugbased people suffering from cvd. in addition, the biosynthesis of agnps of the ud plant occurred, giving spherical 44–48 nm in diameter that has a positive effect on lipid profiles and acts as antioxidants and it was safe in the dose of 15 mg/kg for the rats. references 1. j. miao, x. zang, x. cui and j. zhang. autophagy, hyperlipidemia, and atherosclerosis. advances in experimental medicine and biology, vol. 1207, pp. 237-264, 2020. 2. n. he and h. ye. exercise and hyperlipidemia. advances in experimental medicine and biology, vol. 1228, pp. 79-90, 2020. 3. r. stocker and j. f. keaney, jr. role of oxidative modifications in atherosclerosis. physiological reviews, vol. 84, no. 4, pp. 1381-1478, 2004. 4. r. m. pérez gutiérrez, a. muñiz-ramirez, a. h. garcia-campoy and j. m. mota flores. evaluation of the antidiabetic potential of extracts of urtica dioica, apium graveolens, and zingiber officinale in mice, zebrafish, and pancreatic β-cell. plants (basel), vol. 10, no. 7, p. 1438, 2021. 5. m. a. mayer, c. höcht, a. puyó and c. a. taira. recent advances in obesity pharmacotherapy. current 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ashti y. hussein, faris a. mustafa department of architecture, college of engineering, salahaddin university-erbil, 44002, erbil, kurdistan region, iraq abstract higher education students’ difficulties, especially students in architecture education, lead students to health problems. the design studio is the heart of architecture education and students spent most of their time with staff. students tend to do various activities in the studio such as demonstrating, model-making, drawing, and lecturing. therefore, the learning environment in the studio should attract students to spend most of their time in the studio happily. this research focused on a mixed method to assess the relationship between students’ well-being and the quality learning environment in the design studio. the students’ self-report assessment for the well-being situation has been used that depends on student’s experience to the design studio. for this reason, architecture design studios at private universities in erbil city have been selected as a case study. the statistical program spss has been used to test hypotheses and analyze data. the findings of this study represent the significant positive relationship between students’ well-being and the quality learning environment in the design studio. furthermore, the study illustrates the significant of the quality learning environment in design studio and its roles in the development of the students’ well-being. keywords: quality learning environment, student’s well-being, architectural design studio, private universities, architecture education introduction conventionally, the establishment of a healthy learning environment is concerned with the improvement of students’ well-being and health.[1] in the past decade concern for students’, well-being has increased due to its impacts on health,[2,3] psychological distress, and learning outcomes.[4] students’ satisfaction and positive attitudes to the learning environment are the critical issue in well-being assessment.[5,6] students’ well-being is linked to the learning outcome, student’s performance, pro-social behaviors, positive relationships, creativity, and health.[7-11] however,[6-12] realized that environmental factors of the learning process are the main pillar of students’ well-being. well-being in a learning environment encompasses the development of the learning process, social relationship with students and peers, increasing students’ cognition, and encouraging creative and critical thinking.[13,14] therefore, the classroom has been described as a physical environment that functions as a social and cultural space that must support students’ needs and learning objectives.[13] the quality learning environment in the literature is described as an adaptable environment that is flexible and obtains all student’s needs.[15,16] creating an optimal learning environment improves effective educational space that enhances students’ performance, motivation, health, and well-being.[17,18] the reference[19] indicated that the learning environment contains the physical environment, social activities, motivational and psychological environment,[20,21] and teaching/learning process,[14] the physical environment of the classroom is considered one of the most important indicators that determine benefits in student learning. the reference[15] claimed that the process of teaching and learning architecture necessitates a comfortable environment that promotes lectures, design projects, and tutorials. because students spend so much time in architectural studios, the setting must be stimulating and inspirational. teaching architecture differs from teaching other built-environment disciplines. architectural education requires not only practical and theoretical features but also an engaging learning environment that encourages social relationships.[22] as the foundation of architectural education, the design studio is regarded as a living-learning environment for architectural students. it is the location where architectural students spend a significant amount of time engaging in various corresponding author: ashti y. hussein, department of architecture, college of engineering, salahaddin university-erbil, 44002, erbil, kurdistan region, iraq. e-mail: ashtyyasin22@gmail.com received: december 31, 2022 accepted: january 25, 2023 published: march 1, 2023 doi: 10.24086/cuesj.v7n1y2023.pp21-28 copyright © 2023 ashti y. hussein, faris a. mustafa. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) hussein and mustafa: students self-report well-being in the architectural design studios 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 21-28 socioeducational activities. as a result, the studio should cater to the students’ demands. the design of the studio should be carefully considered to promote student productivity, creativity, and enjoyment.[23] the environment of the design studio is one of the aspects which affect and stimulate the learning ability of students and also have a greater impact on the effective outcome of the students.[24,25] as a result, qle is dependent on its ability to meet students’ requirements and keep their knowledge, as well as students’ proclivity to study. the process of creating a quality educational environment can be sped up by adopting a clear methodology and a set of well-defined principles and important components of social processes, physical space, and learning technologies. students in good learning environments develop logical thinking and higher achievement without a doubt.[26,27] creating ideal learning and teaching environments are an art that promotes student performance and motivation while also allowing students to collaborate and improve their cognitive functions.[18] a healthy learning environment relieves pupils of physical discomfort, allows them to focus on schoolwork, and encourages students to think logically. students in good learning settings surely achieve more. a poor learning environment, on the other hand, is frequently drab in color, inadequate in lighting, noisy in the surroundings, and lacking in air ventilation. students in inadequate learning environments have numerous physical challenges. the purpose of this study is to shed the light on the relationship between students’ well-being and quality learning environment in the design studio. for this reason, private universities in erbil have been selected as case studies. the mixed methodology has been used to analyze the data collected from the questionnaire and checklist. the statistical analysis in this study is descriptive, correlation, one-way analysis of variance (anova), and t-test. hence, the finding of this study improves the learning environment of design studios and tends to define the improvement factors of well-being. literature review various studies studied the learning environment in the design studio and its impacts on students’ outcomes and performance. in his study, el zaza[26] attempted to employ post-occupancy evaluation of the learning environment to find elements that affect students’ satisfaction in the design studio. according to the author, lighting, thermal comfort, indoor air quality, and workplace environment are the primary characteristics related with students’ happiness with the indoor learning environment and affect students’ efficiency. they distributed semi-structured questionnaires to 60 students, and 55 of them were valid. they employed descriptive analysis to examine the questionnaire. the first segment covered building elements, while the last section used a five-point likert scale to gauge student satisfaction. the findings revealed that the workstation, indoor air quality, and thermal comfort contributed to students’ pleasure; however, artificial light had no effect. the finding showed that the students’ satisfaction has been achieved through the workspace, indoor air quality, and thermal comfort while that of artificial light has not been properly met. the result highlighted the factors of the learning environment that affect students’ comfort are the classroom, school location, school facility, school climate, and technology. obeidat and al-share[15] focused on a quality learning environment in their study, and due to a dearth of research in this sector, they chose a design studio classroom as a case study. they investigated students’ opinions of the learning environment to establish students’ requirements and goals for a quality learning environment in architectural studios. a survey method was employed to acquire 94 student responses. this study’s findings reflect earlier research indicating the physical environment that has a direct impact on space users’ pleasure. the data imply that lighting, noise, glare, air quality, temperature, seat comfort, and layout options are all important environmental aspects in achieving an ideal educational environment. besides, muniandy et al.[17] indicated that architectural design studios have different physical settings regarding the culture; therefore, they focused on the physical environment of architecture studios and how it contributes to the social environment at malaysia university. a questionnaire survey with likert-scale components and semi-structured interviews has been used. the influence of these physical indicators on students’ milieu was investigated under maslow’s theory of human motivation. furthermore, ibrahim et al.[22] highlighted a number of researches that focused on architecture curriculum and teaching methods; thus, they concentrated on the constructed environment of architecture design studios and its effects on student happiness. the design of learning settings, such as classes and studios, has a significant impact on student pleasure and performance. to assess student happiness, a survey was given to six architectural departments. each section is evaluated in terms of spatial organization, accessibility, and physical studio qualities. furthermore, the effect of the learning environment in student well-being has been investigated.[28] the developed indicators will allow the teachers to clarify the situation, draw conclusions, and improve the organizational culture. the goal of the research is to find out which indicators reflect the student’s well-being and how to measure them. the study associated well-being with the following factors: self-acceptance, positive relationship, autonomy, competence, and self-regulation of behavior. in addition, gonzález-zamar et al.,[13] for the bibliometric review focused on higher education university classrooms and students’ well-being and motivation. the purpose of their study was to analyze the research trends on the impact of the university educational space on the well-being, motivation, and social interaction of the student, considering the physicalenvironmental, socioperceptual, and motivational attributes. regarding the factors that define well-being, self-acceptance, mastery of the environment, personal growth, self-efficacy, positive relationships with other individuals, autonomy, and having a purpose that makes sense stand out in life. according to stanton et al.,[29] despite the identification of learning settings as a possible setting for developing and promoting well-being, little research has been conducted to investigate students’ perceptions of well-being in learning environments. to investigate students’ conceptions and experiences of wellbeing in learning environments, a semi-structured focus group and interview technique were utilized. the findings shed light on multiple pathways through which learning experiences contribute to student well-being and provide insight into how courses might be designed and delivered to improve student health, learning, and engagement. the findings also look into hussein and mustafa: students self-report well-being in the architectural design studios 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 21-28 the relationship between happiness, contentment, and deep learning. research problem organizations such as the world health organization concentrate on research to create a healthy learning environment. the previous research focused on well-being and student performance, as well as students’ knowledge of wellbeing. however, research on the learning environment focuses on the architecture department’s curriculum. furthermore, there has been few research on the learning environment of design studios and the well-being of students. as a result, the focus of this study is on the impact of a high-quality learning environment on students’ well-being in architectural design studios at erbil’s private institutions. it also confirms the presence of superior learning environment components in the design studio. research aim this study aims to define the availability of a quality learning environment in design studios due to students’ needs and wellbeing. moreover, this study tries to highlight the dimensions of well-being and their definitions in the learning environment of design studios. the dimensions of well-being are physical wellbeing, psychological well-being, and social well-being. research objectives the followings are the main objectives of the research: 1. to assess the existence of quality learning environment factors in the design studio 2. to establish the correlation between dependent and independent factors 3. to define the variables of quality learning environment that have the most impact on students’ well-being. theoretical framework the assessment of well-being can be addressed objectively depending on the quality of life indicators and norms called objective well-being.[1] on another hand, it can include an individual’s life experience and self-report about the perception of the learning environment named subjective well-being (swb).[30] the difference between the two types of well-being showed in figure 1. furthermore, lower objective standards of life may not always imply lower levels of swb and vice versa. this research utilizes students’ self-report assessment of the learning environment. the quality of students’ well-being in this research concerning the well-being dimensions is psychological well-being, physical well-being, and social well-being. the term psychological well-being refers to psychological functioning such as the feelings toward the capabilities of doing work, self-development, and feeling positive toward personal development.[28,31,32] however, the term physical wellbeing is based on the physical health of students.[33] moreover, in the learning environment, physical well-being is based on the comfortable learning environment regarding the lighting, temperature, and thermal environment.[34,35] social well-being is based on the social environment in the design studio, which includes the relationship with friends and peers, and having a supportive and collaborative environment to engage with others.[9,23] the layout arrangement of the design studio has a significant impact on the physical, and social environment, and teaching style. the flexibility of the arrangement layout encourages collaboration, creativity, livable natural setting, and responds to the changes in need in the design studio such as demonstration, group work, lecturing, and individual working.[14,20] for this purpose, furniture should be light in weight and movable[15]. “table 1” shows the possible arrangement layout in architectural studios. arrangement layout affects human behavior, sense of belonging, sense of community, and teaching style. anthropometrics and ergonomics of furniture is a subject of furniture dimension to fulfill students’ needs and the human body. the drafting table in the design studio should have at least (1.2* 0.80 m), with stools that are defined as the best seat type in the design studio to control model-making and drawing activities.[36,37] the spacing between seats defines the privacy of seats and establishes the movement pattern inside the studio.[38,39] the lighting condition in the design studio has unique characteristics. both natural and artificial lighting is preferred to be available in the design studio. the artificial lighting in the design studio should be evenly distributed.[40-42] for the natural lighting condition, it is preferred to have a window on both sides to avoid glare for drawing and laptop work.[22] moreover, north-facing windows are the best window orientation in a design studio.[43] thermal comfort is defined as the most important indoor environment quality parameter that affects satisfaction and comfort (ibrahim et al., 2019). thermal comfort factors are air temperature, humidity, and air velocity. the perception of thermal comfort varies among cultures.[44] temperatures between 68 and 74°f–20 and 24°c– are described as comfort levels. furthermore, 50% relative humidity is defined as an acceptable value for educational spaces in most cultures.[21] thermal comfort improves physical and psychological satisfaction.[17] thermal dissatisfaction reduces students’ concentration and creates physical stress.[37] besides these factors of quality learning environment, a view of the outside landscape is defined as a source to improve students’ motivation and help them relax.[14,20] the design studio has to be placed near the landscape context to observe a tree, garden, and natural environment. besides, the size of the window defines the connection between the indoor and outdoor environment. the esthetic value in the design studio is defined as the quality of wall color or the existence of the works of the pillars of the architecture to promote creativity.[21,22,36,44] table 2 demonstrates the research framework. figure 1: well-being aspects focusing on objective condition and subjective experience of well-being[30] hussein and mustafa: students self-report well-being in the architectural design studios 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 21-28 table 1: arrangement typology in a design studio (researcher, adapted from[14,22,26,47,48]) 1. office-like (cubical) 2. raw method 3. clustered 4. small groups 5. u-shaped or circular figure 2: case studies (researcher) privacy in architectural studios is establishing adequate space for individuals to experience all types of activities comfortably without being isolated from others.[14,45] space between seats,[22] furniture arrangement, and personal storage are defined as variables to satisfy students’ privacy within the studio, students feel more comfortable being physically and visually secure. therefore, shelves or cubbies can be added to obtain privacy. moreover, the area per user within a design studio is estimated between (4 and 4.5) m2 to be able to function well.[26] personal storage for the students and instructors provides a sense of belonging and improves selfesteem. the territoriality in the workspace environment is a sense of ownership of the physical elements.[46] methodology this study relied on a mixed methodology for the data collection and analysis process. in the qualitative approach, the researcher walkthrough has been used for the nine quality learning environment factors. in the quantitative process, the research questionnaire for the nine quality learning environment factors and three well-being dimensions has been applied. the statistical analyses for this study are descriptive analysis, correlation analysis between dependent and independent variables, the analysis of the existence of the variables using t-test analysis, and the anova to define the availability of factors in design studios. scope of the study the department of architecture at erbil university is among the college of engineering. the universities are classified as table 2: research framework (researcher) variables indicators quality learning environment flexibility easy of changing furniture layout anthropometrics and ergonomics of furniture suitable furniture with human joints lighting adequate artificial and natural lighting temperature the comfortable temperature inside the studio view to landscape quality of outside view from studios esthetic quality of wall color and paintings indoor circulation dimension of the movement paths in the studio territoriality personal limitations in the studio privacy adequate and private space per student well-being physical well-being having the physical health being in a comfortable environment doing activities in the studio and having a feeling of joy and excitement psychological well-being having a purpose for what i o in the studio being able to improve personal skills feeling motivated and encouraged to work in the studio social well-being being accepted by others have a good relationship with students have a good relationship with peers having a supportive environment to collaborate and group work hussein and mustafa: students self-report well-being in the architectural design studios 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 21-28 table 3: demographic of the study (researcher) university current number of students sample size studio the sample that i took tishk international university-erbil 377 129 tiu-1 61 tiu-2 68 university of kurdistan-hawler 60 22 ukh-1 13 ukh-2 9 cihan university-erbil 152 47 cu-e 47 catholic university erbil 25 13 cue 14 total 614 211 211 table 4: descriptive statistics for the variables (researcher) variables mean sd % flexibility 3.39 0.87 67.77 anthropometrics and ergonomics of furniture 3.05 0.95 61.04 lighting 3.59 1.42 71.85 temperature 3.18 0.93 63.55 view to landscape 2.88 0.95 57.54 esthetic 2.89 0.85 57.82 indoor circulation 3.09 0.99 61.90 territoriality 3.00 1.04 60.09 privacy 2.89 0.91 57.87 physical well-being 2.96 0.70 59.16 psychological well-being 3.21 0.78 64.17 social well-being 3.46 0.74 69.29 table 7: correlation analysis between the dependent and independent variable (researcher) correlation well-being qle pearson correlation 0.608** sig. (2-tailed) 0.000** ** correlation is significant at the 0.01 level (2-tailed) table 6: anova analysis for studios (researcher) variable studio mean sd f-value p-value qle tiu-1 3.07 0.48 2.651 0.024 tiu-2 3.31 0.64 cu-e 2.96 0.55 ukh-1 3.26 0.66 ukh-2 2.21 0.94 cue 3.22 0.50 well-being tiu-1 3.12 0.66 2.738 0.020 tiu-2 3.45 0.49 cu-e 3.09 0.53 ukh-1 3.21 0.58 ukh-2 3.15 0.76 cue 3.38 0.40 anova: analysis of variance, tiu: tishk international university-erbil, ukh: university of kurdistan hewler, cue: catholic university erbil, cu-e: cihan university-erbil governmental, international, and local private universities. this study used private universities as a case study, that are (1) tishk international university-erbil (tiu), (2) the university of kurdistan hewler (ukh), (3) cihan university-erbil (cu-e), and (4) catholic university erbil (cue). from the universities of tiu and ukh, two studios per university have been selected as case studies. as a result, the six studios from four private universities are studied. figure 2 illustrates the case studies. sample size a statistician was contacted to get the statistically appropriate ratio sample size. using the sample size equation, the total sample size of the questionnaire is 211, with a total population of 614, as equal to 34.36% of the total students. the selected sample size achieved a confidence level of 95% confidence level and a 5% error margin. [table 3] demonstrates the demography of the research data collection. results and discussion the quantitative analysis of the questionnaire data represents the descriptive mean of the quality learning environment table 5: t-test results for the existence of variables according to students’ questionnaire (researcher) variables t-value p-value flexibility 56.456 0.000** anthropometrics and ergonomics of furniture 46.68 0.000** lighting 36.76 0.000** temperature 49.571 0.000** view to landscape 43.783 0.000** esthetic 49.524 0.000** indoor circulation 45.377 0.000** territoriality 41.981 0.000** privacy 46.258 0.000** physical well-being 61.733 0.000** psychological well-being 59.712 0.000** social well-being 68.357 0.000** hussein and mustafa: students self-report well-being in the architectural design studios 26 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 21-28 figure 3: the mean of the quality learning environment and well-being for studios (researcher) figure 4: mean of quality learning environment for studios according to the checklist (researcher) table 8: simple regression in general for dependent and independent variables (researcher) independent dependent constant ß 0 f r² qle 1.28 t (7.248) p-value (0.000)** 0.620 t (11.064) p-value (0.000)** 122.4 (0.000)** 41.6% table 9: mean of quality learning environment for studios according to the checklist (researcher) variable studio mean qle tiu-1 3.41 tiu-2 3.53 cu-e 2.71 ukh-1 3.39 ukh-2 2.80 cue 3.29 factors in private universities [table 4]. the majority of the research questionnaire agreed on the existence of quality natural and artificial lighting with the greatest satisfaction mean of (3.59), and a percentage of satisfaction of (71.85%). nevertheless, the view to landscape environment has the least mean value of (2.88), with a percentage of satisfaction of (57.54%). the percentage of satisfaction with quality learning environment variables and well-being dimensions are at the medium level; therefore, there is no perfect existence of the quality learning environment factors to improve students’ well-being in architectural design studios. to analyze the significance of the existing variables in the design studio, the t-test analysis was adopted for the variables. the significant (p = 0.05), accordingly, p-value is highly significant for all variables. therefore, the existence mean of variables is significant in all studios, as shown in [table 5]. the one-way anova was applied to demonstrate the mean difference among the case studies. as shown in [table 6], tiu-2 has the highest mean value of quality learning environment factors with a total value of (3.31) and its students have the greatest well-being value with a total of (3.45). however, ukh-2 has the lowest quality learning environment value of (2.21), and cu-e has the lowest student well-being value of (3.09). as a result, the design studio of tishk international university has the highest quality learning environment and well-being value [figure 3]. to define the relationship between the dependent variable (well-being dimensions) and independent variable (quality learning environment variables), this research conducted the pearson correlation analysis. the range of correlation coefficients is from −1.00 to +1.00. several −1.00 denotes a perfect negative correlation, whereas a value of +1.00 denotes a perfect positive correlation. a two-tailed statistical significance test with a range of 0.05–0.01 was performed on the correlation technique. according to [table 7], the correlation analysis represents the perfect positive correlation between well-being and quality learning environment with a pearson correlation value of (0.608). as the value of significance which is the collection of all factors value is smaller than 0.01 and 0.05, the correlation is significant. from this, we can conclude that the development of quality learning environment variables can improve students’ well-being. moreover, the increase in students’ well-being in the learning environment encourages students to spend most of their time in the design studio happily with greater achievements and performance. the regression analysis in this study clarifies the impact of independent variables (quality learning environment) influences the dependent variable (well-being). the regression analysis in [table 8] indicated that quality learning environment variables cause 41.6% changes in students’ well-being. moreover, the result is highly significant because p < 0.01 and 0.05. on another hand, the qualitative researcher checklist in [table 9] represents the mean value of quality learning environment variables in the six architectural design studios. according to the checklist, tiu-2 has the highest mean with a total value of (3.53), and then, it is followed by tiu-1, ukh-1, and cue with values of (3.41, 3.39, and 3.29). the architectural design studios with the least mean value are ukh-2 and cu-e with values of (2.8, 2.71), as shown in (figure 4). hussein and mustafa: students self-report well-being in the architectural design studios 27 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 21-28 conclusion the present study justified the impacts of quality learning environment variables such (as flexibility, anthropometric and ergonomic, natural and artificial lighting, temperature, esthetics, view to outside, indoor circulation, territoriality, and privacy) on students’ well-being. the well-being dimensions in the present study are defined to be physical well-being, psychological well-being, and social well-being. the mixed methodology has been used for the data collection method. the qualitative method in this study depended on collecting data from photos and researcher walkthroughs, while the quantitative method represents the data collected from students’ questionnaires. the spss program has been used to statistically analyze the data to get the research objectives. the result indicated that tishk international university studios have the highest quality learning environment and the greatest students’ well-being perception. thus, the t-test analysis represented the existence of the quality learning environment variables in design studios in private universities. this study has significant value in the field of scientific research due to the investigation of the correlation between two group variables that have not been described yet. moreover, the development of quality learning environment factors improves the students’ well-being. furthermore, the development of students’ wellbeing encourages a healthier learning environment that attracts students to spend most of their time in the design studio with greater achievements and outcomes. references 1. s. oliveira, e. griffin, d. cash and e. marco. health and wellbeing in design studio briefs-architecture and engineering graduating students’ motivations and approaches. building 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administration, vol. 47, pp. 381-399, 2009. 44. s. arif and m. ilyas. creating a quality teaching learning environment. international journal of learning, vol. 18, pp. 51-70, 2012. 45. a. s. mutaqi. architecture studio learning: strategy to achieve architects competence. in: shs web of conferences. edp sciences, france, p. 04004, 2018 46. m. g. modell and c. m. gray. searching for personal territory in a human-computer interaction design studio. journal for education in the built environment, vol. 6, pp. 54-78, 2011. tx_1~abs:at/add:tx_2~abs:at 60 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 60-66 research article prevalence of anemia, iron deficiency anemia and its sociodemographic factors among pregnant women in garmian province, kurdistan region of iraq mustafa y. rashid1, adnan l. abdul raheem1, gulzar s. hama amin1, shwan s. weli2, rahman k. faraj3, ali j. muhialdin4, seerwan a. raheem1 1department of biology, university of garmian, kurdistan region, iraq, 2kalar public hospital, kurdistan region, iraq, 3department of chemistry science, college of science, university of garmian, kurdistan region, iraq, 4department of animal production, college of agricultural engineering sciences, university of garmian, kalar, kurdistan region, iraq abstract anemia is the most common hematologic abnormality in pregnancy. maternal anemia is associated with adverse fetal, neonatal, and childhood outcomes. this study aims to determine the prevalence of anemia, and iron deficiency anemia (ida), the severity of the condition, and study the effect of some sociodemographic factors on pregnant women in garmian province. the study was conducted among 157 pregnant women in garmian province between 17 and 49 years old. participants completed a questionnaire that included sociodemographic characteristics, disease, and gestational age. a hematological evaluation, including a complete blood count and serum for ferritin testing. results of this study have shown that the prevalence of anemia and ida were 34.4% and 15.3%, respectively. almost three-quarters of the pregnant women had mild anemia, while 31% of the participants had moderate anemia, and about 60% of the participants were diagnosed with normocytic anemia. the second trimester had the highest prevalence, with 51.9% for the anemic and 45.8% for ida participants, while the first trimester showed the lowest prevalence, with 14.8% for the anemic and 12.5% for ida participants. age, occupation, gravidity with anemia, and ida did not make a significant difference. moreover, there was no significant difference in blood indices between anemic and ida participants. the serum ferritin level was unaffected by the pregnancy trimesters. keywords: anemia, hematological parameters, iron deficiency anemia, pregnant women, severity of anemia introduction anemia in maternity is defined by a low hemoglobin (hb) concentration of <11 g/dl. furthermore, the low red blood cell (rbc) count, and decreased packed cell volume.[1,2] pregnancy-related anemia is a highly unusual finding in hematological tests. it is thought to have an unfavorable effect on fetuses, newborns, and children. anemia in the mother will increase the likelihood of a transfusion during birth. iron deficiency (id) is the most significant problem with anemia during pregnancy.[2] iron plays an important role in a wide range of cellular metabolism, including oxygen transport by rbc hb, electron transport, and dna synthesis. almost three of the four parts of the body’s total iron are contained in heme, hb, and myoglobin, with the remaining parts stored as ferritin and hemosiderin.[3] id is the insufficiency of iron in the body below normal levels to maintain all physiological functions. id anemia (ida) develops when the bodily iron levels drop too low to maintain healthy rbc synthesis.[4] in the absence of infection, serum ferritin is a susceptible and widely used test for the diagnosis of id, which shows decreased serum ferritin with <15 ng/ml in the age of 5 years or older, both in males and females.[5] it has been shown that ida has been associated with hazardous neonatal mortality, premature delivery, low birth weight (lbw), child cognitive growth, and motor development in pregnant women.[6] id is the most common cause of anemia, it causes around 50% of all types of anemia, and this is the most widespread nutritional deficiency globally.[7] id is the probable cause of about 75% of all anemia cases during pregnancy.[8] id is a major dietary problem throughout the world. a pregnant corresponding author: mustafa y. rashid, department of biology, university of garmian, kurdistan region, iraq. e-mail: mustafa.yunis@garmian.edu.krd received: april 15, 2023 accepted: may 23, 2023 published: june 15, 2023 doi: 10.24086/cuesj.v7n1y2023.pp60-66 copyright © 2023 mustafa y. rashid, adnan l. abdul raheem, gulzar s. hama amin, shwan s. weli, rahman k. faraj, ali j. muhialdin, seerwan a. raheem. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/ rashid, et al.: prevalence of anemia, iron deficiency anemia among pregnant women 61 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 60-66 woman’s diet is inadequate to satisfy her iron requirements.[9] women need 4.4 milligrams of iron daily during pregnancy,[10] which is approximately double their normal requirement. in addition, they need more iron than usual due to the increased need for iron during pregnancy.[11] when the mother’s rbc mass increases along with fetal growth, the iron requirements increase. women need to take additional iron supplements daily during their pregnancy. taking these supplements significantly lowers the risk of developing anemia during pregnancy.[12] anemia is considered a global health problem that influences many countries with a big concern for public health besides communal and financial development.[13,14] it affects people at different stages of life but is more common in pregnant women and children.[13] globally, anemia affected around 29% of non-pregnant women and about 38% of pregnant women aged 15–49 in 2011. in 2019, anemia was 29.6% in nonpregnant women and 36.5% in pregnant women.[15,16] there is a wide range of social, cultural, and environmental factors that affect the prevalence of anemia during pregnancy. anemia is widespread in many countries worldwide. it is highly influenced by nutrition, infectious diseases, genetics, and strong menstrual bleeding on a biological level.[17] it has been shown that the risk of anemia is increased by deficiencies of vitamins a and b12, c, d, e, folate, zinc, riboflavin, and copper, and infection with hookworm, schistosomiasis, and hiv.[17,18] furthermore, it has been shown that there is a potential relationship between helicobacter infection and anemia.[19] however, normal hemodilution occurs during pregnancy and ranges between 20 and 24 weeks, and hb levels vary throughout pregnancy.[11] it is well known that hb naturally decreases throughout the second trimester. this decrease is related to a larger increase in plasma levels compared to rbc mass. the relative hemodilution of blood viscosity brought on by this process promotes placental blood flow.[20] some women have little or no iron stored when they become pregnant, and this problem can cause severe id during pregnancy. according to the who recommendations on the prevention, control, and treatment of anemia in women as part of prenatal treatment, daily oral iron and folic acid supplementation is advised to lower the risk of lbw, maternal anemia, and id. according to the united nations multiple micronutrient preparation, supplements may include additional vitamins and minerals in addition to iron and folic acid to address any potential maternal micronutrient deficits. any attempts to prevent and control anemia should be supported by a diet rich in bioavailable iron.[21] this study aims to determine the prevalence of anemia, ida, the severity of the condition, and some associated sociodemographic characteristics in pregnancy in the garmian area. materials and methods study design this study was performed on 157 pregnant women aged 17–49 years, who attended both garmian medical city and zanko medical laboratory during august 2021 and october 2022 in kalar city/kurdistan region of iraq. blood samples have been collected from the patients to perform hematological assessments anonymously. data collection the data were collected from pregnant women by using a questionnaire form. the questionnaire was created to investigate disease and sociodemographic factors. the pregnant women were divided based on gravidity into primigravida and multigravida, and on their occupations into employees and nonemployees, with ages ranging from ≤29, 30–39, to 40–49 years. laboratory assessment venous blood (5 ml) was collected from the participants in this study and placed in an edta tube for a complete blood count (cbc) test, and a gel tube for a serum ferritin test. in zanko medical laboratory, the blood samples were collected, and the cbc test was performed on a fully automated hematology analyzer (medionic m51). the collected blood was centrifuged by an (lc-04l centrifuge). the serum ferritin test was assessed by (cobas e411 analyzer) after serum preparation by centrifugation of blood sample at 3500 rpm for 10 min. each test was done immediately after collecting the blood samples. in garmian medical city, blood samples were collected, and a cbc test was analyzed by a fully automated hematology analyzer (swelab alfa plus) for the cbc test. centrifugation of blood samples was performed at 3500 rpm/10 min by (an 80–1 electric centrifuge). serum ferritin was tested with a cobas e411 analyzer after serum preparation. all tests were performed immediately following the collection of blood samples. differential value and statistical analysis haemoglobin and serum ferritin criteria were used for the diagnosis of anemia and ida based on who guidelines.[1,5] pregnant women who measured hb <11 g/dl and serum ferritin <15 ng/ml were defined as ida. the severity of anemia is categorized according to who thresholds,[1] including mild anemia, where hb levels range between 10 and 10.9 g/dl, and moderate anemia, where blood hb levels range between 7 and 9.9 g/dl. the present findings were compared with how the who categorizes anemia and ida in the community as significant for public health based on prevalence estimates derived from hb and serum ferritin levels in the blood.[1,5] anemia was divided into two categories based on mean cell volume (mcv) levels.[22] statistical analysis was evaluated using microsoft excel 2014 for the achieved data. the chi-square test was used to determine the difference between the different groups. a p < 0.05 was considered statistically significant. ethics statement the studies involving human participants were reviewed and ethically approved by the scientific and ethical of the biology department, university of garmian. the participants provided their written informed consent to participate in this study. participants were guaranteed anonymity and confidentiality for their responses, and they were assured of having the rashid, et al.: prevalence of anemia, iron deficiency anemia among pregnant women 62 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 60-66 ability to withdraw their responses for any reason at any time, without any implications. results in the present study, out of a total of 157 pregnant women in the garmian/kurdistan region of iraq of different ages ranging from (17 to 49) years, 124 were from kalar, 6 from kifry, 21 from rizgary, 3 from bawanour, and 3 from sarqala were involved in this study. based on hb levels, the prevalence rate of anemia among the total number of women was 54 (34.4%) with a mean hb of 10.1 ± 0.83, and (65.6%) non-anemic women with a mean hb of 12.14 ± 0.79, as shown in table 1. furthermore, 54 (34.4%) pregnant women had an id with a mean serum ferritin of 8.92 ± 2.8 ng/ml, and 103 (65.6%) pregnant women did not have id with a mean serum ferritin of 42.64 ± 32.44 ng/ml, [table 2]. in addition, 24 (15.3%) of pregnant women were diagnosed with ida with a mean hb of 10 ± 1.05 g/dl, and the mean of serum ferritin was 7.51 ± 2.88 ng/ml, [table 3]. in the present study, out of 54 anemic patients, 37 (69%) of patients had mild anemia with hb level ranges between 10 and 10.9 g/dl, while 17 (31%) of the patients had moderate anemia with hb level ranges between 7 and 9.9 g/dl. severe anemia was not observed among the patients, [figure 1]. moreover, microcytic anemia was observed in 21 (38.9%) pregnant women, while 33 (61.1%) of pregnant women were diagnosed with normocytic anemia, as shown in figure 2. results of this study have shown that anemia and ida were most prevalent among pregnant women in their second trimester (51.9%) and (45.8%), respectively, compared to the first and third trimesters. however, the first trimester showed the lowest prevalence rate of anemia and ida, with (14.8%) and (12.5%), respectively, as shown in table 4. in the present study, women ≤29 years old showed the highest prevalence of anemia with 55.6% and ida at 62.5%, while 40.7% of the anemic patients and 29.2% of the ida patients were ranged between 30 and 39 years old, and the women with age group between 40 and 49 years old showed the lowest prevalence of anemia and ida with 3.7% and 8.3%, respectively. there was no significant difference between age, occupation, and gravidity with anemia and ida with p = 0.1, 0.5, and 0.3, respectively, [table 5]. the gestational age concerning blood indices, which include these tests; rbc, haematocrit, mean cell hemoglobin (mch), mch concentration (mchc), and red cell distribution width (rdw), have shown that there is no significant difference between anemic pregnant women and ida patients [table 6]. discussion maternal anemia plays an essential role in elevating perinatal and neonatal mortality.[23] moreover, the consequences of ida on the physical and mental development of newborns and the productivity of adults are a major concern.[24] the present results showed that 65.6% of pregnant women were table 2: iron deficiency and non-iron deficiency prevalence among pregnant women (n=157) prevalence number percentage mean sd iron deficient 54 34.4 8.92 2.8 non-iron deficient 103 65.6 42.64 32.44 %: percentage of iron deficient and non-iron deficient pregnant women, sd: standard deviation table 3: prevalence of iron deficiency anemia in pregnant women (n=157) prevalence number percentage mean sd iron deficiency anemia hb 24 15.3 10 1.05 serum ferritin 24 15.3 7.51 2.88 hb: hemoglobin, %: percentage of iron deficiency anemia among women according to their hb and ferritin levels table 1: prevalence of anemia among pregnant women (n=157) prevalence number percentage mean sd anemic 54 34.4 10.1 0.83 non-anaemic 103 65.6 12.14 0.79 %: percentage of anemic and non-anemic pregnant women, sd: standard deviation figure 2: the distribution of types of anemia among anaemic patients figure 1: the distribution of the severity of anemia among the anemic patients rashid, et al.: prevalence of anemia, iron deficiency anemia among pregnant women 63 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 60-66 non-anemic, while 34.4% were anemic. based on the level of hb in the blood, indicates a moderate public health concern.[1] these results are consistent with a previous study that showed that the mean hb level ranges to <11 g/dl.[25] the prevalence of anemia among pregnant women was 34.4% with id and 65.6% with non-id, id may occur due to multi conditions such as bleeding, stomach and intestinal ulcer, and menstrual period and may occur with an inadequate supply of different nutrients, especially for the pregnant women who need more iron supply to replace the amount of lost iron each day, since most of the participants in this study are unemployed, and dietary awareness of the women might play an important role in developing id.[26] this result of id is considered a moderate public health issue based on the blood hb level and ferritin level according to who based on the prevalence estimated from serum ferritin levels <15 μg/l.[5] about 69% of the anemic participants had mild anemia, and 31% had moderate anemia; the present findings are concomitant with other researchers with mild anemia of 72.9%, 32.5%, and 28.1%, respectively.[27,28] these varieties may relate to dietary diversity in different areas worldwide. they could be attributed to differences in socioeconomic, dietary habits, and health-seeking behaviors between cities with a community of relatively different lifestyles, feeding practices, and social norms.[29,30] such as the consumption of red meat and leafy green vegetables that are an essential source of heme iron. many studies have revealed that pregnant women who eat meat occasionally were 2.1 times at higher risk of being anemic than pregnant women who ate meat one or more times per week.[25,31] other studies showed that drinking tea and coffee after meals which are a main inhibitor of non-heme iron absorption due to phenolic compounds in these beverages, might increase the risk of developing anemia.[29,31] all these factors are considered a significant association with anemia in pregnancy.[25,32,33] in addition, insufficient use or consumption of hematopoietic agents required for normal erythropoiesis, including iron, b12, and folic acid, appears to be a prevalent vitamin deficiency worldwide.[29,34] anemia etiologies can be distinguished into microcytic, normocytic, and macrocytic anemia using the mcv value. in the results of the study, microcytic and normocytic anemia are diagnosed in anemic pregnant women; macrocytic anemia is not. ida is the most probable cause of microcytosis, including other factors such as premenopausal, menstrual, or blood loss without proper iron supplementation, thalassemia, sideroblastic anemia, and anemia caused by chronic diseases.[22] most normocytic anemias are side effects of other diseases; they reflect a primary blood disorder. this is due to the anemia of chronic disease (inflammation, cancer), renal failure, endocrine failure (hypothyroidism, hypopituitarism), acute blood loss, marrow failure, and hemolysis.[35,36] according to the study’s results, 38.9% of anemic subjects had microcytic anemia, and 61.1% had normocytic anemia; these results are comparable with those of the study done by bayarkhuu and tsogbadrakh,[37] in which 37.07% of pregnant women were affected by microcytic anemia, 46.55% by normocytic anemia, and 3.2% by macrocytic anemia. in a study conducted by singh et al.[38] at king george’s medical university on 440 reproductive-age females from the tharu tribe living in the balrampur district along the indo-nepal border, microcytic, normocytic, and macrocytic anemia were found in 53%, 44.88%, and 2.12% of the subjects, respectively. the result of gestational age showed that there was no significance between ida and anemic participants in relation to blood indices, and this finding is in concomitant with other study conducted in saudi arabia, which reported that there was no significant difference between anemia and blood indices associated with gestational age, and stated that a lower rate of spontaneous vaginal delivery was significantly associated with anemia, and this might be attributed to the fact that anemia was mostly mild and most participants were taking iron supplements.[39] a study by liu et al.[40] also reported that preconception mild anemia increases the riskiness of lbw and small for gestational age (sga) by 17% and 14%, respectively, table 4: the association between anemic and ida among pregnant women in gestational age prevalence gestational age first trimesters second trimesters third trimesters no. percentage no. percentage no. percentage anemic=54 8 14.8 28 51.9 18 33.3 ida=24 3 12.5 11 45.8 10 41.7 ida: iron deficiency anemia, %: percentage of anemic and ida women according to their trimesters table 5: the association of sociodemographic characteristics among anemic and ida pregnant women variable anemia ida p-value no. % no. % age group ≤29 30 55.6 15 62.5 0.106609∗ 30–39 22 40.7 7 29.2 40–49 2 3.7 2 8.3 total=54 (100) total=24 (100) occupation non employees 43 79.6 22 91.7 0.519276∗ employee 11 20.4 2 8.3 total=54 (100) total=24 (100) gravidity primigravida 23 42.6 13 54.2 0.343961∗ multimigravida 31 57.4 11 45.8 total=54 (100) total=24 (100) ida: iron deficiency anemia, ∗there is no significant difference between the different groups and the p>0.05 rashid, et al.: prevalence of anemia, iron deficiency anemia among pregnant women 64 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 60-66 but moderate and severe anemia had no significant association with lbw and sga. in addition, maternal anemia and high hb levels before pregnancy contribute to the increase of these risks of sga, birth, preterm birth, and perinatal death. moreover, a study by patra et al.[41] has stated that women with severe anemia during the third trimester of pregnancy have high maternal mortality (6.2%) and perinatal mortality (60%). results of this study have shown that different ages, occupations, and gravidity did not produce significant influence both on anemia and ida; no significance was found in gestational age concerning blood indices between the two groups. the result of this study was different from other studies regarding primigravida and multigravida and parity, and these studies indicate that frequent pregnancies are associated with anemia because frequent pregnancies do not give enough time to restore depleted iron stores.[41,42] some risk factors that lead to ida are multiple pregnancies and adolescent pregnancy, as well as delivery with hemorrhage.[43,44] many studies showed the role of blood indices and are recommended as the first step in evaluating pregnancyassociated anemia. the mcv, mch, mchc, rbc count, and rdwcv% correlate with the ferritin levels and can help diagnose ida.[45-47] at the same time, this study’s result is in contrast with these studies and showed no significant influence in blood indices of the gestational age between the anemic women and ida women. however, this study result was similar to a study done by sultana et al.[48] showed that some hematological parameters, such as mcv, mch, and mchc, have low reliability in detecting ida during pregnancy. these red cell indices may be mean values, which cannot express the small variation of red cell size that occurs in the early stage of id; moreover, that may relate to an increase in plasma volume leading to hormonal disturbances and id.[48-50] conclusion anemia and ida during pregnancy continues to be a major health problem worldwide as well as in this study area. threequarters of pregnant women had mild anemia, and about twothirds were diagnosed with normocytic anemia. the second trimester had the highest prevalence of anemia and ida. there was no significant effect of blood indices and sociodemographic characteristics on anemia and ida. the limitations of our study were that it was limited to women’s specialized hospital, and some information relating to the study parameter was missing. we suggest hematological examination for hb and iron status from the beginning of pregnancy. references 1. who. haemoglobin concentrations for the diagnosis of anemia and assessment of severity. 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m. mcdonagh. routine iron supplementation and screening for iron deficiency anemia in pregnancy: a systematic review for the us preventive services task force. annals of internal medicine, vol. 162, no. 8, pp. 566-576, 2015. table 6: the association between anemic and ida pregnant women of gestational age in relation to blood indices variable gestational age f p-value anemic=54 ida=24 first trimester second trimester third trimester first trimester second trimester third trimester mean±sd mean±sd mean±sd mean±sd mean±sd mean±sd rbc 4.27±0.69 3.89±0.35 3.89±0.62 4.53±0.14 3.8±0.3 3.7±0.35 0.706 0.49∗ hct 31.3±1.68 31.6±2.91 31.8±1.9 30.5±2.23 30.9±3.77 31.8±2.3 0.190 0.82∗ mch 24.05±5.14 26.2±3.49 26.7±3.94 20.43±2.05 26.4±3.8 27.8±1.9 1.328 0.27∗ mchc 31.69±1.33 31.96±0.94 32.02±1.06 30.3±0.25 32.3±1.02 32.2±1.19 2.354 0.102∗ %rdw 14.3±1.47 14.7±1.85 14.4±2.08 14.4±0.4 13.9±1.19 13.7±1.4 0.149 0.86∗ rbc: red blood cell, hct: haematocrit, mch: mean cell hemoglobin, mchc: mean cell 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of economic, faculty of economics and administrations, nawroz university, kurdistan region, iraq, 2department of statistics, faculty of economics and administrative sciences, van yuzuncu yil university, van, turkey abstract in this article, two machine learning methods such as classification and clustering are used for decision tree (dt), artificial neural network (ann), and k-nearest neighbors algorithms. the datasets were used to evaluate the effectiveness of the clustering method and the data mining tool. weather data were used to compare algorithms and methods in the study. this study showed that the best model was dt according to accuracy and precision measures but the best model according to f-measure and receiver operating characteristic curve area measures was ann. waikato environment for knowledge analysis, a data mining tool, is utilized in this paper to carry out the clustering. keywords: algorithms, classification, clustering, machine learning, decisions tree introduction data analysis science is one of the disciplines that was created to investigate natural phenomena and address current or upcoming issues. data analysis science aims to shed light on future studies by employing techniques and theories that are appropriate for data structures with a small number of observations to describe the subject with a certain probability. every science discipline in nature has a lot of data, and access to these data is becoming simpler every day. it always raises the question of how much of the collected data can be used or how significant it is. while going about their regular lives, humans not only receive information but also consume it. due to the extensive use of data, classification was required to make the data meaningful and produce new data. a set of data is classified when it is categorized based on algorithmdetermined distinguishing characteristics. data cannot always be categorized manually because millions of data types related to a field are created in just 1 day. the extremely convenient operation of the algorithms makes the classification of data very easy. the classification of data using many algorithms simultaneously can result in a variety of outcomes. which algorithm is used to classify the data set effectively relies on the data set. since not all algorithms will give the same accuracy to every data set, it is possible that the techniques employed to describe the data will also be erroneous for that data set. machine learning and classification algorithms are prominent in this direction.[1] data analysis was done using the open-source waikato environment for knowledge analysis (weka) program, which is a tried-and-true data mining tool with a graphical user interface and regular terminal programs. this program has helped to classify the data by rebalancing and decreasing its dimension. the weka project seeks to offer a full range of machine learning tools and preprocessing methods to academics. this allows applications to rapidly test and contrast various machine learning process strategies on new datasets.[2] literature review in soofi and awan,[3] discussed the fundamental classification strategies in this paper. later, he covered some of the main classification technique subcategories, including bayesian networks, decision tree (dt), k-nearest neighbor (knn), and support vector machine (svm), along with their advantages, disadvantages, prospective applications, problems, and potential solutions. this study’s objective is to offer a thorough evaluation of several machine learning classification methods. cihan university-erbil scientific journal (cuesj) corresponding author: guhdar a. ahmed, department of economic, faculty of economics and administrations, nawroz university, kurdistan region, iraq. e-mail: guhdar.abdulaziz@gmail.com received: march 3, 2023 accepted: may 19, 2023 published: june 10, 2023 doi: 10.24086/cuesj.v7n1y2023.pp52-59 copyright © 2023 guhdar a. ahmed, yıldırım demir. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0) cihan university-erbil scientific journal (cuesj) mulla and demir: the use of clustering and classification methods in machine learning 53 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 both experts and newbies to the field of machine learning will benefit from this research’s efforts to strengthen the conceptual underpinnings of classification techniques. in this study sharma,[4] the many kinds of existing categorization algorithms were thoroughly explained in this work. sharma will mostly compare and discuss in-depth the key 7 categories of categorization algorithms here. the comparison will mostly be focused on each system’s assumptions, benefits, and disadvantages. sharma will address some of the most popular classification algorithms used, different scoring schemes to gauge their effectiveness, multiclass classification strategies, and finally model selection techniques in this paper. the study sisodia and sisodia,[5] have covered the prediction of diabetes using nave bayes, dts, and svms as classification algorithms. the major goal of this study is to create a model that can accurately predict the likelihood of developing diabetes. weka tool was utilized for data classification, with the pima indian diabetes dataset serving as the primary dataset. this research work by ambigavathi and sridharan,[6] the purpose of this work was to examine several clustering techniques from both theoretical and experimental angles. trial findings revealed which category had the best algorithm using a set of physiological data. a variety of internal as well as stability measures were used to validate the effectiveness of each clustering technique in machine learning. this research concluded by highlighting future directions for highdimensional health-care data sets using a suitable clustering technique. in this study lorena et al.,[7] in this research, we investigated resampling techniques and metrics that may be obtained from training datasets to characterize the complexity of the classification issues. recent literature also looked at and addressed their approaches, opening up the possibility of new areas for study. the extended complexity library r package, which includes a number of complex measurements and is finished and publicly accessible, was also defined. this research work by liu et al.,[8] centered on addressing issues with the class imbalance and identifying attribute correlations. kaggle offers two datasets for fraud detection that may be used to create classifiers and assess the effects of various data processing methods. through their approach, they were aware of the latest fraud detection discoveries, learned more about various data processing techniques, and created several sorts of classifiers. they supported the need of addressing class imbalance and examining attribute connections. finally, they looked at the relationships between each attribute and marginally better performance. their experimental findings clearly demonstrated that addressing class imbalance had a beneficial effect on unbalanced data sets and that examining the link between qualities would also be helpful. in zheng,[9] investigation on the application of synthetic minority oversampling technique (smote) with various classifiers. to build predictive models on the heart disease data, the smote method was used for comparison. regular smote, borderline-smote, svm-smote, and k-means smote were combined with four classification algorithms under the classical and neyman pearson (np) paradigms. the performance of these models was compared. according to their findings, the svm-smote and borderline-smote perform better than other smote variations, and the np classification is better at successfully controlling type i error. materials and methods weather forecasting: the machine learning approach before the development of models, people made predictions for a given location based on observable data collected over time. for example, they used a basic approach known as climatology to predict the weather. rough estimations of long-term trends and cumulative values of variables such as temperature or precipitation may be generated by applying basic statistics to recorded weather events over long enough periods. with the development of tools to accurately monitor the atmosphere and disseminate these readings across geographically far places, weather maps became available around the start of the 20th century. with the advent of weather forecasting techniques that anticipated the movement and effects of these pressure systems in the atmosphere, these maps first started to show the position and shape of lowand high-pressure systems. the use of statistical models in weather forecasting was first advocated in the 1950s.[10] malone claimed that “statistics must ultimately play some part” in atmospheric simulation at the same time that the first numerical weather prediction (nwp) models were being built. the author developed a method for forecasting the sea-level pressure field using multiple linear regression. despite the optimism malone expressed in the early 1950s, machine learning and statistical methods have actually been applied as a complement to nwp rather than as a replacement for it. other approaches have not yet been able to match the power of physical equations to simulate the evolution of the atmosphere along the spatial and temporal dimensions. we discuss examples of machine learning approaches being used to address various issues in the field of weather forecasting in this section. the following categories of challenges, in which various machine learning approaches have been effectively employed to enhance our understanding of the processes in the atmosphere, can be identified depending on the task to be solved and the nature of the underlying data. kinds of machine learning supervised, unsupervised, and reinforcement learning methods were used to classify machine learning algorithms: figure 1 depicts the categorization in a visual manner.[11] supervised learning for machine learning, supervised learning is crucial. averaging input and output is the aim of supervised learning. the input data are a list of many interesting items, each of which is referred to as an attribute or an example. the output is a manager’s assessment or conclusion. in classification, a form of supervised learning, averaging (or a discriminant function) is used to distinguish between several groups of occurrences. a list of the different classes is included in the output, mulla and demir: the use of clustering and classification methods in machine learning 54 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 sometimes referred to as the class label in machine learning. the term used to describe the discriminant function is classifier or model. a training set is a set of instances for which the class label has been determined. during classification, a model is developed using a set of parameters to create a mapping between instances in the training set and labels in the training set. to identify or recognize previously undiscovered scenarios, the trained model may be used. supervised learning is the main methodology used in real-world machine learning. in supervised learning, the function that converts input variables (x) to output variables are learned using an algorithm (y). y = f(x) the objective is to anticipate the output variables (y) using new input data by as nearly approximating the mapping function as is practical (x). since an algorithm learns from the training dataset is akin to a teacher guiding the learning process, it is known as supervised learning. when the algorithm operates satisfactorily, learning comes to an end. the algorithm iteratively makes predictions on the training data and is corrected by the teacher as we are aware of the appropriate responses. learning comes to a conclusion when the algorithm works satisfactorily.[12] unsupervised learning it is a machine learning technique used when training data is not classified or labeled. to describe a hidden structure, unsupervised learning examines systems’ capacity to infer a function from unlabeled data. the system examines the data and may utilize datasets to infer hidden structures from unlabeled data even when it is unable to select the ideal output. unsupervised learning occurs when there are just input variables and no matching output variables (x). unsupervised learning simulates the underlying distribution or structure of data to understand it better. these are referred to as unsupervised learning as there are no correct answers and no teachers, in contrast to the supervised learning that was previously discussed. it is up to the algorithms to find and display the fascinating structure of the data. issues with grouping and association abound in unsupervised learning. in contrast to association rule learning problems, where you are looking for rules that adequately describe significant portions of your data, such as people who buy x also tend to buy y, clustering problems are looking for the inherent groupings in the data, like grouping customers based on their purchasing behavior. unsupervised learning techniques include the a priori methodology for learning association rules and k-means for classifying problems.[12] what kinds of data can be mined? any sort of data may be used in data mining, as long as it is pertinent to the application for which it is intended. the most basic sorts of data for mining applications are databases. it is also possible to mine other forms of data, including data streams, ordered/sequence, graph or networked, geographical, text, and multimedia data. data mining will undoubtedly continue to include new data kinds as they emerge. machine learning models in classification method when semantic classes for particular items are already known, classification refers to the process of determining those classes based on those objects’ properties. it involves putting fresh observed samples into a specific class by comparing the sample’s characteristics to an already existing class. by building a model using earlier data, it may then decide on the unobserved cases. therefore, it finds a model (or function) that clarifies and divides groups of data or ideas. it looks at the relationship between the values of the variables for each row and the label assigned to that row using data mining techniques. various classification methods exist, including naïve bayes (nb), logistic regression, svm, knn, dt, and artificial neural network (ann). as a result, the extracted data may be used to predict a label for a new row when it is supplied to the classifier based on the variable values that define it. these methods use different ways to express these interactions. because these correlations change between datasets, it is critical that the classifiers be trained using labeled training data. classification is one of the findings and predicting process a result from a set of data. the algorithm uses a collection of variables and a training set with the property of each variable to predict the outcome. the outcome is often referred to as a target or forecast variable.[13] this article discussed dt, ann, knn. dt it is known that one of the models used in machine learning is dts.[14] a non-parametric, intricate, and computationally costly sorting technique is the dt approach, often known as the recursive partitioning algorithm. the main idea is to keep subdividing the subsample into subgroups until it allows for decision-making, beginning with dividing the sample responses into fresh subsamples that are as similar to one another and distinct from them as practicable. while the nodes reflect the sub-samples, the root node represents the total sample. figure 2 shows a dt that categorizes weather conditions for those as good and bad. the dt models are built employing the greedy method, and the best split is that which minimizes the weighted drop in impurity after evaluating a large number of variable splits at each node: �i l l r rs t i t p i t p i t,� � � � � � � � � � � (1) figure 1: overview of machine learning techniques mulla and demir: the use of clustering and classification methods in machine learning 55 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 where p l and p r indicate the percentage of observations connected to the node t are sent to the left child node t l or right t r he relevant chide node. knn a machine learning method for categorizing data is the nearestneighbors approach, also known as pattern recognition.[15] this classifier’s objective is to select a metric to compare the separation between any two applicants over the whole set of application data.[16,17] proposed using the following metric: d x y x y i dww x y t t, ��{ �� �( �� �( ��)}� � � �� � � � 1 2 (2) where x and y are in the feature space points, i, d is a distance between parameters, w is a specific direction in the measurement space and is the identity matrix. ann ann are mathematical representations of the way the human brain works.[18] with neural networks, practically any nonlinear relationship between input and objective variables may be simulated thanks to their adaptability. despite the numerous proposed architectures, the multilayer perceptron is the one that this article concentrates on (mlp). neurons for each input variable are located in the hidden layer of an mlp, which typically comprises three layers (in our case, one neuron). each neuron processes the information it receives and transmits the results to the neurons in the layer beneath it. each of these neural connections receives a weight during training. using the weighted inputs and its bias term as an example, the logistic function, b i (1) results in the computation of the hidden neuron’s output: h f b w xi j i n ij i�� � � ( )� � � � � � � � � � � � �1 1 (3) w ij stands for the weight connecting input where w is a weight matrix. the weight j represents the input connection to hidden neuron i. machine learning models in clustering method clustering is one of machine learning’s unsupervised learning strategies.[19] when no class labels are available to analyze the datasets, the clustering strategy is critical for breaking down the massive volume of data into smaller groupings of data. each data point is classified and grouped into a distinct cluster using the clustering approach, and each cluster comprises a collection of data points. moreover, data points within the same cluster should have comparable attributes, whereas those inside a separate cluster should have significantly different features.[20] partitioning around medoids, also referred to as k-medoids and hierarchical k-means. the main operations of these algorithms are separated into the following groups. k-means clustering algorithm the primary function of the simple and frequently used clustering method k-means is to classify the provided unlabeled dataset. the main goal of this method is to identify clusters that are comparable to each other, as indicated by the variable k. in this approach, the cluster is described statistically using the mean or centroid. not usually part of the collection, a centroid is a data point that symbolizes the center of a cluster. after that, the k clusters are constructed by grouping the n data points into them, with each data point joining the cluster with the closest centroid. the euclidean distance between each data point n and the cluster centroid is then calculated precisely. a cluster’s data points are always assigned to the place with the shortest euclidean distance from the centroid point. when there are no available data points to assign, an early grouping is considered. the technique is then repeated until the “c” centroids stop migrating and the new “c” centroids are identified.[21] hierarchical clustering hierarchical, also known as hierarchical cluster analysis, is a particular kind of unsupervised. with the use of a tree-based framework, hierarchical cluster analysis aims to merge multiple clusters made up of comparable unlabeled data points. the clusters of data points formed by the last branch of the tree are not alike. in addition, the majority of the data points inside a particular cluster coincide with those within other clusters in the data set. this approach uses a diagram, which is a diagram that resembles a tree and is based on the hierarchy. agglomerative hierarchical clustering, also known as agnes (agglomerative nesting), and divisive hierarchical clustering, also known as diana, are two categories of hierarchical clustering techniques (divisive analysis). this algorithm is an identical duplicate of itself. table 1 compiles several strategies based on various criteria.[21] machine learning application software using the software is a collection of one or more apps created for end users. applications can be found in web browsers, email clients, word processors, spreadsheets, accounting software, music players, file viewers, simulators, console games, and picture editors. machine learning software designed by professionals is generally available. the following are a few of the few notable pieces of software: to conduct data mining tasks, the program employs a variety of machine learning algorithms. manually apply the algorithms to a dataset or simply invoke them in your figure 2: diagram showing decision tree mulla and demir: the use of clustering and classification methods in machine learning 56 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 java code. data pre-processing, classification, regression, clustering, association rules, and visualization are among weka’s capabilities. at the university of waikato in new zealand, weka was upgraded. this tool is an addition to the gnu general public license-licensed free program “data mining: practical machine learning tools and methods”.[22] the measure that used in weka confusion matrix a representative representation of the above-mentioned parameters in matrix form is given in table 2, considering that advanced visualizations would be helpful accuracy to evaluate each created subset in this measurement, classification data mining methods have been offered as a fitness function. the formulation of measuring the accuracy of each subset is described as follows: accuracy= tp+tn tp+tn+fp+fn *100 false positive (fp) reflects the improper placement of the negative example in the positive class, whereas true negative (tn) represents the proper placement of the negative example. true positive (tp) represents the proper classifications of the positive example. false negative (fn) is the term used to describe a positive sample that was mistakenly placed in the negative category. precision the precision is calculated with the following equation and the denominator consists of the total predicted positive. precision= tp tp+fp it is immediately apparent that precision relates to how precise/accurate your model is in terms of the proportion of successful predictions. precision is a helpful statistic to judge whether there are high costs associated with false positives. as an example, email spam detection. when an email that is not spam (actual negative) is wrongly classified as spam, it is known as a false positive in email spam detection (predicted spam). the email user may overlook important emails if the spam detection model’s precision is poor. f-measure in statistical analysis of binary classification, the f-score or f-measure is a measure of a test’s accuracy. even if you read a lot of other literature on precision and recall, you cannot ignore the other measure, f-measure, which is a function of those two variables. the formula is as follows: f-measure=2*� precision*recall precision+recall� f-measure is necessary when seeking to balance precision and recall. what distinguishes f-measure and accuracy from one another then? if we need to find a balance between precision and recall and there is an uneven class distribution, f-measure would be a better statistic to utilize because false negatives and false positives often have business expenses (physical and intangible). as we have previously seen, a huge number of true negatives, which are typically overlooked in corporate settings, can significantly contribute to accuracy (a large number of actual negatives). receiver operating characteristic (roc) area a graph that shows how well a classification model works at each classification threshold is called a receiver operating characteristic curve, or roc area. on this curve, two parameters are plotted: • true positive rate (tpr) • false positive rate (fpr) tpr is a synonym for recall and is therefore defined as follows: tpr=� tp tp+fn fpr is defined as follows: fpr=� fp fp+fn in a roc curve, tpr vs. fpr for various classification criteria are displayed. as the classification threshold is lowered, more items are categorized as positive, which increases the quantity of both false positives and true positives. data we used a dataset about the predicting whether it will rain or not using some weather conditions. and we obtained this table 1: summary of clustering algorithms algorithm size of dataset type of data complexity computation speed modifications corrections cluster shape result interpretation k-means large numerical o (nkd) fast flexible none convex easy k-medoids small categorical o (n^2 dt) moderate difficult none convex difficult hierarchical large numerical o (n) slow flexible none convex easy table 2: confusion matrix predicted true class true positive false negative false positive true negative mulla and demir: the use of clustering and classification methods in machine learning 57 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 dataset from the kaggle side it was shown in this link. https:// www.kaggle.com/datasets/ananthr1/weather-prediction. about dataset: using coloms: • precipitation • tempreture min • tempreture max • wind we will forecast the weather as follows: • drizzle • rain • snow • sun • fog results and discussion three distinct categorization methods were employed for the data being studied during this study. all three algorithms dt, ann, and knn were used in the classification process. we utilized k=1 and divided the dataset into 10 cross-validations for the knn algorithms. the quantity of data used for reducederror pruning when we used dt methods was defined according to the trimming confidence factor, which was 0.25. all three classifications and clustering algorithms were employed, and the training rate was 66% and the testing rate was 34%. we separated into five groups for k-mean methods, used 10 seeds, and decided to use false for debug and false for display stander division. the effectiveness of the three classification methods in table 3 was evaluated using the accuracy precision, f-measure, and roc region metrics. as shown in table 3 for the classification method, the highest performance depending on accuracy and precision was 84.8, 78.7 obtained in dt, but the higher performance according to the f-measure and roc area was obtained in ann. on the other hand, the best model for dt, ann, and knn algorithm in this study was obtained in roc area. figure 3 shows one of the classification methods, so it can be decided based on figure 3. there are 5 decisions from figure 3. the dt region measurement was measured for testing the efficiency of the dt algorithm as shown in figure 3. • the decision 1: precipitation to sun. • the decision 2: precipitation to temperature min to rain. • the decision 3: precipitation to temperature min to wind to snow. • the decision 4: precipitation to temperature min to wind to temperature max to rain. • the decision 5: precipitation to temperature min to wind to temperature max to snow. the k-means region measurement was measured for testing the efficiency of the clustering algorithm as shown in table 4. while we study the output report, we can see 5 classes (class 0, class 1, class 2, class 3, class 4). in the report appear as number of clusters selected by cross-validation: 5 you can find the centroid of the entire population in the first column. the centroids for clusters 0, 1, 2, 3, and 4 may be found, respectively, in the second, third, fourth, and fifth columns. the centroid coordinate for each row’s respective dimension is provided. it is important to note that this step of the process is locating the centroids. the centroids are not unique and are the output of a particular run of the algorithm; a different run could produce a different set of centroids. there is a “clusters prior” probability for each cluster. in the estimators, each conceivable attribute value is represented by a number, which is treated sequentially. classes when we used k-mean algorithms: • class 0 has total 232 things, out of which majority of things (16%) data sets. • class 1 has total of 449 things, out of which majority of things (31%) data sets. • class has total 355 things, out of which majority of things (24%) data sets. • class 3 has total 258 things, out of which majority of things (18%) data sets. • class 4 has total 167 things, out of which majority of things (11%) data sets. classes when we used hierarchical algorithms: • class 0 has total 53 things, out of which majority of things (4%) data sets. • class 1 has total of 641 things, out of which majority of things (44%) data sets. • class has total 640 things, out of which majority of things (44%) data sets. • class 3 has total 26 things, out of which majority of things (2%) data sets. • class 4 has total 101 things, out of which majority of things (7%) data sets. table 3: the performance of the classification algorithms using for weather data summary accuracy precision f-measure roc area dt 84.8 78.7 80.7 89.5 ann 82.5 74.4 84.9 91.7 knn 67.3 67.8 67.6 74.5 dt: decision tree, ann: artificial neural network, knn: k-nearest neighbor figure 3: decision tree visualization mulla and demir: the use of clustering and classification methods in machine learning 58 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 we examined two clustering techniques from table 5 and separated both of them into five clusters in order to identify some distinct things. at the time, k-mean was preferred over hierarchical methods. fewer iterations in the hierarchy than the k-mean. on the other hand, when using hierarchical, we had zero root mean square errors (rmse), k-mean algorithms experienced a 1707.63 (rmse). conclusion the application of data mining to enrollment management is a relatively recent innovation. at the moment, category, and straightforward numerical data are most frequently employed in data mining. data mining will increasingly incorporate complicated data types in the future. by taking into account more variables and how they interact with one another, any model that has been created may also be reinforced. new techniques for discovering the most exciting features, data mining research will lead to the development of the data. as models are developed and implemented, they can be used as a tool for enrollment management. this study showed that the best model was dt according to accuracy and precision measures but the best model according to f-measure and roc area measures was ann. weka, a data mining tool, is utilized in this paper to carry out the clustering. the k-mean approach can cluster large data sets efficiently, and as the number of clusters rises, so does its efficiency. the k-mean method outperforms the hierarchical clustering algorithm. the performance of k-means method increases as the root mean square errors (rmse) drops and the rmse falls as the number of clusters. all the techniques have some noise or ambiguity in certain data when clustered. using massive datasets significantly raises the quality of all algorithms. the k-means algorithm is quite susceptible to dataset noise. this noise interferes with the algorithm’s ability to cluster data into the proper clusters and affects the algorithm’s output. when working with huge datasets, the k-means method produces good clusters more quickly than alternative clustering techniques. for noisy data, the hierarchical clustering technique is more sensitive. weka only enables sequential single-node execution, which leads to its inability to handle very large datasets. as a result, there is little similarity between clusters when data are grouped or clustered, and significant similarity within clusters. one of the clustering methods is the k-means algorithm, which gathers data based on their traits and qualities and runs the clustering process by shortening the distances between the data centers.[5] the finally used dt, nb and svm to make design of model in pima dataset but here we used dt, ann, knn in weather dataset. references 1. m. baran. maki ̇ne öğrenmesi̇ yöntemleri̇yle çoklu eti̇ketli̇ veri̇leri̇n sınıflandırılması. (sivas cumhuriyet üniversitesi, sosya bilimler enstitüsü, turkey, 2020. 2. m. hall, e. frank, g. holmes, b. pfahringer, p. reutemann and i. wietten. the weka data mining software: an update. acm sigkdd explorations newsletter, vol. 11, pp. 10–18, 2009. 3. a. a. soofi and a. awan. classification techniques in machine learning: applications and issues. journal of basic and applied sciences, vol. 13, pp. 459–465, 2017. 4. v. sharma. survey of classification algorithms and various model selection methods. journal of machine learning research, vol. 1, pp. 1–48, 2000. 5. d. sisodia and d. s. sisodia. prediction of diabetes using classification algorithms. procedia computer science, vol. 132, pp. 1578–1585, 2018. 6. m. ambigavathi and d. sridharan. analysis of clustering algorithms in machine learning for healthcare data. in: advances in computing and data sciences. vol. 1244. springer, berlin, pp. 117–128. 7. a. c. lorena, l. p. f. garcia, j. lehmann, m. c. p. souto and t. k. ho. how complex is your classification problem? a survey on measuring classification complexity. acm computing surveys, vol. 52, pp. 1–34, 2018. 8. d. liu, r. sun and h. ren. efficient fraud detection classification: class imbalanceand attribute correlations. the frontiers of society, science and technology, vol. 2, pp. 96–103, 2020. 9. x. zheng. smote variants for imbalanced binary classification: heart disease prediction. university of california, california, 2020. 10. t. f. malone. application of statistical methods in weather prediction. proceedings of the national academy of sciences, vol. 41, pp. 806–815, 1955. 11. l. liu and j. l. priestley. a comparison of machine learning algorithms for prediction of past due service in commercial credit. in: grey literature from phd candidates. digitalcommons kennesaw state university, georgia, 2018. 12. g. nakhaeizadeh and c. c. taylor. machine learning and statistics : the interface. wiley, united states, 1997. table 4: clustering method performance attribute full data (1461) 0 (232) 1 (449) 2 (355) 3 (258) 4 (167) precipitation 3.02 4.06 7.61 0.00 0.00 0.36 temp-max 16.40 18.90 10.60 26.00 16.80 7.52 temp-min 8.23 11.70 5.34 13.70 7.63 0.25 wind 3.24 2.89 4.06 2.85 3.06 2.60 weather rain rain rain sun sun sun table 5: comparison result of clustering algorithms with weather dataset using weka tool hierarchical k-mean name 5 5 no of clusters 1461 1461 cluster instances 18 no of iterations 1707.637908 root mean square errors 9.9 s 0.02 s time is taken to build model 0 0 unclustered instances weka: waikato environment for knowledge analysis mulla and demir: the use of clustering and classification methods in machine learning 59 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 52-59 13. g. a. a. mulla, y. demir and m. m. hassan. combination of pca with smote oversampling for classification of high-dimensional imbalanced data. beu journal of science, vol. 10, pp. 858–869, 2021. 14. m. a. habara. credit risk modelling in a developing economy: the case of libya. griffith university, australia, 2009. 15. l. saitta and f. neri. learning in the “real world”. machine learning, vol. 30, pp. 133–163, 1998. 16. l. c. thomas. a survey of credit and behavioural scoring: forecasting financial risk of lending to consumers. international journal of forecasting, vol. 16, pp. 149–172, 2000. 17. w. e. henley and d. j. hand. a k-nearest-neighbour classifier for assessing consumer credit risk. journal of the royal statistical society. series d, vol. 45, pp. 77–95, 1996. 18. j. a. k. suykens and j. vandewalle. least squares support vector machine classifiers. neural processing letters, vol. 9, pp. 293–300, 1999. 19. p. nerurkar, a. shirke, m. chandane and s. bhirud. empirical analysis of data clustering algorithms. procedia computer science, 125, pp. 770–779, 2018. 20. s. b. tambe and s. s. gajre. cluster-based real-time analysis of mobile healthcare application for prediction of physiological data. journal of ambient intelligence and humanized computing, vol. 9, pp. 429–445, 2017. 21. p. d. kumar, t. amgoth and c. s. r. annavarapu. machine learning algorithms for wireless sensor networks: a survey. information fusion, vol. 49, pp. 1–25, 2019. 22. data mining, machine learning and predictive analytics software minitab. minitab, 2020. available from: https://www.minitab. com/en-us/products/spm [last accessed on 2023 jun 07]. tx_1~abs:at/add:tx_2~abs:at 39 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 research article the protective rolls of taraxacum officinale against carbon tetrachloride infarction in the liver farhang a. mustafa, khabat a. ali department of biology, college of education, salahaddin university-erbil, 44002, erbil, kurdistan region, iraq abstract carbon tetrachloride (ccl4) is a liquid that is colorless, transparent, inflammable, and volatile. its central carbon atom is surrounded by four cl- atoms. as a result, the objective of this effort was to estimate the anti-oxidant properties of silver nanoparticles (agnps) made utilizing aqueous plant extracts from taraxacum officinale leaves. a green synthesis of agnps using a synergistic aqueous extract from t. officinale leaves was tested against liver damage in rats caused by ccl4. the rats were randomly distributed into seven groups: group 1: control group, group 2: olive oil group, group 3: agnps-treated group (100 mg/kg bw.), group 4: agnps-treated group (200 mg/kg bw.), group 5: ccl4 + olive oil group, group 6: (ccl4 + olive oil) + 100 mg/kg of agnps-treated group, and group 7: (ccl4 + olive oil) + 200 mg/kg of agnpstreated group one a week for 6  weeks. this study demonstrated a sustainable method for synthesizing agnps utilizing t.  officinale leaf (tol) extract. to characterize the synthesized t. officinale leaf-silver nanoparticles (tol-agnps), various microscopic and spectroscopic methods were used. the effectiveness of the biosynthesized tol-agnps against ccl4 was tested to assess their antioxidant potential. the antioxidant properties of synthetic tol-agnps were also evaluated. histopathological research showed that all groups treated with nanoextract had less severe inflammatory responses. our findings demonstrated that agnps synthesized using the leaves of t. officinale possess a potential anti-oxidant activity against ccl4-induced liver injury in rats. keywords: carbon tetrachloride, hepatoprotictive effect, oxidative stress, silver nanoparticles, taraxacum officinale introduction carbon tetrachloride (ccl4) is a liquid that is colorless, transparent, inflammable, and volatile. it features four clatoms surrounding the carbon atom at its center. in addition to happening naturally, it may also occur as a result of a variety of chemical reactions. it has an atmospheric halflife of between 30 and 100 years due to its chemical stability.[1] ccl 4 is quickly absorbed by the body by inhalation, ingestion, and cutaneous absorption.[2] according to estimates, the typical adult daily ccl 4 intake is 0.1 ml. this poisonous substance distributes throughout the body, with maximum concentrations through the hepatic, kidney, brain, fat, muscle, and blood after exposure throguh ingestion, inhalation, or dermal absorption.[3] to establish the acute toxicity of ccl 4 , several animal tests have been conducted. for example, studies on rats have demonstrated that the fatal dosage (ld 50 ) is reach after severe oral ingestion, and that the body weight varies from 4.7 to 14.7 ml/kg based on dietary circumstances and supplements that are supplied.[4] a well-known example of a chemical liver injury is ccl 4 induced liver damage. a powerful liver toxin called ccl 4 has been linked to cell death, inflammation, and oxidative stress in histopathological studies.[5] medications used to treat liver problems have inadequate therapeutic efficacy and may have negative effects.[6] as a result, interest in alternative or complementary therapies as well as research into novel drugs has surged.[7] with its distinctive properties and wide-ranging uses in industries including agriculture, food, and medicine, nanotechnology has become one of the most important and appealing areas of research.[8] nanoparticles have well-known uses in the biomedical sector as antibacterial, antioxidant, and antitumor agents due to characteristics including their tiny size, high surface area to volume ratio, visual, magnetic, chemical, and physical capabilities.[9] corresponding author: farhang a. mustafa, department of biology, college of education, salahuddin universityerbil, 44002, erbil, kurdistan region, iraq. e-mail: farhang.mustafa@student.su.edu.krd received: january 14, 2022 accepted: march 28 2023 published: may 20, 2023 doi: 10.24086/cuesj.v7n1y2023.pp39-47 copyright © 2023 farhang a. mustafa, khabat a. ali. this is an openaccess article distributed under the creative commons attribution license (ccby-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) mustafa and ali: rolls of taraxacum officinale against liver infarction 40 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 due to their versatile theranostic properties, nanoparticles of noble metals: including (silver, gold, platinum, copper, zinc, titanium, and magnesium) have attracted a lot of attention for use in biomedical applications.[10] however, although physical and chemical processes are employed to make nanoparticles, they are linked to injurious substances that are dangerous to handle. as an alternative, attention is being drawn to plantmediated production of metal nanoparticles because of their quick time requirements, low cost, and environmental friendliness.[11] proteins, polysaccharides, flavonoids, terpenoids, tannins, alkaloids, amines, ketones, and aldehydes are examples of bioactive secondary metabolites that are found in plants. these substances are also used as lowering, stabilizing, and capping agents in the alteration of metal ions to metallic nanoparticles, producing desired nanoparticles with specific properties.[12] silver nanoparticles (agnps) have overtaken other biosynthesized metal nanoparticles in the past 20 years due to their distinctive biological, chemical, and physical properties.[13] numerous studies have demonstrated that a lower concentration of agno 3 has improved chemical stability, catalytic activity, biocompatibility, and other desirable properties as well as inherent therapeutic potential despite the fact that silver is lethal at higher concentrations.[14] according to reports, agnps may have antioxidant, anticancer, and antibacterial properties.[15] an especially noteworthy benefit of agnps over bulk metals and their salts is their controlled, delayed release of silver.[16] the new-age bionanoformulations’ guiding principle is the integration of traditional medicine with nanotechnology. the perennial plant taraxacum officinale, sometimes known as dandelion, is a member of the asteraceae family, according to classification of t. officinale.[17] kingdom : plantae phylum : magnoliophyta class : magnoliopsida subclass : asteridae order : asterales family : asteraceae subfamily : cichorioideae genus : taraxacum species : officinale subject : taraxacum officinale it is interesting that this herb has a lengthy history of usage as a medicine. medicinal uses for the root and young tips predominate.[18] liver, pancreas, and stomach functions are all enhanced by dandelion. it is employed to treat rheumatism, hepatitis, anemia, and liver cirrhosis,[19] prebiotic, analgesic, anti-hyperglycemic, anti-oxidative, anti-carcinogenic, and anti-inflammatory properties.[20] materials and methods collection of plant and leaf extract preparation the dried t. officinale leaf (tol), weighing 500 g, was obtained from erbil’s (pirmam roads) in september 2021 and was dried at room temperature in the shade. the plant was identified by professor dr. abdulla shakur of salahaddin university-college erbil’s of education. in the department of biology, distilled water was combined with 25 g of the powdered plant material (1:10, sample to solvent ratio) and sonicated for 60 min (power sonic 405/lab tech) while maintaining a temperature of 40 c. the resulting extract was then concentrated using a rotating evaporator with a regulated vacuum after being filtered using whatman no. 1 filter paper with a buchner funnel. the concentrated extract was kept at 4°c in an airtight container after being air-dried. silver nanoparticle synthesis 300 ml of warmed distal water was added to 10 g of the dried plant powder to make the nanoparticle. after 10 min of cooking using a magnetic stirrer, this mixture was filtered through whatman no.1 filter paper and kept at 4°c for storage. then, using a magnetic stirrer, 100 ml of the obtained extract was to be added to 1000 ml of 1 mm aqueous agno 2 and heated for 60 min at 40°c. ultraviolet-visible (uv-vis) spectra were used to identify the production of agnps following centrifugation of the reduced mixture at 5000 rpm for 30 min. the resulting particle was then dissolved in deionized water and the supernatant was thrown away. to get rid of any materials that were adsorbs on the surface of the agnps, the same washing procedure was carried out 2–3 times with deionized water and ethanol.[21] characterization of agnps the most crucial, straight forward, and fundamental method to validate the creation of synthesized nanoparticles is uv-vis spectroscopy. uv-vis spectra between 200 and 800 nm were captured using (agilent technologies, santa clara, ca, usa). an fourier transform-infrared (ft-ir) spectrophotometer was used to conduct the analysis to analyze the functional group that the surface of the generated agnps included (perkin elmer spectrum 100 ft-ir, 710 bridgeport, ct, usa). the ft-ir spectra were scanned in the transmittance mode at a resolution of 4 cm-1 with wave numbers ranging from 4000 to 400 cm-1.[22] an x-ray diffractometer used 2 scans from 30 to 80° at 0.04°/min with a constant value of 2 s to capture the x-ray diffraction (xrd) patterns of the agnps (rigaku, japan). sigma 300 (carl zeiss, germany) was employed together with edx analysis to evaluate the cell surface characteristics and identify the elemental structure of produced agnps. on a fresh coverslip, a modest quantity of agnps (10 l) was drop-cast before being dried. following a gold sputter, the samples were examined using an scanning electron microscopy (sem) and an energy dispersive x-ray (edx).[23] animal housing and breeding the albino rats were kept in a room with a temperature of 22 ± 2°c and a 12/12-h light/dark cycle. each rat weighed between 160 and 200 g, in the animal house of education college/salahaddin university, the rats were kept 1 week for acclimatization. experimental design fifty-six female wistar albino rats (weighing 160–200 g) are used in the present study. the animals have been housed in mustafa and ali: rolls of taraxacum officinale against liver infarction 41 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 conventional conditions (22 ± 2°c, 45 ± 5% moisture, and 12 h light-dark cycles). throughout the trial of the experiment, the rats received conventional rat feed and had an unlimited access to tap water and libitum. rats were divided into seven groups at random (n = 8). group 1: rats in the control group received distilled water and a regular meal. group 2: rats received weekly intraperitoneal injections of 1 ml/kg bw olive oil for a period of 6 weeks. group 3: for 6 weeks, rats in the agnps-treated group received daily oral gavage doses of 100 mg/kg bw of agnps. group 4: for 6 weeks, rats in the agnps-treated group received 200 mg/kg bw of agnps orally. group 5: for 6 weeks, rats in the ccl 4 + olive oil group received a single subcutaneous injection of the mixture (1:1) mg/kg. bw of ccl 4 and olive oil. group 6: ccl 4 + olive oil + agnps-treated group: rats were given a single dose of the mixture (ccl 4 + olive oil) subcutaneously once per week at a dose of (1:1) mg/kg.bw for 6 weeks, followed by the administration of agnps daily administered by oral gavage in a dose of 100 mg.kg b. for 6 weeks. group 7: ccl 4 +olive oil + agnps-treated group: rats were given a single dose of the mixture (ccl 4 + olive oil) subcutaneously once per week at a dose of (1:1) mg/kg. bw for 6 weeks, followed by the administration of agnps daily by oral gavage at a dose of 200 mg.kg body weight for 6 weeks. collection of tissues and blood samples at the end of the study period, rats were given general anesthesia before being decapitated and slaughtered. blood samples were then taken by heart puncture and drawn for biochemical analysis. a little amount of blood — about 5 ml — was collected in a gel tube, left to remain at room temperature for 30 min, and then gently centrifuged for 15 min at 3000 rpm. additionally, liver tissues were gathered, removed, and cleaned in sterile saline. all of the obtained liver tissues underwent autopsy and were divided into pieces. then, until the histological investigation, the tissue was promptly preserved in 10% formalin saline. determination of liver function test using kits and an enzymatic-colorimetric approach with an auto-analyzer, the liver function tests alanine aminotransferase (alt), aspartate aminotransferase (ast), and alkaline phosphatase (alp) were conducted (cobas kenza 450 tx fully automated biochemical analyzer, france). oxidative stress evaluation with the use of solarbio kits from china, malondialdehyde (mda), superoxide dismutase (sod), and glutathione (gsh) were measured. the variance seen among absorbance at 532 nm, 450 nm, and 600 nm is used to compute the mda content.[24] an absorption at 560 nm shows that sod activity declines as the interaction solution’s blue color deepens. sod can eliminate oxygen and prevent the synthesis of methionine.[25] gsh will interact with 2,5-dithiobis-(2-nitrobenzoic acid) to create 2-nitro-5-mercaptobenzoic acid and glutathione disulfide (gssg). the highest absorbance of the 2-nitro-5mercaptobenzoic acid in the yellow material is 412 nm.[25] all absorbencies identified by a microplate reader.[25] histopathological examination liver tissue samples from various groups were autopsied and afterward histopathologically analyzed.[26] all samples were immediately placed in 10% buffered formalin for 24 h, rinsed in ordinary saline, and then dehydrated using successively diluted alcohol solutions (70%, 95%, 100%). the paraffin wax was then used to embed tissue pieces that had been cleared in xylene. using a microtome, pieces of paraffin blocks with a thickness of 6 microns were cut, the tissue slices were placed on glass slides, deparaffinized with xylene, stained with hematoxylin and eosin (h and e), cleared with xylene, and mounted with dpx. scores were given to the histological alterations.[27] statistical analysis the normality and lognormality tests were run after the one-way analysis of variance test (one-way anova) using the graphpad prism (version 8.0). the findings were represented by the mean and standard deviation (sd). the differences between the groups were considered statistically significant when “p” < 0.05. results and discussion uv-vis spectroscopy synthesized agnps’ uv-visible spectroscopic profile and color change. figure 1 shows that after 30 min of incubation at room temperature in the dark, the color of the reaction mixture seemed to shift from light brown to dark brown. in actuality, under the same conditions, the agno 3 solution without plant extract remained colorless. the shift in hue was seen as proof that agnps were forming as a result of the reduction of ag+ ions.[28,29] agnps are present because the absorbance of agnps from 200 to 800 nm displays a maximum between 440 and 450 nm. the surface plasmon resonance features of spherical and aggregate agnp generation are illustrated by the prominent absorption peak at 434 nm.[30] after 1 day and 60 days, uv-vis spectroscopy was used to determine the agnps’ temporal stability. the presence of the distinctive at 434 nm peak in the produced agnps confirms their stability [figure 1]. as a consequence of this finding, we may conclude that the produced agnps exhibited great water stability because a little decrease in absorbance was noted at figure 1: the results of ultraviolet/visible spectra of taraxacum officinale leaf-silver nanoparticles mustafa and ali: rolls of taraxacum officinale against liver infarction 42 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 434 nm. recent studies on the green synthesis of agnps using plant extracts from salvia hispanica and pomegranate leaves, among others, yielded findings that were almost identical to those of this kind of study.[29,31-33] this indicates that t. officinale leaves contain phytochemicals that can function as capping and reducing agents when effectively extracted. ft-ir spectroscopy analysis using ft-ir spectroscopy, it was possible to identify the functional group that took part in the green synthesis of agnps as reduction and coating agents from tol extract. figure 2 shows the tol extract’s ft-ir spectral bands and manufactured agnps made from the leaf extract. the produced agnps showed sharp transmittance maxima at 3439, 1762, 1384, 1136, 875, and 825 cm-1 [figure 2]. with a little variation in peak positions, the ft-ir spectra of t. officinale aqueous leaf extracts and synthetic agnps were quite comparable. this blatant resemblance shows that some of the remaining phytochemical moieties that are present in the tol extract are present on the surface of the produced agnps. the peaks in the ft-ir spectra clearly show that t. officinale aqueous leaf extracts function as decreasing and steadying agents. strong peaks at 3439 cm-1 were indicative of the hydroxyl and amine (n-h) functional groups’ o-h stretching vibration type. the stretching vibrations of the hydroxyl (o-h) and amine (n-h) groups were thought to be responsible for the absorbance maxima between 3000 and 3600 cm-1. while proteins, peptides, and amino acids are assumed to create n-h stretching, polyphenols display o-h stretching vibration instead.[34] the phenolic group of compounds included in leaf extract acts as strong capping and reducing agents when silver nitrate is reduced, resulting in the formation of agnps. due to the high concentration of secondary metabolites that dissolved in the sample, two additional strong peaks at 2088 and 2078 cm-1 were seen. these peaks indicated the existence of alkynes with triple bond c≡c stretched vibration. the peaks at 1762 cm-1 are caused by the amide i band and the -c=cstretching vibration band. the 1384 cm-1 peak was assumed to be caused by the -c-nstretching band and the amide i band of proteins in the leaf extract.[35] the band of amide i connected to the carbonyl group’s (c=o) stretch mode when it was joined to the amide linkage. the ch out of plane bending vibrations was attributed to the maxima at 1136 and 875 cm-1. for stability and to avoid agglomeration, the proteins may be coated or encapsulated around the produced agnps.[36] thus, it became clear from the ft-ir analysis that proteins and bioactive substances, including polyphenols, found in the tol extracts play a significant role in the production of agnps.[32,36,37] xrd analysis t. officinale leaf-silver nanoparticles (tol-agnps) crystal structure was ascertained by powder xrd, as demonstrated in [figure 3]. the planes (111), (200), (220), and (311) are correspondingly represented by ѳ2 = 34°, 38.1°, 44.2°, 64.5°, and 77.6°, where bragg’s reflections were discovered. x-ray diffractograms were used to identify four diffraction peaks (111), (200), (220), and (311) to determine the composition of the created agnps. these peaks fit ag planes according to the typical face-centered cubic structures of ag (joint committee on powder diffraction standards; jcpds no. 04-0783). the formed agnps’ crystalline arrangement was further sustained by the (111) reflection peak’s high intensity in comparison to the other reflections. the findings confirm those of earlier studies, which showed identical agnp diffraction peaks.[38,39] sem and edx analysis using sem methods, the morphological properties of greenly synthesized agnps were identified. the majority of the spherical shape in sem images of produced agnps, with an average size of 50 nm, was observed [figure 4]. the results obviously showed that the synthetic agnps substantially figure 2: the results of fourier transform-infrared spectrum of taraxacum officinale leaf-silver nanoparticles mustafa and ali: rolls of taraxacum officinale against liver infarction 43 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 comprise tiny grains of material that have clumped together to form crystals with almost uniform spherical shapes and smooth surfaces. similar findings have already been published.[34,37,40] the elemental analysis of the material has been shown in figure 4 by the edx spectrum of the produced agnps. silver peaks can be detected in the spectrum, indicating that silver ions are one of the ingredients used to make synthetic agnps. the weight percentage and atomic percentage of the ag peak were 41 and 3.21, respectively. au, na, and cu, among other peaks, were seen. these components were derived from the tol extract biomolecules that were attached to the surface of the agnps.[35,41,42] examination of biochemical analysis the injection of ccl 4 markedly (p = 0.05) improved the activity of the ast and alp a liver blood marker enzymes as matched to the control group. although there has been a little rise in the alt level, it is not statistically significant. table 1 showed that when tol-agnps were used in comparison to the ccl 4 group, increases in the production of these enzymes were considerably reduced (p = 0.05) by 100 mg/kg bw and 200 mg/kg bw, respectively. the results of the present study showed that ccl 4 -induction in rats considerably raised the levels of ast and alp and marginally but insignificantly raised the levels of alt. as a result of the acute hepatocyte damage and impaired membrane integrity brought on by ccl 4 , the hepatocytes’ enzymes leak out.[43] the abnormal elevations in alt, ast, and alp were, however, dramatically reduced following treatment with tol-agnps, coming to those in the control. these findings support prior research in that tol-agnps have the capacity to defend against ccl 4 -induced hepatocyte damage.[44] that reported the protective consequences of polyphenolic compounds in tolagnps against ccl 4 -induced liver cirrhosis. estimation of oxidative stress biomarkers table 2 provides mda levels. the findings showed that the mda levels in the ccl 4 group were substantially higher (p = 0.05) than those in the control group. in comparison to the ccl 4 group, the t. officinale agnps treated groups at doses of 100 mg/kg bw and 200 mg/kg bw considerably (p = 0.05) lowered the mda level. rats treated with ccl 4 had considerably lower (p = 0.05) sod activity and marginally higher (statistically nonsignificant) gsh levels than the control group. administration of 100 mg/kg bw and 200 mg/kg bw of t. officinale agnps in comparison to ccl 4 rats resulted in a rise in lipid peroxidation, a decrease in the activities of antioxidant enzymes, and a substantial increase (p = 0.05) in gsh levels.[43] throughout their lives, both humans and animals are exposed to hepatotoxic substances. unfortunately, there is no effective medication for liver disease prevention or therapy. consequently, academics, owners of domestic animals, and the general public are becoming more interested in complementary and supportive treatments.[45] free radicals and oxidative stress from hazardous chemicals that cause lipid peroxidation, such as ccl 4 , lead to liver damage.[26] mda is a significant biomarker of lipid peroxidation.[6] the findings of this investigation confirm the harmful effects of ccl 4 .[46] it was corroborated by histopathological findings, an elevated figure 4: the photos of the scanning electron microscope and energy dispersive x-ray of the taraxacum officinale leaf-silver nanoparticles figure 3: x-ray diffraction analysis of the taraxacum officinale leafsilver nanoparticles mustafa and ali: rolls of taraxacum officinale against liver infarction 44 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 mda level, and a declining gsh level. similar to the findings of the research by gad et al.[47] groups 6 and 7 dramatically reduced mda levels, comparable to the outcomes shown in group 5.[48] sod and gsh levels were higher than in the ccl 4 group. it was discovered that groups 6 and 7 were effective for hepatoprotection. compared to the ccl 4 group, the gsh and sod levels in this group rose. these findings align with the body of literature.[26] compared to the ccl 4 group, the la group’s mda levels dropped while sod and gsh levels rose, which is consistent with the literature.[49] the findings confirm the effectiveness of several t. officinale agnps as antioxidants.[50] histopathological examination the histopathologic results of the liver tissue of the animals in the control group revealed usual liver architecture with normal central vein and normal arrangement of hepatocytes [figure 5a and b]. tissue sections from the tol-agnp-treated group alone showed a normal structure equivalent to the control to the control groups [figure 5c and d]. in the ccl 4 -treatment groups, many changes are occurs like (vein wall disruption, necrosis around the vein), cholestasis (bile out in channel), inflammatory cells infiltration, hepatocyte necrosis, fatty liver changes, arteria sclerosis, sinusoid wideness (dilatation), as shown in [figure 5e-h]. however, the histological sections of the concurrently treated groups with tol-agnps at an oral dosage of 100 mg/kg revealed that tol-agnps maintained hepatocyte arrangement, no fatty liver changes and mild perivascular hepatocytic infiltration were seen, as well as a noticeable decline in penetration of the inflammatory cells., as shown in [figure 5g-j], respectively. liver damage is a major health issue and a global public health burden.[51] the assertion was substantiated by necrosis and inflammatory cell infiltration discovered during the histological investigation of microphotographs of liver sections.[52] in this investigation, ccl 4 injection led to considerable inflammatory cell infiltration, passive hyperemia, hepatocyte degradation, and the emergence of fat vacuoles across the whole liver. the liver damage generated by ccl 4 was reduced by protecting the hepatocyte basal membrane and reducing inflammatory cell infiltration with a fat accumulation of agnps, as in earlier studies.[52] in our investigation, administering agnps decreased inflammation and hepatocellular disintegration while maintaining the structure of the remark cords. histopathological data revealed that tol was the most effective hepatoprotective species, as previously reported by karkos et al.[53] when compared to normal control rats, animal given ccl 4 had higher blood levels of alt, ast, and alp (p = 0.05), according to the study’s findings. a sudden increase in serum transaminases indicated that ccl 4 caused serious toxicity. necrosis and inflammatory cell infiltration seen during the histological analysis of microphotographs of liver sections supported the claim.[52] in this work, rats who were drunk table 2: the effect of ccl 4 and tol-agnps on assessment of oxidative stress groups experimental parameters mda gsh sod control 1.19±0.38b 71.02±9.64a 72.32±5.14a olive oil 1.32±0.37b 76.86±62.64a 78.42±6.86b agnps 100 mg/kg 1.36±0.72b 79.71±28.14a 89.16±2.76bc agnps 200 mg/kg 1.29±0.64b 81.86±9.29a 84.51±1.46a ccl 4 +olive oil 2.76±0.83a 59.63±11.96a 63.58±5.33a ccl 4 +olive oil+agnps 100 mg/kg 1.25±0.64b 70.70±13.47a 79.22±14.30b ccl 4 +olive oil+agnps 200 mg/kg 2.08±0.78a 66.66±23.07a 82.40±6.21b the same letter in the columns means a non-significant differences among parameters at p<0.05a, b, c. tol-agnps: taraxacum officinale leaf-silver nanoparticles, ccl 4 : carbon tetrachloride, mda: malondialdehyde, gsh: glutathione, sod: superoxide dismutase table 1: the effect of ccl 4 and tol-agnps on indices of hepatotoxicity groups experimental parameters ast (u/l) alt (u/l) alp (u/l) control 88.02±7.724b 28.36±4.893a 106.8±7.155b olive oil 108.12±24.456a 30.04±9.371a 101.38±10.164b agnps 100 mg/kg 101.76±14.285a 29.58±10.262a 110.2±27.797b agnps 200 mg/kg 126.32±45.265a 25.5±10.321a 111.2±20.104b ccl 4 +olive oil 149.92±8.320a 33.82±4.223a 155.8±33.789a ccl 4 +olive oil+agnps 100 mg/kg 101.56±17.928a 24.72±2.192a 95.6±10.502b ccl 4 +olive oil+agnps 200 mg/kg 93.42±28.241b 26.98±7.487a 82±25.700b the same letter in the columns means a non-significant differences among parameters at p<0.05a, b, c. ast: aspartate aminotransferase, alt: alanine aminotransferase, alp: alkaline phosphatase, tol-agnps: taraxacum officinale leaf-silver nanoparticles, ccl 4 : carbon tetrachloride mustafa and ali: rolls of taraxacum officinale against liver infarction 45 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (1): 39-47 with ccl 4 had their raised liver function levels brought back to normal levels by administering agnps of t. officinale at a dosage of 100 mg/kg b.w. the findings of this investigation confirm the harmful effects of ccl 4 [46] were verified by elevated mda levels, declining gsh levels, enzyme activity, and histopathological results. similar to the findings of the gad et al.[47] mda levels were dramatically reduced in the agnps group. these findings align with the body of literature.[26] the results support the antioxidant efficiency of selected plant species. conclusion in this study, green biosynthesized agnps, tol-agnps, were found to have an antioxidant potential on rat liver injury. one of the greatest methods of understanding the underlying structure of hepatotoxicity and development, in addition to, complete therapeutic options, is to use a rodent model for liver injury. according to the present study, such antioxidants, antiinflammation, and enhanced potency of tol-agnps against ccl 4 toxicity were achieved through the lowering amounts of the biochemical parameters as well as restoring their amounts to standard levels across all nano-extract treatment groups. in addition, the decrement in damages caused by ccl 4 in animals treated was revealed by microscopic proofs that showed considerable antioxidant activity. references 1. v. unsal, m. cicek and i̇. sabancilar. toxicity of carbon tetrachloride, free radicals and role of antioxidants. reviews on environmental health, vol. 36, no. 2, pp. 279-295, 2021. 2. l. w. d. weber, m. boll and a. stampfl. hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. critical reviews in toxicology, vol. 33, no. 2, pp. 105-136, 2003. 3. m. srinivasan, r. rukkumani, a. r. sudheer and v. p. menon. ferulic acid, a natural protector against carbon tetrachlorideinduced toxicity. fundamental and clinical pharmacology, vol. 19, no. 4, pp. 491-496, 2005. 4. d. scholten, j. trebicka, c. liedtke and r. weiskirchen. the carbon tetrachloride model in mice. laboratory animals, vol. 49, no. 1 suppl, pp. 4-11, 2015. 5. x. liang, k. wu, m. liu and b. yang. adverse impact of carbon tetrachloride on metabolic function in mice. journal of cellular biochemistry, vol. 120, no. 7, pp. 11973-11980, 2019. 6. f. altinok-yipel, i. o. tekeli, s. ozsoy, m. guvenc, s. sayin and m. yipel. investigation of hepatoprotective effect of some algae species on carbon tetrachloride-induced liver injury in rats. archives of physiology and biochemistry, vol. 126, no. 5, pp. 463467, 2020. 7. m. a. toori, b. joodi, h. sadeghi, h. sadeghi, m. jafari, m. s. talebianpoor, f. mehraban, m. mostafazadeh and m. ghavamizadeh. hepatoprotective activity of aerial parts of otostegia persica against carbon tetrachloride-induced liver damage in rats. avicenna journal of phytomedicine, vol. 5, no. 3, pp. 238-246, 2015. 8. f. erci, r. cakir-koc and i. isildak. green synthesis of silver nanoparticles using thymbra spicata l. var. spicata (zahter) aqueous leaf extract and evaluation of their morphologydependent antibacterial and cytotoxic activity. artificial cells, nanomedicine, and biotechnology, vol. 46, no. 1, pp. 150-158, 2018. 9. c. tanase, l. berta, n. a. coman, i. roșca, a. man, f. toma, a. mocan, l. jakab-farkas, d. biró and a. mare. investigation of in vitro antioxidant and antibacterial potential of silver nanoparticles obtained by biosynthesis using beech bark extract. figure 5: histopathological examination demonstrating the effects of taraxacum officinale leaf-silver nanoparticles (tol-agnps) against ccl 4 on rat’s liver tissues following addition of (agnps). (a and b) the control group: showed normal histological structure: central vein (red arrow), hepatocytes and sinusoids (white arrow), normal nucleus (blue arrow), normal kupffer cells (green arrow), (h and e, ×10, ×40, respectively); (c and d) the tol-agnps -treated group: the section through the liver of tol-agnps alone group showed, maintained normal liver architecture, no significant histopathological changes were seen in which: normal central vein (red arrow), normal sinusoids (white arrow), normal hepatocytes (blue arrow), and normal binuclear (green arrow), (h and e, ×10, ×40, respectively); (e and f) ccl 4 + olive oil group: the liver section through the ccl 4 group showed: fatty changes (red arrow), vein wall disruption (white arrow), necrosis around the vein (blue arrow): sinusoid wideness, sinusoidal cell hypertrophy, and apoptosis (green arrow), (h and e, ×10 and ×40 respectively); (g and h) ccl 4 + olive oil group showed epithelia cell pyknosis of bile duct (red arrow), waste nuclement in the bile duct (white arrow), mild perivascular inflammation, periductal inflammatory cells infiltration (blue arrow), fatty liver changes (green arrow), (h) showed fatty liver changes (red arrow), inflammatory cell infiltration (white arrow), arteria sclerosis, vascular congestion and vascular hyaline changes (blue arrow), and sinusoid (dilatation) (green arrow), (h and e, ×10, ×40, respectively); (i and j) ccl 4 and olive oil + agnps of t. officinale treated group showed maintained hepatocyte arrangement: mild sinusoid dilation (red arrow), normal sinusoid (white arrow), vascular congestion (blue arrow), vascular dilation (green arrow), mild portal chronic inflammation (black arrow), normal sinusoid (white arrow), mild sinusoid dilation (blue arrow), karyomegally, mild periductal chronic inflammation, and vascular dilation (green arrow) (h and e, ×10, ×40, respectively) d jh c ig b f a e mustafa and ali: rolls of taraxacum officinale against liver infarction 46 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42-46, 2004. 51. l. pimpin, h. cortez-pinto, f. negro, e. corbould, j. v. lazarus, l. webber and n. sheron and easl hepahealth steering committee. burden of liver disease in europe: epidemiology and analysis of risk factors to identify prevention policies. journal of hepatology, vol. 69, no. 3, pp. 718-735, 2018. 52. w. jiang, m. h. guo and x. hai. hepatoprotective and antioxidant effects of lycopene on non-alcoholic fatty liver disease in rat. world journal of gastroenterology, vol. 22, no. 46, p. 1018010188, 2016. 53. p. karkos, s. c. leong, c. d. karkos, n. sivaji and d. a. assimakopoulos. spirulina in clinical practice: evidencebased human applications. evidence-based complementary and alternative medicine, vol. 2011, p. 531053, 2011. tx_1~abs:at/add:tx_2~abs:at 17 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 research article applying the bayesian technique in designing a single sampling plan dler h. kadir1,2, abdulrahim k. rahi3 1department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq, 2department of business administration, cihan university-erbil, kurdistan region, iraq, 3department of business administration, dijlah university college, iraq abstract the bayesian sampling plans for production inspection are considered a technique of sampling inspection techniques for determining the characteristics of the sampling plan based on the assumption that the rate of defectives is a random variable that varies from one production batch to the next, resulting in a probability distribution f(p) that could be determined based on experience and the available quality information available. as part of this study, the parameters of a single bayesian sampling plan (n,c) were derived using the betabinomial distribution and compared with those of other single sampling plans. researchers have identified (ala company for soft drinks), which handles product quality control. one hundred and twenty production batches were selected, and the size of the batch and the number of defective items were used to determine the proportion of defective items, given that the variable varies randomly from one production batch to the next. bayesian and decision-making models can be implemented to create a single sampling inspection process that is close to the actual quality level. the researchers discovered that when the decision-making model was used, the sample size was minimal compared to other inspection plans, leading to a low inspection cost. keywords: statistical quality control, average sample number, acceptance quality level, operating characteristic, bayesian sampling plans introduction among the statistical tools used to control and monitor the quality of production are control charts and sampling inspection plans. regarding sample testing plans, they are an accurate and appropriate method to obtain an estimate of the presence of one or more characteristics among the produced units. this is achieved by examining a small percentage of the production, which is randomly selected to determine whether to accept or reject production based on the results of the sample drawn. there are two methods of examination using the sampling method: the discriminatory examination method (by attributes), where the produced units are classified into defective and good units, or by the variable examination method (by variables) based on a standard such as height or weight. the examination may be based on the researcher’s or examiner’s experience, in addition to any other available information about the production process (prior information). this approach leads to the use of bayesian theory and decision theory, and the adoption of bayesian estimation for the quality parameter of production, depending on the loss function, to avoid making wrong decisions regarding the production process.[1,2] due to the effort, cost, and time required to conduct a comprehensive examination, as well as cases known to statisticians where the above examination can be applied, the sampling examination method was used to evaluate the performance of the quality control department in the company. since most of the staff working in this department are not statistical experts and are not familiar with statistical methods in the field of quality control, they rely on the accumulated experience of their members, examiners, and laboratory workers. for the aforementioned reasons, this research aims to shed light on the methods of sample examination and their use in estimating sample size and designing sampling examination plans that can minimize spoilage and reduce costs by making the right decision to accept or reject the produced batch. corresponding author: dler h. kadir, department of statistics, college of administration and economics, salahaddin university-erbil, kurdistan region f.r. iraq. e-mail: dler.kadir@su.edu.krd received: april 01, 2023 accepted: july 10, 2023 published: august 05, 2023 doi: 10.24086/cuesj.v7n2y2023.pp17-25 copyright © 2023 dler h. kadir, abdulrahim k. rahi. this is an open-access article distributed under the creative commons attribution license (cc by-nc-nd 4.0). cihan university-erbil scientific journal (cuesj) kadir and rahi: bayesian technique in sampling plan 18 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 this research aims to investigate the method of sampling and decision-making regarding the acceptance or rejection of batches. this will be achieved through the design of a discriminatory sampling examination plan using the bayesian method and applying it to the production of pepsi ala soft drinks company to make informed decisions on whether to accept or reject the produced batch.[3,4] bayesian sampling plans one of the methods of sample examination is the bayesian sampling plan, which is an alternative to the comprehensive (classic) examination, as well as the single, double, and sequential sampling plans. however, bayesian sampling plans differ from single, double, and sequential sampling plans in that they deal with specifying plan parameters based on the percentage of defects in the product with random variation. this percentage changes from one production batch to another, as it has a probability distribution that can be determined from previous experience and available information on quality.[1,5] bayesian plans have emphasized the importance of utilizing available information on quality, and this information is referred to as prior distributions. the prior distributions are used to obtain the post-quality posterior distribution, assuming that the type of sampling distribution being studied may be binomial with parameters (n,p), poisson with parameter (λ), or normal with parameters (µ,σ). bayesian plans are of great importance in examining products, and it is necessary to provide a brief explanation of this topic to reach a bayesian sampling plan (n, c) that reduces the total cost function of quality control. the cost function is the sum of inspection costs, costs of rejecting non-defective units (good), and costs of accepting defective units (bad).[6] bayesian plans concept for product inspection bayes’ plans have been named by this name in relation to the scientist (thomas bayes) (1702–1761), where bayes is the first to use the prior distribution for defective percentages in statistical inference, and since 1960, attention began to focus on bayes’ plans to test the product. bayesian plans are named after the scientist thomas bayes (1702–1761), who was the first to use prior distributions for defective percentages in statistical inference. since the 1960s, attention has been focused on bayesian plans for testing products. in 1964, the scientist hald (1981) was able to develop a model for the total cost function of quality control. through this model, the parameters of the sampling plans are determined when the quality is fixed or the random variable has a prior distribution (prior distribution).[7,8] in 1968, the scientist hald presented a model for bayesian plans to examine products. this model is used to obtain the parameters of the individual bayesian plan (n, c) by reducing the standard cost function of quality control, which is: r n n c n p p n n p p q p w p dp p p p p w p dps m r r pr , , ( ( ) ( ) ( ) � � � � � �� � � � �� � � � � � 1 ��� � � � � � � � � � � 0 pr (1) where (r = p+q), denotes sample costs. p m -: the standard cost in cases of rejection and acceptance. p s : the average cost of examination per unit in both cases of rejection and acceptance, and that: p s ≥ p r ≥ p m , 0 < p r < 1, 0 < p s < 1 on equation (1) if the value of (p) is very small, we use the poisson distribution, that is,: b x np e np x np x , ! � � � � � � (2) and we change z p p m np m np r r r= = =,� ,�� the form of the function will be: b c np b c mz p z, ,� � � � � � � � (3) total cost function for quality control the scientist hald (1981) developed a model that included the total costs of examination, acceptance, and rejection. the model aims to determine the individual sampling plan (n, c) for examining batch n of the product of quality p by applying filtering examination. he expressed these costs in the following formula: h x x p n n c ns ns n n a x x a x c, , , , , ,� � � � � �� � � �� � �1 2 1 2 (4) h x x p n n c ns ns n n r x x r x c, , , , , ��� � � � � �� � � �� � �1 2 1 1 (5) the examination and repair costs, in addition to the costs resulting from accepting the quantity (n-n) remaining after drawing the sample, represent the first part. the examination and repair costs, in addition to the costs resulting from rejecting the quantity (n–n), represent the second part. equating equation (4) with equation (5) results in: p x x n x r a a rr � � � � � � 1 1 2 2 the average cost in both cases of acceptance and rejection is equal to: in the event of acceptance n(s1+s2p)+(n–n)(a 1 +a 2 p) in case of refusal n(s 1 +s 2 p)+(n–n)(r 1 +r 1 p) and by entering the probability of acceptance of the product, p(p) and the probability of rejection, q(p) since:p p p x c b x n p b c n pr x c � � � �� � � � � � � � 0 , , ( , , ) q p p p p x c b x n p e c n pr x c n � � � � � � � �� � � � � � � � � �1 1 1 , , ( , , ) thus, the cost rate (p) is obtained in the cases of acceptance and rejection as follows: kadir and rahi: bayesian technique in sampling plan 19 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 k p n s s p n n a a p p p r r p q p � � � �� � � �� � � � � � �� � 1 2 1 2 1 2{( ) ( )} k p nk p n n k p p p k p q ps a r� � � � � � �� � � � � � � � � � �� � (6) where q(p) = 1–p(p) becomes clear from equation (6), then: k p nk p n n k p p p k p p ps a r� � � � � � �� � � � � � � � � �� �( ( )1 � � � � �� � � � � � � � � � � �� �nk p n n k p p p k p k p p ps a r r ( ) � � � � �� � � � � � � � � � � � � � � � � �� � � � �� nk p n n k p p p n n k p n n k p p p s a r r( ) ( ) ( ) � � � � �� � � � � � � � � � � � � � � � � � � �� � �nk p n n k p p p nk p nk p n n k p p p s a r r r( ) ( ) �� �� � � � � � � � �� � � � � � � �� � � � � � � � � nk p nk p n n k p p p n n k p p p nk p s r a r r k p n k p k p n n k p k p p p nk ps r a r r� � � � � � � � � �� � � � � � ��� �� � � � � �[ ] (7) the values of k r (p), k a (p), k s (p) express the rates of examination, acceptance, and rejection costs per unit, respectively.[9-12] finding the single bayesian plan (n, c) this method relies on the iterative method for determining the parameters of the single sampling plan (n, c). the method involves minimizing equation (7) k(p) subject to certain conditions of the operating curve function (oc). we find the smallest value of the function at k(p) (c = 0), then at (c = 1), then at (c = 2), and so on. once the absolute minimum value is obtained, we stop and fix the parameters of the single sampling plan (n, c) at this value. it is important to note that the cost function model shown in equation (7) assumes a constant level of quality, but the quality level may change from one production batch to another due to random and attritional reasons that lead to qualitative deviations. therefore, it is necessary to estimate the level of quality in the future by taking advantage of all available information about previous tests and information about the production process, which is supposed to be within control limits. furthermore, natural qualitative changes that occur due to market competition should also be taken into account. it is worth noting that incorporating previous information available on quality levels, as well as estimated information from samples, leads to more accurate decisions regarding quality level estimates. the scientist hald has emphasized this point and relied on the following expected total cost function k relative to the previous distribution of defective ratios f(p). k k p f p dp� � � � � �� � � bayesian plan relative to the beta distribution the beta distribution is considered one of the important statistical distributions in bayesian plans. in this distribution, the percentage of defects is a random variable with a beta distribution, and its parameters can be estimated using the moment method. the probability function of the random variable p with (α, β) parameters takes the following form: f p b p p p o w , , , ( ) �������� . � � � � � �� � � � � � � � � � �1 1 0 1 0 1 1 in many cases, it cannot be assumed that p remains constant from one batch to another. for example, consider a machine used to produce a specific unit. after producing a batch, the machine is checked and put back into operation. for each batch, p can be determined according to certain distributions, which are referred to as the initial distributions of p. for this reason, the beta distribution was chosen as the initial distribution of p with (α, β) parameters. the expected value of the random variable for the beta distribution is α/ (α+β). this means that (β) must be greater than (α), and when estimating (α, β), it must be rounded to the nearest integer. if the estimated value lies between 0 and 1; then, it must be rounded to the expected value of p, to the nearest integer. this method allows us to use standard tables to determine the sampling plan and also shows us why the plans are not clearly sensitive to small changes in the parameters without taking into account the appropriate initial distributions.[13-15] direct formulas for determining individual base plan parameters by decision model to define the parameters of the single bayesian economic statistical (bes) plan (n, c) for product inspection, a formula for the expected risk must be developed and specified directly for the parameters of the sampling plan. we can rely on the definition of risk provided by guthrie and johns, who defined it as the sum of examination costs plus the loss resulting from wrong decisions. it is well-known that examination costs depend on the volume. it represents the loss resulting from accepting defective units and the loss resulting from rejecting good units. the expected risk formula, under the conditions of binomial sampling, takes into account the distributions of continuous defective percentages, which are as follows: r f p n c n s r p s r a r p p q p f p dpr pr [ ( ), , ] {( ) ( ) ( ) ( ) ( ) ( ) } � � � � � � � 2 2 1 1 2 2 0�� � � � � � � � �� � �� � n r p r a r p p f p dp n n n r a p f r pr r { ( ) ( ) ( ) ( ){ 2 1 2 2 0 1 1 1 2 1 ppr� � (8) when neglecting the upper limits, equation (8) is reduced to the following form: kadir and rahi: bayesian technique in sampling plan 20 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 r f p n c an bn c n n n [ ( ), , ] ( )� � � � 1 (9) so: a s r s r r a p p f p dpp r pr � � � � � � ��[( ) ( ) ( ) ( ) ( )2 2 1 1 2 2 0 b r r a r p p f p dpp r pr � � � � ��2 1 2 2 0( ) ( ) ( ) c r a p f p pr r� � � � 1 2 11 1( )( ) ( ) ( ) ( ) ( , ) ( , )p p f p dp ib p ibpr pr pr r pr � � � ��0 1� � � � when we derive equation (9) with respect to n and equate the derivative to zero, we obtain the optimal value of n (n*). to find the sample size necessary to examine batch n of the product, assuming binomial sampling and that the defective percentages change from one production batch to another, we can use the following formula: the random variable has a prior distribution equal to beta with (α, β) parameters, and the optimal value of n can be obtained from the following equation. n r a p p n b s r p s r r a pib r r* ( ) ( ) ( , )[ ) ( ) ( ) ( � � � � � � � � � 1 1 1 2 2 1 1 2 2 1 2 � � � � ppr r pr p ib( , ) ( , ))]� � � �� � � � � � � � � � � � � � � � � �1 1 2 when the values of (α, β) are integers, the relationship between the incomplete beta function and the cumulative binomial function can be derived. this relationship can be used to extract the following: ib c p ppr x x r x r x� � � � � � � � �, ( )� � � � � � � � � � � �� 1 1 11 ib c p ppr x r x r x� � � � � � � � � ��� � � � � � � � � � ��1 1 1 1, ( ) the number of acceptance can be obtained from the following relationship: c n� �� �� 2 3 θ∘: critical quality level. direct formulas for determining single bayes plan parameters from hald model determining the parameters of individual bayesian plans (n, c) for product inspection requires lengthy iterative calculations to find the values (n, c) that achieve the smallest expected total cost or the smallest standard cost. research in this area has focused on finding formulas that efficiently and quickly obtain optimal parameter values. continuous studies and research have led to formulas used directly for large production batches where the quality of the batches is a random variable with a prior distribution f(p), which is continuous and differentiable at points adjacent to a point (p = pr). the discontinuous distribution of defective percentages was discussed by the scientist hald in 1965, and direct formulas were developed by hald in 1968, which were supported by auxiliary tables. these formulas can be used to extract bayesian plans in distributions such as gamma-poisson and beta-binomial.[8,16-19] it is then transformed into the standard cost function (r, n, n, c), which, in turn, requires defining each of the standard loss rates for acceptance d a , rejection d r , and examination d s as follows: d k k a ra a m� � � ( ) ( )2 2 (10) d k k a rr r m� � � ( ) ( )2 2 d k k a rs s m� � � ( ) ( )2 2 if all units of the product are classified correctly, the value of k m is the smallest cost per unit, that is, in the case of acceptance when (p ≤ p r ) and rejection d r when (p > p r ) (). in other words, km represents. k a a p f p dp r r p f p dpm pr pr � �� � � � � �� � � �� �1 20 1 2 1 k a a p f p dp r r p f p dp r r p f p dp m pr pr � �� � � � � �� � � � � �� � � � � � � 1 2 0 1 2 0 1 1 2 0 k k a r p p f p dpm r r pr � � � �� � � ��( )1 2 0 among them, we find that: d p p f p dpr r pr � �� � � ��0 the value of d a ,d s in terms of d r , as: d d p pa r r� � � d d s r s r p a rs r� � � � � � �{( ) ( ) } ( )1 1 2 2 2 2 k k a r dm r r� � �( )�2 2 so it is: nk k a r dm r r� � �� �2 2 � (11) equations (11), (10) are then substituted into equation (12) and the equation of the standard cost function defined in equation (11) is obtained. kadir and rahi: bayesian technique in sampling plan 21 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 r n n c k n n c k a r m, , , , � � � � � � �2 2 (12) r n n c nds n n d n, ,� � � � �� � � � d n d p p b c n p f p dpr r� � � � �� � � � � �� , ,0 1 depending on the equation � � � � � 1 2 2 2 2 � � � p q a r b k k r r s m ( ) ( , )( ) � � � � � � � � 1 2 2 1 3 11 2 3 1 3 1 1 � � � � � � � � � � �� � � � , , ( ) p q p qr r r r the value of n necessary to check the batch (n) is the value defined by the following relation: n n* � �� �1 2 as for the number of acceptance of c, it is extracted from the following relationship: c n pr * *� � �1 whereas: β β α= − −1 ˆ ˆ 1 2r r p q to find the value of the standard cost function r 0 (n) corresponding to the optimal sampling plan ( n* , c* ), which we will symbolize r 0 (n), the following equation will be relied on: r n n ds and d k k a rs s s 0 1 2 2 2 2 2� � � � �� � � � � * �� �� ( ) � � therefore, the value k(p) of the expected total cost of quality control is equal to: k p r n a r nkm� � � � � �� � �0 2 2 results the process of improving product quality requires relying on modern scientific methods. by utilizing modern practical methods and adhering to standard specifications, products can be made suitable and conform to the desired specifications of consumers. this not only elevates the status of the production facility in local and global markets but also enhances the value of these products in these markets. therefore, it is essential to establish quality control requirements fully. this includes prioritizing standard and manufacturing specifications for input, process, and output elements. one of these requirements is to identify and provide a scientific method for examining materials and products to ensure the reduction of damage and to ensure the regular flow and handling of circulation during production processes in the facility. application of the decision-making model based on the decision-making model, a set of bayesian economic-statistical (bes) plans was developed to test the product, depending on the previous distribution of defective percentages (beta-prior). the parameters of the individual bes plan were determined according to the decision-making model to obtain the values of n and the acceptance number c. the inspection plans for this product are shown in table 1, taking into account the levels of quality and sizes of production batches. the parameters of the sampling plan required to check the daily production of the 1.5-l (pepsi ala) product of quality (x p = 0.005349) and value (p r = 0.00617) were extracted using this model. the values obtained were (n,c) = (1495, 14), and the expected risk value for the sampling plan was also determined to be (1495, 14). the value of r{f(p), n, c} was found to be equal to $43,210. hald model application since the distribution of the defective percentages f(p) is one of the continuous and derivable distributions at point (p = p r ), a set of bays plans will be extracted from the direct formulas created by (hald), as the set of bays plans necessary to check the product, which is defined from equation (14), must be calculated before that each of: examination cost rate per unit (1) k s s ps � � � � � �� � �1 2 0 0011 0 03 0 005349 0 00126. . . . �$ rejection cost rate per unit (2) k r r p kr s� � �1 2 examination cost rate per unit k a a ps � �1 2 (3) ( )+ =0 0.208 0.005349 0.0011 $ as well as the values of the ingredients (4, 5, 6, and 7): (4) � p p f p dpr pr �� � � ��0 � � � � �p ib pibr pr pr� � � �, ( , )1 � � �� � �� �� � � 0 006179 0 789972 0 005349 0 734239 0 000954 . . . . . (5) k k a r p p f p dpm r r pr � � � ��( ) ( ) ( )2 2 0 � � � �� �� � �0 00126 0 178 0 0009544 0 0010901. . . . ) (6) � � � � � 1 2 2 2 2 � � � p q a r b k k r r s m ( ) ( , )( ) �� �1 2 148 25059� . whereas: (7) α β= = = = =2 20.0061,    0.208,    0.03,    25, ˆ    4648ˆrp a r � � � � � � � � 2 2 1 3 11 2 3 1 3 1 1 � �� � � �� � � � � � �� � � � ( ) p q p qr r r r kadir and rahi: bayesian technique in sampling plan 22 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 table 1: bayesian plans to test the product according to the distribution of beta-prior extracted from the decision-making model quality level 0.005 quality level 0.004 quality level 0.003 quality level 0.002 quality level 0.001 batch size acceptance number sample size acceptance number sample size acceptance number sample size acceptance number sample size acceptance number sample size c n c n c n c n c n 6 669 5 527 4 449 3 398 7 753 10,000 6 701 5 553 4 471 4 418 7 790 11,000 7 733 5 578 4 492 4 436 8 825 12,000 7 763 5 602 4 512 4 454 8 859 13,000 7 791 6 624 5 532 4 471 8 891 14,000 8 819 6 646 5 550 4 488 9 923 15,000 8 846 6 667 5 569 4 504 9 953 16,000 8 872 6 688 5 586 5 519 9 982 17,000 8 897 6 708 5 603 5 534 9 1011 18,000 9 922 7 727 6 620 5 549 10 1038 19,000 9 946 7 746 6 636 5 563 10 1065 20,000 9 969 7 765 6 651 5 577 10 1092 21,000 9 992 7 783 6 667 5 591 11 1117 22,000 9 1014 7 800 6 682 5 604 11 1143 23,000 10 1036 8 817 6 696 6 617 11 1167 24,000 10 1058 8 834 6 711 6 630 11 1191 25,000 10 1079 8 851 7 725 6 642 11 1215 26,000 10 1099 8 867 7 739 6 654 12 1238 27,000 11 1119 8 883 7 752 6 666 12 1261 28,000 11 1139 8 899 7 766 6 678 12 1283 29,000 11 1159 8 914 7 779 6 690 12 1305 30,000 11 1178 9 929 7 792 6 701 13 1327 31,000 11 1197 9 944 7 804 6 712 13 1348 32,000 11 1215 9 959 8 817 7 724 13 1369 33,000 12 1233 9 973 8 829 7 734 13 1389 34,000 12 1251 9 987 8 841 7 745 13 1410 35,000 12 1269 9 1001 8 853 7 756 14 1430 36,000 12 1287 9 1015 8 865 7 766 14 1449 37,000 12 1304 10 1029 8 876 7 776 14 1469 38,000 13 1321 10 1042 8 888 7 787 14 1488 39,000 13 1338 10 1055 8 899 7 797 14 1507 40,000 13 1355 10 1069 8 910 7 807 15 1526 41,000 13 1371 10 1082 9 921 7 816 15 1544 42,000 13 1387 10 1094 9 932 8 826 15 1562 43,000 13 1403 10 1107 9 943 8 836 15 1581 44,000 14 1419 11 1120 9 954 8 845 15 1598 45,000 14 1435 11 1132 9 964 8 854 15 1616 46,000 14 1450 11 1144 9 975 8 864 16 1634 47,000 14 1466 11 1156 9 985 8 873 16 1651 48,000 14 1481 11 1168 9 995 8 882 16 1668 49,000 14 1496 11 1180 9 1005 8 891 16 1685 50,000 kadir and rahi: bayesian technique in sampling plan 23 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 table 2: bayesian plans to test the product extracted from the (hald) model quality level 0.005 quality level 0.004 quality level 0.003 quality level 0.002 quality level 0.001 batch size acceptance number sample size acceptance number sample size acceptance number sample size acceptance number sample size acceptance number sample size c n c n c n c n c n 10 1162 8 798 7 671 3 588 6 529 10,000 11 1222 8 839 7 707 3 620 6 557 11,000 11 1278 8 879 8 741 3 650 7 584 12,000 11 1333 9 917 8 773 3 679 7 611 13,000 12 1385 9 954 8 804 3 706 7 636 14,000 12 1436 9 989 8 835 3 733 7 660 15,000 12 1485 9 1023 8 864 3 759 7 683 16,000 12 1532 10 1057 9 892 3 784 7 706 17,000 13 1578 10 1089 9 920 3 808 8 728 18,000 13 1623 10 1120 9 946 3 832 8 750 19,000 13 1667 10 1151 9 972 3 855 8 771 20,000 14 1709 10 1181 9 998 3 878 8 791 21,000 14 1751 10 1210 9 1022 3 900 8 811 22,000 14 1791 11 1238 9 1047 3 921 8 830 23,000 14 1831 11 1266 10 1070 3 942 8 849 24,000 15 1870 11 1293 10 1094 3 963 8 868 25,000 15 1908 11 1320 10 1116 3 983 8 886 26,000 15 1945 11 1346 10 1139 3 1003 9 904 27,000 15 1982 11 1372 10 1161 3 1022 9 922 28,000 15 2018 12 1397 10 1182 3 1041 9 939 29,000 16 2054 12 1422 10 1203 3 1060 9 956 30,000 16 2088 12 1446 11 1224 3 1078 9 973 31,000 16 2123 12 1470 11 1245 3 1096 9 989 32,000 16 2156 12 1494 11 1265 3 1114 9 1006 33,000 17 2190 12 1517 11 1285 3 1132 9 1022 34,000 17 2222 13 1540 11 1304 3 1149 9 1037 35,000 17 2255 13 1563 11 1323 3 1166 10 1053 36,000 17 2287 13 1585 11 1342 3 1183 10 1068 37,000 17 2318 13 1607 11 1361 3 1200 10 1083 38,000 18 2349 13 1629 12 1380 3 1216 10 1098 39,000 18 2380 13 1650 12 1398 3 1232 10 1113 40,000 18 2410 13 1672 12 1416 3 1248 10 1127 41,000 18 2440 13 1693 12 1434 3 1264 10 1142 42,000 18 2469 14 1713 12 1452 3 1280 10 1156 43,000 18 2499 14 1734 12 1469 3 1295 10 1170 44,000 19 2527 14 1754 12 1486 3 1310 10 1184 45,000 19 2556 14 1774 12 1503 3 1325 10 1197 46,000 19 2584 14 1794 12 1520 3 1340 10 1211 47,000 19 2612 14 1813 13 1537 3 1355 11 1224 48,000 19 2640 14 1833 13 1553 3 1370 11 1237 49,000 19 2667 14 1852 13 1570 3 1384 11 1251 50,000 kadir and rahi: bayesian technique in sampling plan 24 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 table 3: bayesian plans to test the product according to the decision-making model and the (hald) model hald model decision-making model batch size pn (c) c n pn (c) c n 0.005998286 10 1162 0.00580307 6 669 10,000 0.00610687 11 1222 0.005768515 6 701 11,000 0.006049403 11 1278 0.005919349 7 733 12,000 0.005994006 11 1333 0.005886681 7 763 13,000 0.006107626 12 1385 0.005856515 7 791 14,000 0.006056638 12 1436 0.00600874 8 819 15,000 0.006008444 12 1485 0.005979344 8 846 16,000 0.005962933 12 1532 0.005951307 8 872 17,000 0.006079027 13 1578 0.005924596 8 897 18,000 0.006035578 13 1623 0.006076854 9 922 19,000 0.005993691 13 1667 0.006050899 9 946 20,000 0.006110937 14 1709 0.006026232 9 969 21,000 0.006070984 14 1751 0.006001765 9 992 22,000 0.006033416 14 1791 0.005978548 9 1014 23,000 0.00599631 14 1831 0.006130671 10 1036 24,000 0.006113404 15 1870 0.006107137 10 1058 25,000 0.006078104 15 1908 0.00608484 10 1079 26,000 0.006044122 15 1945 0.006063756 10 1099 27,000 0.006010518 15 1982 0.00621547 11 1119 28,000 0.00597818 15 2018 0.006194081 11 1139 29,000 0.006094842 16 2054 0.00617284 11 1159 30,000 0.006064192 16 2088 0.006152794 11 1178 31,000 0.006032961 16 2123 0.006132879 11 1197 32,000 0.006003807 16 2156 0.00611413 11 1215 33,000 0.006119773 17 2190 0.006264815 12 1233 34,000 0.006091371 17 2222 0.00624578 12 1251 35,000 0.006062356 17 2255 0.00622686 12 1269 36,000 0.006034483 17 2287 0.006208054 12 1287 37,000 0.006007724 17 2318 0.006190397 12 1304 38,000 0.006123612 18 2349 0.006339673 13 1321 39,000 0.006096696 18 2380 0.006321743 13 1338 40,000 0.006070874 18 2410 0.006303915 13 1355 41,000 0.006045269 18 2440 0.006287227 13 1371 42,000 0.006020722 18 2469 0.006270627 13 1387 43,000 0.005995538 18 2499 0.006254115 13 1403 44,000 0.006111111 19 2527 0.006401838 14 1419 45,000 0.006086596 19 2556 0.006385069 14 1435 46,000 0.006063111 19 2584 0.006369427 14 1450 47,000 0.006039808 19 2612 0.006352826 14 1466 48,000 0.006016683 19 2640 0.006337342 14 1481 49,000 0.00599455 19 2667 0.006321932 14 1496 50,000 and i extracted my value (λ 1 ,λ 2 ), so the value of n necessary to check the batch is n = 39388, which is the value specified by the following relationship: n n* � �� �1 2 n units* . . �� � � � � � �12 17582 39388 55 4084 2361 c n pr * * �= as for the acceptance number c, it is extracted from the relationship, so that β β α= − −1 ˆ ˆ 1 2r r p q thus, the value of the acceptance number corresponding to the sample size (n = 2361) is equal to: �1 4648 0 0061 25 0 9939 0 5� � �� � �� �� � �. . . �1 3 0053� . accordingly: c units* . . �� � � � �2361 0 061 3 0053 17 therefore, the single sampling plan necessary to check the production rate is n = 39388, and extracted from the (hald) model is (2361.17), and this plan means that the examination of a random sample of (2361) is invalid. if the number of defective (damaged) units in the sample is equal to (17) champion or less all units are accepted, otherwise, the batch is rejected. as for the total cost of quality control resulting from the sampling plan (2361.17), it will be extracted based on the smallest standard cost r 0 (n) achieved in the optimal sampling plan (2361.17), as r n ds0 1 2 22� � �� �* � � ds k k a r s m� � � ( ) ( )2 2 ds = 0 000954. r0 2 2361 148 2506 55 4084 0 00954� � � � �[ . . ]( . ) r0 4 4162= . therefore, the value of the expected total cost k(p) of quality control is equal to: k p r a r nkm� � � �� � �0 2 2 � � � � � �4 4162 0 178 49388 0 001091. . . = 42 9731. �$ table 2 includes all the results of bayesian plans to test the 1.5-l liquid pepsi product extracted from the (hald) model according to the previous distribution (beta), classified according to quality levels x p = 0.001(0.001)0.005 and batch sizes n = 10,000(1000)50,000. calculating the value of the defective fraction in the unexamined quantities according to the decision-making models and the (hald) model, a group of bayesian plans have been extracted to test the product. after this extraction, it is necessary to determine the expected value of the expected fraction of the defective kadir and rahi: bayesian technique in sampling plan 25 http://journals.cihanuniversity.edu.iq/index.php/cuesj cuesj 2023, 7 (2): 17-25 fraction in the unexamined quantities (n–n) which will be accepted based on the acceptance of the sample, and then, we will depend on the value of the average of the subsequent distribution for the defective lineage (e(p/x)) when (x = c), that is, (pn(c)) and it has become clear to us that the subsequent distribution f(p/x) is also a house with features (α+β+n, x+α), and therefore, it is e p x p x x nn |� � � � � � � � � � � � and when x = c is p c c nn � � � � � � � � � the following table 3 includes a comparison of bis plans according to the (hald) model and the decision-making model and at the level of quality (x p = 0.005349) and the values of (pn(c)) and for each of the plans of the decision-making model and the (hald) model. table 3 shows that the expected value of the fraction of defective items in the accepted quantities (n–n) is 0.6% according to both the decision-making model and the (hald) model. this value corresponds to the permissible percentage of defective items (ltpd) approved by pepsi company (ala) for soft drinks. the correspondence of the average value of (pn(c)) with the value of (ltpd) indicates the efficiency of the bis plans, which take into account all available information about the quality when estimating the quality of subsequent production batches. this congruence underscores the importance and efficiency of the bis plans. moreover, the actual production quality level ( xp = 0 005349. ) shows that the sample size for different batch sizes is smaller compared to other sampling inspection plans, which reduces examination costs and total costs. conclusion 1. the appropriate probability distribution to represent the defective percentages of the actual production is the betabinomial distribution with a rate of (0.005349) 2. after applying the bayes model in designing the sampling inspection plan, it was found that the parameters of this plan are (n = 1495) and (c = 14), and for the quality level of the actual production ( xp = 0 005349. ), we find that the sample size for the different batch sizes is small compared to other sampling plans, which means reducing examination costs and therefore the total costs. references 1. d. h. kadir and a. r. k. rahi al-harthy. application of bayesian technique for ala pepsi softdrink company in sampling plan design. university of sulaimani, iraq, 2007. 2. a. m. q. muhammad. a study of restricted bayesian acceptance sampling examination plans for quality control with practical application. msc dissertation, college of administration and economics. university of baghdad, 1999. 3. b. ahmed and h. yousof. a new group acceptance sampling plans based on percentiles for the weibull fréchet model. statistics, optimization and information computing, vol. 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