IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Astudy About Some Physiological Parameters In Smokers L. H. A. Al-Azzawy , A. G. S. Al-Qaicy Departme nt of Gommunity Health ,College of Health and Medical Technology, Foundation of Technical Education. Received in:18 April 2010 Accepte d in:14 December 2010 Abstract Smokin g has been accepted as a risk factor for many chronic diseases, including cardiovascu lar diseases, resp iratory diseases, cancer, u lcers and ost eoporosis. Tobacco smoke contains many oxidants and free r adicals t hat can cause dama ge to lipids, p roteins, DNA, carbohydrates and other biomolecu les. In vivo, antioxidant nutrients which include vitamin C, selen ium ( Se), zinc (Zn) and copp er (Cu) p lay a crucial role in defendin g against oxidant damage. The p resent st udy was designed to invest igate the influence of cigarette smoking on serum Zn, Cu ,PC V,W.B.Cs., and BM I. Eighty healthy men (40 smokers and 40 non-smokers) from Baghdad, the cap ital of Iraq, volunteered to p articip ate in this st udy . Two overnight fasting blood samples were collected fro m all volunteers. Serum concentrations of trace elements were measured by atomic absorp tion sp ectrop hotometer. The statistical method of t-test and ANOVA was used to identify differences between the cigarette ’ s smoker and non-smoker group . M ean body mass index (BM I) was different for the smok er and non-smoker group . M ean of PCV for smoker was significantly (p <0.01) higher than those of non-smokers. The mean count of W.B.Cs., was similar for both smokers and non-smok ers The serum zinc lev els of smokers were significantly (P<0.05) lower than those of non-smokers. Smokers had significantly no difference in serum copp er concentration comp ared with non-smokers. Key w ords: Zn, Cu, Smokers, PCV, W.B.C. Introduction Cigarett e smoking h as been implicated as a si gnificant risk factor for the establishment and p rogression of several diseases, including emp hy sema, chronic bronchitis, cardiovascular diseases and cancer [1] . Tobacco smoke contains numerous comp ounds, many of which are o xidants and p ro-oxidants cap able of p roducin g fr ee radicals and enhancing the o xidative st ress in vivo[2] . Each p uff of a tobacco contains 10 14 oxidants in the tar p hase and 10 15 in the gas p hase[3] . The high p roduction of these multiple types of oxidant and reactive oxy gen sp ecies (ROS) associated with smoking may exceed the cap acity of the oxidant defense system, resulting in oxidative dama ge to select p roteins, lipids and DNA[4,5] . Consistent with a condition of IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 oxidative st ress, concentrations of serum antioxidant vitamins, such as ascorbic acid and vitamin E, have b een reported to be lower in chronic smokers than in non- smokers[6] . Also, several invest igators have reported that zinc and selenium concentrations are typically reduced in adu lt smokers, whereas copp er concentrations are typ ically increased [7-9]. Changes in Zn and Cu homeost asis in smokers have been p ostulated to contribute to some of the long-term negative effects associated with smoking and hyp ertension [8] . However, the available data are inconsist ent regarding the effect of smoking on trace elements. Researchers demonst rated that one of the p rominent risk factors for increased lip id p eroxidation is smoking [10 , 11] It has b een reported that smoking is associated with elevations in p lasma LDL- cholesterol and biolo gically modified lip op roteins such as oxidized low density lip op rotein (OXLDL), and decreases in plasma HDL- cholesterol levels[12] . Ot her indications suggest that smokin g leads to elevated p acked cell volu me (PCV)[13,14] . Gen erally, there are conflicting results regardin g the effect of cigarett e smoking on trace elements and PCV; therefore, the three objectives of this study were to invest igate the effect of smoking on some seru m trace elements (Zn, and Cu); evaluate the effect of smoking on the PCV, W.B.Cs., and d etermine the influence of cigarett e smoking on body mass index of healthy , adult Iraqi males Materials and methods Collection of blood and laboratory methods: The volunteers were asked not to smoke for more than 10 hours before samplin g to exclud e the effects of acute smoking on the blood p arameters st udied. All reagents were at least of analytical gr ade and sup p lied by Sigma-Aldrich chemicals ( Sigma-Aldrich Ltd., Ont., Canada) unless indicated otherwise. All glasses or p last ic wares used for tracin g elements determin ation were cleaned by soaking overnight in 10% (v/v)hydrochloric acid, followed by thorough rinsing with de- ionized distilled water and dry ing. Aqueous solutions were made up in de-ionized distilled water. Two blood samples were collected fro m all the volunteers. The first blood sa mples were centrifuged at 3000 g for ten minutes at room temp erature and then serum was st ored frozen until analyzed for the serum zinc, copp er. The second blood samples were used for PCV, and W.B.Cs. measurement. Hemoly zed samples were exclud ed from the analysis. Copp er and zinc concentrations were determined by using an air/acety lene flame atomic absorp tion sp ectrop hotometer. Copp er, zinc, hollow cathode lamps (Thermo Electron, Cambridge, UK)were op erated at 20mA for copp er, at 15mA for zinc, . At omic absorp tion measurements were made at a wavelen gth of 324.8nmfor copp er, at 213.9nm for zinc . Stock atomic absorp tion standard solutions of copper or zinc (Sigma chemical, St. Louis, U SA)containing from 0.05 to 1 mg/ml wer e diluted with10% (v/v) glycerol to obtain a st andard curve for external calibr ation. A 10% (v/v) glycerol solution was used as a blank solution as instructed by the manufacturer. Serum samples were diluted (1:4)with 0.05% Triton-X 100 in 0.125%(v/v) nitric acid [15]. M ethod of Walker[16 ] usin g hematocrit centrifuge was p erformed to measure PCV, while method using h emocytometer (imp roved neubaur chamber) was used to calculate W.B.C.[17 ].M ethod of Romro et.al., was used to calculate BM I[18 ] IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Results Data in table (1) have shown si gnificant differences in mean concentration of Zn level between smokers and non-smoker (92 µg /dl, 105.5 µ g/dl at p <0.05) while no st atistical significant differences wer e noted in mean concentration of Cu level of both smokers and non- smokers (167.5 µ g/dl, 169.5 µ g/dl at p >0.05). Table(2)demonstrates highly significant differences in mean PCV amon g smokers and non-smokers (44%,39% at p <0.01 )resp ectively, although there were no si gnificant differences in mean count of W.B.Cs. for both smokers and non-smokers ( 6180 cell/mm 3 ,6230 cell/mm 3 at p >0.05 ) . Highly differ ences wer e found in BM I between smokers and non-smokers (20.273k g/m 2 , 23.66kg/m 2 at p <0.01) .These results are shown in table (3). Discussion Some of the chemicals in cigarett e ’ s smoke generate a lar ge number of free radicals, which may be related to the etiology of cancer and various diseases [19, 20]. Recent st udies have demonstrated that antioxidants, includ ing vitamin C and some trace elements such as selenium, zinc and copper, p rotect the body against reactive oxy gen sp ecies (ROS) [21] . Furt hermore, it has been reported that higher blood lev els of antioxidants might be associated with a lower risk of cancer; however, until now t here was a lack and conflicting results about the effect of cigarette smoking on seru m trace elements, serum lip id p rofile, hyp ertension and body weight of the male Saudi p op ulation. Therefore, the p resent study is designed to evaluate the effect of c igarette smokin g on body mass index, serum trace element concentrations (zinc, and copper), PVC and W.B.C in healthy Iraqi men. Smoking and trace element s: In our st udy , the lower serum zinc concentrations in the smokers are consistent with the results of Uz et al.[22] who found significant decreases in serum Zn levels of smokers comp ared with non-smokers and attributed the lower serum levels of Z n in smokers t o deficient absorp tion of Zn caused by a tobacco chelation effect. In t he other view cigarette smokin g represents a source of substantial exp osure to cadmiu m over a prolonged period. Each st ick of cigarette contains 1-2 ug of cadmium of which about 10% is absorbed by the lung The very significantly decreased Zn level in smokers appears to confirm the known mutual antagon ism between zinc and cad mium. Elsenh ans[23] and Schümann[24] One p ossible exp lanation for this could be due to t he est ablished comp etition between Zn and Cd for binding sites on metallothionein. This suggests the strong p ossibility therefore, that increase in cad mium level diminishes z inc lev els in smokers. As the results indicated that even though d ietary intake of minerals in smokers was adequate , the habitual diet is not able to maintain the p lasma Zn concentration in the normal ran ge , in the other hands, Z n is cofactor of antio xidant enzy mes , t herefore low level of Z n can r esult in health risk, be cause it p lay s a p reventive role in degener ative dise ase [27] However, M archeggian i et al.[26] found no differ ences in Zn levels in smokers as compared to non-smokers. In the same resp ect, Kocy igit et a l.[9] found that p lasma zinc was st atistically unaffected by smoking. Inst ead, as regards copp er no differen ces were found in the two groups of smokers comp ared with the non-smokers. Our data confirms the results of Vargas et.al.,[27], y et disagrees t he results obt ained by Khalid S.[28 ]. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Cigarette smoking and PCV, W.B.C.: The Data in table (2) rev ealed a h ighly sign ificant increase in mean of PCV for smokers comp ared with that for non- smokers at (p < 0.01).Our findin gs are generally in agreement with those p reviously rep orted [29,30]. M ark man, however ,rep orted a promp t fall in p acked cell volume in smokers who were not allowed to smoke after their admission to hosp ital for various reasons [31] He att ributed this fall to t he exp osure to carbon monoxide ( carbon monoxide r educes p lasma volume) and changes of circulating catecho lamine’s induced by smoking. The same table has shown that there was no significant difference in mean count of W.B.Cs. for both smokers and non smokers. Cigarette smoking and body mass i ndex: In the present study high differences in body mass index were observed between smokers and non- smokers. The association between BM I and smoking is controversial in epidemiological resear ch, since some authors have observed an inverse association [32, 33] unlike others showing no such association [34] mean while, st ill other r esearchers have observed a positive association [35]. Our results are similar to t he results of Kim et a l [36], who found t hat t he slightly lower body weight of smokers was p robably secondary to a lower calorie intake in the smoking group comp ared to the non-smoking group . Kim et al. reported that cigarette smoking was not associated with a reduction in height. at p <0.05. Conclusion The p resent st udy has revealed a depressed antioxidant nutritional st atus (i.e. serum zinc), and changing in other p arameters like PCV and BM I. These effects may be due to the increase in the free radical load incurr ed by smoking which may initiate oxidative damage and the deterioration process. Re ferences 1.Rahman, I. and M acNee, M . (1996). "Oxidant/antioxidant imb alance in smok ers and chronic obstructive pulmonary disease". Thorax 51:348- 350. 2.Prest on, A.M . 1991. Cigarette smoking nutritional imp lication. Pro g. Food Nut r. Sci. 15:193-247. 3.Bluhm, A.L., Weinst ein, J. and Sousa, A.J. (1971). "Free-radicals in tobacco smoke". Nature 229:500-507. 4.Cross, C.E., O’Nell, A.C. and Rezinick, Z.A. (1993). "Cigarette smoke oxidation of human p lasma const ituents". Ann. NY. Acad. Sci. 686:72- 89. 5. Yoshie, Y. and Ohshima, H. (1997). "Sy nergistic induction of DNA st rand breakage by cigarette tar and nitric o xide". Carcino genesis 18:1359- 1363. 6.Abou-Seif, M .A. (1996). "Blood antioxidant st atus and urine sulfate and thiocy anate levels in smok ers". J. Biochem. To xico l. 11:133-138. 7.Dubick, A.M . and Keen, C.L. (1991). "Influence of nicotine on tissue trace element concentrations and tissue antioxidant defense". B iol. Trace Elem. R es. 31:97-109. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 8.Keen, C.L., Clegg, M .S. and Ferrell, F. (1987). "Hyp ertension induced alterations in copper and zinc metabolism: a link to vascular disease". In Sorenson, J.R. (ed.). Biolo gy of Copp er Comp lexes. Human Press, Clifton, NH, pp . 141-149. 9.Kocy igit, A., Erel, D. and Gur, S. (2001). "Effects of tobacco smoking on p lasma selenium, zinc, copp er and iron concentrations and related antio xidative enzy me activities". Clinical B iochemist ry 34:629-633. 10.Nadiger, H.A., M athew, C.A. and Sadasivudu, B. (1987). "Serum malondialdehyde (TBA reactive substance) levels in ci garette smoke". At herosclerosis 64:71-73. 11.Nielsen, F., M ikkelsen, B.B., Nielsen, J.B., Andersen, H.R. and Granjean, P. (1997). "Plasma malondialdehyde as biomarker for o xidative st ress: reference interval and effects of life-sty le factors". Clin. Chem. 43:1204-1214. 12.Craig, W.Y., Palomäki, G.E. and Haddow, J.E. (1989). "Cigarett e smoking and serum lip id and lip op rotein concentrations: An analysis of p ublished data". Br. M ed. J. 298:784-788. 13.Terres, W., Becker, P. and Rosenberg, A. (1994). "Changes in cardiovascular risk p rofile during the cessation of smoking". Am. J. M ed. 97:242-249. 14.Gibson, R.S. (1990). "Princip les of Nutritional Assessment". 1 st ed., Oxford University Press, Oxford, pp . 163-208. 15.Keilchiro,F.,Pulido,R.,M ckay ,B. and Vallee,L. (1964) "measurement of trace metals by atomic absorption sp ectrometry ".Anal.Chem.,Vol.36, No.13, pp 2407-2411 . 16.Walker,H.,Kenneth,H.and Dallas,H.W.(1990)"Clinical M ethods, The hist ory , p hy sical and laboratory examination".third edition,Butterworths publishers,Butterworths,pp .0- 409. 17.M itchell , S.L. , Barbar a,J.B. and Imelda , B. (1993) " Practical hematolo gy "tenth edition , Elsevier publisher, p p .0-450 18. http://en.wikipedia.org/wiki/Body _mass_index - cite_ref-12 Ro mero -Corral , Somers A. , Sierra –Jhnson V. K., Thomas ,J., Collazo-Clavell, R. J., Korinek , M . L., Allison,J., Bastsis ,T.G., Sert-Kuniyoshi,J.A.,,F.H.andLopez-Jimenez,F. (2008)"Accuracy of body mass index in d iagnosin g obesity in the adult general p op ulation". International Journal of Obesity,(June 2008).. 32,No.6 ,pp . 959–956. doi:10.1038/ijo. http://www.ncbi.nlm.nih.gov /pubmed/18283284 19.International Agency for Research on Cancer (1986). "Tobacco smoking. IARC M onographs on the Evaluation of the Car cino genic R isk of Chemicals and Humans",. 38, Lyons, France. 20.Lee, B.M ., Lee, S.K. and Kim, H.S. (1998). "Inhibition of oxidative DNA danger, 8- OHdG, and carbony l contents in smokers treated with antioxidants (vitamin E, v itamin C, β-carotene and red ginseng)". Cancer Letters 132:219-227. 21.Stry ker, W.S., Kap lan, L.A., Stein, E.A., Stamp fer, M .J., Sober, A. and Willett , W.C. (1988). "The relation of diet, cigarett e smoking and alcohol consumption to p lasma beta- carotene and alp ha-tocop herol levels". Am. J. Epidemiol. 127:283-296. 22.Uz , E., Semsettin, S., Ibrahim, F.H., Var, A., Sadik, S. and Omer, A. (2003). "The relationship between serum trace element chan ges and visua l function in h eavy smokers". Acta ophthalmologica Scandinavia 81:161- 164. 23.Elsenhans, B, Schümann, K. and Fort h W. (1991)"Toxic metals: interacting with essential metals". In :Rowland, I.D.( ed). Nutrition, Toxicity and Cancer. CRC Press, Boca Raton, An Arbor,Boston, London , pp 223-258. 24.Schümann, K. (1993). "Interaction between iron and lead. In: Metal- Metal Interactions". Elsenhans,B, Fort h, W. and Schümann,K. (eds).Bertelsmann Foundation Publishers, Güt ersloh,Germany, pp 56-71. 25.M archeggian i, F., Stella, F., Batt istelli, S., Stoceh i, O. and Troccoli, R. (1990). "Serum levels of Cd, Pb, Cu and Zn in cigarette smokers". Boll. Soc. Ital. Biol. Sp er. 66:921-928. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 27.Vargas,M . Eugenia,M .Ninfa,B.,Alfonso, etal.( 2007) “Influencia del habito de fu mar sobre las concentraciones sericas de zinc,cobre y selenio en adultos jovenes”.AVFT,jun.,.26,No.1,p 28.Khalid S. Al- Numair. (2006) ."The influ ence of cigarette smoking on v itamin C, trace elements and lip id p rofile of healthy , Saudi adult males", Journal of Food, Agriculture & Environment.4 (3&4) : 79-83 29.Dintenfass, L.( 1975) “Elevation of blood viscosity aggregation of r ed cells,hematocrit values and fibr inogen levels in cigarett e smoking”M ed.J.Aust..1,p p .20-617 . 30. Galea, G. & Davidson,R.J.( 1985) “ Hematological and hemorheological chan ges associated with cigarett e smoking “Clin Pathol.. 38 ,No.9 p p .978-984. 31.M arkman ,P.F.(1981) " PCV on cessation of smoking" JAMA,. 45,p p .245-1314. 32.Pekkarian, H. G.Hanuineno,G.Tiau, H.Jin, R.(2002) "Comparison of dietery and non- dietery risk factors in over weight and normal weight Chinese adults".Br J Nutr ,. 88 ,pp .7-91. 33.Eck, L.H.Hackett-Renner, C. and Kles ges, L.M .(1992) "Impact of diabeteic st atus ,dietery intake,physical activity ,and smoking st atus on body mass index in NHANES II". Am J Clin Nutr ,.56 , p p .33-329. 34.Coldtz , G.A. Giovanuncia , E .Rimm, E.B. Stamp fer, R. B. and Sp eizer, F.E..(1991) "Alcohol intake in related to diet and ob esity in women and men" Am J Clin Nutr ,.54 ,pp .49-55 . 35.M arti ,B.,Tuomilehto, J., Korhonen, H.J. ,Kartovaara, L. and Vartiainen, E.(1989) " Smokin g andleanness:evidence for chan ge in finland " BM J ,. 90 ,pp .298- 1287. 36 .Ki m, S.H., Ki m, J.S., Shin, H.S. and Keen, C.L. (2003). "Influence of s moking on markers of oxi dative stress and serum mineral conce ntrations in teenage girls in Korea". Nut rition 19:24 0- 24 3. Tables Table (1): Comparison among me an conc. Of Zn, Cu of both smokers and non- smokers. Samp les No. of p articipants M ean conc. Of Z n* (µg/dl) M ean conc. of Cu**(µ/dl) P- value Smokers 40 92 167.5 S* P<0.05 Non- smokers 40 105.5 169.5 N.S** P>0.05 Total 80 S*= Sign ificant Difference N.S**= Non-Significant Difference Table (2): Comparison of mean Of PCV and mean count of W.B.C in smoker and non-smokers Sample No. of part icipant Mean of P CV* ( %) Mean count of W.B.C** (cell/mm 3 ) P - value Smo kers 40 44 61 80 P <0.01 HS* Non-smokers 40 39 62 30 P >0.05 NS** Total 80 PCV: Packed C ell Volume W.B.C : White Blood Cell. HS*: Hi ghly Significant S*: Significant. NS**:Non-Si gnificant. IBN AL- HAITHAM J. FOR PURE & APPL. SCI. VOL.24 (3) 2011 Table (3) M ean of BM I of both smokers and non- smokers. Samples No. of participate Mean of BMI* (Kg/m 2 ) P - Value Smo kers 40 20.273 H.S** P <0.01 Non-smokers 40 23.66 BM I*: Body M ass Index H.S**: Hi ghly Significant 2011 (3) 24المجلد للعلوم الصرفة والتطبیقیة ابن الهیثم مجلة دراسة بعض المعاییر الفسیولوجیة في المدخنین احمد غازي صبار القیسي، لمى حسین علي العزاوي . التقني التعلیم هیئة , والطبیة الصحیة التقنیات كلیة , المجتمع صحة قسم ساننی 18:في البحث استلم 2010 2010 االول كانون 14:في البحث قبل الخالصة مدخنا "شخصا 40 الدراسة هذه شملت.المدخنین لألشخاص الفسلجیة التغیرات بعض لتحدید الدراسة هذه أجریت باستعمال الدم مصل في والنحاس الزنك مستوى قیس سنة 22 أعمارهم معدل كان و المدخنین غیر من "شخصا 40و باستخدام المرصوصة الخالیا وحجم البیضاء الدم كریات مجموع من كل حسبت بینما, الذري االمتصاص تقنیة الوزن قیاس بعد خاصة معادلة باستعمال الجسم كتلة مقیاس وحسب,التوالي على الهیماتوكریت وجهاز الهیموسایتومیتر .مضبوط بشكل والطول ل غیر المدخنین كان مستوى الزنك والنح س في مص 16و 105.5(ا یسیلیتر \مایكروغرام 9.5 ى ) د عل م المدخنین ,التوالي ، . على التوالي ) 167.5و 92(بینما كانت نسبة الزنك، والنحاس في د وحجم الخالیا المرصوصة ي ) 6180و 44(وللمدخنین ,على التوالي ) 6230و 39(ومجموع كریات الدم البیضاء لغیر المدخنین كان على التوال للمدخنین 20.273لغیر المدخنین 23.66مقیاس كتلة الجسم كان . ر تحلیل اإلحصائي للنتائج - استعمل اختبا .ت واختبار تحلیل المتغیرات في ال مجموع كریات الدم البیضاء, حجم الخالیا المرصوصة , مدخنین , نحاس , زنك : الكلمات المفتاحیة