946 | 1Department of Phar- macology, Government Medical College and Sir Takhtasinhji General Hos- pital, Bhavnagar-364001 (Gujarat), India. 2Department of Pathol- ogy, Government Medical College and Sir Takhtas- inghji General Hospital, Bhavnagar-364001 (Gujarat), India. Divyesh R. Mandavia,1 Mahendra K. Patel,1 Jayshree C. Patel,1 Ashish P. Anovadiya,1 Seema N. Baxi,2 Chandrabhanu R. Tripathi1 Anti-Urolithiatic Effect of Ethanolic Extract of Pedalium Murex Linn. Fruits on Ethylene Glycol-Induced Renal Calculi Corresponding Author: Chandrabhanu Rajkishor Tripathi, MD CM 31/13, Shantinagar 2, Kalvibid, Bhavna- gar-364002, Gujarat, India. Tel: +91 9825951678 E-mail: cbrtripathi@ yahoo.co.in Received July 2012 Accepted November 2012 Purpose: To evaluate effect of ethanolic extract of Pedalium murex Linn. fruits on experimental model of calcium oxalate nephrolithiasis. Materials and Methods: Thirty-six male Wistar albino rats were randomly divided in 6 groups. Normal controls received distilled water for 28 days. Other five groups received ethylene glycol (1% v/v) in distilled water for 28 days. Pedalium murex ethanolic extract was given 200 mg/kg and 400 mg/kg orally in distilled water for 28 days in prophylactic groups (III and IV) and from 15th to 28th days in treatment groups (V and VI). The urea, creatinine, random blood sugar, aspar- tate aminotransferase, alanine aminotransferase, alkaline phosphatase, bilirubin and calcium were measured on 28th day. 24 hr urinary oxalate and volume were measured on day 0 and 28. On day 28, kidneys were removed, weighed and subjected to histopathological examination. Calcium oxa- late crystallization was evaluated by renal histopathology and in-vitro method of mineralization. All parameters were analyzed by Kruskal-Wallis or one-way ANOVA with post-hoc test. Results: Pedalium murex showed significant improvement in renal function and kidney weight in prophylactic groups as compared to ethylene glycol controls. It did not show any effect on urinary oxalate, urine volume and any other serological parameters. Calcium oxalate crystallization was significantly reduced in all the Pedalium murex treated groups (P < .05). Calcium oxalate and phos- phate mineralization were also inhibited by 33% and 57%. Conclusion: Ethanolic extract of Pedalium murex fruits possess significant activity for prevention of renal calculi. Keywords: calcium oxalate; nephrolithiasis; polyethylene glycols; Pedalium murex Linn.; kidney calculi; pathology ENDOUROLOGY AND STONE DISEASE Endourology and Stone Disease 947Vol. 10 | No. 3 | Summer 2013 |U R O LO G Y J O U R N A L INTRODUCTION Renal calculi are the third leading cause among uri-nary diseases. In the United States, incidence of renal calculi was 1116 per 1,000,000 population in the year 2000 among adults.(1,2) Recurrence of calculi is a serious problem. Recurrence rate is 30 to 40 % at 5 years as seen in observational study.(3) Studies have shown that effective treatment like dietary modification or medication can reduce the recurrence rate significantly.(4-6) So recur- rence of renal stone is partially preventable. Seventy to eighty percent of calculi are made up of calcium oxalate and phosphate. The most common abnormality among stone formers is hyperoxaluria. Consumption of high dietary oxa- late is a major risk factor for stone formation.(7,8) At present, management of stone depends mainly on surgical treatment i.e. extracorporeal shock wave lithotripsy, percutaneous lithotripsy, and transureteral lithotripsy.(9) These surgeries are expensive with higher complication rate than medical management, and do not affect the recurrence of stones. So there is need for medical treatment of renal stone that has curative as well as preventive action on stone formation. Medicinal plants are used for various chronic disorders world- wide. Pedalium murex (P. murex) Linn. (Name in vernacular language – Hindi, is Bada Gokhru) is a member of Pedalia- cae. It has been used as an important medicinal herb in India. According to Indian medicinal literature, it has aphrodisiac, antitussive, appetizer properties and useful in vesical calculi, urinary discharge, gonorrhea etc.(10-14) Fruits of P. murex Linn. are used traditionally for treatment of genito-urinary disorders, infertility, impotency, intestinal colic, diabetes. A few studies had been done for its antimicrobial, aphrodisiac, nephroprotective, hypolipidemic and anti-inflammatory ac- tivities.(15-19) Though it is used as a folk medicine for renal calculi, its effectiveness and mechanism as anti-urolithiatic agent is still unknown. We have done this study to explore its effect and mechanism for prevention and treatment of renal calculi. MATERIAL AND METHODS Animals All the experiments were performed after prior approval from Institutional Animal Ethics Committee (IAEC), Gov- ernment Medical College, Bhavnagar, Gujarat, India. Male Wistar albino rats were procured from the central animal house of the institute. They were housed in standard transpar- ent polycarbonate cages and kept in a 12 hr light-dark cycle under controlled room temperature (24 ± 2° C) and humidity. Animals were given standard laboratory diet and allowed to acclimatize at least three days before starting experiments. The animal handling was performed according to the Good Laboratory Practice (GLP) guidelines. Drugs and Chemicals P. murex Linn. 10% ethanolic fruit extract (Tulsi Amrit Pvt. Ltd., Indore, Madhya Pradesh, India), ethylene glycol (Fisher Scientific Co., Mumbai, India), oxalate kit (Trinity Biotech, Ireland), sodium phosphate (Aldrich, India), sodium oxalate and calcium acetate (Alfa Aesar, Hyderabad, India) were used in this study. Acute toxicity studies of ethanolic fruit extract of P. murex Linn. showed that it was safe to administer it up to 2000 mg/ kg in rat.(17) Based on these studies, we have taken 1/10th of the highest safe dose for the present study. We have done the study with two incremental doses of 200 and 400 mg/kg of P. murex Linn. ethanolic extract to see its dose dependent action. Study Design Thirty six male Wistar albino rat (250 - 350 g) were randomly divided into six equal groups. Group I received distilled wa- ter instead of tap water and served as normal control. Group II to VI received ethylene glycol 1% v/v in distilled water for 28 days. Group II served as ethylene glycol control. Group III and IV animals received P. murex ethanolic extract in 200 mg/kg and 400 mg/kg orally in distilled water for 28 days, respectively and served as prophylactic groups. Group V and VI animals did not receive P. murex for first 14 days. These groups received P. murex ethanolic extract in 200 mg/kg and 400 mg/kg orally in distilled water from 15th to 28th day, re- spectively and served as treatment groups. Outcome measures Biochemical parameters Blood samples were collected in plain vaccuates from the retro-orbital plexus under ketamine (50 mg/kg intra peritone- ally) and xylazine (10 mg/kg intra peritoneally) anaesthesia Effect of Pedalium murex Linn. on Ethylene Glycol | Induced Renal Calculi 948 | 24 hr after the last dose of drug. Serological parameters for renal and hepatic functions were measured. Urinary parameters Twenty-four hr. urine specimens were collected on day 0 and 28 of the study by keeping each rat in separate metabolic cage (B.I.K. Industries, Mumbai, India). Urine volume was measured. It was acidified and kept under refrigeration (2 - 8° C). Urinary oxalate was measured by oxalate kit within 7 days of collection of sample by spectrophotometer.(20) Histopathological parameters The animals were sacrificed soon after blood collection un- der the continued effect of anesthesia. Both kidneys were re- moved and kept in formaldehyde (10% v/v) for at least 24 hours. Then 5 mm thick sections were taken and enclosed in paraffin. They were cut into 5 µm thin sections, stained with hematoxylin-eosin (H & E) and evaluated under optical light binocular microscope. Calcium oxalate crystal deposi- tions were calculated in 10 microscopic fields (159 × 10-9 m2 each) and other changes e.g. necrosis, leukocyte infiltration and tubular dilatations were also noted. In-vitro method of mineralization To evaluate the inhibition of calcium oxalate and phosphate mineralization by P. murex Linn,; we used simultaneous flow static model (S.S.M.) described by Farook et al.(21) We used P. murex Linn. 200 mg/ml, calcium acetate (0.1 M) and sodium oxalate (0.1 M) (for calcium oxalate) or sodium phosphate (for calcium phosphate) in three separate burettes (25 ml) and were allowed to fall simultaneously into a 250 ml beaker with a slow and steady pace. After 30-40 min, the mixture was kept in a hot water bath for 10 min, cooled to room tem- perature and the precipitate was collected into a pre-weighed centrifuge tube. Supernatant fluid was discarded. Then, these tubes were dried in a hot air oven at 120° C, cooled to room temperature and weighed till constant weight is achieved. Weight of the precipitates (ppt.) was calculated. Then per- centage inhibition was calculated by following formula: Inhibition (%) = (Weight of ppt. in blank set - Weight of ppt. in experimental set x 100) / Weight of ppt. in blank set Statistical analysis Endourology and Stone Disease Table 1. Effect of Pedalium murex Linn. on various biochemical parameters Parameter Random blood sugar (mg/dL) Blood urea (mg/dL) Serum creatinine (mg/dL) Alanine ami- notrnasferase (IU/L) Aspartate ami- notrnasferase (IU/L) Alkaline phos- phatase (IU/L) Serum bilirubin (mg/dL) Serum calcium (mmol/L) Group I (Normal control) 110.17 ±21.04 34.83 ± 4.51 0.65 ± 0.15 75.17 ± 14.93 247.83 ± 20.05 280.33 ± 42.45 0.68 ± 0.22 1.08 ± 0.05 Group II (EG control) 82.67 ± 11.27 258.50 ± 85.75* 2.90 ± 0.63* 154.33 ± 94.94 272.33 ± 112.00 343.67 ± 95.36 0.80 ± 0.27 1.21 ± 0.07 Group III (PM 200 mg/kg – 28 days) 176.33 ± 21.43 48.00 ± 1.57 0.78 ± 0.17 ** 74.50 ± 7.62 148.17 ± 39.70 300.33 ± 50.72 0.32 ± 0.03 1.20 ± 0.03 Group IV (PM 400 mg/kg – 28 days) 58.00 ± 11.08 35.83 ± 11.60 ** 0.82 ± 0.27 49.00 ± 3.52 141.00 ± 32.76 186.33 ± 17.05 0.22 ± 0.02 1.28 ± 0.04 Group V (PM 200 mg/kg – 15- 28 days) 151.83 ± 38.81 82.00 ± 22.22 0.98 ± 0.25 89.50 ± 4.71 103.83 ± 45.11 282.33 ± 44.29 0.27 ± 0.03 1.19 ± 0.02 Group VI (PM 400 mg/kg – 15 - 28 days) 241.17 ± 51.63 53.00 ± 3.82 0.77 ± 0.09 46.67 ± 8.66 225.50 ± 80.34 132.17 ± 15.18* 0.32 ± 0.10 1.14 ± 0.08 All values are expressed as mean ± SEM. n=6 for all groups. *P < .05 as compared to group I, **P < .05 as compared to group II by Kruskal-Wallis test followed by Dunn’s multiple comparisons. Keys: PM, Pedalium murex Linn. EG, Ethylene glycol. 949Vol. 10 | No. 3 | Summer 2013 |U R O LO G Y J O U R N A L Data were expressed as Mean ± Standard Error of Mean (SEM). All the quantitative variables (urinary parameters for day 0 and 28, biochemical parameters, kidney weight and calcium oxalate crystal depositions) were compared by one- way ANOVA or Kruskal-Wallis test followed by post hoc test as per Gaussian distribution of the data. All the statisti- cal analysis was done using GraphPad InStat 3.0 (Demo ver- sion). P < .05 was considered as the significant difference. RESULTS Biochemical parameters Ethylene glycol administration increased the values of blood urea and serum creatinine significantly as compared to nor- mal control. P. murex Linn. showed the significant improve- ment in renal parameters in prophylactic groups (P < 0.05). Renal parameters were non-significantly improved in treat- ment groups. Random blood sugar, ALT, AST, ALP, serum bilirubin and serum calcium were not significantly affected by either ethylene glycol or P. murex Linn. (Table 1). Urinary parameters There was no statistically significant difference in urinary volume and oxalate in all the groups on day 0. Urinary oxa- late was significantly increased after ethylene glycol admin- istration. P. murex Linn. did not show significant effect on urinary oxalate. Urine volume was not significantly affected by ethylene glycol or P. murex Linn. (Table 2). Kidney weight Ethylene glycol significantly increased the kidney weight as compared to normal control. P. murex Linn. restored kid- ney weight in prophylactic group in lower dose. (P < .05 for group II vs. III) In other groups, kidney weights were im- proved but they were not statistically significant (Table 3). Histopathological examination Ethylene glycol caused significant calcium oxalate crystal depositions in the renal tubules. These crystals were found in proximal tubules, loop of Henle, distal tubules and associated with significant leukocyte infiltration, necrosis, hemorrhage and tubular dilatation. These crystals were large polygonal in shape, heterogeneous in distribution and pattern (Figure). Treatment with P. murex showed significant reduction in num- ber of crystal depositions all the groups (P < .05). There was no significant difference between the P. murex treated groups. Calcium oxalate crystals were small and found in lesser area in group III, IV, VI as compared to group II. Tubular dilatation and leukocyte infiltration were found to lesser extent in these groups but necrosis was not detected (Table 3). In vitro mineralization P. murex Linn. 200 mg/ml showed 33% inhibition of calcium oxalate and 57% inhibition of calcium phosphate crystalliza- tion. DISCUSSION Table 2. Effect of Pedalium murex Linn. on urinary parameters Urine oxalate (mmol/l) Day Group I (Normal control) Group II (EG control) Group III (PM 200 mg/kg – 28 days) Group IV (PM 400 mg/kg – 28 days) Group V PM 200 mg/kg – 15- 28 days) Group VI (PM 400 mg/kg – 15 - 28 days) Day 0 0.48 ± 0.14 0.35 ± 0.15 0.28 ± 0.07 0.56 ± 0.32 0.85 ± 0.39 1.68 ± 0.26 Day 28 0.18 ± 0.05 1.53 ± 0.19 * 1.74± 0.41* 1.58 ± 0.46 1.52 ± 0.23 1.86 ± 0.42 * Urine Volume (ml) Day Group I (Normal control) Group II (EG control) Group III (PM 200 mg/kg – 28 days) Group IV (PM 400 mg/kg – 28 days) Group V PM 200 mg/kg – 15-28 days) Group VI (PM 400 mg/kg – 15 - 28 days) Day 0 17.58 ± 4.34 11.75 ± 1.82 14.50 ± 3.62 13.67 ± 2.54 16.17 ± 3.0 13.83 ± 2.52 Day 28 33.33 ± 4.22 24.00 ± 5.13 21.83 ± 5.7 30.67 ± 5.87 32.00 ± 8.21 12.83 ± 7.44 All values are expressed as mean ± SEM. n=6 for all groups. *P <.05 as compared to group I by Kruskal-Wallis test followed by Dunn’s multiple comparisons. Keys: PM, Pedalium murex Linn. EG, Ethylene glycol. Effect of Pedalium murex Linn. on Ethylene Glycol | Induced Renal Calculi 950 | Endourology and Stone Disease Urolithiasis is a multifactorial, urological disorder in which super saturation of urine with oxalate plays a key role in pathogenesis. Presently, medical management of renal stone consists of lifestyle modification, calcium channel blocker, diuretics, citrate and magnesium-rich diet.(22) This therapy does not affect recurrence of stone, so traditional medicinal plants have been tried for the prevention of recurrence. We have found increased level of urinary oxalate and cal- cium oxalate crystal depositions in ethylene glycol control group (P <.05). Ethylene glycol administration leads to de- velopment of nephrolithiasis by producing hyperoxaluria. It also increases urinary calcium and phosphate. Studies have shown that hyperoxaluria causes proximal renal tubular dam- age and shedding of brush border cells. It would become a site for calcium oxalate monohydrate (COM) crystal attachment with renal papilla.(23,24) Urinary calcium and super saturation of oxalate develop into nucleation, aggregation and forma- tion of renal stone. So rat model of ethylene glycol is useful for evaluation of renal papillary stone. In present study, all the P. murex treated groups showed significant reduction in calcium oxalate crystal depositions as compared to ethylene glycol control but urinary oxalate was not affected (Table 3). This may be due to lack of diuretic activity of the extract (Ta- ble 2). This suggests direct action of P. murex extract on cal- cium oxalate crystallization in kidney. In addition, P. murex extract has shown significant inhibition of calcium oxalate and phosphate mineralization as seen by in vitro model. We also found significant improvement in renal function (urea and creatinine) in prophylactic groups (P < .05). It sug- gests that P. murex extract can prevent renal damage caused by hyperoxaluria. Ethylene glycol administration also leads to hypertrophy of renal papilla and increased kidney weight probably by inflammation and fluid accumulation.(25) Res- toration of renal function is also associated with improve- ment in kidney weight in P. murex treated rats in prophylac- tic group in low dose (P < .05). Previous study of P. murex fruit extract showed significant improvement in acidic phos- phatase (ACP), ALT, ALP, and lactate dehydrogenase (LDH) in urine, serum and kidney homogenate in ethylene glycol- induced renal damage.(26) These enzymes are non-specific and may increase in many other pathological conditions of liver, kidney and muscles. We have considered more specific parameters for renal calculi like renal function test, kidney weight, urinary oxalate, volume and calcium oxalate crystal depositions in kidney. Calcium oxalate crystal deposition is associated with severe oxidative stress to renal tissue. It leads to lipid peroxidation of membranes by generation of reactive oxygen species like hydroxyl, superoxide ions.(27) It causes accumulation of non protein nitrogenous (NPN) compounds like urea and creati- nine in blood as found in our study. Several antioxidants like vitamin E, vitamin C, flavonoids and phenolic compounds Table 3. Effect of Pedalium murex Linn. on body weight, kidney weight, calcium oxalate crystal depositions Groups Body Weight (g) Kidney weight (g) No. of calcium oxalate crystals in 10 microscopic fields Group I (Normal control) 286.67 ± 12.56 1.19 ± 0.05 0.333 ± 0.211 Group II (EG control) 300.00 ± 8.16 2.03 ± 0.09* 17.667 ± 1.961# Group III (PM 200 mg/kg – 28 days) 263.33 ± 15.42 1.32 ± 0.14** 4.833 ± 1.887## Group IV (PM 400 mg/kg – 28 days) 263.33 ± 17.26 1.50 ± 0.24 6.833 ± 2.915** Group V (PM 200 mg/kg – 15- 28 days) 255.00 ± 15.86 1.65 ± 0.1* 6.500 ± 2.579** Group VI (PM 400 mg/kg – 15 - 28 days) 293.33 ± 17.26 1.64 ± 0.24 1.833 ± 0.872## One way ANOVA F (df ) P = 0.199 1.601 (5, 30) P = 0.0166 3.322 (5, 30) P < 0.0001 9.619 (5, 30) All values are expressed as mean ± SEM. n=6 for all groups. *P < .05 as compared to group I, **P < .05 as compared to group II, #P < .001 as compared to group I, ##P < .001 as compared to group II by one - way ANOVA followed by Tukey-Kramer multiple comparison tests. Keys: PM, Pedalium murex Linn. EG, Ethylene glycol. 951Vol. 10 | No. 3 | Summer 2013 |U R O LO G Y J O U R N A L Effect of Pedalium murex Linn. on Ethylene Glycol | Induced Renal Calculi are found to be effective in prevention of oxidative dam- age and deterioration of renal function.(28,29) Studies have shown that P. murex fruit extract possess strong nitric oxide, 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide, and hydroxyl radical scavenging activities.(30) Phytochemi- cal analysis of ethanolic extract P. murex Linn. fruits show that it contains high levels of flavonoids, glycosides, steroids, phenols, terpenoids, saponins and tannins.(31) Among these components, flavonoids and tannins possess significant anti- oxidant property.(32) Saponins and steroids possess antibacte- rial and antioxidant properties.(33) In present study, inhibition of crystallization may have de- creased oxidative stress to the tissue. Antioxidants present in the extract also may have decreased lipid peroxidation–in- duced renal tubular damage and may contribute to its anti- urolithiatic action. Overall, P. murex fruit extract shows sig- nificant improvement of renal function, kidney weight, and calcium oxalate crystal depositions in prophylactic groups (III and IV) more than treatment groups (V and VI). Reason for this finding can be hypothesized as crystallization is on- going process associated with continued oxidative damage. P. murex fruit extract may not reverse oxidative damage that as already occurred in 14 days treatment with ethylene glycol in group V and VI. So, renal function and kidney weight have not improved significantly in treatment groups. In addition, group VI had lowest calcium oxalate crystal depositions and group V had also similar rate of crystal depositions with pro- phylactic groups which is suggestive of solubilizing effect of the P. murex on already formed crystals. It suggests that P. murex fruit extract may have substantial action on process of crystallization. Further studies are needed to confirm above hypothesis. Nowadays, there is increased concern about toxicity profile of phytotherapy. In present study, we have found that P. mu- rex fruit extract does not affect hepatic function and blood sugar significantly even in ethylene glycol treated rats. It is safe to use for prevention of renal stone. Further clinical stud- ies are required to evaluate efficacy and safety in human be- ings. CONCLUSION Pedalium murex Linn. ethanolic fruit extract has significant anti-urolithiatic activity for prevention of renal calculi prob- ably by affecting calcium oxalate crystallization. There was no dose dependent increment in the effects. CONFLICT OF INTEREST None declared. REFERENCES 1. Hadjzadeh MA, Khoei A, Hadjzadeh Z, Parizady M. Ethanolic extract of nigella sativa L seeds on ethylene glycol-induced kidney calculi in rats. Urol J. 2007;4:86-90. 2. Romero V, Akpinar H, Assimos DG. Kidney stones: A global picture of prevalence, incidence, and associated risk factors. Rev Urol. 2010;12:e86-96. 3. Johnson CM, Wilson DM, O’Fallon WM, Malek RS, Kurland LT. Renal stone epidemiology: a 25- year study in Rochester, Minnesota. Kidney Int. 1979;16:624-31. 4. Borghi L, Schianchi T, Meschi T, et al. Comparison of two di- ets for the prevention of recurrent stones in idiopathic hy- percalciuria. N Engl J Med. 2002;346:77-84. 5. Ettinger B, Pak CY, Citron JT, Thomas C, Adams-Huet B, Vangessel A. Potassium-magnesium citrate is an effective prophylaxis against recurrent calcium oxalate nephrolithi- asis. J Urol. 1997;158:2069-73. Figure . Histopathological images of kidney sections after hema- toxylin & eosin staining under light microscope from (A) normal control showing normal renal tubules and glomeruli (10 × mag- nification), (B) ethylene glycol control showing calcium oxalate crystals (arrows), (C) group treated with P. murex 200 mg/kg for 28 days, (D) group treated with P. murex 400 mg/kg for 28 days, (E) group treated with P. murex 200 mg/kg from 15th – 28th days, (F) group treated with P. murex 400 mg/kg from 15th – 28th days (Images from B to F are of 40 × magnification). 952 | 21. Farook NAM, Dameem GAS, Alhaji NMI, Sathiya R, Muni- yandi J, Sangeetha SJ. Inhibition of Mineralization of Uri- nary Stone Forming Minerals by Hills Area Fruit. E-J Chem. 2004;1:137-41. 22. ShekarKumaran MG, Patki PS. Evaluation of an Ayurvedic formulation (Cystone) in urolithiasis: a double blind, place- bo-controlled study. European Journal of Integrative Medi- cine 2011;3;23-8. 23. Kuo RL, Lingeman JE, Evan AP, et al. Urine calcium and vol- ume predict coverage of renal papilla by Randall’s plaque. Kidney Int. 2003;64:2150-4. 24. Kim SC, Coe FL, Tinmouth WW, et al. Stone formation is pro- portional to papillary surface coverage by Randall’s plaque. J Urol. 2005;173:117-9. 25. de Water R, Noordermeer C, van der Kwast TH et al. Calcium oxalate nephrolithiasis: effect of renal crystal deposition on the cellular composition of the renal interstitium. Am J Kid- ney Dis. 1999;33:761-71. 26. Ananta Teepa KS, Kokilavani R, Balakrishnan A, Gurusamy K. Effect of ethanolic fruit extract of Pedalium murex linn. in ethylene glycol induced urolithiasis in male wistar albino rats. Anc Sci Life. 2010;29:29–34. 27. Thamilselvan S, Hackett RL, Khan SR. Lipid peroxidation in ethylene glycol induced hyperoxaluria and calcium oxalate nephrolithiasis. J Urol 1997;157:1059-63. 28. Khan SR. Hyperoxaluria-induced oxidative stress and anti- oxidants for renal protection. Urol Res. 2005;33:349-57. 29. Thamilselvan S, Menon M. Vitamin E therapy prevents hy- peroxaluria-induced calcium oxalate crystal deposition in the kidney by improving renal tissue antioxidant status. BJU Int. 2005;96:117-26. 30. Sermakkani M, Thangapandian V. Phytochemical screen- ing for active compounds in Pedalium murex L. Rec Res Sci Tech. 2010;2;110-4. 31. Patel DK, Kumar R, Prasad SK, Hemalatha S. Pedalium murex Linn (Pedaliaceae) fruits: a comparative antioxidant activity of its different fractions. Asian Pac J Trop Med. 2011;1:395- 400. 32. Polterait O. Antioxidants and free radical scavengers of nat- ural origin. Current org chem. 1997;1:415-40. 33. Mandal P, SinhaBabu SP, Mandal NC. Antimicrobial activ- ity of Saponins from Acacia auriculiformis. Fitoterapia. 2005;76:462-5. 6. Ettinger B, Tang A, Citron JT, Livermore B, Williams T. Rand- omized trial of allopurinol in the prevention of calcium oxa- late calculi. N Engl J Med. 1986;315:1386-9. 7. Finkielstein VA, Goldfarb DS. Strategies for preventing cal- cium oxalate stones. CMAJ. 2006;174:1407-9. 8. Leonetti F, Dussol B, Berthezene P, Thirion X, Berland Y. Di- etary and urinary risk factors for stones in idiopathic calcium stone formers compared to healthy subjects. Nephrol Dial Transplant. 1998;13:617-22. 9. Miller NL, Lingeman JE. Management of kidney stones. BMJ. 2007;334:468-72. 10. Nadkarni KM. Indian Materia Medica. 3rd Ed. Vol 2. Bombay; Popular Prakashan: 1982. 11. Mhaskar KS, Blatter E, Caiur JF. In: Kritikar and Basu’s illus- trated Indian Medicinal Plants, their usage in Ayurveda and Unani medicines, Vol 8, Indian Medicinal Science Series No 93, PID; New Delhi, 2000, p. 2555-59. 12. Shukla, YN, Khanuja SPS. Chemical, pharmacological and botanical studies on Pedalium murex. J Med Arom Pl Sci. 2004;26:64-9. 13. Sinha RK. Herbal remedies of street vendors for some urino- genital diseases. Anc Sci Life. 1992;11:187-92. 14. Singh NP, Panda H. Meditional herbs with their formulation. New Delhi: Daya Publishiag House; 2005. 15. Nalini K, Ashokkumar D, Venkateswaran V. Antimicrobial Ac- tivity of Petroleum Ether and Methanol Extracts of Pedalium murex Leaves. IJPFR 2011;1:1-10. 16. Balamurugan G, Muralidharan P, Polapala S. Aphrodis- iac activity and curative effects of Pedalium murex (L.) against ethanol-induced infertility in male rats. Turk J Biol. 2010;34:153-63. 17. Balasubramanian MN, Muralidharan P, Balamurugan G. Anti hyperlipidemic activity of Pedalium murex (Linn.) fruits on high fat diet fed rats. Int J Pharmacol. 2008;4:310-13. 18. Adikay S, Latha JP, Koganti B. Effect of fruits of Pedailum mu- rex against cadmium chloride-induced nephrotoxicity in rats. Int J Drug Dev & Res. 2010;2:40-6. 19. Muralidharan, P, Balamurugan G. Analgesic and anti inflam- matory activities of aqueous extract of Pedalium murex linn. Biomedicine. 2008;28:84-7. 20. Li MG, Madappally MM. Rapid Enzymatic Determination of Urinary Oxalate. Clin Chem. 1989;35:2330-3. Endourology and Stone Disease