Restorative effect of Cadamba Fruit Extract on Arsenic-Induced Nephrotoxicity in Albino Mice

Arsenic is one of the most important global environmental toxicants and its exposure in humans comes mainly from consumption of drinking water contaminated with inorganic arsenic. In clinical trials it is considered as a first choice cancer chemotherapeutic against certain leukemia and has potential against a variety of other cancers, including solid tumors. Specifically, arsenic trioxide (As2O3,) is used in the treatment of acute promyelocytic leukemia and it greatly improves the clinical outcome even in refractory or multiple relapsed cases . But, toxic side effects of arsenicals are often a major concern; including the potential for fatal hepatotoxicity .The liver is a major target organ for both arsenic metabolism and toxicity. Arsenic induced hepatic injury is known to be exerted through excess production of reactive oxygen species, namely superoxide (O2), hydroxyl OH), and peroxy (ROO) radicals and hydrogen peroxide (H2O2). The harmful expressions of arsenic are primarily due to an imbalance between pro-oxidant and antioxidant homeostasis in physiological system and also due to its fascination to bind sulfhydryl groups of proteins and thiols of glutathione (GSH). Thus, an agent able to reduce the toxic potential of arsenic in liver cells would clearly to be useful ccompounds for arsenical chemotherapy. Neolamarckia cadamba primarily consist of indole alkaloids, terpenoids, sapogenins, saponins, terpenes, steroids, fats and reducing sugars. The bark also consists of tannins and an astringent principle; which is due to the presence of an acid similar to cincho-tannic acid. A new pentacyclic triterpenic acid isolated from the stem bark Neolamarckia cadamba named cadambagenic acid (18α-olean-12ene-3β-hydroxy 27, 28-dioic acid) (Fig. 1), along with this acid quinovic acid (Fig. 2) and β- sitosterol (Fig. 3) have also been isolated The isolation and structure of 3β-dihydrocadambine and 3β-isodihydrocadambine (Fig. 8) alkaloids reported from the leaves of Neolamarckia cadamba with molecular formula (C37H44N15O2). A new saponin named saponin B (C48H76O17) reported from Neolamarckia cadamba (Miq.). Neolamarckia cadamba also contain an acid called chlorogenic acid (CGA) (Fig. 9) . Recently some worker isolated two novel triterpenoidsaponins, phelasin A and phelasin B from the bark of Neolamarckia cadamba (Roxb.) Miq. Two novel monoterpenoidindole alkaloids, aminocadambine A (C24H27N305) (fig. 10) and

worker biosynthetically synthesized glucosidicindole alkaloid cadambine from its biological precursor secologanin which is the main precursor of various indole alkaloids Three monoterpenoidgluco-indole alkaloids, 3β-isodihydrocadambine, cadambine and 3α-dihydrocadambine isolated from Nauclea cadamba (Roxb.). The flowers of Neolamarckia cadamba yield an essential oil and the main constituents of oils are linalool, geraniol, geranyl acetate, linalyl acetate, α-selinene, 2-nonanol, β-phellandrene, α-bergamottin, p-cymol, curcumene, terpinolene, camphene and myrcene. Two triterpenoid glycosides, glycosides A and B were isolated from the bark of Neolamarckia cadamba and defined as 3-o-(α-L-rhamnopyranosyl)-quinovic acid-28-o-(β-D-glucopyranosyl) ester and 3-o-(β-D- glucopyranosyl)-quinovic acid-28-o-(β-D- glucopyranosyl) ester respectively and eight different alkaloids also obtained from Neolamarckia cadamba named cadambine, CFJ 83, isomalindan, cadamine, 2 derivs. HFP34, GZM28, malindan, dihydro cadambine (Fig. 12), 2derivs. GPX71, GPX73, isomalindan, isodihydro cadambine, 2 derivs. GPX51, GPX53, malindan. The seeds of Anthocephalusindicus composed of water-soluble polysaccharides D-xylose, D-mannose and D-glucose in the molar ratio 1:3:5. Almost all parts of the plant Neolamarckia cadambais used in the treatment of various diseases. Decoction of leaves is used as gargle in aphthae or stomatitis and in the treatment of ulcers, wounds, and metorrhea. Bark of the plant is used in fever, inflammation, cough, vomiting, diarrhoea, diabetes, burning sensation, diuresis, wounds, ulcers and in the treatment of snake-bite. Arsenic trioxide (4 mg/kg/day b.wt) was administered to produce nephrotoxicity in rats followed by administration of Cadamba fruit extract (100 mg/kg/day p.o. 80 days) to attenuate the arsenic mediated nephro-toxicity. The kidney function tests were assayed and were found with elevated levels of urea, uric acid, and creatinine. Furthermore, their free radical assessment like lipid peroxidation levels were assayed which was found significantly high. Moreover the histopathological evaluations of kidney showed severe changes in mice treated with arsenic trioxide. But, after administration of aqueous extract of Cadamba fruit extract, it caused significant restoration in serum levels of urea, uric acid, creatinine, and lipid peroxidation levels and also in histological changes. Thus, it is evident from study that Cadamba fruit extract possesses antidote and acts effectively against arsenic induced nephrotoxicity. Cadamba fruit extract is a naturally occurring antioxidant that plays an important role by inactivating harmful free radicals produced through normal cellular activity and from various stressors thus species (ROS). Therefore cadamba fruit extract is used to ameliorate the toxic effects of arsenic.

In the present study, fresh fruit of cadamba were collected from plant of Neolamarckia cadamba from premises of Mahavir Cancer Institute $Research Centre ,Patna,India. The identity of the medicinal plant was confirmed by Dr. S.K RAY . The collected fruit of cadamba were shade dried and were grinded to fine powder. The aqueous extract dose was calculated after LD50 estimation which was found to be 1500mg kg-1 body weight and the final dose was fixed to 100mg kg-1 body weight. Animals: Thirty male swiss albino mice (28g to 32g) were obtained from animal house of Mahavir Cancer Institute & Research Centre, Patna, India . Food and water to mice were provided ad libitum (prepared mixed formulated feed by the laboratory itself). Animals were maintained in colony rooms with 12 hrs light/dark cycle at 22 ± 2o C. Experimental protocol The mice were divided into four groups of six rats each, a normal control group, a cadamba fruit extract control which received 100 mg/kg b.wt of cadmba fruit extract, one As2O3 (4 mg/kg b.wt) administered group and a combination group treated with 4 mg/kg b.wt of As2O3 and 100 mg/kg b.wt of cadamba fruit extract. 0.2% DMSO solution was used as vehicle for cadamba fruit extract administration. Experimental groups received this via oral intubation daily for a period of 80 days. At the end of the experimental period animals were decapitated, blood was collected and centrifuged at 3 000 rpm for 20 minutes; the clear serum obtained was used for the determination of marker enzymes. kidney was removed immediately, washed in ice cold 0.15M NaCl and blotted on a filter paper. Then the tissue was weighed and homogenized by using Teflon glass homogenizer (1/10th weight/volume) in ice cold tris-HCl buffer (0.2M, pH 7.4). The homogenate was centrifuged at 10 000g for 20 min at 4 C and the supernatant was used for the estimation of lipid peroxidation and various enzymatic and non enzymatic assays. Plasma specimens were obtained and used for determination of creatinine , urea and uric acid using an automatic analyser (Reflotron plus system , roche , Germany). Kidney homogenates were obtained using a tissue homogenizer . the homogenates (1:10 w/v) were prepared using a 100 mM KCL buffer (7:00 pH ) containing EDTA 0.3 mM

used for the biochemical assays of glutathione (GSH) and superoxide dismutases (SOD) levels using GSH and SOD assays kits (sigma-Aldrich com.) according to the manufacture‘s instruction with some modification . also , kidneys were quickly removed , immersed in 10% formalin , dehydrated and embedded in paraffin , sectioned at 4 µm, stained with hematoxylin and cosin (H&E) and evaluated by light microscopey. Images representative of typical histological profile in control and all treated groups were captured with the aid of motic imaging software.

The calculations and statistical analysis were carried out using the statistical package for social sciences (SPSS) for windows version 12.0 software. All data were represented as mean ±standard deviation (SD). Data e=were subjected to one-way analysis of variance (ANOVA) followed by student‘s t-test .statistical probability of p < 0.05 was considered to be significant.

Histopathological examination of the kidney specimens showed severe alterations in mice exposed to heavy metals . Renal tubular dilatations with congestion of blood vessels with hemorrhage and degeneration of renal corpuscles were notedcompared to the normal structure of control group .Administration of cadamba flower extract protected the kidney exposed to arsenic trioxide as evidenced by appearance of normal structures of kidney, specially renal corpuscles .Additionally, the histopathological evaluation of only cadamba flower extract treated mice showed normal renal corpuscle (A–H) Histological changes of kidney in each group. (A) Normal structure of renal corpuscle in control mice (1000×). (B) Renal cortex and medulla structure of arsenic trioxide related mice (100×). (C–E) Renal cortex structures of arsenic trioxide treated mice (400×). (F) Renal corpuscle structure of arsenic trioxide treated mice (1000×). (G) Renal corpuscle structure of arsenic trioxide pluscadamba flower extract treated mice (1000×). (H) Renal corpuscle structure of cadamba flower extract treated mice (1000×). P < 0.05: Student‘s t-test (significance levels shown for difference between control and treated groups). P < 0.05: Student‘s t-test (significance levels shown for difference between mice exposed to Arseic trioxide and arsenic trioxide plus cadamb). P < 0.05: Student‘s t-test (significance levels shown for difference between mice exposed to arsenic trioxide and cadamba).

Plasma creatinine level was found to be 0.36 +/- 0.04 mg/dL in control. When control was treated with arsenic trioxide for 80 days, the plasma creatinine level was found to be increased upto 155.56% i.e. 0.92 +/- 0.05 mg/dL from 0.36 +/- 0.04 mg/dL. When this arsenic treated mice was further treated with cadamba extract, a significant decrease of 59.78% in plasma cretinine level was observed. When arsenic trioxide and cadamba extract were given to control simultaneously for 80 days, the plasma creatinine level was found to increase by 5% which is very less as compare to 155.56% increase when mice was treated with arsenic trioxide alone. Hence, it was observed that administration of cadamba extract reduces the toxic effect of arsenic trioxide on the kidney of mice. Plasma urea level was found to be 13.96 +/- 1.37 mg/dL in control. When control was treated with arsenic trioxide for 80 days, the plasma urea level was found to be increased upto 82.38% i.e. 25.46 +/- 3.02 mg/dL from 13.96 +/- 1.37 mg/dL. When this arsenic treated mice was further treated with cadamba extract, a significant decrease of 42.22 % in plasma urea level was observed. When arsenic trioxide and cadamba extract were given to control simultaneously for 80 days, the plasma urea level was found to increase by 6.52 % which is very less as compare to 82.38 % increase when mice was treated with arsenic trioxide alone. Hence, it was observed that administration of cadamba extract reduces the toxic effect of arsenic trioxide on the kidney of mice. Plasma uric acid level was found to be 1.89 +/- 0.12 mg/dL in control. When control was treated with arsenic trioxide for 80 days, the plasma uric acid level was found to be increased upto 62.96 % i.e. 3.08 +/- 0.32 mg/dL from 1.89 +/- 0.12 mg/dL. When this arsenic treated mice was further treated with cadamba extract, a significant decrease of 43.18 % in plasma uric acid level was observed. When arsenic trioxide and cadamba extract were given to control simultaneously for 80 days, the plasma uric acid level was found to increase by 1.59 % which is very less as compare to 62.96 % increase when mice was treated with arsenic trioxide alone. Hence, it was observed that administration of cadamba extract reduces the toxic effect of arsenic trioxide on the kidney of mice. Kidney GSH value of control mice was measured to 3.43 +/- 0.49. When control was treated with arsenic trioxide for 80 days, the kidney GSH value was found to be decreased upto 28.28 % i.e. 2.46 +/- 0.48 GSH level was observed. When arsenic trioxide and cadamba extract were given to control simultaneously for 80 days, the kidney GSH value was found to decrease by 3.50 % which is very less as compare to 28.28 % decrease when mice was treated with arsenic trioxide alone. Hence, it was observed that administration of cadamba extract reduces the toxic effect of arsenic trioxide on the kidney of mice. Kidney SOD value of control mice was measured to 4.59 +/- 0.37. When control was treated with arsenic trioxide for 80 days, the kidney SOD value was found to be decreased upto 40.09 % i.e. 2.75 +/- 0.66 mg/dL from 4.59 +/- 0.37 mg/dL. When this arsenic treated mice was further treated with cadamba extract, a significant increase of 74.18 % in kidney SOD value was observed. When arsenic trioxide and cadamba extract were given to control simultaneously for 80 days, the kidney SOD value was found to decrease by 4.14 % which is very less as compare to 40.09 % decrease when mice was treated with arsenic trioxide alone. Hence, it was observed that administration of cadamba extract reduces the toxic effect of arsenic trioxide on the kidney of mice.

The present study showed that cadamba flower extract has protective effect on heavy metals-induced renal and testicular oxidative stress and injuries. our study therefore suggests that cadamba flower extract may be a useful preventive agent against the effect of the studied heavy metals at least partly due to its antioxidant properties. There are three parts of the study. First part is to know the effect of arsenic trioxide on the kidney of the swiss albino mice. In second phase of our study, these arsenic treated mice were treated with cadamb fruit extract to find out the remedial action of cadamba fruit on the arsenic affected organs of the mice. This part of the study confirmed that cadamb fruit can be used in treatment of arsenic induced toxicity. The final part of the study comprises the determination of combine effect of arsenic trioxide and cadamb extract on the organs of Albino mice. The study concludes that when the people who are bound to intake arsenic trioxide due to its presence in water given cadamb fruit, can get relief from the toxic effects of arsenic. In most of the areas of Bihar, Uttar Pradesh and Madhya Pradesh, arsenic trioxide is present in significant amount in groundwater. People living in these areas are bound to drink this water. Although

by the economically weaker section of the population living in these areas. Cadamba fruit is available in these areas in abundant and is in reach of every person. Therefore, the study was oriented in the direction of finding the most economical remedy for the most common and severe health problems of Bihar, Uttar Pradesh and Madhya Pradesh etc. Now we discuss about the major cause of sufferings for the people of these states i.e. arsenic and the diseases caused by this.

Kamata J.P., Boloora K.K., Devasagayam T.P.A. and Venkatachalam S.R. (2000). Antioxidant properties of Asparagus racemosus against damage induced by γradiation in rat liver mitochondria. Journal of Ethnopharmacology 71: pp. 425-435. Kamiya K., Hamabe W., Tokuyama S., Hirano K., Satake T., Kumamoto-Yonezawa Y., Yoshida H. and Mizushina Y. (2009). Inhibitory effect of anthraquinones isolated from the Noni (Morindacitrifolia) root on animal A-, B- and Y-families of DNA polymerases and human cancer cell proliferation. Food Chemistry 118: pp. 725-730. Kapadia G.J., Tokuda H., Konoshima T. and Nishino H. (1996). Chemoprevention of lung and skin cancer by Beta vulgaris (beet) root extract. Cancer Letters 100: pp. 211-214. Kaur P., Singh B., Kumar S. and Kaur S. (2008). In vitro evaluation of free radical scavenging activity of Rubiacordifolia L. Journal of Chinese Clinical Medicine 3: pp. 278-284. Kavitha K. and Manoharan S. (2006). Anticarcinogenic and antilipidperoxidative effects of Tephrosiapurpurea (Linn) Pers. in 7, 12-dimethylbenz(a)anthracene (DMBA) induced hamster buccal pouch carcinoma. Indian Journal of Pharmacology 38: pp. 185-189. Kinghorn A.D., Farnsworth N., Soejarto D., Cordell G., Swanson S., Pezzuto J., Wani M., Wall M., Oberlies N., Kroll D., Kramer R., Rose W., Vite G., Fiarchild C., Peterson R. and Wild R. (2003). Novel strategies for the discovery of plant-derived anticancer agents. Pharmaceutical Biology 41: pp. 53-67. Kulkarni M.P. and Juvekar A.R. (2008). Effect of Alstoniascholaris (Linn.) R. Br. on stress and cognition in mice. Indian Journal of Experimental Biology 47: pp. 47-52. effects of ethanolic neem leaf extract on prostate cancer cell line (PC-3). Journal of Ethnopharmacology 105: pp. 246-250. Kumaraswamy MV and Satish S (2008). Free radical scavenging activity and lipoxygenase inhibition of Wood for diafructicosa Kurz and Betulautilis Wall. African Journal of Biotechnology 7: pp. 2013-2016. Lee Y.K., Lee W.S., Hwang J.T., Kwon D.Y., Surh Y.J. and Park J. (2009). Curcumin exerts antidifferentiation effect through AMPKα-PPAR-γ in 3T3-L1 adipocytes and antiproliferatory effect through AMPKα-COX-2 in cancer cells. Journal of Agricultural and Food Chemistry 57: pp. 305-310. Liu X., Zhao M., Wua K., Chai X., Yu H., Tao Z. and Wang J. (2012). Immunomodulatory and anticancer activities of phenolics from emblica fruit (Phyllanthusemblica L.). Food Chemistry 131: pp. 685-690. Mahishi P., Srinivasa B.H. and Shivanna M.B. (2005). Medicinal plant wealth of local communities in some villages in Shimoga district of Karnataka, India. Journal of Ethnopharmacology 98: pp. 307-312. Malik A., Afaq F., Sarfaraz S., Adhami V.M., Syed D.N. and Mukhtar H. (2005). Pomegranate fruit juice for chemoprevention and chemotherapy of prostate cancer. Proceedings of the National Academy of Sciences of the United States of America 102: pp. 14813-14818. Mondal S., Mondal N.B. and Mazumder U.K. (2007). In vitro cytotoxic and human recombinant caspase inhibitory effect of Annona reticulata leaves. Indian Journal of Pharmacology 39: pp. 253-254. Mossman T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65: pp. 55-63. Nair P.K.R., Melnick S.J., Wnuk S.F., Rapp M., Escalon E. and Ramachandran C. (2009). Isolation and characterization of an anticancer catechol compound from Semecarpusanacardium. Journal of Ethnopharmacology 122: pp. 450-456. Nam N., Lee C., Hong D., Kim H., Bae K. and Ahn B. (2003). Antiinvasive, antiangiogenic and antitumour activity of Ephedra sinica extract. Phytotherapy Research 17: pp. 70-76.

growth and angiogenesis by a medicinal herb: Ocimumgratissimum. International Journal of Cancer 121: pp. 884-894. Newman D.J., Cragg G.M. and Snader K.M. (2003). Natural products as sources of new drugs over the period 1981-2002. Journal of Natural Products 66: pp. 1022-1037. Onay-Ucar E., Karagoz A. and Arda N. (2006). Antioxidant activity of Viscum album ssp. album. Fitoterapia 77: pp. 556-560. Padmavathi B., Rath P.C., Rao A.R. and Singh R.P. (2005). Roots of Withaniasomnifera inhibit forestomach and skin carcinogenesis in mice. Evidence-Based Complementary and Alternative Medicine 2: pp. 99-105. Pezzuto J.M. (1997). Plants-derived anticancer agents. Biochemical Pharmacology 53: pp. 121-133. Pietarinen S.P., Willfor S.M., Ahotupa M.O., Hemming J.E. and Holmbom B.R. (2006). Knotwood and bark extracts: strong antioxidants from waste materials. Journal of Wood Sciences 52: pp. 436-444. Prasad K.N., Ashok G., Raghu C., Shivamurthy G.R., Vijayan P. and Aradhya S.M. (2005). In vitro cytotoxic properties of Ipomoea aquatica leaf. Indian Journal of Pharmacology 37: pp. 397-398. Prince P.S.M. and Menon V.P. (1999). Antioxidant activity of Tinosporacordifolia roots in experimental diabetes. Journal of Ethnopharmacology 65: pp. 277-281. Raghu C., Ashok G., Dhanaraj S.A., Suresh B. and Vijayan P. (2004). In vitro cytotoxic activity of Lantana camara Linn. Indian Journal of Pharmacology 36: pp. 94-95. Rajendran P., Ekambaram G. and Sakthisekaran D. (2008). Effect of mangiferin on benzo(a)pyrene induced lung carcinogenesis in experimental Swiss albino mice. Natural Product Research 22: pp. 672-680. Rajkapoor B., Jayakar B. and Murugesh N. (2004). Antitumor activity of Indigoferaaspalathoides on Ehrlich ascites carcinoma in mice. Indian Journal of Pharmacology 36: pp. 38-40. Rao S.K., Rao P.S. and Rao B.N. (2008). Preliminary investigation of the radiosensitizing activity of guduchi (Tinosporacordifolia) in tumor Rastogi S., Shukla Y., Paul B.N., Chowdhuri D.K., Khanna S.K. and Das M. (2007). Protective effect of Ocimum sanctum on 3-methylcholanthrene, 7, 12dimethylbenz (a) anthracene and aflatoxin B1 induced skin tumorigenesis in mice. Toxicology and Applied Pharmacology 224: pp. 228-240. Reddy D.B., Reddy T.C.M., Jyotsna G., Sharan S., Priya N., Lakshmipathi and Reddanna P. (2009). Chebulagic acid, a COX-LOX dual inhibitor isolated from the fruits of Terminalia chebulaRetz., induces apoptosis in COLO-205 cell line. Journal of Ethnopharmacology 124: pp. 506-512. Reddy V.V.S. and Sirsi M. (1969). Effect of Abrusprecatorius L. on experimental tumors. Cancer Research 29: pp. 1447-1451. Rohini G. and Devi C.S.S. (2008). Bacopamonniera extract induces apoptosis in murine sarcoma cells (S-180). Phytotherapy Research 22: pp. 1595-1598. Samuelsson G. (1999). Drugs of Natural Origin. A Textbook of Pharmacognosy. 4th Ed., Stockholm, Swedish Pharmaceutical Press. Sharma P.R., Shanmugavel M., Saxena A.K. and Qazi G.N. (2008). Induction of apoptosis by a synergistic lignan composition from Cedrusdeodara in human cancer cells. Phytotherapy Research 22: pp. 1587-1594. Sharma PR, Mondhe DM, Muthiah S, Pal HC, Shahi AK, Saxena AK and Qazi GN (2009). Anticancer activity of an essential oil from Cymbopogonflexuosus. ChemicoBiological Interactions 179:160-168. Shoeb M. (2006). Anticancer agents from medicinal plants. Bangladesh Journal of Pharmacology 1: pp. 35-41. Shukla Y., Prasad S., Tripathi C., Singh M., George J. and Kalra N. (2007). In vitro and in vivo modulation of testosterone mediated alterations in apoptosis related proteins by [6]-gingerol. Molecular Nutrition and Food Research 51: pp. 1492-1502. Simard F., Legault J., Lavoie S., Mshvildadze V. and Pichette A. (2008). Isolation and identification of cytotoxic compounds from the wood of Pinusresinosa. Phytotherapy Research 22: pp. 919-922.

of kalimusli (Curculigoorchioides). International Journal of Green Pharmacy 2: pp. 34-36. Sivakumar R. and Alagesaboopathi C. (2008).Studies on cytotoxicity and antitumor screening of red and white forms of Abrusprecatorius L. African Journal of Biotechnology 7: pp. 3984-3988. Skehan P., Storeng R., Scudiero D., Monks A., McMahon J., Vistica D., Warren J.T., Bokesch H., Kenney S. and Boyd M.R. (1990). New colorimetric cytotoxicity assay for anticancer-drug screening. Journal of National Cancer Institute 82: pp. 1107-1112. Son J.K., Jung S.J., Jung J.H., Fang Z., Lee C.S., Seo C.S., Moon D.C., Min B.S., Kim MR and Woo MH (2008). Anticancer constituents from the roots of Rubiacordifolia L. Chemical and Pharmaceutical Bulletin 56: pp. 213-216. Stoilova I., Krastanov A., Stoyanova A., Denev P. and Gargova S. (2007). Antioxidant activity of a ginger extract (Zingiberofficinale). Food Chemistry 102: pp. 764-770. Sultana S., Ahmed S. and Jahangir T. (2008). Emblica officinalis and hepatocarcinogenesis: A chemopreventive study in Wistar rats. Journal of Ethnopharmacology 118: pp. 1-6. Sundararajan P., Dey A., Smith A., Doss A.G., Rajappan M. and Natarajan S. (2006). Studies of anticancer and antipyretic activity of Bidenspilosa whole plant. African Health Sciences 6: pp. 27-30. Surh Y. (2003). Cancer chemoprevention with dietary phytochemicals. Nature Reviews Cancer 3: pp. 768-780. Swamy S.M.K. and Tan B.K.H. (2000). Cytotoxic and immunopotentiating effects of ethanolic extract of Nigella sativa L. seeds. Journal of Ethnopharmacology 70: pp. 1-7. Szewczuk V.D., Mongelli E.R. and Pomilio A.B. (2006). In vitro anthelmintic activity of Melia azedarach naturalized in Argentina. Phytotherapy Research 20: pp. 993-996. Tan G., Gyllenhaal C. and Soejarto D.D. (2006). Biodiversity as a source of anticancer drugs. Current Drug Targets 7: pp. 265-277. Tannin-Spitz T., Bergman M. and Grossman S. (2007a). Cucurbitacin glucosides: Antioxidant Communications 364: pp. 181-186. Tannin-Spitz T., Grossman S., Dovrat S., Gottlieb H.E. and Bergman M. (2007b). Growth inhibitory activity of cucurbitacin glucosides isolated from Citrulluscolocynthis on human breast cancer cells. Biochemical Pharmacology 73: pp. 56-67. Tiwari A.K., Srinivas P.V., Kumar S.P. and Rao J.M. (2001). Free radical scavenging active components from Cedrusdeodara. Journal of Agricultural and Food Chemistry 49: pp. 4642-4645. Tomosaka H., Chin Y., Salim A.A., Keller W.J., Chai H. and Kinghorn A.D. (2008). Antioxidant and cytoprotective compounds from Berberis vulgaris (Barberry). Phytotherapy Research 22: pp. 979-981. Unnikrishnan M.C. and Kuttan R. (1998). Cytotoxicity of extracts of spices to cultured cells. Nutrition and Cancer 11: pp. 251-257. Venukumar M.R. and Latha M.S. (2002). Antioxidant activity of Curculigoorchioides in carbon tetrachloride induced hepatopathy in rats. Indian Journal of Clinical Biochemistry 17: pp. 80-87.

Research Scholar, School of Life Science, OPJS University, Churu, Rajasthan