From the ancient period cow's urine has been used as a medicine. In Veda, cow's urine was compared to the nectar. In Susrut, several medicinal properties of cow's urine have been mentioned and are known to cause weight loss, reversal of certain cardiac and kidney problems, indigestion, stomach ache, edema, etc. However, the literature and scripture did not mention the antigenotoxic properties of cow's urine. Methods In the present investigation, the antigenotoxic/ antioxidant properties of cow's urine distillate and redistillate were studied in vitro. The antioxidant status and volatile fatty acid levels were determined. Actinomycin-D (0.1 μmol/L) and hydrogen peroxide (150 μmol/L) were used for inducing DNA strand break with 0.1% DMSO as negative control. Dose for the antigenotoxic effect of cow's urine was chosen from the dose response study carried out earlier. Results Both actinomycin-D and H2O2 caused statistically significant DNA unwinding of 80% & 75% respectively (P<0.001) as revealed by fluorimetric analysis of DNA unwinding (FADU), and the damage could be protected with the redistilled cow's urine distillate (1, 50 & 100 μL) in simultaneous treatment with genotoxic chemicals. Conclusion The redistillate of cow's urine was found to possess total antioxidant status of around 2.6 mmol, contributed mainly by volatile fatty acids (1500 mg/L) as revealed by the GC-MS studies. These fatty acids and other antioxidants might cause the observed protective effects.

OBJECTIVETo study the anticlastogenic effect of redistilled cow's urine distillate (RCUD) in human peripheral lymphocytes (HLC) challenged with manganese dioxide and hexavalent chromium.

METHODSThe anticlastogenic activity of redistilled cow's urine distillate was studied in human polymorphonuclear leukocytes (HPNLs) and human peripheral lymphocytes in vitro challenged with manganese dioxide and hexavalent chromium as established genotoxicants and clastogens which could cause induction of DNA strand break, chromosomal aberration and micronucleus. Three different levels of RCUD: 1 microL/mL, 50 microL/mL and 100 microL/mL, were used in the study.

RESULTSManganese dioxide and hexavalent chromium caused statistically significant DNA strand break, chromosomal aberration and micronucleus formation, which could be protected by redistilled cow's urine distillate.

CONCLUSIONThe redistilled cow's urine distillate posseses strong antigenotoxic and anticlastogenic properties against HPNLs and HLC treated with Cr+6 and MnO2. This property is mainly due to the antioxidants present in RCUD.

Animals ; Antimutagenic Agents ; pharmacology ; Antioxidants ; pharmacology ; Cattle ; urine ; Cells, Cultured ; Chromium ; antagonists & inhibitors ; toxicity ; DNA Damage ; Humans ; Hydrogen-Ion Concentration ; Lymphocytes ; drug effects ; Manganese Compounds ; antagonists & inhibitors ; Mutagenicity Tests ; Mutagens ; toxicity ; Oxides ; antagonists & inhibitors ; toxicity ; Urine ; chemistry

OBJECTIVETo study the modulatory effect of distillate of Ocimum sanctum (traditionally known as Tulsi) leaf extract (DTLE) on genotoxicants.

METHODSIn the present investigation, we studied the antigenotoxic and anticlastogenic effect of distillate of Tulsi leaf extract on (i) human polymorphonuclear leukocytes by evaluating the DNA strand break without metabolic activation against mitomycin C (MMC) and hexavalent chromium (Cr+6) and (ii) human peripheral lymphocytes (in vitro) with or without metabolic activation against mitomycin C (MMC), hexavalent chromium (Cr+6) and B[a]P by evaluating chromosomal aberration (CA) and micronucleus assay (MN). Three different doses of DTLE, 50 microL/mL, 100 microL/mL, and 200 microL/mL were selected on the basis of cytotoxicity assay and used for studying DNA strand break, chromosomal aberration and micronucleus emergence. The following positive controls were used for inducing genotoxicity and clastogenicity: MMC (0.29 micromol/L) for DNA strand break, chromosomal aberration and 0.51 micromol/L for micronucleus assay; Potassium dichromate (Cr+6) 600 micromol/L for DNA strand break and 5 micromol/L for chromosomal aberration and micronucleus assay; Benzo[a]pyrene (30 micromol/L) for chromosomal aberration and 40 micromol/L for micronucleus assay. The active ingredients present in the distillate of Tulsi leaf extract were identified by HPLC and LC-MS.

RESULTSMitomycin C (MMC) and hexavalent chromium (Cr+6) induced statistically significant DNA strand break of respectively 69% and 71% (P<0.001) as revealed by fluorometric analysis of DNA unwinding. Furthermore, the damage could be protected with DTLE (50 microL/mL, 100 microL/mL, and 200 microL/mL) on simultaneous treatment. Chromosomal aberration and micronucleus formation induced by MMC, Cr+6 and B[a]P were significantly protected (P<0.001) by DTLE with and without metabolic activation.

CONCLUSIONDistillate of Tulsi leaf extract possesses antioxidants contributed mainly by eugenol, luteolin and apigenin as identified by LC-MS. These active ingredients may have the protective effect against genotoxicants.

Adult ; Benzopyrenes ; toxicity ; Cell Survival ; drug effects ; Chromium ; toxicity ; Chromosome Aberrations ; drug effects ; DNA ; metabolism ; DNA Damage ; drug effects ; Humans ; Lymphocytes ; drug effects ; Mass Spectrometry ; Mitomycin ; toxicity ; Mutagens ; toxicity ; Ocimum ; chemistry ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry

OBJECTIVEBacoMind (BM) is a standardized extract of Bacopa monnieri, which belongs to the family Scrophulariaceae and is a creeping annual plant found throughout the Indian subcontinent. It has been used by Ayurvedic medicinal practitioners in India for almost 3000 years and is classified as a medharasayana, a substance which improves memory and intellect. With the widespread traditional use as well as scientific validation of Bacopa monnieri for nootropic activity, a bioactive-rich unique phytochemical composition-BacoMind was developed from B. monnieri for use as a cognition and memory enhancing agent. The present study aimed to investigate the in vitro toxicity of this formulation of BacoMind on human lymphocytes and to rule out its possible contribution to mutagenicity.

METHODSIn the present investigation the active ingredients present in BM were identified and quantified by high performance liquid chromatography (HPLC) and high performance thin-layer chromatography (HPTLC). Antioxidant and anticlastogenic properties of BM were studied in vitro with and without metabolic activation. Doses of BM were chosen on the basis of mitotic index (MI) and cytokinesis-block proliferation index (CBPI). Clastogenicity assays were performed at 31.2 microg/mL, 62.5 microg/mL, and 125 microg/mL, while the Salmonella reverse mutation assay (Ames test) was performed at doses of 61.72, 185.18, 555.55, 1666.67, and 5000.00 microg/plate.

RESULTSHPLC and HPTLC analysis of BM revealed the presence of bacoside A3, bacopaside I, bacopaside II, jujubogenin isomer of bacopasaponin C, bacosine, luteolin, apigenin, bacosine, and beta-sitosterol D glucoside. BM demonstrated significant antioxidant activity. The number of chromosomal aberrations and the frequency of micronuclei induced by BM were not statistically significant up to a dose of 62.5 microg/mL. A subsequent dose of 125 microg/mL prior to metabolic activation induced mild clastogenicity, but it was found to be biologically insignificant as this effect was not seen post metabolic activation. BM also demonstrated a dose-dependent protection against the clastogens used in this study using the above tests for clastogenicity. Maximum protection was observed in presence of metabolic activation. Moreover, BM demonstrated no mutagenic effect on the tested strains, as observed in the Ames test.

CONCLUSIONBM protected human lymphocytes against various clastogens. BM also exhibited high antioxidant activity which might be responsible for the observed protective effects against the clastogens since the used clastogens are known to induce their clastogenic effects via production of oxidative radicals.

Antimutagenic Agents ; adverse effects ; pharmacology ; Bacopa ; chemistry ; Biotransformation ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Chromosome Aberrations ; Humans ; Lymphocytes ; drug effects ; Plant Extracts ; adverse effects ; pharmacology