Podophyllum hexandrum ameliorates endosulfan-induced genotoxicity and mutagenicity in freshwater cyprinid fish crucian carp.

CONTEXT: Medicinal plants continue to act as a repository for novel drug leads with novel mechanisms of action. Podophyllum hexandrum Royale (Berberideceae) treats diverse conditions in folk medicine. OBJECTIVE: The antimutagenic potential of P. hexandrum was evaluated against endosulfan-induced clastogenicity in a piscine model by cytogenetic endpoints. MATERIALS AND METHODS: Podophyllum hexandrum rhizomes were subjected to successive solvent extraction. Fish were exposed to hexane, chloroform, ethyl acetate, methanol and aqueous extracts (15 mg/L each) of plant and endosulfan (0.05 mg/L) alone followed by their combination for antimutagenicity estimates. Chromosomal aberrations (CA) were made from kidney cells and micronuclei (MN) slides from peripheral blood erythrocytes at 48, 72 and 96 h. Antioxidant activity was analyzed by the DPPH assay. Phytochemical analyses were carried out using chromatographic and spectroscopic techniques. RESULTS: Endosulfan induced significant (p < .05) MN, authenticated by scanning electron microscopy, and CA in a time-dependent manner. However, methanol and ethyl acetate extracts revealed ameliorating effects. The column eluted methanolic fraction-2 (ME-F2) showed highest reduction profile of 83 and 84% in CA and MN, followed in its extent (73 and 72%) by ethyl acetate fraction-4 (EE-F4). ME-F2 and EE-F4 showed three and six major peaks when analyzed by GC-MS. To explore possible mechanism of action, ME-F2 showed potent antioxidant potential and strong correlation (R2 = .900) with antimutagenic activity, whereas EE-F4 seemed to act through a different mechanism. DISCUSSION AND CONCLUSION: This study confirms the antimutagenic potential of the subject plant with the identification of some novel compounds, justifying their use in folk medicine, and their corresponding benefit to mankind.

Massive land system changes impact water quality of the Jhelum River in Kashmir Himalaya.

The pristine aquatic ecosystems in the Himalayas are facing an ever increasing threat from various anthropogenic pressures which necessitate better understanding of the spatial and temporal variability of pollutants, their sources, and possible remedies. This study demonstrates the multi-disciplinary approach utilizing the multivariate statistical techniques, data from remote sensing, lab, and field-based observations for assessing the impact of massive land system changes on water quality of the river Jhelum. Land system changes over a period of 38 years have been quantified using multi-spectral satellite data to delineate the extent of different anthropogenically driven land use types that are the main non-point sources of pollution. Fifteen water quality parameters, at 12 sampling sites distributed uniformly along the length of the Jhelum, have been assessed to identify the possible sources of pollution. Our analysis indicated that 18% of the forested area has degraded into sparse forest or scrublands from 1972 to 2010, and the areas under croplands have decreased by 24% as people shifted from irrigation-intensive agriculture to orchard farming while as settlements showed a 397% increase during the observation period. One-way ANOVA revealed that all the water quality parameters had significant spatio-temporal differences (p < 0.01). Cluster analysis (CA) helped us to classify all the sampling sites into three groups. Factor analysis revealed that 91.84% of the total variance was mainly explained by five factors. Drastic changes in water quality of the Jhelum since the past three decades are manifested by increases in nitrate-nitrogen, TDS, and electric conductivity. The especially high levels of nitrogen (858 ± 405 µgL(-1)) and phosphorus (273 ± 18 µgL(-1)) in the Jhelum could be attributed to the reckless application of fertilizers, pesticides, and unplanned urbanization in the area.

Assessment of endosulfan induced genotoxicity and mutagenicity manifested by oxidative stress pathways in freshwater cyprinid fish crucian carp (Carassius carassius L.).

Over the past few decades, endosulfan, one of the polychlorinated pesticides still in use, has received considerable attention of a number of international regulations and restriction action plans worldwide. This study aimed to evaluate the cytogenetic effects of endosulfan using robust genotoxicity assays, along with the oxidative stress pathways in order to understand biochemical mechanism, in Carassius carassius L. The LC50-96 h (95% confidence limits) value of endosulfan was 0.070 (0.046-0.093) ppm; and on its basis three test concentrations (sub-lethal I: 0.052, II: 0.035 and III: 0.017 ppm) were selected for 35 d in vivo exposure. The mean concentration of endosulfan in aquaria was always constant, when analyzed by dispersive liquid-liquid micro extraction (DLLME) followed by GC-MS. Autopsy was done on days 1, 2, 3, 4, 7, 14, 21, 28 and 35 of endosulfan exposure; the micronucleus formation (MN), authenticated by scanning electron microscopy, and chromosomal aberrations (CA), were induced significantly (p<0.05) in all the treated groups, including positive control cyclophosphamide (4 ppm), when compared to negative control. Similarly lipid peroxidation (LPO) was induced significantly with the maximal at higher concentration (SL-I) on 4th day (722.45%; p<0.01). Antioxidant biomarkers like glutathione reduced, superoxide dismutase and catalase also fluctuated significantly (p<0.01) in all treatment groups. Collective findings demonstrated that genotoxic effects were invariably accompanied and correlated with increased oxidative stress and disturbance of antioxidant enzymes; and the MN and CA assays are useful tools in determining potential genotoxicity of aquatic xenobiotics and might be appropriate as a part of monitoring program.

Pharmacological and toxicological evaluation of Urtica dioica.

CONTEXT: Medicinal plants are a largely unexplored source of drug repository. Urtica dioica L. (Urticaceae) is used in traditional medicine to treat diverse conditions. OBJECTIVE: The present study describes the antidiabetic, antiinflammatory, antibacterial activity, and toxicological studies of Urtica dioica. MATERIALS AND METHODS: U. dioica leaves were subjected to solvent extraction with hexane, chloroform, ethyl acetate, methanol, and aqueous, respectively, and screened for antidiabetic (300 mg/kg bw by glucose tolerance test; GTT), antiinflammatory (200 mg/kg bw by rat paw edema assay) and antibacterial activities [by disc-diffusion and minimum inhibitory concentration (MIC) assays]. Toxicological studies were carried on Artemia salina and Wistar rats; phytochemical analyses were carried out, using chromatographic and spectroscopic techniques. RESULTS: The aqueous extract of U. dioica (AEUD) significantly (p < 0.001; 67.92%) reduced the blood glucose level during GTT in Wistar rats with an effective dose of 300 mg/kg bw in dose-dependent studies. High-performance liquid chromatography with photodiode-array detection (HPLC-DAD) analysis showed the presence of hydroxycinnamic acid derivatives and flavonoids in AEUD. Hexane Fraction-2 (HF2) exhibited both antiinflammatory activity (48.83% after 3 h), comparable to that of indomethacin (53.48%), and potent antibacterial activity with MIC values ranging from 31.25-250 µg/mL against all the tested strains. Gas chromatography-mass spectrometry (GC-MS) analysis showed fatty acid esters and terpenes as the major constituents of HF2. Toxicity tests showed higher safety margin of all the solvent extracts with LC(50) > 1000 µg/mL each on A. salina. DISCUSSION AND CONCLUSION: Our results showed that the U. dioica leaves are an interesting source of bioactive compounds, justifying their use in folk medicine, to treat various diseases.