Huma Ali

Sensitivity of freshwater pulmonate snail Lymnaea luteola L., to silver nanoparticles

Toxicity of nanoparticles depends on many factors including size, shape, chemical composition, surface area and surface charge. Silver nanoparticles (AgNPs) are likely to enter the aquatic ecosystems because of their multiple applications and pose a health concern for humans and aquatic species. Therefore, we used a freshwater snail Lymnaea luteola L (L. luteola) to investigate the acute toxicity and genotoxicity of AgNPs in a static-renewal system for 96 h. AgNPs caused molluscicidal activity in L. luteola, with 96-h median lethal concentrations (LC50) (48.10 μg L(-1)). We have observed that AgNPs (36 μg L(-1)) elicited a significant (p<0.01) reduction in glutathione, glutathione-s-transferase and glutathione peroxidase with a concomitant increase in malondialdehyde level and catalase in digestive gland of L. luteola. However, a significant (p<0.01) induction in DNA damage was observed by the alkaline single cell gel electrophoresis in digestive gland cells treated with AgNPs for 24 and 96 h. These results demonstrate that silver nanoparticles are lethal to freshwater snail L. luteola. The oxidative stress biomarkers and comet assay can successfully be used as sensitive tools of aquatic pollution biomonitoring.

In vitro antimicrobial activity of flavanoids of Ocimum sanctum with synergistic effect of their combined form

Abstract Objective To evaluate the antibacterial activity of flavanoids, Orientin and Vicenin, obtained from leaves of Ocimum sanctum , have also been compared by their combine sample. Methods Aqueous extract of fresh leaves of Ocimum sanctum was assessed for the isolation and purification of different flavanoids. The antibacterial activity of Orientin, Vicenin and Combine sample of both these flavanoids was evaluated according to well diffusion method against some bacteria causing Urinary Tract Infection (UTI) in human. Results The result indicated that the combined sample or synergistic activity of both individual flavanoids showed positive result against Escherichia coli , Proteus , Staphylococcus aureus , Staphylococcus cohni and Klebsialla pneumonia with zone of inhibition 20.12, 20.75, 20.95, 19.55 and 20.1 mm at concentration of 400 mg/ml respectively. But the individual flavanoids showed the positive result against only limited microorganism. Conclusions The finding of the present study provides the evidence that this flavanoid sample is used as an antibacterial agent. This is also beneficial to use this combine sample of different flavanoids of Ocimum sanctum for medication and other purposes.

Isolation and evaluation of anticancer efficacy of stigmasterol in a mouse model of DMBA-induced skin carcinoma

Stigmasterol (99.9% pure) was isolated from Azadirachta indica and its chemopreventive effect on 7,12-dimethylbenz[a]anthracene (DMBA)-induced skin cancer was investigated in Swiss albino mice. Skin tumors were induced by topical application of DMBA and promoted by croton oil. To assess the chemopreventive potential of stigmasterol, it was orally administered at a concentration of 200 mg/kg and 400 mg/kg three times weekly for 16 weeks. Reduction in tumor size and cumulative number of papillomas were seen as a result of treatment with stigmasterol. The average latency period was significantly increased as compared with the carcinogen-treated control. Stigmasterol induced a significant decrease in the activity of serum enzymes, such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and bilirubin as compared with the control. Stigmasterol significantly increased glutathione, superoxide dismutase, and catalase as compared with the control. Elevated levels of lipid peroxide and DNA damage in the control group were significantly inhibited by administration of stigmasterol. From the present study, it can be inferred that stigmasterol has chemopreventive activity in an experimental model of cancer. This chemopreventive activity may be linked to the oxidative stress of stigmasterol. The antigenotoxic properties of stigmasterol are also likely to contribute to its chemopreventive action.

Extraction Optimization of Tinospora cordifolia and Assessment of the Anticancer Activity of Its Alkaloid Palmatine

Objective. To optimize the conditions for the extraction of alkaloid palmatine from Tinospora cordifolia by using response surface methodology (RSM) and study its anticancerous property against 7,12-dimethylbenz(a)anthracene (DMBA) induced skin carcinogenesis in Swiss albino mice. Methods. The effect of three independent variables, namely, extraction temperature, time, and cycles was investigated by using central composite design. A single topical application of DMBA (100 μg/100 μL of acetone), followed 2 weeks later by repeated application of croton oil (1% in acetone three times a week) for 16 weeks, exhibited 100 percent tumor incidence (Group 2). Results. The highest yield of alkaloid from Tinospora cordifolia could be achieved at 16 hours of extraction time under 40°C with 4 extraction cycles. Alkaloid administration significantly decreases tumor size, number, and the activity of serum enzyme when compared with the control (Group 2). In addition, depleted levels of reduced glutathione (GSH), superoxide dismutase (SOD), and catalase and increased DNA damage were restored in palmatine treated groups. Conclusion. The data of the present study clearly indicate the anticancer potential of palmatine alkaloid in DMBA induced skin cancer model in mice.

Ecotoxicity of single-wall carbon nanotubes to freshwater snail Lymnaea luteola L.: Impacts on oxidative stress and genotoxicity.

Mammalian studies have raised concerns about the toxicity of carbon nanotubes, but there is very limited data on ecogenotoxicity to aquatic organisms. The aim of this study was to determine eco‐geno toxic effects of single walled carbon nanotubes (SWCNTs) in fresh water snail, Lymnea luteola (L. luteola). A static test system was used to expose L. luteola to a freshwater control, 0.05, 0.15, 0.30, 0.46 mg/L SWCNTs for up to 4 days. SWCNTs changed a significant reduction in glutathione, glutathione‐S‐transferase, and glutathione peroxidase with in hepatopancreas of L. luteola. Lipid peroxidation (LPO) and catalase showed dose‐ and time‐dependent and statistically significant increase in hepatopancreas during SWCNTs exposure compared with control. However, a significant (p < 0.01) induction in DNA damage was observed by the comet assay in hepatopancreas cells treated with SWCNTs. These results demonstrate that SWCNTs are ecogenotoxic to freshwater snail L. luteola. The oxidative stress and comet assay can successfully be used as sensitive tools of aquatic pollution biomonitoring.

Susceptibility of the freshwater pulmonate snail Lymnea luteola L. to copper oxide nanoparticle

Copper oxide nanoparticles (CuONP) are among the most widely used engineered nanoparticles and thus likely to permate the environment predominantly in sediments. The present study was designed to examine the adverse effects of CuONP in freshwater snail Lymnea luteola L. (L. luteola) exposed for 5 days. Induction of oxidative stress in digestive gland was evidenced by a decrease in reduced glutathione (GSH) levels, and activities of glutathione peroxidase (GPx) and glutathione-S-transferase (GST) whereas lipid peroxidation levels were increased at CuONP 7 or 21 µg/L. Superoxide dismutase activity was numerically higher at lower concentration of CuONP at 1 day but significantly decreased at 5 days. Catalase activity was reduced at 2 days but elevated at lower concentration of CuONP at 5 days. DNA impairment was noted in L. luteola based upon comet assay findings and expressed in terms of % tail DNA and olive tail moment. Results indicate that interaction of CuONP with snail produces toxicity, which is mediated by oxidative stress.

Regulation of apoptosis through bcl-2/bax proteins expression and DNA damage by nano-sized gadolinium oxide

Gadolinium oxide (Gd2O3) nanoparticles (GNPs) are applied in industrial products, for example, additives, optical glass, and catalysis. There are various suggestions of metal nanoparticles paradigm but the underlying basic mechanism about the toxicity of metal nanoparticles, for example GNPs, remains unclear. This experiment was done to measure the effective toxicity of GNPs (10, 25, 50, and 100 µg/mL) over 24 and 48 h and to evaluate toxicity mechanism in human neuronal (SH-SY5Y) cells. GNPs produced reactive oxygen species (ROS), as evaluated by 2′, 7′-dichlorodihydrofluorescein diacetate. Due to incorporation into cells, GNPs generated ROS in a concentration- and time-dependent manner. To determine the toxicity of GNP mechanism related to ROS, we also found chromosome condensation and dysfunction of mitochondrial membrane potential (MMP) after exposure of GNPs. Furthermore, the increased cell apoptosis rate and DNA fragmentation were closely related to the increased dose and exposure duration of GNPs in SH-SY5Y cells. The reduction in MMP with a simultaneous increase in the expression of bax/bcl2 gene ratio indicated that mitochondria-mediated pathway involved in GNPs induced apoptosis. Thus, our finding has provided valuable insights into the probable mechanism of apoptosis caused by GNPs at in vitro level.

Isolation and evaluation of biological efficacy of quercetol in human hepatic carcinoma cells

Quercetol is a polyphenolic molecule present in vegetables and fruits, and is beneficial to human and animal health. The current work aimed to test cytotoxic and apoptotic effects of quercetol on HepG2 cells. Quercetol was isolated from Ocimum sanctum and characterized by gas chromatography–tandom mass spectrometry (GC-MS/MS), nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. Quercetol (50–600 μg/mL) was examined for cytotoxic activity by tetrazolium salt and neutral red uptake tests and comet assay for genotoxicity, using HepG2 cells, over 24 hours. Data from 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and neutral red uptake tests demonstrated quercetol-induced cytotoxicity in HepG2 cells in a concentration-dependent manner. With 4′,6-diamidino-2-phenylindole staining, a significant induction of chromosomal condensation was observed at 300 μg/mL of quercetol. DNA fragmentation analysis showed that quercetol produced cell death in HepG2 cells in a concentration-dependent manner. Thus, our study suggests that an environmentally relevant concentration of quercetol, which was a chemically standardized extract from O. sanctum, induced cell death and DNA damage in HepG2 cells.

Quercetin attenuates the development of 7,12-dimethyl benz（a）anthracene（DMBA） and croton oil-induced skin cancer in mice

Abstract To evaluate the chemopreventive potential of quercetin in an experimental skin carcinogenesis mouse model. Skin tumor was induced by topical application of 7, 12-dimethyl Benz (a) anthracene (DMBA) and Croton oil in Swiss albino mouse. Quercetin was orally administered at a concentration of 200 mg/kg and 400 mg/kg body weight daily for 16 weeks in mouse to evaluate chemopreventive potential. Skin cancer was assessed by histopathological analysis. We found that quercetin reduced the tumor size and the cumulative number of papillomas. The mean latent period was significantly increased as compared to carcinogen treated controls. Quercetin significantly decreased the serum levels of glutamate oxalate transaminase, glutamate pyruvate transaminase, alkaline phosphatase and bilirubin. It significantly increased the levels of glutathione, superoxide dismutase and catalase. The elevated level of lipid peroxides in the control group was significantly inhibited by quercetin. Futhermore, DNA damage was significantly decreased in quercetin treated mice as compared to DMBA and croton oil treated mice. The results suggest that quercetin exerts chemopreventive effect on DMBA and croton oil induced skin cancer in mice by increasing antioxidant activities.

Detection of oxidative stress and DNA damage in freshwater snail Lymnea leuteola exposed to profenofos

Extensive production and use of organophosphate pesticide in agriculture, has risen concerned about its ecotoxicity and risk assessment of insecticides, which are more important. Therefore, the present investigation was aimed to study the induction of oxidative stress and DNA damage by organophosphate insecticide profenofos (PFF) in freshwater snail Lymnea luteola (L. luteola). The median lethal value (96 h LC50) of PFF was estimated as 1.26 mg/L for L. luteola in a semi-static system and on the basis of LC50 value three concentrations viz., 0.126 (1/10 of LC50, Sublethal I), 0.63 (1/2 of LC50, Sublethal II) and 0.84 mg/L (2/3 of LC50, Sublethal III) were determined. Snails were exposed to above-mentioned concentrations of PFF along with solvent control (acetone) and negative control for 96 h. The haemolymph was collected at 24 and 96 h of after treatment. In heamolymph of PFF exposed snail, lipid peroxide, glutathione reduced glutathione S transferase and superoxide dismutase activities at the tested concentrations significantly differ from those in the control. The genotoxicity induced in hemocytes of treated snails was measured by alkaline single cell gel electrophoresis assay. The data of this experiment demonstrated significantly enhancement of oxidative stress and DNA damage in the treated snails as compared to controls. Also, we observed statistically significant correlations of ROS with DNA damage (% tail DNA) (R2 = 0.9708) for 24 h and DNA damage (R2 = 0.9665) for 96 h.Results of the current experiment can be useful in risk evaluation of PFF among aquatic organisms. The study confirmed the use of comet assay for in vivo laboratory experiments using freshwater snail for selecting the toxic potential of industrial chemicals and environmental contaminants.