Permethrin induced cytotoxicity of rat splenocytes: Protective effect of N-acetylcysteine

ABSTRACT The present study was designed to determine the molecular mechanism associated with permethrin induce cytotoxicity. Rat splenocytes were incubated with increasing concentration of permethrin (0-39 ug/ml) for 6 to 24 h. Cytotoxic effect of permethrin was evaluated by MTT assay. To assess the underneath mechanism of cytotoxicity, different biochemical indices of cell death namely Annexin V binding assay, DNA fragmentation, and levels of caspase 3 were analyzed. To evaluate the oxidative stress, glutathione depletion and melondialdehyde levels were analyzed. MTT assay revealed that permethrin induces cytotoxicity in dose-dependent way. In Annexin-V binding assay, above 7.8 µg/ml concentration, significant necrosis of cells was noticed and consistent with DNA fragmentation assay. A significant dose and time dependent depletion of cellular glutathione (GSH) and increased MDA levels were observed and consistent with the percentage of cells undergoing apoptosis. Co administration of N-acetycysteine mitigates permethrin- induced apoptosis, showing the role of oxidative stress in apoptosis induction. The present study gives experimental evidence that emphasizing the role of oxidative stress in permethrin-induced cytotoxicity in rat splenocytes in vitro

Permethrin induced cytotoxicity of rat splenocytes: Protective effect of N-acetylcysteine

11-18Permethrin is a synthetic insecticide, extensively used in pest control. Exposures to permethrin have been attributed to increased cell death. The mechanism for its toxicity is still not clear. Hence, in the present study we determined the molecular mechanism associated with permethrin induce cytotoxicity. Rat splenocytes were incubated with increasing concentration of permethrin (0-39 ug/Ml) for 6 to 24 h. Cytotoxic effect of permethrin was evaluated by MTT assay. To assess the mechanism of cytotoxicity, different biochemical indices of cell death, namely annexin V binding assay, DNA fragmentation assay, and levels of caspase 3 were analyzed. To evaluate the oxidative stress, glutathione depletion and melondialdehyde levels were analyzed. MTT assay revealed that permethrin induces cytotoxicity in dose-dependent way. In annexin-V binding assay, above 7.8 µg/mL concentration, significant necrosis of cells was noticed and consistent with DNA fragmentation assay. A significant dose and time dependent depletion of cellular glutathione (GSH) and increased MDA levels were observed and consistent with the percentage of cells undergoing apoptosis. Co administration of N-acetycysteine mitigates permethrin- induced apoptosis, showing the role of oxidative stress in apoptosis induction. The present study demonstrated the role of oxidative stress in permethrin-induced cytotoxicity in rat splenocytes in vitro

ASSESSMENT OF CYP2D6*10 POLYMORPHISM WITH POST HERPETIC NEURALGIA PATIENTS UNDERGOING TRAMADOL TREATMENT

objective: To evaluate association of CYP2D6*10 polymorphism with respect to demographic characteristics (age at onset, genders and weight), numerical rating scale (NRS) for measuring pain intensity in relation with resting and movement associated pain and adverse drug effects of PHN patients receiving tramadol therapy. Methods: Total 246 patients of PHN (148 males and 98 females) were selected who fulfilled the inclusion/exclusion criteria. Clinicians were recorded numerical rating scores (at rest and with movement), and note down adverse drug side effects during the time of study. All samples were analyzed for CYP2D6*10 polymorphism using PCR-RFLP method. results: We observed genotype distribution of CYP2D6* 10 did not vary significantly with age at onset [non-responders (p=0.317) and responders (p=0.260)], genders[ non-responders (p=0.317) and responders (p=0.949)], and weight [non-responders (p=0.298) and responders (p=0.279)] and also did not find significant role with respect to resting (p=0.428) and movement associated type of pain (p=0.178). In addition, CYP2D6*10 was not associated with adverse effects such as somnolence (p=0.135), dizziness (p=0.178), local site reactions (p=0.535), headache (p=0.502), hypotension (p=0.567) and nausea and vomiting (p=0.268) of analgesic therapy. Therefore we conclude that, CYP2D6*10 may not be a predictor of treatment outcomes of patients with PHN receiving tramadol

Xeno-Estrogenic Pesticides and the Risk of Related Human Cancers

In recent decades, “environmental xenobiotic-mediated endocrine disruption”, especially by xeno-estrogens, has gained a lot of interest from toxicologists and environmental researchers. These estrogen-mimicking chemicals are known to cause various human disorders. Pesticides are the most heavily used harmful xenobiotic chemicals around the world. The estrogen-mimicking potential of the most widely used organochlorine pesticides is well established. However, their effect is not as clearly understood among the plethora of effects these persistent xenobiotics are known to pose on our physiological system. Estrogens are one of the principal risk modifiers of various disorders, including cancer, not only in women but in men as well. Despite the ban on these xenobiotics in some parts of the world, humans are still at apparent risk of exposure to these harmful chemicals as they are still widely persistent and likely to stay in our environment for a long time owing to their high chemical stability. The present work intends to understand how these harmful chemicals may affect the risk of the development of estrogen-mediated human cancer

A preliminary study of cross-amplified microsatellite loci using molted feathers from a near-threatened Painted Stork (Mycteria leucocephala) population of north India as a DNA source

Abstract Objective In continuation of an earlier study in which we reported the cross-amplification of Wood stork microsatellites on the DNA obtained from molted feathers of Painted stork (Mycteria leucocephala), here we investigated the nature of cross-amplified microsatellites and the effect of non-invasive samples on cross-amplification success. In a limited manner, we also addressed the genetic diversity and differentiation in a north Indian population of the Painted Stork examined over three nesting seasons. Results Among the nine cross-amplified loci, only 5 were polymorphic. Three and 6 loci exhibited low ( 80), respectively. For 36 of 145 samples most of the loci failed to amplify. For genetic diversity, only 3 loci could be used since others exhibited low amplification and linkage disequilibrium. Probability of identity (0.034) was not low enough to develop a confidence that the similar genotypes originate from the same individual. Forty-two unique genotypes were identified. In 3 loci, a low to moderate level of genetic diversity (mean He = 0.435) was reported. Non-significant Fst (0.003, P = 0.230), G’stH (0.005, P = 0.247) and Dest (0.003, P = 0.250) values indicate a lack of structuring in temporally distributed populations of Delhi Zoo. The limitations and uniqueness of this study are discussed

Expression analysis of apolipoproteins AI & AIV in hepatocellular carcinoma: A protein-based hepatocellular carcinoma-associated study

Background & objectives: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer mortality. The objective of this study was to find out the differential expression of apolipoproteins (ApoAI and ApoAIV) in HCC and cases of liver cirrhosis and chronic hepatitis (controls) without HCC and to compare ApoAI and ApoAIV expression with alpha-foetoprotein (AFP), the conventional marker in HCC. Methods: Fifty patients with HCC and 50 controls comprising patients with liver cirrhosis (n=25) and chronic hepatitis (n=25) without HCC were included in this study. Total proteins were precipitated using acetone precipitation method followed by albumin and IgG depletion of precipitated protein using depletion kit. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The expression changes of ApoAI and ApoAIV were confirmed by western blotting using specific primary and secondary polyclonal antibodies followed by densitometric protein semi-quantitative estimation. ApoAI, ApoAIV and AFP were measured in the plasma samples by ELISA method. Results: Semi-quantitative densitometric image analysis of the western blot images and the comparison between HCC patients with those without HCC (control) revealed differential expression of ApoAI and ApoAIV. Levels of ApoAI were significantly higher in patients with HCC compared to controls without HCC (0.279±0.216 vs 0.171±0.091 and 0.199±0.014; P <0.001). Levels of ApoAIV were significantly lower in patients of HCC compared to controls without HCC (0.119±0.061 vs 0.208±0.07 and 0.171±0.16; P <0.01). ELISA assays of apolipoproteins (ApoAI and ApoAIV) revealed similar results of expression of ApoAI and ApoAIV as detected in western blotting densitometric image analysis. Interpretation & conclusions: Increased expression of ApoAI and decreased expression of ApoAIV in HCC patients compared to controls without HCC revealed the abnormalities in HCC. These molecules need to be studied further for their use as potential biomarkers in the future diagnostic tools along with other conventional biomarkers for screening of HCC cases. It needs further analysis in higher number of patient population

Cross-species amplification of microsatellite markers in <i>Mycteria leucocephala</i> Pennant 1769: Molted feathers as successful DNA source

1011-1016DNA from molted feathers is being increasingly used for genetic studies on birds. However, the DNA obtained from such non-invasive sources is often not of enough quantity and quality for isolation of new microsatellite markers. The present study examined the potential of shed feathers of near threatened Painted Stork as a source of its DNA for cross-species amplification of microsatellites. Thirty-one shed feathers of varying conditions (‘good’ and ‘deteriorated’) and sizes (‘large’, ‘<i style="mso-bidi-font-style: normal">intermediate’ and ‘small’) collected in a north Indian population were used to isolate DNA by a standard isopropanol method and 11 microsatellite markers already developed in the Wood Stork were screened for amplification. Nine plucked feathers from two dead Painted Storks were also used to compare the DNA yield and amplification success. The DNA yield of feathers varied significantly in relation to the calamus size and condition. Among molted feathers, ‘<i style="mso-bidi-font-style: normal">good’ and ‘large’ samples provided more DNA than ‘deteriorated’ and ‘small’ ones, respectively. ‘Large’ plucked feathers yielded more DNA than ‘large’ molted feathers. DNA was almost degraded in all the samples and ratio of absorbance at 260/280 nm varied from 1.0 to 1.8, indicating impurity in many samples. Independent of DNA yields, all microsatellites were cross-amplified in all kinds of feathers, with >80% success in different feather categories. It is concluded that the shed feathers can be successfully used to isolate DNA in the Painted Stork and for cross-species amplification of microsatellites

THE IMPACT OF PHARMACOGENETICS ON ADVERSE DRUG REACTIONS TO PREDICT THE EFFICACY OF TRAMADOL MONOTHERAPY FOR THE TREATMENT OF POST HERPETIC NEURALGIA PATIENTS

Objective: To evaluate the potential role of tramadol treatment with respect to CYP2D6 polymorphism in reducing the incidence of adverse drug reactions.Methods: The study comprised 246 patients of PHN receiving tramadol treatment. Adverse drug events during the time of the study were recorded by the physician. All samples were analyzed for CYP2D6 (*2, *4 and *10) polymorphism using PCR-RFLP method.Results: The CYP2D6 polymorphism did not find significantly among onset at age, genders and weight in non-responders and responders. The CYP2D6*4 polymorphism was significantly associated with somnolence (p=0.009), dizziness (p=0.007), local site reactions (p=0.015), headache (p=0. 039), and nausea and vomiting (p=0. 017). The dizziness (p=0. 029) and headache (p=0. 004) were found associated with CYP2D6*2 both the groups. No associations were observed between adverse events compared with CYP2D6*2 and CYP2D6*10 polymorphisms (p&gt;0.05).Conclusion: CYP2D6*4 polymorphism may be as important drug toxicity marker predictors of experiencing adverse drug reactions as PHN patients undergoing tramadol treatment. Â

Microwave radiation induced oxidative stress, cognitive impairment and inflammation in brain of Fischer rats

889-896Public concerns over possible adverse effects of microwave radiation emitted by mobile phones on health are increasing. To evaluate the intensity of oxidative stress, cognitive impairment and inflammation in brain of Fischer rats exposed to microwave radiation, male Fischer-344 rats were exposed to 900 MHz microwave radiation (SAR = 5.953×10-4 W/kg) and 1800 MHz microwave radiation (SAR = 5.835×10-4 W/kg) for 30 days (2 h/day). Significant impairment in cognitive function and induction of oxidative stress in brain tissues of microwave exposed rats were observed in comparison with sham exposed groups. Further, significant increase in level of cytokines (IL-6 and TNF-) was also observed following microwave exposure. Results of the present study indicated that increased oxidative stress due to microwave exposure may contribute to cognitive impairment and inflammation in brain. </span

Xenobiotic-induced Immune Alterations: Implications in Health and Disease

7-15Immune function may be significantly altered following occupational, inadvertent or therapeutic exposure to chemically diverse xenobiotics. The environmental chemicals like pesticides, halogenated hydrocarbons, polychlorinated dibenzofurans, organic solvents, asbestos, silica, heavy metals etc. may interact with both cellular and humoral components of the immune system which can result in altered immune status that in turn may lead to decreased resistance to infection, certain forms of neoplasia or in some cases exacerbate allergy or autoimmunity. Recent advances in pharmacogenomics and toxicogenomics have contributed a lot to delineate the mechanism of interaction of xenobiotics with the biological system at the cellular and molecular level. However, detection of immune changes on exposure to immunotoxic agents is highly complex, especially in humans due to several confounding factors like age, sex, race gender, co- existence of disease, food habits, smoking etc. Thus, establishing a quantitative relationship between immunotoxicological data and risk assessment, following xenobiotic exposure is still a challenge. The present article reviews the immune alterations caused by exposure to variety of xenobiotics, and their implications in health and disease