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Glycogen synthase kinase 3 (GSK3) is a serine/threonine kinase which in the presence of ATP in its ATP-binding pocket transfers a phosphate to a primed substrate. GSK3β is an isoform of GSK3 which has been projected as a potent therapeutic target in human diseases including cancers and metabolic syndrome. Incidentally, cardiovascular disease is a common cause of non-cancer related deaths in prostate cancer (PCa) patients, mainly due to the effects of androgen-deprivation therapy (ADT), a mainstay for PCa treatment. Several small molecular inhibitors of GSK3 are either ATP-competitive (bind to the ATP-binding pocket), or non-ATP-competitive inhibitors (binding to the substrate-binding site of the enzyme). In this study, 2-β-D-glucopyranosyl-1,3,6,7-tetrahydroxy-9H-xanthen-9-one (βDGT), a natural xanthonoid present in many plant species, is reported to bind to the ATP-binding pocket of GSK3β and inhibit its activity, as demonstrated by the molecular docking and molecular dynamics simulation analysis and experimental validation

Identification of new DPP-IV inhibitors by integrating validated in silico approach is being presented herein. Novel hits were identified by combining pharmacophore and structure based virtual screening of ZINC and Knowledge Base in house database followed by ADME profiling, consensus docking studies. Six potential hits were identified and analysed for their synthetic accessibility score, novelty analysis and pan assay interference compounds filtration. Out of six, three hits viz., ZINC25060187, ZINC53746227 and KB-10 were analysed for stability studies using Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) and Molecular Dynamics (MD) simulation. The simulation studies of the identified hits revealed that these hits have good selectivity and stability in DPP-IV binding pocket. Important interactions with amino acids viz., Tyr547, Glu205 and Glu206 similar to co-crystallized ligand were also observed. One of the hits viz., KB-10 was synthesized and evaluated for its biological potential. The compound KB-10 showed good DPP-IV inhibition in both in vitro and in vivo studies with IC50: 22.69 µM. This study supports the fact that these techniques hold potential for efficient screening of compounds with unknown affinity for DPP-IV that could serve as candidates for therapeutic development. Communicated by Ramaswamy H. Sarma

The continuous emergence of resistance to the available drugs poses major constraints in the development of effective therapeutics against malaria. Malaria drug resistance has been attributed to be the manifestation of numerous factors. For example, mutations in the parasite transporter protein acetyl-CoA transporter (Pfact) can remarkably affect its uptake affinity for a drug molecule against malaria, and hence enhance its susceptibility to resistance. To identify major contributors to its loss of functionality, we have thoroughly scrutinized eight such recently reported resistant mutants, via in-silico tools in terms of alterations in different properties. We performed molecular dynamics simulations of the selected Pfact mutants to gain deeper insights into the structural perturbation and dynamicity. Comparison of residue interaction network map of mutants with that of Wild type (WT) protein suggests structural changes as a result of the mutation(s) that translate into the weakening of intra-protein interactions, especially around the drug binding pocket. This, in turn, diminishes the affinity of drug molecules towards the binding site, which was validated by docking analysis. Finally, collating all the observations, we have delineated R108K mutant to deviate the most from WT protein, which, intriguingly suggests that replacing an amino acid with another of similar nature can even translate into greater functional effects as those with dissimilar substitutions.Communicated by Ramaswamy H. Sarma.

The current theoretical perception of enzymatic activity is highly reliant on the determination of the activation energy of the reactions, which is often calculated using computationally demanding quantum mechanical calculations. With the ever-increasing use of bioengineering techniques that produce too many variants of the same enzyme, a fast and accurate way to study the relative efficiency of enzymes is currently in high demand. Here, we propose the local electric field (LEF) of the enzyme along the reaction axis as a descriptor for the enzymatic activity using the example of chorismate mutase in its native form and several variants (R90A, R90G, and R90K/C88S). The study shows a direct correlation between the calculated enzymatic EF and the enzymatic activity for all the complexes. MD simulations of the Michaelis complex and the transition state analog (TSA) show a stabilizing force on the TSA due to the enzymatic EF. QM/MM and QM-only DFT calculations in the presence of an external electric field (EEF) oriented along the reaction axis show that the electric field can interact with the dipole moment of the TS, thereby stabilizing it and thus lowering the activation energy.

The current theoretical perception of enzymatic activity is highly reliant on the determination of activation energy of the reactions which is often calculated using a computational demanding quantum mechanical calculation. With the ever-increasing use of bioengineering techniques that produce too many variants of the same enzyme, a fast and accurate way to study the relative efficiency of enzymes is a need of time. Here, we propose the electric field (EF) of the enzyme along the reaction axis as a descriptor for the enzymatic activity using an example of Chorismate Mutase in its native and several variants (R90A, R90G, and R90K/C88S). The study shows a linear correlation between the calculated enzymatic EF and enzymatic activity of all complexes. The MD simulations of the Michaelis complex and the transition state analog (TSA) show a stabilizing force on TSA due to the enzymatic EF. The QM/MM and QM-only DFT calculations with the presence of External Electric Field (EEF) oriented along the reaction axis show that the electric can increase the dipole moment of the TS, thereby, stabilizing it and thus lowers the activation energy.

Cytochrome P450 peroxygenases use hydrogen peroxide to hydroxylate long-chain fatty acids by bypassing the use of O 2 and a redox partner. Among the peroxygenases, P450 OleT uniquely performs decarboxylation of fatty acids and production of terminal olefins. This root taken by P450 OleT is intriguing, and its importance is augmented by the practical importance of olefin production. As such, this mechanistic choice merits elucidation. To address this puzzle we use hybrid QM/MM calculations and MD simulations for the OleT enzyme as well as for the structurally analogous enzyme, P450 BSβ . The study of P450 OleT reveals that the protonated His85 in the wild-type P450 OleT , plays a crucial role in steering decarboxylation activity by stabilizing the corresponding hydroxoiron(IV) intermediate (Cpd II). In contrast, for P450 BSβ in which Q85 replaces H85, the respective Cpd II species is unstable and it reacts readily with the substrate radical by rebound, producing hydroxylation products. As we show, this single-site difference creates in P450 OleT a local electric field (LEF), which is significantly higher than that in P450 BSβ . In turn, these LEF differences are responsible for the different stabilities of the respective Cpd II/radical intermediates, and hence for different functions of the two enzymes. P450 BSβ uses the common rebound mechanism and leads to hydroxylation, whereas P450 OleT proceeds via decarboxylation and generates terminal olefins. Olefin production projects the power of a single residue to alter the LEF and the enzyme’s function!

This study identifies and validates hexokinase type 4 (HK4), an isozyme of hexokinase in the liver and pancreas, as an important target of C2-β-D-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone (βdGT), a xanthone glucoside suggested to have antidiabetic property. In the study, we applied the computational pipeline of molecular docking followed by the molecular dynamics simulations to shortlist potential βdGT protein targets. The analysis of protein dynamics and the binding free energy (Δ

A series of 1-(1-benzyl-2-methyl-5-((1-phenyl-1H-1,2,3-triazol-4-yl)methoxy)-1H-indol-3-yl)ethanone and ethyl 1-benzyl-2-methyl-5-((1-phenyl-1H-1,2,3-triazol-4-yl)methoxy)-1H-indole-3-carboxylate derivatives were designed based on bioisosteric replacement of previously reported antitubercular agent (IND-07). Twenty ligands were successfully synthesized and some of them were found to have good in vitro activity (MIC 10 μM) against the H

Purpose In vivo phenotyping of CYP isoforms involved in the metabolism of anti-HIV and antitubercular drugs is important to determine therapeutic dose levels in HIV/AIDS-TB coinfections. In this study, we used a cocktail of bupropion, losartan and dapsone for in vivo phenotyping of CYP2B6, CYP2C9 and N-acetyltransferase-2 (NAT2) in plasma. CYP2B6 is the main catalyst of anti-HIV efavirenz, while NAT2 is involved in antitubercular drug isoniazid metabolism. CYP2C9 has a significant association with antitubercular drug-induced reactions. The activity level of these isoforms has a significant bearing on therapeutic dose in rapid and poor metabolizers. Methods Briefly, a cocktail of probe drugs was administered to human volunteers and the drugs and metabolites were determined by an inhouse LC–MS/MS method in 250 μl plasma. The mobile phase and drug/metabolite extraction methods were optimized before analysis. Retention time, Cmax and tmax were calculated from the same sample and the values were used for phenotyping the isoforms. Results Retention time of drugs and metabolites was calculated. The method was sensitive (4.5–8.2 %CV) and no interfering peak was observed in any batch. %Accuracy of the calibrator and QC was 85–115%. %CV of storage stability testing was within FDA approved limits. Cmax and tmax were comparable to the values reported for individual drugs. Conclusions This study advocates the use of a cocktail of bupropion, losartan and dapsone for in vivo phenotyping of CYP2B6, CYP2C9 and NAT2, which is important in determining therapeutic dose levels of anti-HIV and anti-TB drugs in HIV/AIDS-TB coinfections.

Zeaxanthin protects the macula from ocular damage due to light or radiation by scavenging harmful reactive oxygen species. In the present study, zeaxanthin product (OmniXan®; OMX), derived from paprika pods (Capsicum annum; Family-Solanaceae), was tested for its efficacy in the rat retina against photooxidation.Forty-two male 8-week-old Wistar rats exposed to 12L/12D, 16L/8D and 24L/0D hours of intense light conditions were orally administrated either 0 or 100 mg/kg BW of zeaxanthin concentration. Retinal morphology was analyzed by histopathology, and target gene expressions were detected with real-time polymerase chain reaction methods.OMX treatment significantly increased the serum zeaxanthin concentration (p 0.001) and ameliorated oxidative damage by increasing the antioxidant enzyme activities in the retina induced by light (p 0.001). OMX administration significantly upregulated the expression of genes, including Rhodopsin (Rho), Rod arrestin (SAG), Gα Transducin 1 (GNAT-1), neural cell adhesion molecule (NCAM), growth-associated protein 43 (GAP43), nuclear factor-(erythroid-derived 2)-like 2 (Nrf2) and heme oxygenase (HO-1) and decreased the expression of nuclear factor-κB (NF- κB) and GFAP by OMX treatment rats. The histologic findings confirmed the antioxidant and gene expression data.This study suggests that OMX is a potent substance that can be used to protect photoreceptor cell degeneration in the retina exposed to intense light.

Fluoride (F–) pollution in groundwater of the Older Alluvial Plain (OAP) of Delhi has been reported as a major problem. About 34% of the groundwater samples collected for this study had F– level beyond the permissible limit; with F– concentration in the range of 0.14–3.15 mg/L (average 1.20 mg/L). In this context, this article for the first time attempts on the genesis of major ions chemistry and F– pollution in groundwater of OAP Delhi by going beyond the statistical analysis to sediment geochemistry, chemical weathering processes and understanding of the processes using stable environmental isotopes (2H and 18O). The XRD of the OAP sediments revealed the dominance of fluor-biotite, albite, calcite, quartz, and chlorite. Whereas, the separated clay revealed the dominance of chlorite, kaolinite, and illite minerals. The saturation index (SI) values indicated that the groundwater chemistry is in the process of further F– enrichment by way of sediment groundwater interaction. With the given mineralogy of the sediments, the dominance of major ions like Na+, K+, Mg2+, Ca2+, Cl– and F– has been attributed to chemical weathering of biotites, phlogopites, albite, and calcite during sediment–water interaction. While the dominance of SO42– has been attributed to anthropogenic sources and confirmed by its association with heavier stable isotopes of hydrogen (δ2H: −50.44 to −40.02‰) and oxygen (δ18O: −7.19 to −5.62‰) indicating evaporative enrichment during isotopic fractionation.

Substituted 1,2,4-triazole nucleus is common in several drugs used in a variety of clinical conditions including infections, hypoglycemia, hypertension and cancer. In this study, we synthesized 1,2,4-triazole and its 16 hydrazone derivatives (B1-B16), characterized them by IR, NMR and Mass spectroscopy, and evaluated their radical scavenging and anti-inflammatory activities in vitro and in vivo. Out of 16 derivatives, five (B1, B5, B6, B9, and B13) demonstrated a significant radical scavenging and anti-inflammatory activity in vitro. B6, which possessed two electron-donating hydroxyl groups, was most active among all. Molecular docking and MD simulation of the complex of B6 with prostaglandin-endoperoxide synthase (PTGS) or cyclooxygenase (COX) showed that B6 occupied celecoxib binding site in COX with high affinity (the binding free energy of the complex with COX-1 was -10.5, and -11.2 kcal/mol with COX-2). Maximum anti-inflammatory activity was also shown by the B6 derivative in vivo, in the rat model of carrageenan-induced inflammation. B6, along with four other derivatives (B1, B5, B9 and B13) exhibited 80-90% free radical scavenging activity. The IC50 values of these compounds were ≥40 µM. Griess nitrite and dichloro-dihydro-fluorescein-diacetate assays suggested a significant inhibition of nitric oxide and reactive oxygen species, especially by B6 and B9. Taken together, out of 16 derivatives, B6 is reported to have highest anti-inflammatory and antioxidant activity at a low dose level, which may be attributed to its two electron-donating hydroxyls. B6 is proposed to be an important scaffold for the synthesis of new drugs against PTGS for use in a myriad of inflammatory and infectious diseases.Communicated by Ramaswamy H. Sarma.

Tuberculosis is an infectious disease that affects millions of population every year. Mtb-DHFR is a validated target that is vital for nucleic acids biosynthesis and therefore DNA formation and cell replication. This paper report identification and synthesis of novel compounds for selective inhibition of Mtb-DHFR and unleash the selective structural features necessary to inhibit the same. Virtual screening of databases was carried out to identify novel compounds on the basis of difference between the binding pockets of the two proteins. Consensus docking was performed to improve upon the results and best ten hits were selected. Hit 1 was subjected to analogues design and the analogues were docked against Mtb-DHFR. From the docking results 11 compounds were selected for synthesis and biological assay against H37Rv. Most potent compound (IND-07) was tested for selectivity using enzymatic assay against Mtb-DHFR and h-DHFR. The compounds were found to have good inhibitory activity (25-200 µM) against H37Rv and in enzyme assay against Mtb-DHFR and h-DHFR the compound was found selective towards Mtb-DHFR with selectivity index of 6.53. This work helped to identify indole moiety as novel scaffold for development of novel selective Mtb-DHFR inhibitors as antimycobacterial agents.

Groundwater pollution is a serious health concern in north-western India. In this study, we have reported very high concentration of fluoride i.e. 18.5 and 16.6 mg/l from Sainiwas locality in Siwani block of Bhiwani district, Haryana, India. The values are much higher than the permissible limit set by WHO and BIS. The evapotranspiration in the area leads to Ca2+ precipitation, which allows an increase in F- content in the groundwater. In addition, the replacement of hydroxyl of secondary clay mineral under alkaline condition is responsible for release of F-. In absence of alternative source, the fluoride polluted groundwater in some of these localities is also used for drinking. Further, the suitability of groundwater for irrigation is also evaluated by various parameters such as Sodium Adsorption Ratio (SAR), Sodium Percentage (Na%), Kelly's Ratio (KR), Magnesium Hazard (MH) and Residual Sodium Carbonate (RSC). It emerges out that in a few localities, groundwater is not suitable for irrigation and with respect to Magnesium Hazard (MH) almost all samples are unsuitable for irrigation. This article highlights groundwater quality of Siwani block in Haryana and proposes for immediate remedial measures.

Pseudomonas stutzeri TS44 is a moderately halotolerant, arsenite-oxidizing bacterium, containing genes for arsenite oxidation, arsenic resistance, and ectoine/hydroxyectoine biosynthesis. This paper reports in silico studies to understand bioremediation to eliminate toxic metal arsenic in water, air and soil by arsenite oxidase (AO), the bacterial enzyme from P. stutzeri TS44 that can be used for a low cost and eco-friendly removal of arsenite from the environment. To understand the activity of AO in elimination of arsenite, sequence analysis was carried out and homologs, orthologs, domains, family, and conserved residues were identified followed by model generation using various homology modeling tools. The generated models were validated for the best quality protein structure and the best model was used for further optimization using energy minimization approach. Molecular docking studies were performed to study the binding interaction of AO with arsenite. The study predicts and validates the 3D structure of P. stutzeri TS44 arsenite oxidase and reports four active site residues (His197, Glu205, Arg421, and His425) from a close structural homolog of AO from A. faecalis (PDB ID: 1G8K_A). The molecular docking studies suggested the formation of a stable complex and in silico site-directed mutagenesis revealed the importance of Arg421, which resulted in a decrease in stability of the complex when mutated. The study implicates P. stutzeri TS44 arsenite oxidase as a non virulent protein for low cost and eco-friendly bioremediation of arsenite.

Soy consumption in human diet has been linked to decreased incidence of a variety of cancers, suggesting a potential role of soy products in cancer prevention and control. Furthermore, a substantial body of evidence in the literature suggests that soy supplementation may improve the efficacy and prevent the adverse effects of cancer chemotherapy and radiation therapy. Isoflavones constitute the predominant anticancer bioactive compounds in soy. Genistein, which is the most abundant and active isoflavone in soy, has a multitude of effects on cancer cells, including inhibition of NF-κB activation and DNA methylation, enhancement of histone acetylation, inhibition of cell growth and metastasis, and antiangiogenic, anti-inflammatory, and anti-oxidant effects. Isoflavones are orally bioavailable, easily metabolized, and usually considered safe. In this article, we review in vitro and in vivo evidence as well as the results of clinical and epidemiological studies on the effects of soy isoflavones, with a focus on sensitization of cancer cells to chemotherapy and radiation while at the same time protecting normal cells from the harmful effects of these treatments.

The dipeptidyl peptidase-IV (DPP-IV) family of receptors possesses a large binding cavity that imparts promiscuity for number of ligand binding which is not common to other receptors. This feature increases the challenge of using computational methods to identify DPP-IV inhibitors, therefore using both pharmacophore and structure-based screening seems to be a reliable approach. Mining of novel DPP-IV inhibitors by integrating both of these in silico techniques was reported. Pharmacophore model (Model_008) obtained from structurally diverse reported compounds was used as a template for screening of MolMall database followed by structure-based screening against PDB ID: 5T4E. After absorption, distribution, metabolism and excretion (ADME) analysis of shortlisted compounds, consensus docking and molecular mechanics/generalized born surface area studies were carried out. The results of the docking studies obtained were comparable to that of the reference ligand. Out of nine hits identified, only one hit (ID MolMall-20062) was available which was procured through exchange program. Molecular dynamic simulation studies of the procured hit revealed its good selectivity and stability in DPP-IV binding pocket and interactions observed with important amino acids viz., Trp629, Lys544 and Arg125. Biological testing of the compound MolMall-20062 showed promising DPP-IV inhibition activity with IC50: 6.2 µM. Compound MolMall-20062 could be taken as a good lead for the development of DPP-IV inhibitors. AbbreviationsADMEabsorption, distribution, metabolism and excretionChEBIchemical entities of biological interestDPP-IVdipeptidyl peptidase IVDISCOtechdistance comparisonsHTVShigh throughput virtual screeningMDmolecular dynamicsMM-GBSAmolecular mechanics‐generalized born surface areaOGTToral glucose tolerance testPBVSpharmacophore-based virtual screeningPDBprotein data bankRMSDroot mean square deviationROCreceiver operating characteristicsSPstandard precisionSBVSstructure-based virtual screeningVSvirtual screeningXPextra precision absorption, distribution, metabolism and excretion chemical entities of biological interest dipeptidyl peptidase IV distance comparisons high throughput virtual screening molecular dynamics molecular mechanics‐generalized born surface area oral glucose tolerance test pharmacophore-based virtual screening protein data bank root mean square deviation receiver operating characteristics standard precision structure-based virtual screening virtual screening extra precision Communicated by Ramaswamy H. Sarma

Fluoride contamination in water is a major problem across the globe, with health hazards such as dental and skeletal fluorosis. Most earlier studies are confined to local or regional scales. As the problem has serious socioeconomic implications, there is a need for a global perspective. Thus, here we review worldwide research for nearly a century on fluoride contamination in water. We investigated the distribution of fluoride contamination in water, its sources, mobilization and association. The major findings are: (1) Anomalous fluoride concentration in groundwater is mainly confined to arid and semiarid regions of Asia and North Africa. (2) The geogenic sources of fluoride in water are mainly fluorine-bearing minerals in rocks and sediments, whereas anthropogenic sources of fluoride in water are mainly pesticides and industrial waste. (3) Fluoride mobilization from geogenic sources is mainly controlled by alkalinity and temperature. (4) Fluoride occurrence in water is associated with ions such as sodium, arsenic chloride and bicarbonate. There are few associations of fluoride in water with calcium and magnesium.

The present study has been carried out to determine the sources and distribution of fluoride (F−) contamination in the water of district Bijapur, Chhattisgarh, central India. For this study, 45 water samples were collected from surface and groundwater from shallow and deeper aquifers. It was observed that 50% of groundwater samples have elevated F− level and varies from 0.1 to 7.1 mg/L, though, F− level in water bodies was below the WHO maximum permissible limit (1.5 mg/L). The major ions of water samples reveal the dominance of Ca–Mg–HCO3 hydrochemical facies. This study suggests F− contaminated water shows a significant correlation with Cl− which further supported by statistical correlation. Analysis of rocks and sediments from the study area suggests leaching of minerals like biotite, hornblende, and richterite which are likely sources for elevated F− level in groundwater. Gibbs diagram reveals the dominance of the evaporative process for contaminated water by F− in the study area. Due to improper awareness, the local tribal population (≈2, 55,000) of the area is consuming F− enriched groundwater and vulnerable to dental and skeletal fluorosis. Thus, surface and uncontaminated water w.r.t F− should be developed for providing safe water to the inhabitants.

An abnormally high apoptosis has been associated with a number of clinical conditions including embryonal malformations and various pathologies such as neuronal degeneration and diabetes. In this study, boron is reported to inhibit apoptosis in hyperapoptosis conditions as demonstrated in a model of hyperapoptosis. Boron is a metalloid which is present in food in small amounts and is suggested here to inhibit apoptosis by stabilizing the mitochondrial membrane structure, thus preventing matrix remodeling and the release of cytochrome c, an apoptosis-inducer protein from the mitochondrion. The protective effect was assessed by measuring the changes in mitochondrial membrane potential, the levels of cytochrome c and downstream activation of caspase 3, besides phosphatidylserine exposure on the cell surface and DNA damage. The study has implication in clinical conditions characterized by hyperapoptosis as seen in certain embryonal malformations and various pathologies.

The presence of elevated nitrate (NO3-) and nitrite (NO2-) concentration in drinking water higher than the standard limits could endanger the health of consumers. For this data article, concentration of NO3- and NO2- was measured in 118 samples collected from 59 active rural wells in Divandarreh County and the non-carcinogenic risk in the adults and children was estimated by Monte Carlo simulation (MCS). The obtained data showed that the average concentration of NO3- and NO2- was ranges from 31.37 ± 18.87 mg/L and 1.45 ± 0.90 mg/L respectively. Based on acquired data, NO3- concentrations were 37 times higher than NO2- with significant p value of < 0.05. The average concentration of NO3- and NO2- was lower than the national standard with p value < 0.05. However, the concentration of NO3- and NO2- in 23.7% and 13.5% of wells was higher than the national standard of Iran. Total target hazard quotient (TTHQ) in the adults and children was 1.78 and 1.54, respectively. Although, the average concentration of NO3- and NO2- in drinking water was lower than the national standard limits, but the non-carcinogenic risk assessment showed that the children and adults are at a significant risk via nitrate and nitrite in the rural Divandarreh County (TTHQ > 1). Keywords: Nitrate, Nitrite, Groundwater contamination, Risk assessment, Rural Iran, Divandarreh County

Natural coagulant is a habitually developed; plants derived coagulant which could be applied in coagulation- flocculation activity of textile wastewater treatment for reducing the concentration of dyes. In this study the capability of Capparis Spinosa in excluding dyes has been investigated. At a number of dosages of the dye the absorbance amount is noticed applying a coupled ray UV spectrophotometer and a standardization curve has been made. A set of examinations were implemented for verifying the dye removal efficiency using a powder extracted from Capparis Spinosa plant. The effects of various factors such as the acidity function, dye concentration, Capparis Spinosa concentration and the settling time have been investigated. PH implies be a remarkable parameter and dye removal efficiency declines as pH rises and settling time decreases after 20 minutes of settling. The findings illustrate that the dye removal efficiency is achieved highest equal to 96% using Capparis Spinosa as a natural coagulant.

Deregulation of PI3K signalling pathway is strongly involved in pathology of cancer and development of resistance in tumour cells. Here, we report that pharmacologically active vasicinone analogue, RLX (7, 8, 9, 10-Tetrahydroazepino [2, 1-b] quinazolin-12-(6H)-on), exhibited potent anticancer activities both in vitro and in vivo. In this study, RLX treatment displayed strong inhibition of proliferation against various cancer cell lines. However, colon cancer cells were found to be the most sensitive towards RLX mediated inhibition of proliferation. The result showed that RLX treatment followed strong concentration dependent inhibition of HCT-116 cell proliferation and colony formation. RLX treatment to HCT-116 was observed to be associated with down-regulation of p110α and p85 subunits of PI3K thereby decreasing the expression of subsequent downstream effector proteins. Interestingly, silencing of PI3K gene by siRNA in combination with RLX confirmed the anti-proliferation effect of RLX against HCT-116 cells and is mediated by the PI3K pathway. We also found that RLX induced sub-G1 arrest and mitochondrial potential loss followed by pFoxO3a(Thr32) nuclear-cytoplasmic translocation inhibition. Moreover, RLX treatment in in vivo models substantially resulted in a tumour growth inhibition. Overall, our findings reveal the functional role of the PI3K/Akt/FoxO3a pathway that gets deregulated in cancer and suggests its simultaneous targeting by RLX thereby further identifying the compound as a potent inhibitor of the PI3K/Akt/FoxO3a pathway under in vitro and tumour regression in vivo.

Background Dried flower bud of Syzygium aromaticum (clove) is rich in eugenol, an antioxidant and antiinflammatory compound that can protect liver against injury. Clove, besides eugenol, also contains other pharmacologically active phytochemicals such as β-sitosterol and ascorbic acid. This study reports the effect of eugenol-rich fraction (ERF) of clove on liver cirrhosis induced by thioacetamide. Methods Cirrhosis of the liver, which predisposes to hepatocellular carcinoma, was induced by administering thioacetamide (0.03%) in drinking water for 16 weeks. Cirrhotic animals were divided into two groups; the treated group was administered ERF for 9 weeks, one week after discontinuation of thioacetamide, while the other group received normal saline for a similar duration of time. Results The treatment with ERF, as determined by histopathology and through a battery of biochemical markers of hepatic injury, oxidative stress and drug metabolizing enzymes, significantly ameliorated the signs of liver cirrhosis. It lowered the elevated levels of alkaline phosphatase, γ-glutamyl transferase and other biochemical changes in liver cirrhosis. Histopathology of the liver corroborated the effect of ERF with biochemical findings. ERF treatment further inhibited cell proliferation, as demonstrated by reduced [(3)H]-thymidine uptake. Conclusions Data provide evidence supporting the protective action of ERF on liver cirrhosis. The study assumes significance because cirrhosis predisposes the liver to cancer, which is characterized by abnormal cell proliferation. ERF in this study is reported to inhibit hepatic cell proliferation and at the same time decrease oxidative stress, which might be the mechanism of protection against liver cirrhosis.

Zinc oxide nano-particles (ZnO NPs), with their unique physico-chemical properties conferred by various size formulations, are extensively used in consumer products. The enormous usage coupled with their release to the environment demands risk assessment of ZnO NPs on health and the environment. Toxicity of ZnO NPs is well understood in comparison to the bulk ZnO. However, toxicity in relation to the NP size is poorly understood. In this context, we examined the adverse effects of different sizes (35 nm, 50 nm and 100 nm) of ZnO NPs in soil nematode C. elegans along with bulk ZnO and ZnCl2. Here, we show that growth, reproduction and behavior of worms were adversely affected by ZnO NPs in a size dependent manner. Further, exposure to ZnO NPs caused modulation of expression/function of genes associated with Insulin/IGF-like signaling pathway and/or stress response pathway in a size dependent manner in exposed worms. The expression of pro-apoptotic gene and suppression of anti-apoptotic genes, together with increased numbers of cell corpses in the germ line, indicated that apoptosis was also dependent on the size of the ZnO NP. Taken together, our study provides evidence that exposure to ZnO NPs disrupts various physiological processes and causes apoptosis in the germ-line even at very low concentration in a size dependent manner. Our finding suggests the inclusion of size as an additional measure for the cautious monitoring of ZnO NP disposal into the environment.

Oxytocin (OT) injections have been indiscriminately used to milk cattle in dairy industries. There is no study available regarding surveillance of OT in market milk samples.OT from milk samples was extracted by precipitation with trichloroacetic acid and passed through the solid phase extraction column. OT was eluted and evaporated to dryness under a gentle stream of nitrogen. The residue was either dissolved in milli Q water or buffer for analysis through HPLC or EIA. The intake assessment of OT through milk was assessed through the Food Frequency Recall method employing a Food Frequency Questionnaire. On the basis of milk consumption and the values of OT in milk, the actual intake of OT was calculated.In the present study, a total of 55 milk samples (39 milkman and 16 branded) were analyzed for occurrence of OT by EIA and UV-HPLC from different locations of Lucknow, Uttar Pradesh (India). OT contamination in milkman samples was found to be 21 pg/mL to 18.9 ng/mL with the mean value of 8.9 ng/mL. The average daily intake of OT in terms of µg/day/person was highest (2.3-2.4 µg/day/person) in 1-3-year age group.Since there is no prescribed level of OT in milk and the intake of OT through this commodity is quite high there is need to implement regulatory laws so that non-physiological OT exposure may not occur in children which may have deleterious effects.

Introduction: Piperine (PIP) is well known multifunctional antioxidant and anti-inflammatory phytochemical that showed neuroprotection. Pre-treatment of UV-R irradiated piperine (UVR-PIP) 10 mg/kg body weight bw, intravenously showed attenuated neuroprotective effects compared to that of PIP (10µM) in PC12, cortical neuronal culture in-vitro and a rat model of focal cerebral ischemia in-vivo. Method: Neurological parameters were evaluated for UVR-PIP and PIP treated against transient focal cerebral ischemia of SD rats through quantification of infarct volume by TTC staining. In-vitro results deal with reduction of cell viability on UVR-PIP treatment in PC12 and cortical neuronal cell. The result of photodegradation of PIP under UV-R irradiation revealed the formation of photoproducts. Estimation of Mitochondrial ROS, antioxidant enzyme and non-enzyme activities in UVR-PIP and PIP treated brain tissue. Results: Results indicated OGD induced primary cortical neuron cultures and PC12 cells showed that SUN-PIP treatment decrease cell viability and increased LDH secretion compared to that of the PIP pre-treatment. In addition, Bcl-2 expression was decreases after the SUN-PIP treatment. Thus, our results demonstrated that SUN-PIP preconditioning failed to increase levels of Bcl-2 and normalized the endogeneous antioxidant, mediating the neuroprotective effects of PIP. UV-R irradiation attenuated the neuroprotective effects of PIP through modulation of the antioxidant and Bcl-2 mediated pathway. Conclusion: Thus, the ability of UV-R to serve as a modulator of this neuroprotective signaling pathway. Piperine loses some of its neuroprotective ability when irradiated by UV radiation and care has to be taken during storage to avoid exposing piperine to the sun.

It is a well-known fact that the Indian groundwater is polluted by fluoride. However, for the first time in India, non-carcinogenic risk assessments and meta-analysis of fluoride exposure to humans were carried out due to consumption of groundwater. In this context, we collected fluoride concentration data in groundwater across India by systematic searches conducted in various international search engines databases. Here, we demonstrated a detailed meta-analysis and meta-regression of fluoride and evaluated health risk assessment. For this purpose, meta-analysis of 63 studies on fluoride in groundwater in India, comprising 57381 samples are included. We found that 1.) The pooled concentration of fluoride in India is around 2.37 mg/L with 95% confident interval (1.46–3.28 mg/L) which is higher than WHO and national standards limit of 1.5 mg/L. 2) The meta-analysis of data suggests that in rural parts of the country, fluoride concentration is 1.85 times higher than urban areas. 3) The concentration of fluoride in groundwater decreased significantly (p 1). The findings are helpful in identifying the affected areas of India and we recommend that the safer options of drinking water should be adopted.

Okadaic acid (OKA) is a potent inhibitor of protein phosphatases 1/2A (PP2A). Inhibition of PP2A leads to hyperphosphorylation of Tau protein. Hyperphosphorylated Tau protein is present in intraneuronal neurofibrillary tangles a characteristic feature of neuropathology of Alzheimer's disease. Intracerebroventricular (ICV) administration of OKA causes neurotoxicity, which is associated with increased intracellular Ca(2+) level, oxidative stress, and mitochondrial dysfunction in the brain areas. The present study explored Tau phosphorylation in OKA-treated rats in relation to memory function, PP2A activity, intracellular Ca(2+), glycogen synthase kinase-3β (GSK-3β) and N-methyl-d-aspartate (NMDA) receptor after 13days of OKA (200ng, ICV) administration in rats, memory was found impaired in the water maze test. OKA-induced memory-impaired rats showed increased mRNA and protein expression of Tau, Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), Calpain and GSK3β in the hippocampus and cerebral cortex. On the other hand, mRNA expression and activity of PP2A was reduced in these brain areas. OKA treatment also, resulted in decrease in mRNA expression of C and N terminals of Tau. Treatment with NMDA antagonist, MK801 (0.05mg/kg, i.p.) for 13days significantly prevented OKA-induced changes in the expression of PP2A, Tau, GSK3β, CaMKII and Calpain. Further, daily administration of anticholinergic drug, donepezil (5mg/kg, p.o.), and the NMDA receptor antagonist, memantine (10mg/kg, p.o.) initiated after OKA administration for 13days significantly attenuated OKA-induced variation in Tau, Tau-C terminal, Tau-N terminal CaMKII, Calpain, PP2A and GSK3β. These results infer that NMDA antagonist MK801 and memantine are effective against OKA-induced neurotoxicity. Therefore, the present study clearly indicates the involvement of NMDA receptor in OKA (ICV)-induced Tau hyperphosphorylation.

There has been a controversy regarding the use of exogenous oxytocin (OT) in milking cattle which may have toxicological consequences during nonphysiological exposure. In the present study, a new sensitive extraction method for OT was developed followed by enzyme immune assay (EIA) or high-performance liquid chromatography (HPLC) analysis. The extraction of OT in milk involves two steps: (1) TCA precipitation of milk proteins and (2) solid-phase extraction (SPE) cleanup process. Without these steps, analysis of OT in milk was not possible. Utilizing EIA as a quantitative tool the limit of detection (LOD) and limit of quantitation (LOQ) were found to be 7.74 and 10.3 pg ml−1, precision in terms of intra- and interday coefficient of variation was below 13 % (%RSD, N = 8), while percent recoveries were between 85 and 92 %. Utilizing UV-HPLC, the LOD, LOQ, precision, and recovery values were found to be 4.1 ng ml−1, 9.8 ng ml−1, 2–10 %, and 84–91 %, respectively. OT was found to be stable against adverse temperature (up to 100 °C) and pH (2 to 10) and simulated gastric fluid digestibility assay. Four milk samples collected from the market were analyzed, which showed that TCA precipitation and SPE steps are mandatory and the results were validated by LC-MS showing mass ion peak at 1 kD.

Arginine silicate inositol (ASI) complex is a composition of arginine, silicon, and inositol that has been shown to have beneficial effects on vascular health. This study reports the effects of an ASI ointment on wound healing in rats. A full-thickness excision wound was created by using a disposable 5 mm diameter skin punch biopsy tool. In this placebo-controlled study, the treatment group’s wound areas were covered by 4 or 10 % ASI ointments twice a day for 5, 10, or 15 days. The rats were sacrificed either 5, 10, or 15 days after the wounds were created, and biopsy samples were taken for biochemical and histopathological analysis. Granulation tissue appeared significantly faster in the ASI-treated groups than in the control groups (P

Chemical mediators of inflammation (CMI) are important in host defense against infection. The reduced capacity of host to induce the secretion of these mediators following infection is one of the factors in host susceptibility to infection. Boron, which has been suggested for its role in infection, is reported in this study to increase lymphocyte proliferation and the secretion of CMI by the lipopolysaccharide (LPS)-stimulated peritoneal macrophages in BALB/c mice. Boron was administered to mice orally as borax at different doses for 10 consecutive days, followed by the stimulation of animals with ovalbumin and isolation of splenocytes for proliferation assay. The lymphocyte subsets were determined by flow cytometry in spleen cell suspension. The mediators of inflammation, TNF-α, IL-6, IL-1β and nitric oxide (NO), were measured in culture supernatant of LPS-primed macrophages isolated from borax treated mice. TNF and ILs were measured by ELISA. NO was determined by Griess test. The expression of inducible nitric oxide synthase (iNOS) in macrophages was studied by confocal microscopy. Results showed a significant increase in T and B cell populations, as indicated by an increase in CD4 and CD19, but not CD8, cells. Boron further stimulated the secretion of TNF-α, IL-6, IL-1β, NO and the expression of iNOS by the LPS-primed macrophages. The effect was dose dependent and most significant at a dose level of 4.6 mg/kg b. wt. Taken together, the study concludes that boron at physiological concentration induces lymphocyte proliferation and increases the synthesis and secretion of pro-inflammatory mediators by the LPS-primed macrophages, more specifically the M1 macrophages, possibly acting through Toll-like receptor. The study implicates boron as a regulator of the immune and inflammatory reactions and macrophage polarization, thus playing an important role in augmenting host defense against infection, with possible role in cancer and other diseases.

Mitochondrial abnormalities have been identified in a large proportion of neurodegenerative diseases. Recently we have reported that intracerebroventricular (ICV) administration of okadaic acid (OKA) causes memory impairment in rat. However involvement of mitochondrial function in OKA induced memory impairment and neuronal damage has not been determined. OKA (200 ng) was administered by ICV route. After 13th day of OKA administration memory function was evaluated by Morris Water Maze test. Following completion of behavioral studies on 16th day, mitochondrial membrane potential, Ca(2+) and reactive oxygen species were evaluated in mitochondrial preparation of cortex, hippocampus, striatum and cerebellum of rat brain. While ATP, mitochondrial activity, lipid peroxidation and nitrite were investigated in synaptosomal preparation of rat brain areas. The activities and mRNA expression of apoptotic factors, caspase-3 and caspase-9, were studied in rat brain regions. The neuronal damage was also confirmed by histopathological study. OKA treated rats showed memory impairment including increased Ca(2+) and reactive oxygen species and decreased mitochondrial membrane potential, ATP and mitochondrial activity in mitochondrial preparation. There was a significant increase in lipid peroxidation and nitrite in synaptosomal preparations. Preventive treatment daily for 13 days with antidementic drugs, donepezil (5 mg/kg, p.o) and memantine (10 mg/kg, p.o), significantly attenuated OKA induced mitochondrial dysfunction, apoptotic cell death, memory impairment and histological changes. Mitochondrial dysfunction appeared as a key factor in OKA induced memory impairment and apoptotic cell death. This study indicates that clinically used antidementic drugs are effective against OKA induced adverse changes at behavioral, cellular, and histological levels and mitochondrial dysfunction.

The purpose of study was to examine the cytotoxic and anti-cancer properties along with addressing the plausible pathway followed by scorpion venom to reduce cell viability in SH-SY5Y and MCF-7 cells. Following exposure of cells with scorpion venom, cytotoxicity was estimated using MTT and lactate dehydrogenase assays. Apoptotic effects were measured by assessment of mitochondrial membrane potential, reactive nitrogen species, DNA fragmentation, and caspase-3 activity whereas antiproliferative effect was assayed using BrdU incorporation. Our results indicate that scorpion venom causes suppression of proliferation by arresting S-phase and induction of apoptosis through increased nitric oxide production, caspase-3 activity and depolarization of mitochondrial membrane. Induction of apoptosis and arrest of DNA synthesis are critical determinant factors for development of anti cancer drugs. These properties may lead to isolation of effective molecule(s) with potential anticancer activity from scorpion venom of Androctonus crassicauda.

Citrus sinensis is a seasonal fruit. Its zester is rich in bioactive phytochemicals, such as limonene, β-sitosterol, and ascorbic acid, which possess pharmacological action. In this study, we report the effect of fraction prepared from dried peel of C. sinensis on biochemical and histopathological changes in rat model of liver cirrhosis. Liver cirrhosis was induced in rats by administering thioacetamide at a concentration of 0.03% in drinking water for 16 weeks. Thioacetamide was discontinued after 16 weeks and from the 18th week rats were given the extract orally for 9 weeks. Following the completion of the treatment, animals were killed and biochemical and histopathological changes associated with liver cirrhosis were evaluated. The treatment was found to reverse the elevated levels of alkaline phosphatase, γ-glutamyl transferase, and other biochemical markers related to oxidative stress and selected drug metabolizing enzymes. Histopathology of the hepatic tissue confirmed the curative effect of the extract, and corroborated with the biochemical findings. HPTLC fingerprinting of the test fraction confirmed the presence of limonene, β-sitosterol, and ascorbic acid, which may partially explain the effect. The extract was also found to possess the anti-proliferative activity, determined by measuring the incorporation of radioactive thymidine by the hepatic DNA. The study indicates the inhibitory action of the test preparation on collagen accumulation in the extracellular matrix, and hence suggests its use as a potential therapeutic agent in liver fibrosis and cirrhosis.

Transition temperature and thermal stability of proteins were studied in the presence and absence of boron. The observed midpoint of thermal denaturation (T m) of cytochrome c (Cyt c) at pH 9.2 was 68.8 °C, which in the presence of boron increased to 71.0 °C. For metmyoglobin, T m increased from 79.7 °C in the absence of boron to 83.5 °C in the presence of boron. Boron caused an increase of 10% in the reversibility of thermal denaturation of cytochrome c when compared with control. Activity measurements of the heat treated proteins and T m suggest an increased thermal stability toward inactivation and denaturation of heme proteins in the presence of boron.

Shear stress and hydrostatic pressure-induced stretch are known to enhance the production of free radicals, causing oxidative stress in the vascular wall and have been implicated in endothelial inflammation and vascular lesions. Production of antioxidant enzymes such as superoxide dismutase, glutathione peroxidase, and catalase probably constitute a key factor in maintaining the redox state of arterial wall in response to hemodynamic forces. In the present study, redox state of arterial wall at constant pulsatile/laminar shear and varying hydrostatic pressure (70 and 150 cm water) was evaluated. The aim was to study the pressure, in combination with different flow patterns (laminar/pulsatile shear stress), induced oxidative stress which presumably cause arterial wall inflammation leading to vascular legions. The level of reduced glutathione (GSH), lipid peroxidation (LPO) and enzymes known to contribute to the redox status of the cell/tissue, have been measured. Present study quantitatively evaluates the hemodynamic forces induced vascular oxidative stress. We individually assessed the activity of Superoxide Dismutase (SOD), catalase, glutathione peroxidase (GPx), xanthine oxidase (XO), and glutathione-s-transferase (GST) under hemodynamic stress. Secondary antioxidant enzymes like glucose-6-phosphate dehydrogenase (G6PD) and glutathione reductase (GR) were also measured. The increased oxidative stress under pulsatile shear stress and high hydrostatic pressure potentially suggests that vascular inflammation is the key to understand initiation of vascular lesions. Higher concentrations of SOD in arterial wall ought to be considered since this explains arterial wall antioxidant defense in vascular pathologies, which potentially involve oxidative stress. In conclusion, pulsatile shear stress in combination with hydrostatic pressure is a weak inducer of antioxidant defense in blood vessels, hence, considered to be atherogenic. The finding is relevant to both the normal and pathophysiologically relevant hemodynamically stress rabbit thoracic aorta.

Bergenia ligulata Wall., family Saxifragaceae, is an Indian folk medicine used for a variety of pharmacological effects. In this study, evidence is provided in animal model to demonstrate the role of aqueous as well as 50% ethanolic extract of B. ligulata in inflammation and as antibacterial agent. Oral administration of the extract at a dose level of 1 gm/kg bw showed anti-inflammatory and free radical scavenging activity as evaluated using pharmacological and biochemical parameters. The effect was studied on biochemical parameters reportedly perturbed in inflammation. While the extract treatment could alleviate the level of succinate dehydrogenase and xanthine oxidase, which increase in inflammation, the level of superoxide dismutase increased following the treatment with the extract as well as the diclofenac. Role of oxygen free radicals/peroxides was evaluated by measuring lipid peroxidation and glutathione. Treatment with the extract could significantly decrease the enhanced level of lipid peroxidation in inflammation, and increased the level of glutathione. Further, the antibacterial activity of various fractions was tested in vitro using cultures of Escherichia coli, Baccillus subtilis, and S. aureus, and the fractions were found to be antibacterial. The antifungal activity was also tested using the culture of Saccharomyces. However, the drug was ineffective in inhibiting fungal growth. Results provide evidence suggesting the anti-inflammatory as well as the antibacterial role of B. ligulata, thus implicating the plant extract in treatment against the bacterial infection and inflammation.

Here in, we demonstrate a rapid method for covalent immobilization of enzymes onto activated polystyrene microtiter plate by application of pressure. Thus, when HRP, taken in an activated well of a polystyrene microtiter plate, is subjected to an optimum pressure of 2.0 × 10 5 Pa for an optimum time of 25 min in a closed chamber, it shows more than 5-fold increase in absorbance value compared to control experiment carried out without applying pressure. Pressure-induced immobilized HRP shows better thermal and storage stability with respect to free enzyme. Reusability study shows that immobilized enzyme retained almost full activity after three successive uses; however activity falls to half after ten cycles of repeated use. The Michaelis–Menten constant ( K m) and maximum reaction velocity ( V max) for pressure-induced immobilized HRP are found to be 55.3 μM and 0.957 mM/min, respectively, whereas for free enzyme the corresponding values are 34.7 μM and 1.089 mM/min. The method is found to be equally effective for immobilization of other enzymes including glucose oxidase, alkaline phosphatase and invertase. As no thermal incubation is required, pressure-induced immobilization could be used for immobilization of heat sensitive biomolecules useful for preparation of biosensors, biochips and protein-microarrays.

Acute hepatic failure is a severe complication induced by certain chemicals, drugs, or virus. Thioacetamide generally has been used for the study of hepatic failure in experimental animal model. The present study was aimed at to examine the role of boron in the pathogenesis of acute hepatic failure in rats. A single intraperitoneal injection of thioacetamide produced severe liver injury, as manifested by elevation in serum aminotransferases, alkaline phosphatase, and hepatic lipid peroxidation. Boron, when administered in the form of boric acid for three consecutive days followed by thioace-tamide, attenuated thioacetamide-mediated changes in the level of these biochemical parameters in a dose-dependent manner. The effect of boron supplementation on the survival rates of rats treated with a lethal dose of thioacetamide was also determined and found to lower the mortality rates in the group of animals supplemented with boron followed by thioacetmide. The effects on biochemical parameters and the survival rates were dependent on the dose of boron administered as boric acid. It is concluded that boron provides protection against the thioacetamide-induced acute hepatic failure in rats in a dose-dependent manner. J. Trace Elem. Exp. Med. 15: 47–55, 2002. © 2002 Wiley-Liss, Inc.

Hepatocarcinogenesis is one of the most prevalent and lethal cancers. We studied the mechanisms underlying the inhibition of diethylnitrosamine (DEN)-induced hepatocarcinogenesis by lycopene in rats. Hepatocarcinogenesis was induced by an intraperitoneal injection of DEN followed by promotion with phenobarbital for 24 successive wk. The rats were given lycopene (20 mg/kg body weight) 3 times a week orally for 4 wk prior to initiation, and the treatment was continued for 24 consecutive wk. Lycopene reduced incidence, number, size, and volume of hepatic nodules. Serum alanine transaminase, aspartate aminotransferase, total bilirubin, and malondialdehyde (MDA) considerably increased and hepatic antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase) and glutathione decreased in DEN-treated rats when compared with the control group. Lycopene significantly reversed these biochemical changes and increased the expression of NF-E-2-related factor-2)/heme oxygenase-1, and it decreased NF-κB/cyclooxygenase-2, inhibiting the inflammatory cascade and activating antioxidant signaling (P0.05). Lycopene also decreased DEN-induced increases in phosphorylated mammalian target of rapamycin (p-mTOR), phosphorylated p70 ribosomal protein S6 kinase 1, phosphorylated 4E-binding protein 1, and protein kinase B (P0.05). Lycopene is an active chemopreventive agent that offers protection against DEN-induced hepatocarcinogenesis by inhibiting NF-κB and mTOR pathways.

Many epidemic outbreaks of Chikungunya fever (CHIKF) have been reported throughout the world including India after its reemergence in 2005. The immuno protective role of envelope proteins during Chikungunya virus (CHIKV) infection has been reported. With the aim of identifying the immunodominant epitopes within the envelope protein we investigated the detailed analysis of fine specificity of antibody response in different individuals during CHIKV infection.The peptides corresponding to the full length of E1, E2 and E3 proteins of S27 strain of CHIKV were synthesized and their seroreactivity with CHIKV positive patients’ sera collected from different epidemic regions of India was determined using indirect ELISA.The data analysis reveals many potent epitopes throughout the length of envelope E2 protein thus displaying it as the most promising antigen for diagnostic purpose. We found that the main IgG isotype response to envelope protein was predominantly of subclass IgG3. Interestingly, most of the epitopes were found to be conserved for detecting IgM, IgG and IgG3 antibody response.Peptides E2P3, E2P7, E2P16 and E2P17 were revealed as the most immunodominant peptides that together can form the basis for designing an accurate, economical and easy to synthesize a peptide-based immunodiagnostic for CHIKV. This study provides new and important insight into the humoral response generated by CHIKV S27 strain during the early phase of infection.

Nardostachys jatamansi is a medically important herb of Indian origin used for centuries in Ayurvedic and Unani systems of medicine for the treatment of various ailments. In the present paper, a 50% ethanolic extract of the rhizomes of N. jatamansi is shown to possess hepatoprotective activity. Pretreatment of rats with the extract (800 mg/kg body wt, orally) for three consecutive days significantly ameliorated the liver damage in rats exposed to the hepatotoxic compound thioacetamide. Elevated levels of serum transaminases (aminotransferases) and alkaline phosphatase, observed in thioacetamide alone treated group of animals, were significantly lowered in N. jatamansi pretreated rats. Pretreatment of the animals with the extract also resulted in an increase in survival in rats intoxicated with LD90 dose of the hepatotoxic drug.

The present study was undertaken to investigate short-term (21 days) effects of oral administration of Azadirachta indica leaf extract and vanadate, separately and in combination, on the activities of antioxidant enzymes in streptozotocin-induced diabetic rats. Vanadate is a remarkable antidiabetic agent and shows insulin mimetic effect. However, severe toxicity is associated with vanadate when used in high concentration while at lower concentration the hypoglycemic property of vanadate is reduced. So, we used a low dose of vanadate in combination with A. indica leaf extract and evaluated their effect on the antioxidant defense system. Streptozotocin-diabetic rats were treated separately with insulin, vanadate (0.6 mg/ml), A. indica, and with combined dose of vanadate (0.2 mg/ml) and A. indica. At the end of the experiment, rats were sacrificed and serum glucose levels and activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were determined in cytosolic fraction of liver and kidney. Diabetic rats showed hyperglycemic condition and alteration in antioxidant enzyme activities. Treatment with antidiabetic compounds resulted in the reduction of glucose levels and restoration of enzyme activities to normal. Results showed that combined treatment of vanadate and A. indica leaf extract was the most effective in normalizing altered antioxidant enzyme system.

The redox cycling contact herbicide paraquat (PQ) causes oxidative damage to pulmonary tissue. PQ is reduced enzymatically to PQ radical in lung where it reacts with molecular oxygen, generating reactive oxygen species (ROS). ROS damage various macromolecules including DNA. However, the ability of paraquat to mediate DNA damage is unknown. In this study, Bam H1 site (5'-GGATCC-3') on pBR322 DNA was chosen as the target sequence for a study of the PQ-mediated DNA damage. The incubation of PQ with plasmid DNA in the presence of freshly prepared rat lung microsomes and NADPH resulted in damage to the restriction site. The PQ-treated DNA was not digested with the endonuclease reflected by the digestion pattern of DNA on agarose gels. The effect was dependent on the dose of PQ. The PQ-mediated damage to DNA was comparable to DNA damage caused by ROS generated through the xanthine-xanthine oxidase system. The results of the present study suggest that ROS generated by PQ in vitro under aerobic conditions may lead to a modification of the restriction site on DNA.