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      • SCISCIESCOPUS

        Monocrotophos Induces the Expression of Xenobiotic Metabolizing Cytochrome P450s (CYP2C8 and CYP3A4) and Neurotoxicity in Human Brain Cells

        Tripathi, V. K.,Kumar, V.,Pandey, A.,Vatsa, P.,Dhasmana, A.,Singh, R. P.,Appikonda, S. H.,Hwang, I.,Lohani, M. Humana Press 2017 Molecular Neurobiology Vol. No.

        <P>Expression of various cytochrome P450s (CYPs) in mammalian brain cells is well documented. However, such studies are hampered in neural/glial cells of human origin due to nonavailability of human brain cells. To address this issue, we investigated the expression and inducibility of CYP2C8 and CYP3A4 and their responsiveness against cyclophosphamide (CPA) and organophosphorus pesticide monocrotophos (MCP), a known developmental neurotoxicant in human neural (SH-SY5Y) and glial (U373-MG) cell lines. CPA induced significant expression of CYP2C8 and CYP3A4 in both types of cells in a time-dependent manner. Neural cell line exhibited relatively higher constitutive and inducible expression of CYPs than the glial cell line. MCP exposure alone could not induce the significant expression of CYPs, whereas the cells preexposed to CPA showed a significant response to MCP. Similar to the case of CPA induced expressions, neural cells were found to be more vulnerable than glial cells. Our data indicate differential expressions of CYPs in cultured human neural and glial cell lines. The findings were synchronized with protein ligand docking studies, which showed a significant modulatory capacity of MCP by strong interaction with CYP regulators-CAR and PXR. Similarly, the known CYP inducer CPA has also shown significant high docking scores with the two studied CYP regulators. We also observed a significant induction in reactive oxygen species (ROS), lipid peroxides (LPO), micronucleus (MN), chromosomal aberration (CA), and reduction in reduced glutathione (GSH) and catalase following the exposure of MCP. Moreover, the expressions of apoptotic markers such as caspase-3, caspase-9, Bax, and p53 were significantly upregulated, whereas the levels of antiapoptotic marker, Bcl2, was downregulated after the exposure of MCP in both cell lines. These findings confirm the involvement of ROS-mediated oxidative stress, which subsequently triggers apoptosis pathways in both human neural (SH-SY5Y) and glial (U373-MG) cell lines following the exposure of MCP.</P>

      • Effect of Extracellular Tyrosinase on the Expression Level of P450, Fpr, and Fdx and Ortho-hydroxylation of Daidzein in Streptomyces avermitilis

        Pandey, B. P.,Lee, N.,Kim, B. G. Springer Science + Business Media 2018 Applied biochemistry and biotechnology Vol.184 No.3

        <P>We have reported that the expression of CYP105D7 in Streptomyces avermitilis produces 112.5 mg L-1 of 7,3',4'-trihydroxyisoflavone (3'ODI) in 15 h of the reaction time, when 7,4'-dihydroxyisoflavone (daidzein) is used as a substrate. Although production is significant, rapid degradation of 3'ODI after 15 h was observed in a whole-cell biotransformation system, suggesting the further modification of 3'ODI by endogenous enzymes. In this present study, the effect of deletion of extracellular tyrosinase (melC2) in S. avermitilis for 3'ODI production as well as the expressions of CYP105D7, ferredoxin (Fdx), and ferredoxin reductase (Fpr) were investigated. The result revealed that daidzein hydroxylation activity in the a dagger melC2 mutant decreased by 40% compared with wild-type S. avermitilis. Further, melC2 deletion significantly affects the messenger RNA (mRNA) expression profile of CYP105D7 and its electron transfer counterparts. Real-time PCR analysis of 9 Fdx, 6 Fpr, and CYP105D7 revealed a significant decrease in mRNA expression level compared to wild-type S. avermitilis. The result clearly shows that the decrease in daidzein hydroxylation activity is due to the lower expression level of CYP105D7 and its electron transfer counterpart in the a dagger melC2 mutant. Furthermore, melC2 deletion prevents the degradation of 3'ODI.</P>

      • SCISCIESCOPUS

        Regioselective hydroxylation of isoflavones by Streptomyces avermitilis MA-4680

        Roh, C.,Seo, S.H.,Choi, K.Y.,Cha, M.,Pandey, B.P.,Kim, J.H.,Park, J.S.,Kim, D.H.,Chang, I.S.,Kim, B.G. Society for Bioscience and Bioengineering, Japan ; 2009 Journal of bioscience and bioengineering Vol.108 No.1

        Screening of bacterial whole cells was performed for regioselective hydroxylation of daidzein and genistein. Among the strains examined, Streptomyces avermitilis MA-4680 showed high ortho-dihydroxylation activity to produce 3',4',7-trihydroxyisoflavone and 3',4',5,7-tetrahydroxyisoflavone from daidzein (4',7-dihydroxyisoflavone) and genistein (4',5,7-trihydroxyisoflavone), respectively. Using 100 mg cells (wet wt.) and 1% (v/v) Triton X100 in 1 ml of total reaction volume, where 100 μl of the substrate solution (0.5 mM in 10% (v/v) mixed solvent of DMSO:MeOH=3:7) was added to 900 μl of potassium phosphate buffer (100 mM, pH 7.2), a 16% molar conversion yield of 3',4',7-trihydroxyisoflavone was obtained from 0.5 mM daidzein after 24 h of reaction time at 28 <SUP>o</SUP>C and 200 rpm. Ketoconazole significantly (ca. 90%) inhibited the ortho-hydroxylation activity of daidzein, suggesting that cytochrome P450 enzymes putatively play roles in regiospecific daidzein hydroxylation. The analysis of the reaction products was determined by gas chromatography/mass spectrometry (GC/MS) and <SUP>1</SUP>H NMR.

      • SCIESCOPUSKCI등재

        Cytokines Expression and Nitric Oxide Production under Induced Infection to Salmonella Typhimurium in Chicken Lines Divergently Selected for Cutaneous Hypersensitivity

        Singh, Rani,Jain, Preeti,Pandey, N.K.,Saxena, V.K.,Saxena, M.,Singh, K.B.,Ahmed, K.A.,Singh, R.P. Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.7

        In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI) was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs) induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h). The differential mRNA expression patterns of IFN-${\gamma}$, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO) was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI) as compared to the low cell-mediated line (LCMI) upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-${\gamma}$ were observed in high response birds (HCMI); but IL-2 was down regulated in this line compared to the low response birds (LCMI). Significantly (p<0.05) higher expression of iNOS, IFN-${\gamma}$ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.

      • KCI등재

        Characterization of Microstructure of HAZs in As-Welded and Service Condition of P91 Pipe Weldments

        C. Pandey,A. Giri,M. M. Mahapatra,P. Kumar 대한금속·재료학회 2017 METALS AND MATERIALS International Vol.23 No.1

        Steels 9-12% Cr, having the high creep rupture strength are advocated for the modern low polluting thermal power plants. In the present investigation, the P91 pipe weldments have been characterized for microstructural responses in as-welded, post-weld heat treatment (PWHT) and ageing conditions. The PWHT of welded samples were carried out at 760 °C for time of 2 h and ageing at 760 °C for 720 h and 1440 h, respectively. The effect of time has been studied on precipitates size, distribution of precipitates and grain sizes present in various zones of P91 steel weldments. The impact toughness and hardness variation of heat affected zone (HAZ) have also been studied in as-welded condition as well as at different heat treatment condition. A significant change was observed in grain size and precipitates size after each heat treatment condition. The maximum impact toughness of HAZ was obtained after PWHT at 760 °C for 2 h. The main phase observed in weld fusion zone in as-welded, PWHT and ageing conditions were M23C6, MX, M7C3, Fe-rich M3C and M2C. The unwanted Z-phase (NbCrN) was also noticed in weld fusion zone after ageing of 1440 h.

      • SCIESCOPUSKCI등재

        Effect of Dietary L-ascorbic Acid (L-AA) on Production Performance, Egg Quality Traits and Fertility in Japanese Quail (Coturnix japonica) at Low Ambient Temperature

        Shit, N.,Singh, R.P.,Sastry, K.V.H.,Agarwal, R.,Singh, R.,Pandey, N.K.,Mohan, J. Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.7

        Environmental stress boosts the levels of stress hormones and accelerates energy expenditure which subsequently imbalance the body's homeostasis. L-ascorbic acid (L-AA) has been recognized to mitigate the negative impact of environmental stress on production performances in birds. The present investigation was carried out to elucidate the effect of different dietary levels of L-AA on production performance, egg quality traits and fertility in Japanese quail at low ambient temperature. Sixty matured females (15 wks) were equally divided into three groups (20/group) based on the different dietary levels of L-AA (0, 250 and 500 ppm) and coupled with an equal number of males (1:1) obtained from the same hatch. They were managed in uniform husbandry conditions without restriction of feed and water at 14 h photo-schedule. Except for feed efficiency, body weight change, feed consumption and hen-day egg production were recorded highest in 500 ppm L-AA supplemented groups. Among the all egg quality traits studied, only specific gravity, shell weight and thickness differed significantly (p<0.05) in the present study. Fertility was improved significantly ($p{\leq}0.01$) to a dose dependent manner of L-AA. The findings of the present study concluded that dietary L-AA can be a caring management practice at least in part to alleviate the adverse effect of cold induced stress on production performance in Japanese quail.

      • SCIESCOPUSKCI등재

        Ram Mating Behaviour under Different Social Conditions

        Patel, M.,Das, N.,Pandey, H.N.,Yadav, M.C.,Girish, P.S. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.1

        The present study was conducted to investigate the attributes of ram sexual behaviour under different social conditions. Six intact rams and eight overiectomized ewes were used in this experiment. Ewes were artificially brought into oestrus. All mating traits were recorded every 10 sec during a 1 h mating session under five social conditions viz., ram exposed to single oestrus ewe, ram exposed to single oestrus ewe and audience ram, ram exposed to multiple oestrus ewes, ram exposed to single oestrus ewe with competitor ram and ram exposed to multiple oestrus ewes with competitor ram. Mean intensity of the mating activities during the 1 h mating session under five social conditions was compared. Either vocalization or leg kicking was found to be the predominant teasing activity under all five conditions. Sniffing differed significantly (p<0.05) between different social conditions. Vocalization, leg kicking, and leg kicking with vocalization did not differ significantly (p<0.05) between different social conditions. Mounts/h in the multiple oestrus ewe condition was higher and differed significantly (p<0.05) from other social conditions. Ejaculations/h for the experimental ram was higher when exposed to multiple oestrus ewes in comparison to other social conditions. Mounts/ejaculation was low in the audience ram condition in comparison to the other four social conditions.

      • SCIESCOPUSKCI등재

        Oxidative Stress and Antioxidant Status during Transition Period in Dairy Cows

        Sharma, N.,Singh, N.K.,Singh, O.P.,Pandey, V.,Verma, P.K. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.4

        The study was conducted on 20 Holstein X Sahiwal cross bred dairy cows, with an average milk production of $2,752{\pm}113.79$ liters in $284{\pm}5.75$ days during a single lactation, that were divided in to two groups of 10 animals. We investigated the oxidative stress and antioxidant status during the transition period in dairy cows. In this study, plasma level of MDA was considered as an indicator of lipid peroxidation and SOD, catalase, GSH and GSHPx as antioxidants. The lipid peroxidation was significantly (p<0.001) higher in cows during early lactation as compared to the cows in advanced pregnancy. A significant positive correlation (r = +0.831, p<0.01) was determined between MDA and catalase in early lactating cows. In early lactating cows, blood glutathione was significantly lower than in advanced pregnant cows. However, early lactating cows showed non-significant negative correlation for all antioxidant enzymes with lipid peroxidation. In conclusion, dairy cows seemed to have more oxidative stress and low antioxidant defense during early lactation or just after parturition than advanced pregnant cows, and this appears to be the reason for their increased susceptibility to production diseases (e.g. mastitis, metritis, retention of fetal membranes etc.) and other health problems.

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