Novel surface antigen based impedimetric immunosensor for detection of Salmonella typhimurium in water and juice samples.

A specific surface antigen, OmpD has been reported first time as a surface biomarker in the development of selective and sensitive immunosensor for detecting Salmonella typhimurium species. The OmpD surface antigen extraction was done from Salmonella typhimurium serovars, under the optimized growth conditions for its expression. Anti-OmpD antibodies were generated and used as detector probe in immunoassay format on graphene-graphene oxide (G-GO) modified screen printed carbon electrodes. The water samples were spiked with standard Salmonella typhimurium cells, and detection was done by measuring the change in impedimetric response of developed immunosensor to know the concentration of serovar Salmonella typhimurium. The developed immunosensor was able to specifically detect S. typhimurium in spiked water and juice samples with a sensitivity upto 10(1)CFUmL(-1), with high selectivity and very low cross-reactivity with other strains. This is the first report on the detection of Salmonella typhimurum species using a specific biomarker, OmpD. The developed technique could be very useful for the detection of nontyphoidal Salmonellosis and is also important from an epidemiological point of view.

Rat testicular mitochondrial antioxidant defence system and its modulation by aging.

Accumulation of oxidative damage caused by reactive oxygen species (ROS) underlies fundamental changes found during aging. In the present study, age related effect on testicular mitochondrial oxidant generation and antioxidant defence profile was investigated in Wistar rats at 3 months (young adults), 12 months (old adults) and 24 months (senescent animals) of age. Mitochondrial oxidative stress parameters viz., lipid peroxidation (LPx), protein carbonylation, hydrogen peroxide (H2O2) generation and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), levels of total, oxidized (GSSG) and reduced glutathione (GSH) were studied to find out their roles in maintenance of mitochondrial glutathione redox pool as a function of age. Increased levels of LPx, H2O2 and decreased GSH content accompanied by a decline in activities of SOD, GPx and GR with advancing age suggest that antioxidant defense profile of testicular mitochondria exhibit age related alterations which might play a critical role in regulating physiological functions of the testis such as steroidogenesis and spermatogenesis.

Differential expression profiles of antioxidant enzymes and glutathione redox status in hyperthyroid rats: a temporal analysis.

Our objective was to elucidate a temporal profile of expression of antioxidant enzymes (AOEs) and glutathione redox status in rat liver under the influence of thyroid hormone (T3). The key AOEs, superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx-1) and glutathione reductase (GR) were characterized at transcriptional, translational and biochemical levels after 24 h, 72 h and 120 h of treatment. In general, catalase and GPx-1 showed opposite responses in both transcription and translation. T3 treatment caused tightly coordinated downregulation of catalase. However, transcriptional changes of other AOEs over the different durations of treatment were not always reflected in their respective protein and/or activity levels. Discordance among transcripts, proteins and biological activities of AOEs suggested differential regulation by T3 at multiple levels. Reduced and oxidized glutathione were depleted in hyperthyroid rats. Though T3 exerted a positive stimulatory effect on glucose-6-phosphate dehydrogenase, it was not sufficient to compensate for massive glutathione depletion and impaired activities of GPx-1, GR and GST, disturbing the cellular redox status in the process. Apparently, while transcriptional induction of AOEs might be adaptive responses in conditions of oxidative stress, yet post-transcriptional regulation appeared to be the predominant mechanism of regulation of AOE expression.

Production of phytase (myo-inositolhexakisphosphate phosphohydrolase) by Aspergillus niger van Teighem in laboratory-scale fermenter.

The growth and production pattern of phytase by a filamentous fungus, Aspergillus niger van Teighem, were studied in submerged culture at varying agitation rates and controlled and uncontrolled pH conditions. Allowing the initial culture to grow under neutral condition with subsequent decline in pH resulted in increased phytase productivity. A maximum of 141 nkat/mL phytase was obtained when the broth pH was maintained at pH 2.5 as compared to 17 nkat/mL units at controlled pH 5.5. The culture morphology and rheological properties of the fermentation broth significantly varied with the agitation rate. The volumetric oxygen transfer coefficient was determined at different phases of fungal growth during batch fermentation using static gassing out and dynamic gassing out methods. The oxygen transfer coefficient (k(L)a) of the fermenter was found to be 125 h(-)(1) at 500 rpm as compared to 38 h(-)(1) at 200 rpm. The oxygen transfer rates at different phases of growth were significantly affected by cell mass concentration and fungal morphology. During the course of fermentation there was a gradual decline of k(L)a from 97 h(-)(1) on day 2 to 63 h(-)(1) on day 6 of fermentation, after which no significant change was observed. The degree of agitation considerably influenced the culture morphology where shear thinning of filamentous fungus was observed with the increase in agitation.

Purification and characterization of two extracellular alkaline proteases from a newly isolated obligate alkalophilic Bacillus sphaericus.

Two novel extracellular serine proteases were purified to homogeneity from the cell-free culture filtrate of an obligate alkalophilic Bacillus sphaericus by a combination of ultrafiltration, ammonium sulfate precipitation and chromatographic methods. The enzymes showed similar substrate specificities, but differed in hydrophobicity and molecular mass. Protease A was a monomeric protease with a relative molecular mass (M(r)) of 28.7 kDa, whereas protease B, with a M(r) of 68.0 kDa, apparently consisted of smaller subunits. The purified protease A had a specific activity on hemoglobin of 5.1 U/mg protein compared to 40.9 U/mg protein in the case of protease B. Both proteases were most active on SAAPF-pNa, a substrate for chymotrypsin-like serine proteases. However, the K(m) values of these two proteases on SAAPF-pNa were higher than that for alpha-chymotrypsin, indicating a lower affinity of proteases A and B for this substrate compared to chymotrypsin. Unlike other Bacillus serine proteases, neither protease A nor B stained with Coomasie blue R-250, even with loading of a large amount of protein, and they stained poorly with the silver staining method. However, NH(2)-terminal amino acid sequencing of protease B revealed a high similarity with subtilisin Carlsberg (67% homology). Almost total inhibition of both proteases by PMSF, but very little/no inhibition by trypsin and chymotrypsin inhibitors (TPCK and TLCK) or thiol reagents (PCMB and iodoacetic acid), further supported the view that the enzyme belonged to the serine protease family.

Studies on alkaline protease produced by Bacillus sp. NG312.

An alkalophilic hyperproducer of alkaline protease, Bacillus sp. NG312, was isolated, and the enzyme showed maximum activity at pH 11.0 and 60 degrees C. The temperature optimum was increased by 10 degrees C in presence of Ca2+. The crude enzyme was found to have half-life of 11 d at 37 degrees C and maximum stability at pH 9.0-10.0. It also exhibited very good stability in presence of detergent components and some locally available commercial detergent powders.