Sestrin2 regulates monocyte activation through AMPK-mTOR nexus under high-glucose and dyslipidemic conditions

The vicious cycle between hyperinsulinemia and insulin resistance results in the progression of atherosclerosis in the vessel wall. The complex interaction between hyperglycemia and lipoprotein abnormalities promotes the development of atherogenesis. In the early phase of atherosclerosis, macrophage-derived foam cells play an important role in vascular remodeling. Mechanistic target of rapamycin (mTOR) signaling pathway has been identified to play an essential role in the initiation, progression, and complication of atherosclerosis. Recently sestrin2, an antioxidant, was shown to modulate TOR activity and thereby regulating glucose and lipid metabolism. But the role of sestrin2 in monocyte activation is still not clearly understood. Hence, this study is focussed on investigating the role of sestrin2 in monocyte activation under hyperglycemic and dyslipidemic conditions. High-glucose and oxidized low-density lipoprotein (LDL) treatments mediated proinflammatory cytokine production (M1) with a concomitant decrease in the anti-inflammatory cytokine (M2) levels in human monocytic THP1 cells. Both glucose and oxidized LDL (OxLDL) in a dose and time-dependent manner increased the mTOR activation with a marked reduction in the levels of pAMPK and sestrin2 expression. Both high-glucose and OxLDL treatment increased foam cell formation and adhesion of THP1 cells to endothelial cells. Experiments employing activator or inhibitor of adenosine monophosphate kinase (AMPK) as well as overexpression or silencing of sestrin2 indicated that high-glucose mediated monocyte polarization and adhesion of monocytes to the endothelial cells were appeared to be programmed via sestrin2-AMPK-mTOR nexus. Our results evidently suggest that sestrin2 plays a major role in regulating monocyte activation via the AMPK–mTOR-pathway under diabetic and dyslipidemic conditions and also AMPK regulates sestrin2 in a feedback mechanism.     

Gaseous ozone: A potent pest management strategy to control Callosobruchus maculatus (Coleoptera: Bruchidae) infesting green gram

Extending the storage life of legumes by protecting it from the Callosobruchus maculatus infestation is a major concern for the producers, processors and exporters. Legume processing industry requires “greener” alternatives to the conventional fumigants. Gaseous ozone has a great potential as an insect management strategy that is suited for this niche. Nevertheless, the efficacy of ozone against C. maculatus is yet unknown. A laboratory study was conducted to test the insecticidal effect of ozone in controlling the infestation of C. maculatus in green gram. We have determined the concentration of ozone exposure time–mortality relationship for all the stages of C. maculatus that were exposed to 500–1,500 ppmv ozone. The percentages of mortality for different stages of C. maculatus increased with the increase in ozone concentration and exposure time. It was documented that adult stage is least tolerant to ozone (500 ppmv for 274.40 min exposure required to kill 90%), whereas the most tolerant stage is pupa (500 ppmv for 1816.54 min is required to kill 90%). The results indicate that gaseous ozone is the attractive alternative to the synthetic fumigants.     

Changes in nuclear proteins of astrocytes as a result of acute ammonia or ethanol exposure

We have used an in vitro system to monitor the effects of high levels of ammonia and ethanol on glial cells. Nuclei were isolated and the protein profiles examined by two-dimensional gel electrophoresis. Acute exposure of rat astrocyte cell cultures to ammonia or ethanol resulted in changes in cellular morphology and the level of some nuclear proteins. Glial fibrillary acidic protein (GFAP) levels remained constant under both treatments. Several nuclear proteins were increased specifically. Only one protein was visually detected which was unique to treatment with ammonia or ethanol. This protein (p2a) appeared only in the presence of ammonia. There were no changes in previously observed astrocyte-associated proteins (Silverman et al. Neurochem Int. 12, 513–518, 1988). Two proteins appeared de novo upon either treatment with either ammonia or ethanol. These latter proteins had a molecular weight and pI profile similar to the major class of nuclear stress proteins (hsp70). However, results from immunoblot experiments clearly demonstrated that hsp70 was not induced in astroycte cultures following exposure to ammonia or ethanol.     

Oxidative stress and the development of diabetic complications - antioxidants and lipid peroxidation in erythrocytes and cell membrane.

Erythrocytes isolated from 131 cases of Non-Insulin Dependent Diabetes Mellitus (NIDDM) were studied for lipid peroxidation, antioxidant defences, and the maximum peroxidisable substrate in the cell membrane. Antioxidant defences are lowered in NIDDM, followed by significant rise in lipid peroxidation products. However, in the erythrocyte membrane, the total polyunsaturated peroxidisable lipids are lower than in normal erythrocytes which may be a causative factor affecting the survival of the cells.     

Polymerase chain reaction and its applications: special emphasis on its role in embryo sexing

The polymerase chain reaction (PCR) has developed into one of the most promising methods for in vitro enzymatic amplification of DNA and has found widespread application in DNA cloning, sequencing and mutagenesis related studies. This innovative technique can selectively amplify a single target DNA molecule a billion-fold in a span of a few hours. Amplification of specific DNA sequences by PCR is useful in identification of sex, novel genes, pathogens and diseases. PCR has facilitated the establishment of evolutionary relationships among species and in revealing structural intricacies of single cells. In this article we review some of the major advances and applications of PCR, especially, its role in embryo sexing.     

Incidence of lab-confirmed dengue fever in a pediatric cohort in Delhi,India

BackgroundOur aim was to estimate the overall and age-specific incidence of lab-confirmed dengue fever using ELISA based assays among children 6 months to 15 years in Delhi.MethodsWe enrolled a cohort of 984 children aged 6 months to <14 years in South Delhi and followed-up weekly for fever for 24 months or till 15 completed years of child-age. Households of the enrolled children were geo-tagged. NS1, IgM and IgG assays were conducted using ELISA method to confirm dengue fever in children with ≥3 consecutive days of fever. Molecular typing was done in a subset of NS1 positive cases to identify the circulating serotypes.Principal findingsWe had a total of 1953 person-years (PY) of follow up. Overall, there were 4208 episodes of fever with peaks during June to November. The overall incidence (95%CI) of fever was 215/100 PY (209 to 222). A total of 74/1250 3-day fever episodes were positive for acute dengue fever (NS1 and/or IgM positive). The overall incidence (95%CI) of acute dengue fever was 37.9 (29.8 to 47.6) per 1000 PY; highest among children aged 5 to 10 years (50.4 per 1000 PY, 95% CI 36.5 to 67.8). Spatial autocorrelation analysis suggested a clustering pattern for the dengue fever cases (Moran’s Index 0.35, z-score 1.8, p = 0.06). Dengue PCR was positive in 16 of the 24 specimens tested; DEN 3 was the predominant serotype identified in 15/24 specimens.ConclusionsWe found a high incidence of dengue fever among under 15-year children with clustering of cases in the community. DEN 3 was the most commonly circulating strain encountered. The findings underscore the need for development of affordable pre-vaccination screening strategy as well as newer dengue vaccines for young children while continuing efforts in vector control.     

Effect of phenol on protein and amino acid content of Xanthomonas oryzae pv. oryzae

Leaf blight disease of rice (Oryza sativa) is caused by the bacterium Xanthomonas oryzae pv. oryzae. Phenol (1 to 4 mM) induced changes in protein profiles of X. o. pv. oryzae and a stress protein with a molecular mass of 69,000 appeared. HPLC analysis indicated occurrence of amino acids such as asparagine, alanine, methionine and cystine in phenol treated cells. Proton NMR analysis also revealed variation on the presence of amino acids in the cells treated with phenol.     

Reversible alteration of morphology in an invertebrate erythrocyte: properties of the natural inducer and the cellular response

The normal shape of the erythrocytes of the bivalves known as blood clams is maintained by a marginal band (MB) of microtubules. When hemolymph (or "blood") is withdrawn from the animal, its erythrocytes change, within minutes, from the normal smooth-surfaced, flattened ellipsoids (N-cells) to spheroids with folded surfaces (X-cells). This alteration can be prevented by rapidly diluting the hemolymph with physiological medium, yielding N-cells for use in studying the transformation to X-cells. Bioassays showed that shape transformation was induced by a hemolymph activity (Hx) and was a function, in part, of cell responsiveness to this activity. Eventually the shape of the cells spontaneously returned to normal, at a rate dependent upon the concentration of the cells and of Hx; recovery was correlated with loss of Hx. The X-cells contained an intact but highly deformed MB, but this was not the effector of the transformation. Erythrocytes made to lack MBs still changed shape, although they did not recover as completely as did the MB-containing controls. When clams were cooled before hemolymph was withdrawn, the concentration of Hx was reduced. Hx was retained after dialysis of hemolymph, and initial filtration and chromatography indicated that its Mr was greater than 500,000. Shape transformation was blocked by EGTA, by serine protease inhibitors, and by sodium azide; the last indicates ATP-dependence. Although the mechanism responsible for shape transformation remains to be determined, the data suggest that the change is triggered by a coagulation-related activity in response to the removal of hemolymph from the animal.     

Development of acetylcholinesterase silica sol-gel immobilized biosensor--an application towards oxydemeton methyl detection

An amperometric silica sol-gel film immobilized biosensor doped with acetylcholinesterase was fabricated in the laboratory finding application in organophosphate detection based on enzyme inhibition. The substrate used was acetylthiocholine chloride and thiocholine released from the enzymatic hydrolysis was electrochemically oxidized giving larger anodic current at 0.5-0.6 V (versus Ag/AgCl reference). The dependencies of the current response on pH, enzyme loading and thermal stability of the developed biosensor were evaluated. The analytical performance of enzyme electrode towards substrate and inhibitor was investigated. Oxydemeton methyl was taken as a model compound for the inhibition studies. Linear calibration for oxydemeton methyl was obtained in the range of 2-200 ppb under the optimized conditions following an incubation time of 20 min. Treatment of the inhibited enzyme with pyridine-2-aldehyde restored 92% of its original activity. The sensor stored at -20 degrees C had a good storage and operational stability retaining 85% of its original activity for 60 successive measurements.     

Perinatal transmission of SHIV-SF162P3 in Macaca nemestrina

We developed a SHIV/macaque model of transmission from infected dams to their infants. Ten pregnant dams were infected intravenously with 100 MID(50) of macaque-titered SHIV-SF162P3 during the second trimester. Nine infants were born; the seven surviving beyond day of birth suckled for 6 months. Four of nine infants were infected (transmission rate = 44.4%), with one infection in utero, and three intrapartum and/or immediately post-birth via suckling. Varying levels of binding and neutralizing antibodies were transplacentally transferred to infants. Passive antibodies were detected in plasma on the day of birth and persisted for 5 weeks. Infants infected at or after birth controlled acute and post-acute viremia. Exposure to maternal SHIV-SF162P3 during birth and suckling in the presence of autologous maternal neutralizing antibodies may have affected transmission or pathogenesis in the infants. This transmission model can allow investigation of key parameters involved in perinatal transmission of HIV.