The Asian tiger mosquito, Aedes albopictus, is a medically important invasive species whose geographic distribution has expanded dramatically during the past 20 years, and one of the key elements of its success is its capacity to survive long distance transport as a diapausing pharate first instar larva, encased within the chorion of the egg. We report that pharate larvae entering diapause are larger and contain 30% more lipid than their nondiapausing counterparts. To improve our understanding of the molecular regulation of lipid metabolism during diapause, we assessed the relative mRNA abundance of 21 genes using qRT-PCR. Elevated expression of lipid storage droplet protein 2 during embryonic development likely contributes to the higher amounts of lipid we noted in diapausing individuals. The conservation of lipids during diapause is reflected in downregulation of genes involved in lipid catabolism, including lipase 2, lipase 3, lipase 4, acyl-CoA dehydrogenase 4, and isovaleryl-CoA dehydrogenase. Two genes involved in fatty acid synthesis and modification, Δ(9)-desaturase, and fatty acyl-CoA elongase, were both upregulated in diapausing pharate larvae, suggesting roles for their gene products in generating unsaturated fatty acids to enhance membrane fluidity at low temperatures and generating precursors to the surface hydrocarbons needed to resist desiccation, respectively. Together, the results point to substantial distinctions in lipid metabolism within the embryo as a consequence of the diapause program, and these differences occur both before the actual onset of diapause as well as during the diapause state.
A cross sectional study was carried out at the department of Forensic Medicine in Dhaka Medical College during the period of January 2008 to December 2009. Data were collected from 3rd copy of the post mortem reports which were preserved in the department of Forensic Medicine with the verbal consent of the doctors who performed autopsy report. During this period total 5114 autopsies were conducted. Out of this 970 cases (19%) were suicidal in nature. It was noticed that all suicidal deaths occurred from 10 years to all age group respectively, but top amongst age group of suicidal deaths occurred in between 21 to 30 years of people. Suicidal deaths are more common in female than male. Suicidal deaths due to hanging is highest, next common causes of death due to organophosphorus compund poisoning. Suicidal deaths by hanging is more in female than male but in poisoning cases male are more lvictimised than female. Objectives of our study are to see the occurrence and methods of suicidal death.
Metabolic labeling with stable isotopes is a prominent technique for comparative quantitative proteomics, and stable isotope labeling with amino acids in cell culture (SILAC) is the most commonly used approach. SILAC is, however, traditionally limited to simple tissue culture regimens and only rarely employed in the context of complex culturing conditions as those required for human embryonic stem cells (hESCs). Classic hESC culture is based on the use of mouse embryonic fibroblasts (MEFs) as a feeder layer, and as a result, possible xenogeneic contamination, contribution of unlabeled amino acids by the feeders, interlaboratory variability of MEF preparation, and the overall complexity of the culture system are all of concern in conjunction with SILAC. We demonstrate a feeder-free SILAC culture system based on a customized version of a commonly used, chemically defined hESC medium developed by Ludwig et al. and commercially available as mTeSR1 [mTeSR1 is a trade mark of WiCell (Madison, WI) licensed to STEMCELL Technologies (Vancouver, Canada)]. This medium, together with adjustments to the culturing protocol, facilitates reproducible labeling that is easily scalable to the protein amounts required by proteomic work flows. It greatly enhances the usability of quantitative proteomics as a tool for the study of mechanisms underlying hESCs differentiation and self-renewal. Associated data have been deposited to the ProteomeXchange with the identifier PXD000151.
The pediatric early warning score (PEWS) tool helps providers to detect subtle clinical deterioration in non-intensive care unit pediatric patients and intervene early to prevent significant adverse outcomes. Although widely used in general pediatrics, limited studies report on its validation; none report on use with burn-injured patients. New York-Presbyterian/Weill Cornell Medical Center modified a general PEWS system to a burn-specific PEWS and integrated its use into standard practice. This study investigated the external validity of the PEWS process in clinical practice. Fifty cases of patients aged 0 to 15.9 years admitted between January 2012 and June 2013, whose length of stay (LOS) more than 3 days were selected for review from this cohort of n equal to 187. Demographics, total PEWS and score changes, and compliance with PEWS documentation and with resultant interventions were reviewed. Continuous variables are presented as mean ± SD, P less than 0.05. Mean age, burn size, and LOS were 3.2 ± 3.3 years, 4.8 ± 5.7%, and 9.8 ± 7.0 days; 26% required grafting, and 50% were male. No mortalities occurred. One thousand six hundred and twelve PEWS from 1745 opportunities were documented (92.4%). For all PEWS (n = 1612) and PEWS greater than 0 (n = 912), means were 0.9 ± 1.2 and 1.6 ± 1.2, respectively. Among the 162 PEWS increase events, intake (54.1%) and output (4.5%) parameters increased most commonly. Of these, 129 PEWS increases (79.6%) were followed by an intervention that most commonly included text notation of score increase (93.7%), physician/physician assistant notification (70.5%), and feeding-tube insertion (25.6%). Patients with PEWS greater than 0 had similar age, LOS, and larger burn size (5.2% vs 1.4%, P < 0.05) than those with PEWS equal to 0. Compliance with PEWS performance and resultant actions based on score increases are high. Data support that even small changes in burn-injury specific PEWS stimulate provider discussion and intervention and support its validation; further studies on its effect on practice are warranted.
Background and Objectives: Metabolic labeling with stable isotopes remains a prominent technique for comparative quantitative proteomics and stable isotope labeling with amino acids in cell culture (SILAC) is the most prominent approach used. However, despite its power the approach traditionally is limited if applied to complex tissue culture regiments as those required for human embryonic stem cells (hESC). Classic hESC culture is based on use of mouse embryonic fibroblasts (MEFs) for conditioning the cell culture medium or as a feeder layer. As a result the possibility of xenogeneic contaminants, contribution of unlabeled amino acids, inter-laboratory variability of MEFs and the trick complexity of the culture system are all concerns when using SILAC and beyond. Methods: We applied high accuracy LC-MS/MS analyses to evaluate the SILAC labeling efficiency of the hESCs cultured in the new SILAC- hESCs culture system. Results: The analysis yielded over 15,00 distinct hESC proteins with more than 99% accuracy of identification as estimated by reverse database searching. The efficiency of labeling was estimated to be higher than 99% for lysine and arginine; moreover, SILAC-labeled hESCs maintained undifferentiated self-renewal status. Conclusions: Here we use an enhanced feeder-free SILAC culture system based on a customized chemically defined SILAC-medium and a modified culture protocol to overcome these limitations and achieve reproducible labeling in a process easily scaled to proteomic protein requirements. The protocol is expected to greatly enhance the usability of quantitative proteomics as a tool for the study of mechanisms underlying hESC differentiation and self-renewal.
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