Modulation of Nuclear Receptor 4A1 Expression Improves Insulin Secretion in a Mouse Model of Chronic Pancreatitis.

OBJECTIVES: Diabetes secondary to chronic pancreatitis (CP) presents clinical challenges due to lack of understanding on factor(s) triggering insulin secretory defects. Therefore, we aimed to delineate the molecular mechanism of ß-cell dysfunction in CP. MATERIALS AND METHODS: Transcriptomic analysis was conducted to identify endocrine-specific receptor expression in mice and human CP on microarray. The identified receptor (NR4A1) was overexpressed in MIN6 cells using PEI linear transfection. RNA-Seq analysis of NR4A1-overexpressed (OE) MIN6 cells on NovaSeq6000 identified aberrant metabolic pathways. Upstream trigger for NR4A1OE was studied by InBio Discover and cytokine exposure, whereas downstream effect was examined by Fura2 AM-based fluorimetric and imaging studies. Mice with CP were treated with IFN-γ-neutralizing monoclonal antibodies to assess NR4A1 expression and insulin secretion. RESULTS: Increased expression of NR4A1 associated with decreased insulin secretion in islets (humans: controls 9 ± 0.2, CP 3.7 ± 0.2, mice: controls 8.5 ± 0.2, CP 2.1 ± 0.1 µg/L). NR4A1OE in MIN6 cells (13.2 ± 0.1) showed reduction in insulin secretion (13 ± 5 to 0.2 ± 0.1 µg/mg protein per minute, P = 0.001) and downregulation of calcium and cAMP signaling pathways. IFN-γ was identified as upstream signal for NR4A1OE in MIN6. Mice treated with IFN-γ-neutralizing antibodies showed decreased NR4A1 expression 3.4 ± 0.11-fold ( P = 0.03), showed improved insulin secretion (4.4 ± 0.2-fold, P = 0.01), and associated with increased Ca 2+ levels (2.39 ± 0.06-fold, P = 0.009). CONCLUSIONS: Modulating NR4A1 expression can be a promising therapeutic strategy to improve insulin secretion in CP.

GeneAI 3.0: powerful, novel, generalized hybrid and ensemble deep learning frameworks for miRNA species classification of stationary patterns from nucleotides.

Due to the intricate relationship between the small non-coding ribonucleic acid (miRNA) sequences, the classification of miRNA species, namely Human, Gorilla, Rat, and Mouse is challenging. Previous methods are not robust and accurate. In this study, we present AtheroPoint's GeneAI 3.0, a powerful, novel, and generalized method for extracting features from the fixed patterns of purines and pyrimidines in each miRNA sequence in ensemble paradigms in machine learning (EML) and convolutional neural network (CNN)-based deep learning (EDL) frameworks. GeneAI 3.0 utilized five conventional (Entropy, Dissimilarity, Energy, Homogeneity, and Contrast), and three contemporary (Shannon entropy, Hurst exponent, Fractal dimension) features, to generate a composite feature set from given miRNA sequences which were then passed into our ML and DL classification framework. A set of 11 new classifiers was designed consisting of 5 EML and 6 EDL for binary/multiclass classification. It was benchmarked against 9 solo ML (SML), 6 solo DL (SDL), 12 hybrid DL (HDL) models, resulting in a total of 11 + 27 = 38 models were designed. Four hypotheses were formulated and validated using explainable AI (XAI) as well as reliability/statistical tests. The order of the mean performance using accuracy (ACC)/area-under-the-curve (AUC) of the 24 DL classifiers was: EDL > HDL > SDL. The mean performance of EDL models with CNN layers was superior to that without CNN layers by 0.73%/0.92%. Mean performance of EML models was superior to SML models with improvements of ACC/AUC by 6.24%/6.46%. EDL models performed significantly better than EML models, with a mean increase in ACC/AUC of 7.09%/6.96%. The GeneAI 3.0 tool produced expected XAI feature plots, and the statistical tests showed significant p-values. Ensemble models with composite features are highly effective and generalized models for effectively classifying miRNA sequences.

Distribution of microplastics in seafloor sediments and their differential assimilation in nearshore benthic molluscs along the south-west coast of India.

Spatial and temporal distribution of microplastics (MPs) in the nearshore seafloor sediments along the Southwest coast of India and their patterns of accumulation in selected infaunal and epibenthic molluscs with diverse feeding strategies were investigated. Along the 300-km coastal stretch, which is one of the most productive and biodiversity rich regions of the eastern Arabian Sea, notable levels of MP contamination in both sediment (617.7 items/kg dry weight) and molluscs (5.39 items/g) was recorded. The concentration of MPs in sediments also varied seasonally, with a higher prevalence during the post-monsoon season. Among the four molluscan groups studied, the highest MP abundance was recorded among scavenging gastropod Pseudominolia biangulosa (9.13 items/g), followed by microcarnivore scaphopod Tesseracme quadrapicalis (5.96 items/g). In comparison, the suspension feeding bivalve, Anadara hankeyana and deposit feeding clam Jitlada philippinarum had lesser accumulation of MPs (2.98 items/g and 3.50 items/g respectively). The majority of MPs in sediments and within molluscs were less than 250 µm in size (89.14%) and were predominantly fibres and fragments. Chemical characterisation of MPs revealed eleven types of polymers dominated by polyethylene (PE) and polypropylene (PP). Present study identified positive correlations between ingested MP polymers and the feeding strategies of molluscs. Higher values for the ecological risk assessment indices (PHI, PLI and PERI) in most of the stations indicated the severity of plastic pollution in the region. Molluscs being a major contributor to the benthic food web is also a connecting link to higher trophic levels. Hence understanding the specificity in the MPs accumulation pattern within this group has far reaching significance in utilizing them as potential bioindicators for pollution studies in marine ecosystems.

Anterior Segment OCT for Detection of Narrow Angles: A Community-Based Diagnostic Accuracy Study.

PURPOSE: To assess the diagnostic accuracy of anterior segment OCT (AS-OCT) screening for detecting gonioscopically narrow angles. DESIGN: Population-based cross-sectional study. PARTICIPANTS: A stratified random sample of individuals aged ≥ 60 years, selected from a door-to-door census performed in low-lying Nepal. TESTING: Participants underwent AS-OCT, posterior segment OCT, and intraocular pressure (IOP) testing in the community. Those meeting referral criteria in either eye were invited to have a comprehensive eye examination including gonioscopy. Referral criteria included (i) the lowest 2.5% of AS-OCT measurements, (ii) retinal OCT results suggestive of glaucomatous optic neuropathy, diabetic retinopathy, or age-related macular degeneration, and (iii) elevated IOP. MAIN OUTCOME MEASURES: Sensitivity and specificity of 5 semiautomated AS-OCT parameters relative to gonioscopically narrow angles, defined as the absence of visible trabecular meshwork for ≥ 180° on nonindentation gonioscopy. RESULTS: Of 17 656 people aged ≥ 60 years enumerated from 102 communities, 12 633 (71.6%) presented for AS-OCT testing. Referral was recommended for 697 participants based on AS-OCT criteria and 2419 participants based on other criteria, of which 858 had gonioscopy performed by a glaucoma specialist. Each of the 5 AS-OCT parameters offered good diagnostic information for predicting eyes with gonioscopically narrow angles, with areas under the receiver operating characteristic curve ranging from 0.85 to 0.89. The angle opening distance at 750 µm from the scleral spur (AOD750) provided the most diagnostic information, providing an optimal sensitivity of 87% (95% confidence interval [CI], 75%-96%) and specificity of 77% (71%-83%) at a cutpoint of 367 µm, and a sensitivity of 65% (95% CI, 54%-74%) when specificity was constrained to 90% (cutpoint, 283 µm). CONCLUSIONS: On AS-OCT, the AOD750 parameter detected approximately two-thirds of cases of gonioscopically narrow angles when test specificity was set to 90%. Although such a sensitivity may not be sufficient when screening solely for narrow angles, AS-OCT requires little additional effort if posterior segment OCT is already being performed and thus could provide incremental benefit when performing OCT-based screening. FINANCIAL DISCLOSURE(S): The authors have no proprietary or commercial interest in any materials discussed in this article.

Trophic fingerprinting of a pristine but rapidly deteriorating downstream region of a Western Ghats River.

Chalakudy River is renowned for its pristine waters and rich ichthyofaunal biodiversity. The downstream area of the river is confronting a series of risks, including pollution, saline water ingression, sand mining, illegal and intensified fishing practices, and invasion of exotic and alien species. A mass balanced ecosystem model was constructed for the downstream region of Chalakudy River (DCR) using Ecopath with Ecosim (EWE), incorporating 12 functional groups to delineate the food web and network flow indices for the period 2020 to 2021. The trophic level (TL) of the ecosystem network ranged from TL-1 (detritus) to TL-3.4 (birds). High fishing pressure is one possible cause for the high ecotrophic efficiency values as evidenced by the fish groups. Both the grazing food chain and detritus food chain (detritivory: herbivory ratio 0.94) contributed more or less equal to the energy transfer between TL. Network analysis of the model indicated a mean transfer efficiency of 12%, with shares from primary producers (14%) and detritus (11%). A mixed trophic impact analysis demonstrated a strong positive impact of primary producers and detritus groups on most of the other ecological groups at higher trophic levels. The DCR model showed a high system throughput (32,464.7 t km-2 year-1), low system omnivory (0.09), low connectance index (0.36), low Finn's cycling index (4.9), and mean path length (2.8), low relative ascendency (37.5%), and high system overhead (62.5%). These indices propound that DCR is an immature and developing ecosystem with moderate strength in reserve to resist external perturbations.

Spatio-temporal evaluation and risk assessment of microplastics in nearshore surface waters post-2018 Kerala deluge along the southwest coast of India.

Spatial and temporal distribution of microplastics along the nearshore surface waters of Kerala after the floods of 2018 was studied. Results indicated a seven-fold increase in its mean concentration (7.14 ± 3.03 items/m3) post deluge. The average abundance was highest during pre-monsoon (8.27 ± 3.09 items/m3). Fibres were the dominant group, with blue and black being the most prevalent colours. Polyethylene and polypropylene were the most commonly found polymers, possibly gaining entry through sewage waste or land-based plastic litter. Highest abundance of microplastic was recorded off Kochi categorising it at Hazard Level I under Pollution Load Index assessment. Similarly high levels of Pollution Hazard Index and Potential Ecological Risk Index were also reported due to the presence of hazardous polymers PVC and PU that can cause concern to marine life. The differential weathering pattern and surface morphology analysis suggested microplastics to be relatively old that had undergone substantial mechanical and oxidative weathering.

Cross-sectional study to describe the severity, bio-chemical associations, and final outcomes of COVID-19-associated rhino-orbital-cerebral mucormycosis in a tertiary hospital of East India.

Purpose: The second wave of coronavirus disease 2019 (COVID-19) pandemic triggered a mucormycosis epidemic in India. Diabetes mellitus and dysregulated immune response were contributors, and rhino-orbital-cerebral mucormycosis (ROCM) was the most common presentation. It is however not known whether bio-chemical parameters at presentation correlate with stage of ROCM or final outcome in terms of vision or mortality. Methods: This retrospective, hospital-based study included all in-patients of mucormycosis with ophthalmic manifestations at presentation admitted during June 1, 2021 to August 31, 2021. It aimed to evaluate the association between severity of infection, serum levels of HbA1c, ferritin, interleukin-6 (IL-6), C-reactive protein (CRP), and D-dimer levels at presentation and outcome. Results: There were altogether 47 eligible cases having a mean age of 48.8 ± 10.9 years with a male:female ratio of 2.6:1; forty-two (89.4%) had pre-existing diabetes, and five (10.6%) had steroid-induced hyperglycemia. The mean HbA1c among diabetics was 9.7 ± 2.1. HbA1c and serum CRP showed an increase over subsequent stages, which was not statistically significant (P = 0.31). IL-6 values for all stages were similar (P = 0.97). Only serum ferritin levels showed a statistically significant increase over stages (P = 0.04). IL-6 was significantly lower (P = 0.03) in patients who survived, whereas CRP levels were significantly lower in patients who had final visual acuity (VA) better than only perception of light (P = 0.03). Conclusion: Uncontrolled diabetes mellitus is a significant association of ROCM. Serum ferritin levels at presentation best correlate with extent of the disease. CRP levels are best to prognosticate cases that will have sufficient VA to carry on activities of daily living, whereas IL-6 levels are best associated with survival.

A Prospective Comparative Study Between Ultrasound-Guided Combined Sciatic-Femoral Nerve Block Versus Spinal Anesthesia for the Patients Undergoing Elective Below-Knee Surgeries.

Introduction The scope of anesthesia has shifted from general anesthesia (GA) and spinal anesthesia (SA) for below-knee surgery to peripheral nerve blocks (PNB). Combined sciatic-femoral nerve block (SFNB) with ultrasound (USG) guidance can be a better format for use.  Objectives The primary objectives were to compare the duration of onset of sensory and motor blockade, total duration of sensory and motor blockade, and time of first analgesic requirement between both groups. Methods A prospective, randomized comparative study was carried out at a tertiary care teaching hospital in Odisha, India, from April 2019 to April 2021 in the Department of Anaesthesiology. Patients admitted for elective below-knee surgeries with American Society of Anesthesiology (ASA) grade II or less were divided into two groups (Group A receiving USG-guided SFNB and group B receiving SA) by computer-generated sampling. The block randomization method was used to ensure equal samples in both groups. Data collection was done using the Magpi software (Magpi, Inc., Washington, D.C., United States) on android-based mobile phones. Data were analyzed using Stata Statistical Software: Release 12 (2011; StrataCorp LP, College Station, Texas, United States) for analysis. Relevant statistical tests were used to compare the results between the groups (independent sample t-test or Wilcoxson signed-rank test). Repeated measures ANOVA (RM-ANOVA) was used to check the hemodynamic stability within the groups. Results Thirty-seven subjects were enrolled in each arm (Group A and Group B). Baseline parameters in both groups were comparable. The most common indication among the study subjects was single or multiple meta-tarsal fractures (20, 27.0%) followed by malleolus (15, 20.3%) and calcaneum fractures (13, 17.6%). Most of the study subjects were from ASA grade I (around 80%). The time of onset of sensory and motor block was found to be more for USG-guided SFNB (8.08±2.11 minutes and 11.35±1.84 minutes, respectively) as compared to the SA group (3.03±0.50 minutes and 4.89±0.52 minutes, respectively) (p<0.001). Total anesthesia and time to first analgesic requirement were, however, more in USG-guided SFNB (349.43±53.49 minutes and 339.73±54.24 minutes, respectively) as compared to the SA group (137.30±34.21 minutes and 137.30±34.21 minutes, respectively) (p<0.001). The mean time to first urination in USG-guided SFNB (178.92±20.92) was significantly less (p<0.001) compared to the SA group (419.19±40.30). There were no adverse events (0%) in USG-guided SFNB while 64.9% of the subjects in the SA group experienced adverse events (p<0.001). The most common adverse events were nausea/vomiting and hypotension (around 50% for both). Hemodynamic stability was present in both the groups of anesthesia subjects, though fluctuations in blood pressure may be seen more frequently in cases of SA. All the subjects in both the groups had achieved a Bromage score of 3 universally. The grand mean score of pain by SA (2.347±0.044) was more (p<0.001) in comparison to that in subjects with USG-guided SFNB (1.961±0.073) and this was significant in both the groups. The mean increase in pain score at 24 hours in comparison to baseline was, however, significantly more (p<0.05) in the SA group (1.784±0.111) in comparison to those receiving USG-guided SFNB (1.324±0.190). Conclusion USG-guided SFNB is a better option for below-knee surgeries as compared to SA.

Profile of donors and corneal tissue obtained through hospital cornea retrieval programme in a recently established eye bank of a tertiary care teaching hospital of Eastern India.

CONTEXT: There is a global need for quality eye banking practices and sensitization of primary care physicians toward corneal donation. AIMS: To evaluate performance of a recently established eye bank (EB) and quality of corneas obtained, and identify areas of improvement during procurement and utilization of donor corneas. SETTINGS AND DESIGN: This retrospective observational study is based on records of corneas collected through hospital cornea retrieval programme (HRCP) in the EB of a tertiary care institution during the first 2 years of its establishment. METHODS AND MATERIAL: Data on demographic characteristics of donors, death-preservation interval, specular microscopy parameters of corneas, indications for utilization, and reasons for non-utilization of corneas were collected. STATISTICAL ANALYSIS USED: Means, standard deviation, range, frequencies, and proportions were analyzed. Spearman's correlation coefficient and Kruskal-Wallis test were applied taking P < 0.05 as significant. RESULTS: The EB retrieved 54 corneas from 27 donors with mean age 42.3 ± 24.2 years. All tissues were preserved in Cornisol®. Majority (50%) of transplantable tissues had an endothelial cell density (ECD) between 2,000 and 2,500 cells/mm2. ECD decreased significantly with increasing age (Spearman's ρ -0.747, P < 0.001; Kruskal-Wallis P < 0.001). Overall utilization rate of tissues was 87.04% (47/54), and utilizable corneas (50/54, 92.6%) were mainly used for optical purposes (34/50, 68%). CONCLUSIONS: Successful HCRP of the recently established EB has shown considerable promise in terms of quality and utilisation of corneas. There is need for active involvement of primary care physicians in contributing to increasing voluntary eye donation through awareness, advocacy, and social mobilization.

On spatial molecular arrangements of SARS-CoV2 genomes of Indian patients

A pandemic caused by the SARS-CoV2 is being experienced by the whole world since December, 2019. A thorough understanding beyond just sequential similarities among the protein coding genes of SARS-CoV2 is important in order to differentiate or relate to the other known CoVs of the same genus. In this study, we compare three genomes namely MT012098 (India-Kerala), MT050493 (India-Kerala), MT358637 (India-Gujrat) from India with NC_045512 (China-Wuhan) to view the spatial as well as molecular arrangements of nucleotide bases of all the genes embedded in these four genomes. Based on different features extracted for each gene embedded in these genomes, corresponding phylogenetic relationships have been built up. Differences in phylogenetic tree arrangement with individual gene suggest that three genomes of Indian origin have come from three different origins or the evolution of viral genome is very fast process. This study would also help to understand the virulence factors, disease pathogenicity, origin and transmission of the SARS-CoV2.

Activated protein C ameliorates chronic graft-versus-host disease by PAR1-dependent biased cell signaling on T cells.

Soluble thrombomodulin plasma concentrations are elevated in steroid-resistant graft-versus-host disease (GVHD), implying endothelial hypofunctioning for thrombomodulin-dependent generation of activated protein C's (APC) anticoagulant, anti-inflammatory, and antiapoptotic functions. Recombinant thrombomodulin or APC administration decreases acute GVHD, manifested by intense inflammation and tissue destruction. Here, we administered recombinant murine wild-type (WT) APC to mice with established chronic GVHD (cGVHD), a less-inflammatory autoimmune-like disease. WT APC normalized bronchiolitis obliterans-induced pulmonary dysfunction. Signaling-selective APC variants (3A-APC [APC with lysine 191-193 replaced with 3 alanines] or 5A-APC [APC with lysine 191-193 replaced with 3 alanines and arginine 229/230 replaced with 2 alanines]) with normal cytoprotective properties, but greatly reduced anticoagulant activity, provided similar results. Mechanistically, WT APC and signaling-selective variants reduced T follicular helper cells, germinal center formation, immunoglobulin, and collagen deposition. WT APC can potentially cleave protease-activated receptor 1 (PAR1) at Arg41 or Arg46, the latter causing anti-inflammatory signaling. cGVHD was reduced in recipients of T cells from WT PAR1 or mutated Gln41-PAR1 donors but not from mutated Gln46-PAR1 donors. These data implicate donor T-cell APC-induced noncanonical cleavage at Arg46-PAR1, which is known to confer cytoprotective and anti-inflammatory activities. Together, these data indicate that APC anticoagulant activity is dispensable, whereas anti-inflammatory signaling and cytoprotective cell signaling by APC are essential. Because a phase 2 ischemic stroke clinical trial did not raise any safety issues for 3A-APC treatment, our studies provide a foundational platform for testing in clinical cGVHD therapy.

PAR1 biased signaling is required for activated protein C in vivo benefits in sepsis and stroke.

Activated protein C (APC) cleaves protease-activated receptor 1 (PAR1) in vitro at R46 to initiate beneficial cell signaling; however, thrombin and APC can cleave at R41. To elucidate PAR1-dependent aspects of the pharmacologic in vivo mechanisms of APC, we generated C57BL/6 mouse strains carrying QQ41 or QQ46 point mutations in PAR1 (F2r gene). Using these strains, we determined whether or not recombinant murine signaling-selective APC mutants would reduce septic death or provide neuroprotection against ischemic stroke when mice carried PAR1-homozygous mutations that prevent cleavage at either R41 or R46. Intercrossing PAR1+/R46Q mice generated expected numbers of PAR1+/+, PAR1+/R46Q, and R46Q/R46Q offspring whereas intercrossing PAR1+/R41Q mice gave decreased R41Q/R41Q homozygotes (resembling intercrossing PAR1+/PAR1-knockout mice). QQ41-PAR1 and QQ46-PAR1 brain endothelial cells showed the predicted retention or loss of cellular responses to thrombin receptor-activating peptide, thrombin, or APC for each PAR1 mutation. In sepsis studies, exogenous APC reduced mortality from 50% to 10% in Escherichia coli-induced pneumonia for wild-type (Wt) PAR1 and QQ41-PAR1 mice (P < .01) but had no benefit for QQ46-PAR1 mice. In transient distal middle cerebral artery occlusion stroke studies, exogenous APC significantly reduced infarct size, edema, and neuronal apoptosis for Wt mice and QQ41-PAR1 mice but had no detectable benefits for mice carrying QQ46-PAR1. In functional studies of forelimb-asymmetry and foot-fault tests at 24 hours after stroke induction, signaling-selective APC was beneficial for Wt and QQ41-PAR1 mice but not QQ46-PAR1 mice. These results support the concept that APC-induced, PAR1-dependent biased signaling following R46 cleavage is central to the in vivo benefits of APC.

Activated protein C light chain provides an extended binding surface for its anticoagulant cofactor, protein S.

Protein S anticoagulant cofactor sensitivity and PAR1 cleavage activity were assayed for 9 recombinant APC mutants.Residues L38, K43, I73, F95, and W115 on one face of the APC light chain define an extended surface containing the protein S binding site.

Prothrombotic skeletal muscle myosin directly enhances prothrombin activation by binding factors Xa and Va.

To test the hypothesis that skeletal muscle myosins can directly influence blood coagulation and thrombosis, ex vivo studies of the effects of myosin on thrombogenesis in fresh human blood were conducted. Addition of myosin to blood augmented the thrombotic responses of human blood flowing over collagen-coated surfaces (300 s-1 shear rate). Perfusion of human blood over myosin-coated surfaces also caused fibrin and platelet deposition, evidencing myosin's thrombogenicity. Myosin markedly enhanced thrombin generation in both platelet-rich plasma and platelet-poor plasma, indicating that myosin promoted thrombin generation in plasma primarily independent of platelets. In purified reaction mixtures composed only of factor Xa, factor Va, prothrombin, and calcium ions, myosin greatly enhanced prothrombinase activity. The Gla domain of factor Xa was not required for myosin's prothrombinase enhancement. When binding of purified clotting factors to immobilized myosin was monitored using biolayer interferometry, factors Xa and Va each showed favorable binding interactions. Factor Va reduced by 100-fold the apparent Kd of myosin for factor Xa (Kd ∼0.48 nM), primarily by reducing koff, indicating formation of a stable ternary complex of myosin:Xa:Va. In studies to assess possible clinical relevance for this discovery, we found that antimyosin antibodies inhibited thrombin generation in acute trauma patient plasmas more than in control plasmas (P = .0004), implying myosin might contribute to acute trauma coagulopathy. We posit that myosin enhancement of thrombin generation could contribute either to promote hemostasis or to augment thrombosis risk with consequent implications for myosin's possible contributions to pathophysiology in the setting of acute injuries.

Apolipoprotein E Receptor 2 Mediates Activated Protein C-Induced Endothelial Akt Activation and Endothelial Barrier Stabilization.

OBJECTIVE: Activated protein C (APC), a plasma serine protease, initiates cell signaling that protects endothelial cells from apoptosis and endothelial barrier disruption. Apolipoprotein E receptor 2 (ApoER2; LRP8) is a receptor known for mediating signaling initiated by reelin in neurons. ApoER2 contributes to APC-initiated signaling in monocytic U937 cells. The objective was to determine whether ApoER2 is required for APC's beneficial signaling in the endothelial cell surrogate EA.hy926 line. APPROACH AND RESULTS: We used small interfering RNA and inhibitors to probe requirements for specific receptors for APC's antiapoptotic activity and for phosphorylation of disabled-1 by Src family kinases and of Akt. When small interfering RNA for ApoER2 or endothelial cell protein C receptor or protease activated receptor 1 was used, APC's antiapoptotic activity was ablated, indicating that each of these receptors was required. In EA.hy926 cells, APC induced a 2- to 3-fold increased phosphorylation of Ser473-Akt and Tyr232-disabled-1, a phosphorylation known to trigger disabled-1-mediated signaling in other cell types. Ser473-Akt phosphorylation was inhibited by ApoER2 small interfering RNA or by inhibitors of Src (PP2), phosphatidylinositol-3 kinase (LY303511), and protease activated receptor 1 (SCH79797). ApoER2 small interfering RNA blocked the ability of APC to prevent thrombin-induced endothelial barrier disruption in TransEndothelial Resistance assays. Binding studies using purified APC and purified immobilized wild-type and mutated ApoER2 ectodomains suggested that APC binding involves Lys49, Asp50, and Trp64 on the surface of the N-terminal LA1 domain of ApoER2. CONCLUSIONS: ApoER2 contributes cooperatively with endothelial cell protein C receptor and protease activated receptor 1 to APC-initiated endothelial antiapoptotic and barrier protective signaling.

Disruptive environmental chemicals and cellular mechanisms that confer resistance to cell death.

Cell death is a process of dying within biological cells that are ceasing to function. This process is essential in regulating organism development, tissue homeostasis, and to eliminate cells in the body that are irreparably damaged. In general, dysfunction in normal cellular death is tightly linked to cancer progression. Specifically, the up-regulation of pro-survival factors, including oncogenic factors and antiapoptotic signaling pathways, and the down-regulation of pro-apoptotic factors, including tumor suppressive factors, confers resistance to cell death in tumor cells, which supports the emergence of a fully immortalized cellular phenotype. This review considers the potential relevance of ubiquitous environmental chemical exposures that have been shown to disrupt key pathways and mechanisms associated with this sort of dysfunction. Specifically, bisphenol A, chlorothalonil, dibutyl phthalate, dichlorvos, lindane, linuron, methoxychlor and oxyfluorfen are discussed as prototypical chemical disruptors; as their effects relate to resistance to cell death, as constituents within environmental mixtures and as potential contributors to environmental carcinogenesis.

Impact of yoga on biochemical profile of asthmatics: A randomized controlled study.

BACKGROUND: Asthma is a chronic inflammatory disorder of the airways. The chronic inflammation causes an associated increase in airway hyperresponsiveness that leads to recurrent episodes of wheezing, breathlessness, chest tightness, and coughing at night or in the early morning. Most of the studies have reported, as the effects of yoga on bronchial asthma, significant improvements in pulmonary functions, quality of life, and decrease in medication use, but none of the studies has attempted to show the effect of yoga on biochemical changes. OBJECTIVE: To evaluate the effect of yoga on biochemical profile of asthmatics. MATERIALS AND METHODS: In the present study, 276 patients of mild to moderate asthma (FEV 1> 60%) aged between 12 to 60 years were recruited from the Department of Pulmonary Medicine, King George's Medical University, U.P., Lucknow, India. They were randomly divided into two groups: Yoga group (with standard medical treatment and yogic intervention) and control group as standard medical treatment (without yogic intervention). At completion of 6 months of the study period, 35 subjects were dropped out, so out of 276 subjects, only 241 subjects completed the whole study (121 subjects from yoga group and 120 subjects from control group). Biochemical assessment was carried out at baseline and after 6 months of the study period. RESULTS: In yoga group, there was significant improvement found in the proportion of hemoglobin and antioxidant superoxide dismutase in comparison to control group and significant decrease was found in total leukocyte count (TLC) and differential leukocytes count in comparison to control group. There was no significant change found in TLC, polymorphs, and monocytes in between group comparison. CONCLUSIONS: Yoga group got significantly better improvement in biochemical variables than control group. Result shows that yoga can be practiced as adjuvant therapy with standard inhalation therapy for better outcome of asthma.

Note: Multi channel Doppler tuned spectrometer to study highly charged ions.

We describe the design and implementation of a multi channel Doppler tuned spectrometer setup to study physics of highly charged ions at high resolution in a direct way. A unique Soller slit assembly coupled with a long one dimensional position sensitive proportional counter enables us to get distinct x-ray peaks at different angles, which allows us to cover large number of angle in one shot. By using this setup, 1s2s (3)S1 - 1s(2) (1)S0 M1 transition in He-like Fe has been resolved from its satellite line 1s2s2p 4P(5/2)° - 1s(2)2s (2)S(1/2) M2 transition in Li-like Fe and measured the lifetime of their respective upper levels with high precision.

Type II phosphatidylinositol 4-kinase ß is an integral signaling component of early T cell activation mechanisms.

The early signaling events in T cell activation through CD3 receptor include a rapid change in intra cellular free calcium concentration and reorganization of actin cytoskeleton. Phosphatidylinositol 4-kinases (PtdIns 4-kinases) are implicated as key components in these early signaling events. The role of type II PtdIns 4-kinase ß in CD3 receptor signaling was investigated with the help of short hairpin RNA sequences. Cross-linking of CD3 receptors on Jurkat T Cells with monoclonal antibodies showed an early increase in type II PtdIns 4-kinase activity and co-localization of type II PtdIns 4-kinase ß with CD3 ζ. Transfection of Jurkat T Cells with shRNAs inhibited CD3 receptor mediated type II PtdIns 4-kinase activation with a concomitant reduction in intra cellular calcium release, suggesting a role for type II PtdIns 4-kinase ß in CD3 receptor signal transduction. Knock-down of type II PtdIns 4-kinase ß with shRNAs also correlated with a decrease in PtdIns 4-kinase activity in cytoskeleton fractions and reduced adhesion to matrigel surfaces. These results indicate that type II PtdIns 4-kinase ß is a key component in early T cell activation signaling cascades.