Molecular regulators of glucose and lipid metabolism in skeletal muscle

Skeletal muscle is a primary site of insulin action and insulin-stimulated glucose transport and occupies a center stage in maintaining whole body glucose and lipid homeostasis. Another key feature of a healthy skeletal muscle is its ability to switch between utilization of lipids and glucose as fuel in response to feeding or fasting respectively. This metabolic flexibility is impaired in skeletal muscle from insulin resistant and type 2 diabetic patients. Key molecules such as AMP Kinase [AMPK] and Pyruvate Dehydrogenase Kinase [PDK] play a crucial role in maintaining metabolic flexibility in a healthy skeletal muscle. These molecules have recently emerged as potential drug targets to combat diabetes. One strategy to activate the AMPK pathway is via altering the expression of AMP-metabolizing enzymes, such as 5’-nucleotidases [NT5C]. The role of cytosolic 5´-nucleotidases was determined in metabolic responses linked to the development of insulin resistance in obesity and type 2 diabetes (Study I). NT5C1A and NT5C2 gene silencing led to increase in the AMP/ATP ratio, increase in phosphorylation of AMPK and ACC, and an increase in palmitate oxidation and glucose transport in mouse and human skeletal muscle. Another strategy to activate the AMPK system would be to lower the threshold of AMPK activation by rendering AMPK more sensitive to its activators. This strategy has been undertaken in this thesis by pre-treating skeletal muscle with methotrexate [MTX] and targeting 5-Amino-4-imidazolecarboxamide Ribonucleotide Transformylase [ATIC] (Study II). MTX is an inhibitor of ATIC, an enzyme involved in de-novo nucleotide biosynthesis. ATIC imposes a metabolic block leading to intracellular ZMP accumulation, lowering the threshold for AMPK activation. Increased glucocorticoid action leads to reduced whole body insulin action and may predispose to type 2 diabetes. Local conversion of cortisone to active cortisol by the enzyme 11β-hydroxysteroid dehydrogenase [11β-HSD1] in target tissues may regulate tissue specific roles of glucocortioids in pathophysiological states. Chronic high dose exposure to cortisone or cortisol reduced glucose metabolism and enhanced lipid metabolism, via induction of pyruvate dehydrogenase kinase 4 [PDK4] expression in myotubes. siRNA-mediated reduction or pharmacological inhibition of 11β-HSD1 prevented the effects of cortisone but not cortisol, on metabolic responses (Study III). Type 2 diabetes mellitus is associated with abnormal substrate metabolism, raising the possibility that alterations in the expression of mitochondrial enzymes controlling lipid uptake and metabolism may be altered (Study IV). Evidence that expression of key enzymes regulating mitochondrial function in skeletal muscle is altered in type 2 diabetes mellitus [T2DM] patients is provided. In summary, activation of AMPK can play a central role in overcoming impairments in skeletal muscle glucose and lipid metabolism and this can be achieved by perturbing the enzymes involved in nucleotide metabolism such as 5’-nucleotidases and ATIC. Alterations in molecular regulators of substrate metabolism such as PDK4, reflect pathogenic condition and could be targeted to restore skeletal muscle energy homeostasis

Experimental Results of the First Two Stages of an Advanced Transonic Core Compressor Under Isolated and Multi-Stage Conditions

NASA's Environmentally Responsible Aviation (ERA) Program calls for investigation of the technology barriers associated with improved fuel efficiency of large gas turbine engines. Under ERA the task for a High Pressure Ratio Core Technology program calls for a higher overall pressure ratio of 60 to 70. This mean that the HPC would have to almost double in pressure ratio and keep its high level of efficiency. The challenge is how to match the corrected mass flow rate of the front two supersonic high reaction and high corrected tip speed stages with a total pressure ratio of 3.5. NASA and GE teamed to address this challenge by using the initial geometry of an advanced GE compressor design to meet the requirements of the first 2 stages of the very high pressure ratio core compressor. The rig was configured to run as a 2 stage machine, with Strut and IGV, Rotor 1 and Stator 1 run as independent tests which were then followed by adding the second stage. The goal is to fully understand the stage performances under isolated and multi-stage conditions and fully understand any differences and provide a detailed aerodynamic data set for CFD validation. Full use was made of steady and unsteady measurement methods to isolate fluid dynamics loss source mechanisms due to interaction and endwalls. The paper will present the description of the compressor test article, its predicted performance and operability, and the experimental results for both the single stage and two stage configurations. We focus the detailed measurements on 97 and 100 of design speed at 3 vane setting angles

Bovine rotavirus pentavalent vaccine development in India

AbstractA bovine rotavirus pentavalent vaccine (BRV-PV) containing rotavirus human-bovine (UK) reassortant strains of serotype G1, G2, G3, G4 and G9 has been developed by the Serum Institute of India Ltd, in collaboration with the National Institute of Allergy and Infectious Diseases (NIAID), USA. The vaccine underwent animal toxicity studies and Phase I and II studies in adults, toddlers and infants. It has been found safe and immunogenic and will undergo a large Phase III study to assess efficacy against severe rotavirus gastroenteritis

Clinico-social parameters of diabetes among patientsutilizing emergency medical services

Background: Diabetes mellitus (DM) is increasing in its potential in developing countries. Rise in diabetic patients calling emergency medical services (EMS) is expected. It calls for thorough assessment of patients with DM utilizing EMS. In the present study, was to assess magnitude of DM among patients utilizing EMS and its clinico-social parameters.Methods: It was hospital record based observational study of patients calling EMS delivered by a tertiary care hospital in Pune, Maharashtra, India during 1st January 2013 to 31st December 2014. Patients with incomplete records were excluded. A person with medical background was trained to extract required information from hospital records. Patients with previously diagnosed DM were considered those who were on diet, oral hypoglycemic agents or taking insulin therapy and newly diagnosed patients with DM were considered those with the value of glycemia on admission >200 mg/dl in first 24 hours. Data analysis was done using SPSS 15.0 software.Results: 45.8% (894/1951) were Patients with DM out of that 78.19% (699/894) were known to have DM while 28.81% (195/1951) were new cases of DM. 5.1% (100/1951) patients had uncontrolled DM. DM was significantly more in >60 years age group and in urban residents (p<0.001). Patients with DM were significantly more to have breathlessness, altered sensorium and dyspnoea as major purpose to call EMS (p<0.001, <0.001 and 0.045 respectively). Other co-morbidities in the form of Hypertension, other cardiovascular abnormalities, COPD, CKD and history of CVA were significantly more among Patients with DM (p<0.001, except for COPD, p=0.027).Conclusions: There was a high burden of patients with DM on EMS. EMS teams should be well trained to diagnose and manage such emergencies. Mass awareness of screening for DM and its proper management will help to decrease such burden

Accelerated protein synthesis via one–pot ligation–deselenization chemistry

Peptide ligation chemistry has revolutionized protein science by facilitating access to synthetic proteins. Here, we describe the development of additive-free ligation-deselenization chemistry at β-selenoaspartate and γ-selenoglutamate that enables the generation of native polypeptide products on unprecedented timescales. The deselenization step is chemoselective in the presence of unprotected selenocysteine, which is highlighted in the synthesis of selenoprotein K. The power of the methodology is also showcased through the synthesis of three tick-derived thrombin-inhibiting proteins, each of which were assembled, purified, and isolated for biological assays within a few hours. The methodology described here should serve as a powerful means of accessing synthetic proteins, including therapeutic leads, in the future

SIRT1 enhances glucose tolerance by potentiating brown adipose tissue function

Objective: SIRT1 has been proposed to be a key signaling node linking changes in energy metabolism to transcriptional adaptations. Although SIRT1 overexpression is protective against diverse metabolic complications, especially in response to high-fat diets, studies aiming to understand the etiology of such benefits are scarce. Here, we aimed to identify the key tissues and mechanisms implicated in the beneficial effects of SIRT1 on glucose homeostasis. Methods: we have used a mouse model of moderate SIRT1 overexpression, under the control of its natural promoter, to evaluate glucose homeostasis and thoroughly characterize how different tissues could influence insulin sensitivity. Results: mice with moderate overexpression of SIRT1 exhibit better glucose tolerance and insulin sensitivity even on a low fat diet. Euglycemic-hyperinsulinemic clamps and in-depth tissue analyses revealed that enhanced insulin sensitivity was achieved through a higher brown adipose tissue activity and was fully reversed by housing the mice at thermoneutrality. SIRT1 did not influence brown adipocyte differentiation, but dramatically enhanced the metabolic transcriptional responses to β3-adrenergic stimuli in differentiated adipocytes. Conclusions: our work demonstrates that SIRT1 improves glucose homeostasis by enhancing BAT function. This is not consequent to an alteration in the brown adipocyte differentiation process, but as a result of potentiating the response to β3-adrenergic stimuli

Accelerated protein synthesis via one–pot ligation–deselenization chemistry

Peptide ligation chemistry has revolutionized protein science by facilitating access to synthetic proteins. Here, we describe the development of additive-free ligation-deselenization chemistry at β-selenoaspartate and γ-selenoglutamate that enables the generation of native polypeptide products on unprecedented timescales. The deselenization step is chemoselective in the presence of unprotected selenocysteine, which is highlighted in the synthesis of selenoprotein K. The power of the methodology is also showcased through the synthesis of three tick-derived thrombin-inhibiting proteins, each of which were assembled, purified, and isolated for biological assays within a few hours. The methodology described here should serve as a powerful means of accessing synthetic proteins, including therapeutic leads, in the future

Multiple novel prostate cancer susceptibility signals identified by fine-mapping of known risk loci among Europeans

Genome-wide association studies (GWAS) have identified numerous common prostate cancer (PrCa) susceptibility loci. We have fine-mapped 64 GWAS regions known at the conclusion of the iCOGS study using large-scale genotyping and imputation in 25 723 PrCa cases and 26 274 controls of European ancestry. We detected evidence for multiple independent signals at 16 regions, 12 of which contained additional newly identified significant associations. A single signal comprising a spectrum of correlated variation was observed at 39 regions; 35 of which are now described by a novel more significantly associated lead SNP, while the originally reported variant remained as the lead SNP only in 4 regions. We also confirmed two association signals in Europeans that had been previously reported only in East-Asian GWAS. Based on statistical evidence and linkage disequilibrium (LD) structure, we have curated and narrowed down the list of the most likely candidate causal variants for each region. Functional annotation using data from ENCODE filtered for PrCa cell lines and eQTL analysis demonstrated significant enrichment for overlap with bio-features within this set. By incorporating the novel risk variants identified here alongside the refined data for existing association signals, we estimate that these loci now explain ∼38.9% of the familial relative risk of PrCa, an 8.9% improvement over the previously reported GWAS tag SNPs. This suggests that a significant fraction of the heritability of PrCa may have been hidden during the discovery phase of GWAS, in particular due to the presence of multiple independent signals within the same regio