PCR based Detection of Food Borne Pathogens

Abstract-Many high-risk pathogens that cause disease in humans are transmitted through various food items. Food-borne disease constitutes a major public health problem. Assessment of the quality and safety of foods is important in human health. Rapid and easy detection of pathogenic organisms will facilitate precautionary measures to maintain healthy food. The Polymerase Chain Reaction (PCR) is a handy tool for rapid detection of low numbers of bacteria. We have designed gene specific primers for most common food borne pathogens such as Staphylococci, Salmonella and E.coli. Bacteria were isolated from food samples of various food outlets and identified using gene specific PCRs. We identified Staphylococci, Salmonella and E.coli O157 using gene specific primers by rapid and direct PCR technique in various food samples. This study helps us in getting a complete picture of the various pathogens that threaten to cause and spread food borne diseases and it would also enable establishment of a routine procedure and methodology for rapid identification of food borne bacteria using the rapid technique of direct PCR. This study will also enable us to judge the efficiency of present food safety steps taken by food manufacturers and exporters

Analysis of SNPs of MC4R , GNB3 and FTO gene polymorphism in obese Saudi subjects

Background: The goal of this study was to analyze the association between the FTO rs17817449 (G>T), G protein beta3 subunit (GNB3) C825T and Melanocortin 4 receptor (MC4R) A822G single nucleotide  olymorphism (SNP) with obesity in Saudi subjects.Methods: The subjects were divided into 2 groups according to BMI: Obese (BMI> 29.9) and non- obese control (BMI<24.9). Genotyping of the target genes were determined by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis (RFLP).Results: We demonstrated the association of the FTO genotype TT with increased weight, BMI and leptin levels in both males and females. However, there was no association of genotype TT with fasting blood glucose, triglycerides and cholesterol levels. Regarding GNB3 rs5443 polymorphism, the likelihood of obesity was linked to the TT genotype which was also associated with increased leptin levels. On the other hand, the SNP of MC4R A822G did not exhibit any significant association with obesity among studied subjects and showed only the presence of homozygous AA genotype.Conclusion: The polymorphism of FTO gene rs17817449 and GNB3 gene rs5443 (C825T) may be a genetic determinant of obesity in Saudi population whereas impact of MC4R Asn274Ser change could not be detected.Keywords: Obesity, FTO gene-polymorphism

Institutional deliveries and stillbirth and neonatal mortality in the Global Network's Maternal and Newborn Health Registry

Background Few studies have shown how the move toward institutional delivery in low and middle-income countries (LMIC) impacts stillbirth and newborn mortality. Objectives The study evaluated trends in institutional delivery in research sites in Belagavi and Nagpur India, Guatemala, Kenya, Pakistan, and Zambia from 2010 to 2018 and compared them to changes in the rates of neonatal mortality and stillbirth. Methods We analyzed data from a nine-year interval captured in the Global Network (GN) Maternal Newborn Health Registry (MNHR). Mortality rates were estimated from generalized estimating equations controlling for within-cluster correlation. Cluster-level analyses were performed to assess the association between institutional delivery and mortality rates. Results From 2010 to 2018, a total of 413,377 deliveries in 80 clusters across 6 sites in 5 countries were included in these analyses. An increase in the proportion of institutional deliveries occurred in all sites, with a range in 2018 from 57.7 to 99.8%. In 2010, the stillbirth rates ranged from 19.3 per 1000 births in the Kenyan site to 46.2 per 1000 births in the Pakistani site and by 2018, ranged from 9.7 per 1000 births in the Belagavi, India site to 40.8 per 1000 births in the Pakistani site. The 2010 neonatal mortality rates ranged from 19.0 per 1000 live births in the Kenyan site to 51.3 per 1000 live births in the Pakistani site with the 2018 neonatal mortality rates ranging from 9.2 per 1000 live births in the Zambian site to 50.2 per 1000 live births in the Pakistani site. In multivariate modeling, in some but not all sites, the reductions in stillbirth and neonatal death were significantly associated with an increase in the institutional deliveries. Conclusions There was an increase in institutional delivery rates in all sites and a reduction in stillbirth and neonatal mortality rates in some of the GN sites over the past decade. The relationship between institutional delivery and a decrease in mortality was significant in some but not all sites. However, the stillbirth and neonatal mortality rates remain at high levels. Understanding the relationship between institutional delivery and stillbirth and neonatal deaths in resource-limited environments will enable development of targeted interventions for reducing the mortality burden. Trial registration The study is registered at clinicaltrials.gov . ClinicalTrial.gov Trial Registration: NCT01073475

Mechanisms of brain wiring by axonal miRNAs: miR-181 and miR-182

The highly complex nervous system is built upon an intricate network of neurons. In order to make a functional network, the establishment of precise connections is crucial. Neuronal networks are established early during development when neurons send out axons that navigate through complex environments to connect to their target. Chemotropic attractant or repellent cues, cell adhesion molecules, morphogens and a wide range of factors secreted or expressed by guidepost cells enable axon guidance. The leading tip of the axon, the GC is important to sense the environment and integrate extracellular signals to navigate precisely. The axonal GC has a large repertoire of mRNAs that are dynamic in nature. Local regulation of transcripts in navigating axons is suspected to ensure precise pathfinding. However, mechanisms involving regulation of expression of these transcripts within GCs are largely unknown. This thesis investigates whether microRNAs, one of the quintessential posttranscriptional regulators, can regulate axon guidance by fine-tuning mRNA expression within subcellular compartments. To explore microRNA roles in axon guidance, Xenopus laevis visual system was used as a model. Profiling axons of retinal ganglion cells revealed the presence of miRNAs within axons. The most abundant axonal miRNAs, the miR-181 family and miR-182, exhibit distinct roles in regulating axon guidance in vivo. Loss of function analyses suggests that both miRNA families are required for accurate axonal targeting but involve different mechanisms. Thus, specific axonal microRNAs locally regulate mRNAs contributing to error-free pathfinding

Structural and Mechanistic Investigation of D-Arginine Dehydrogenase and L-Arginine Dehydrogenase from Pseudomonas Aeruginosa

Pseudomonas aeruginosa has a two-enzyme D-arginine conversion system that enables it to utilize D-arginine as a nutrient. The coupled-enzyme system consists of D-arginine dehydrogenase (PaDADH) and L-arginine dehydrogenase (PaLADH). PaDADH has been characterized structurally and mechanistically and established as an FAD-dependent enzyme that catalyzes the oxidation of D-arginine to iminoarginine, which can be non-enzymatically hydrolyzed to ketoarginine and ammonia. A tyrosine 249 to phenylalanine variant of PaDADH (PaDADH-Y249F) was expressed and purified as a mixture of two cofactors, one protein population with regular FAD and one with a green-colored modified FAD. The chemically modified green FAD was investigated through various spectroscopic techniques. The yellow and green protein fractions were investigated structurally to uncover any clues about the modification. Molecular Dynamics and QM/MM were also performed with the wild-type and the variant enzymes to investigate the contribution of protein dynamics and flavin electronics into making the 6-hydroxylation reaction possible in the variant enzyme. PaDADH-Y249F was also expressed and crystallized in the presence of D-arginine. The structure of this incubated enzyme was similar in overall topology to the wild-type enzyme, except for ambiguous electron density around the N5 and C6 atoms of the isoalloxazine moiety. The FAD modified at the N5 position was present in the variant enzyme and the green modified FAD. The presence of two modified flavin cofactors in a single crystal structure is unusual. The second modified FAD was characterized via accurate mass analysis and investigated spectroscopically. While a lot is known about PaDADH, PaLADH has yet to be characterized kinetically. PaLADH is the second enzyme in the coupled-enzyme D-arginine conversion system and catalyzes the reaction of ketoarginine and ammonia to L-arginine using NAD(P)H. PaLADH can also catalyze the reverse reaction converting L-arginine to ketoarginine and ammonia using NAD(P)+ and was characterized using a stopped-flow spectrophotometer. The forward reaction of PaLADH was investigated as the product of the coupled-enzyme reaction by providing the nicotinamide substrate and D-arginine for PaDADH, with PaLADH also being present in the same reaction mix. We also hypothesize that PaLADH and PaDADH might form a protein complex for converting D-arginine to L-arginine, and the complex was probed using analytical ultracentrifugation and size-exclusion chromatography

Price discovery and convergence in the Indian commodities market

Purpose – The purpose of this paper is to examine whether futures markets play a dominant role in the price discovery process. The rate of convergence of information from one market to another is analyzed to infer the efficiency of futures as an effective hedging tool. Design/methodology/approach – The paper uses a two-regime threshold vector autoregression (TVAR) and a two-regime threshold autoregression for six commodities. The regimes (or states) are defined around the expiration week of the futures contract. Findings – This paper finds evidence for price discovery process happening in the futures market in five out of six commodities. However, the rate of convergence of information is slow, particularly in the non-expiration weeks. For copper, gold and silver, the rate of convergence is almost instantaneous during the expiration week of the futures contract affirming the utility of futures contracts as an effective hedging tool. In case of chickpeas, nickel and rubber the convergence worsens during the expiration week indicating the non-usability of futures contract for hedging. Originality/value – This paper extends the framework developed by Garbade et al. by superimposing a two-regime TVAR model to quantify the price discovery process. It is the first paper to analyze the differential impact of price discovery and convergence during expiration week (compared to non-expiration weeks) for the Indian commodities market.Commodity markets, Economic convergence, Futures markets, India, Prices

Inhaled statins to combat COVID-19 – prophylactic and treatment approach

The coronavirus disease 19 (COVID-19) is a highly transmittable and pathogenic viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged in Wuhan, China and spread around the world (WHO, 2020). The genome of the SARS-CoV-2 has been reported over 80% identical to the previous human coronavirus (SARS-like bat CoV) [1]. As of May 2020, more than 5 million people have been affected worldwide with deaths amounting to 333000, the numbers increasing at an alarming rate day by day