Prevalence, virulence genes and Antimicrobial Resistance of Shiga-toxigenic E.coli in diarrhoea patients from Kitale, Kenya

Introduction: Shiga toxin-producing Escherichia coli (STEC) are among the most important causes of food-borne diseases. They cause illnesses ranging from mild diarrhea to more severe conditions that may progress to hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). The burden of STEC in patients with diarrheal illness in Kitale county referral hospital, Trans-Nzoia County had not been established.Objectives: To determine the prevalence of STEC, its associated virulence genes and antimicrobial resistance among patients seeking treatment for diarrhoeal illness at Kitale County Referral Hospital.Methods: Stool samples from patients seeking treatment for diarrheal illness and had consented to participate in the study were collected and cultured for enteric bacteria. Suspect E.coli isolates were further identified using conventional biochemical methods. Conventional multiplex PCR targeting Shiga toxins (stx1, stx2, hlyA and attaching and effacing mechanisms (eaeA) were used to detect STEC virulence markers responsible for the Pathogenicity of STEC infection among other E.coli pathotypes.Results: A total of 295 participants were enrolled; median age 120 months (IQR: 36-312). 39 %( 115) were children aged <5yearsof whom 54% (160) were females. The prevalence of pathogenic E.coli was 19%56/295 and STEC was the most prevalent among E.coli pathotypes at5.4%16/295. The Stx2 gene and the Stx1/Stx2/hlyAcombination were the most prevalent in the STEC strains. The virulence genes (Stx1, Stx2, eaeA* and HlyA*)were observed in 13, 19, 9 and 14 in STEC isolates respectively.The most common gene was Stx2 and combinations of (Stx1+Stx2+hlyA)genes. Antimicrobial resistance to commonly prescribed antibiotics: chloramphenicol, ampicillin 10μg, erythromycin15μg, gentamicin10μg, ciprofloxacin 5μg, tetracycline 30μg, Trimethoprim/Sulfamethoxazole 25 μg, Cefotaxime 30 μg, furazolidine (8μg) and nalidixic acid 30 μg. were observed for all E.coli isolates except one (1.8%; 95% CI=0.1-9.6%). No isolates among STEC showed resistance to Furazolidine drug. However, Trimethoprim / Sulphurmethoxazole) was the drug which exhibited the highest resistance at (94%, 95% CI 70 to 99%).Conclusion and recommendation: Prevalence of STEC was 5.4%, (Stx1/Stx2/hlyA) virulence genes combination was the most common. High resistance to commonly prescribed antibiotics were observed in E.coli isolates and may be an existing problem that needs to be further research investigation.Keywords: Shiga-Toxigenic Escherichia coli (STEC), antimicrobial resistance, Kitale County referral hospitalAfr J Health Sci. 2017; 30(2):105-11

Comprehensive functional profiling of long non-coding RNAs through a novel pan-cancer integration approach and modular analysis of their protein-coding gene association networks

Background Long non-coding RNAs (lncRNAs) are emerging as crucial regulators of cellular processes in diseases such as cancer, although the functions of most remain poorly understood. To address this, here we apply a novel strategy to integrate gene expression profiles across 32 cancer types, and cluster human lncRNAs based on their pan-cancer protein-coding gene associations. By doing so, we derive 16 lncRNA modules whose unique properties allow simultaneous inference of function, disease specificity and regulation for over 800 lncRNAs. Results Remarkably, modules could be grouped into just four functional themes: transcription regulation, immunological, extracellular, and neurological, with module generation frequently driven by lncRNA tissue specificity. Notably, three modules associated with the extracellular matrix represented potential networks of lncRNAs regulating key events in tumour progression. These included a tumour-specific signature of 33 lncRNAs that may play a role in inducing epithelial-mesenchymal transition through modulation of TGFβ signalling, and two stromal-specific modules comprising 26 lncRNAs linked to a tumour suppressive microenvironment and 12 lncRNAs related to cancer-associated fibroblasts. One member of the 12-lncRNA signature was experimentally supported by siRNA knockdown, which resulted in attenuated differentiation of quiescent fibroblasts to a cancer-associated phenotype. Conclusions Overall, the study provides a unique pan-cancer perspective on the lncRNA functional landscape, acting as a global source of novel hypotheses on lncRNA contribution to tumour progression

Optimal alpha-Chymotrypsin-Catalyzed Synthesis of N-Ac-Phe-Gly-NH2

N-Acetyl-phenylalanine-glycinamide (N-Ac-Phe-Gly-NH2), a type of dipeptide derivative, was synthesized from N-acetyl phenylalanine ethyl ester and glycinamide and catalyzed by alpha-chymotrypsin, a protease, in a biphasic system. Response surface methodology with a four-factor, five-level central composite rotatable design was employed to evaluate the effects of selected parameters that included incubation time, reaction temperature, enzyme activity, and pH level on the yield of the dipeptide derivative. The results indicated that pH significantly affected the yield of N-Ac-Phe-Gly-NH2 In a ridge max analysis, the optimum condition for this synthesis included an incubation time of 30.9 min, a reaction temperature of 35.8 degrees C, an enzyme activity of 159.2 U, and a pH of 8.98. The predicted and the actual (experimental) yields were 98.0 and 95.1%, respectively

Optimal covalent immobilization of alpha-chymotrypsin on Fe3O4-chitosan nanoparticles

This study investigated the immobilization of alpha-chymotrypsin onto magnetic Fe3O4-chitosan (alpha-chymotrypsin-Fe3O4-CS) nanoparticles by covalent binding. The response surface methodology (RSM) with a 3-factor-3-level Box-Behnken experimental design was employed to evaluate the effects of the manipulated variables, including the immobilization time, temperature, and pH, on the enzyme activity. The results indicate that the immobilized temperature and pH significantly affected enzyme activity. In a ridge max analysis, the optimal condition for alpha-chymotrypsin immobilization included a reaction temperature of 21.7 degrees C, a pH of 7.6, and an incubation time of 1.1 h. The predicted and the experimental immobilized enzyme activities were 354 and 347 +/- 46.5 U/g-support, respectively, under the optimal condition. Besides, the synthesis reactions of the dipeptide derivative using the free and immobilized alpha-chymotrypsin were compared. The yields of the dipepticle derivative via the free or immobilized alpha-chymotrypsin catalyzed were almost the same. The alpha-chymotrypsin-Fe3O4-CS nanoparticles exhibited a good acid-resisting ability and the less reaction time was required for dipeptide synthesis. After twelve repeated uses in dipeptide synthesis, the immobilized alpha-chymotrypsin still retained over 60% of its original activity. The magnetic alpha-chymotrypsin-Fe3O4-CS nanoparticles can be easily recovered by magnetic field will have potential application in industry. (C) 2012 Elsevier By. All rights reserved

Mapping subnational HIV mortality in six Latin American countries with incomplete vital registration systems

Background Human immunodeficiency virus (HIV) remains a public health priority in Latin America. While the burden of HIV is historically concentrated in urban areas and high-risk groups, subnational estimates that cover multiple countries and years are missing. This paucity is partially due to incomplete vital registration (VR) systems and statistical challenges related to estimating mortality rates in areas with low numbers of HIV deaths. In this analysis, we address this gap and provide novel estimates of the HIV mortality rate and the number of HIV deaths by age group, sex, and municipality in Brazil, Colombia, Costa Rica, Ecuador, Guatemala, and Mexico. Methods We performed an ecological study using VR data ranging from 2000 to 2017, dependent on individual country data availability. We modeled HIV mortality using a Bayesian spatially explicit mixed-effects regression model that incorporates prior information on VR completeness. We calibrated our results to the Global Burden of Disease Study 2017. Results All countries displayed over a 40-fold difference in HIV mortality between municipalities with the highest and lowest age-standardized HIV mortality rate in the last year of study for men, and over a 20-fold difference for women. Despite decreases in national HIV mortality in all countries—apart from Ecuador—across the period of study, we found broad variation in relative changes in HIV mortality at the municipality level and increasing relative inequality over time in all countries. In all six countries included in this analysis, 50% or more HIV deaths were concentrated in fewer than 10% of municipalities in the latest year of study. In addition, national age patterns reflected shifts in mortality to older age groups—the median age group among decedents ranged from 30 to 45 years of age at the municipality level in Brazil, Colombia, and Mexico in 2017. Conclusions Our subnational estimates of HIV mortality revealed significant spatial variation and diverging local trends in HIV mortality over time and by age. This analysis provides a framework for incorporating data and uncertainty from incomplete VR systems and can help guide more geographically precise public health intervention to support HIV-related care and reduce HIV-related deaths