Introduction: Community-acquired infections due to extended-spectrum beta-lactamase (ESBL) producing Escherichia coli are rising worldwide, resulting in increased morbidity, mortality, and healthcare costs, especially where poor sanitation and inadequate hygienic practices are very common. Objective: This study was conducted to investigate the prevalence and characterization of multidrug-resistant (MDR) and ESBL-producing E. coli in drinking water samples collected from Rohingya camps, Bangladesh. Methods: A total of 384 E. coli isolates were analyzed in this study, of which 203 were from household or point-of-use (POU) water samples, and 181 were from source water samples. The isolates were tested for virulence genes, ESBL-producing genes, antimicrobial susceptibility by VITEK 2 assay, plasmid profiling, and conjugal transfer of AMR genes. Results: Of the 384 E. coli isolates tested, 17% (66/384) were found to be ESBL producers. The abundance of ESBL-producers in source water contaminated with E. coli was observed to be 14% (27/181), whereas, 19% (39/203) ESBL producers was found in household POU water samples contaminated with E. coli. We detected 71% (47/66) ESBL-E. coli to be MDR. Among these 47 MDR isolates, 20 were resistant to three classes, and 27 were resistant to four different classes of antibiotics. Sixty-four percent (42/66) of the ESBL producing E. coli carried 1 to 7 plasmids ranging from 1 to 103 MDa. Only large plasmids with antibiotic resistance properties were found transferrable via conjugation. Moreover, around 7% (29/384) of E. coli isolates harbored at least one of 10 virulence factors belonging to different E. coli pathotypes. Mahmud et al. ESBL-Producing E. coli in Drinking Water Conclusions: The findings of this study suggest that the drinking water samples analyzed herein could serve as an important source for exposure and dissemination of MDR, ESBL-producing and pathogenic E. coli lineages, which therewith pose a health risk to the displaced Rohingya people residing in the densely populated camps of Bangladesh.
Excessive body fat and the related dysmetabolic diseases affect both developed and developing countries. The aim of this study was to investigate the beneficial role of a bacterial culture supernatant (hereafter: BS) of Lactobacillus and Bifidobacterium and their potential mechanisms of action on white-fat browning and lipolysis. For selection of four candidates among 55 Lactic acid producing bacteria (LAB) from human infant faeces, we evaluated by Oil Red O staining and Ucp1 mRNA quantitation in 3T3-L1 preadipocytes. The expression of browning and lipolysis markers was examined along with in vitro assays. The possible mechanism was revealed by molecular and biological experiments including inhibitor and small interfering RNA (siRNA) assays. In a mouse model, physiological, histological, and biochemical parameters and expression of some thermogenesis-related genes were compared among six experimental groups fed a high-fat diet and one normal-diet control group. The results allow us to speculate that BS treatment promotes browning and lipolysis both in vitro and in vivo. Moreover, the BS may activate thermogenic programs via a mechanism involving PKA-CREB signaling in 3T3-L1 cells. According to our data, we can propose that two LAB strains, Bifidobacterium longum DS0956 and Lactobacillus rhamnosus DS0508, may be good candidates for a dietary supplement against obesity and metabolic diseases; however, further research is required for the development as dietary supplements or drugs.
BackgroundSafe water is essential for life but unsafe for human consumption if it is contaminated with pathogenic microorganisms. An acceptable quality of water supply (adequate, safe and accessible) must be ensured to all human beings for a healthy life.MethodsWe collected and analyzed a total of 12,650 drinking water samples, for the presence of Escherichia coli and faecal coliforms, from a large habitation of the displaced Rohingya population comprising of about 1.16 million people living within 4 km2.ResultsWe found that 28% (n = 893) water samples derived from tubewells were contaminated with faecal coliforms and 10.5% (n = 333) were contaminated with E. coli; also, 73.96% (n = 4644) samples from stored household sources (at point of use—POU) were found contaminated with faecal coliforms while 34.7% (n = 2179) were contaminated with E. coli. It was observed that a higher percentage of POU samples fall in the highest risk category than that of their corresponding sources.ConclusionsFrom our findings, it appears that secondary contamination could be a function of very high population density and could possibly occur during collection, transportation, and storage of water due to lack of knowledge of personal and domestic hygiene. Hence, awareness campaign is necessary, and the contaminated sources should be replaced. Further, the POU water should be treated by a suitable method.
Lipid accumulation in white adipose tissue is the key contributor to the obesity and orchestrates numerous metabolic health problems such as type 2 diabetes, hypertension, atherosclerosis, and cancer. Nonetheless, the prevention and treatment of obesity are still inadequate. Recently, scientists found that brown adipose tissue (BAT) in adult humans has functions that are diametrically opposite to those of white adipose tissue and that BAT holds promise for a new strategy to counteract obesity. In this study, we evaluated the potential of sinapic acid (SA) to promote the thermogenic program and lipolysis in BAT. SA treatment of brown adipocytes induced the expression of brown-adipocyte activation-related genes such as Ucp1, Pgc-1, and Prdm16. Furthermore, structural analysis and western blot revealed that SA upregulates protein kinase A (PKA) phosphorylation with competitive inhibition by a pan-PKA inhibitor, H89. SA binds to the adenosine triphosphate (ATP) site on the PKA catalytic subunit where H89 binds specifically. PKA-cat-1 gene-silencing experiments confirmed that SA activates the thermogenic program via a mechanism involving PKA and cyclic AMP response element-binding protein (CREB) signaling. Moreover, SA treatment promoted lipolysis via a PKA/p38-mediated pathway. Our findings may allow us to open a new avenue of strategies against obesity and need further investigation. [BMB Reports 2020; 53(3): 142-147] BMB Rep. 2020; 53(3): 142-147 www.bmbreports.org
Introduction: In Bangladesh, human sludge from dry pit latrines is commonly applied directly to agricultural lands as manure. This study was conducted to investigate the presence of antibiotic resistance, virulence factors and plasmid contents of E. coli strains isolated from sludge samples. Methodology: E. coli were isolated from human feces from closed pit latrines and identified by culture method. Antibiotic susceptibility patterns of the isolates were determined by Standard Kirby-Bauer disk diffusion method. Pathogenic genes and antibiotic resistance genes of ESBL producing isolates were determined by PCR assay. Results: Of the 34 samples tested, 76.5% contained E. coli. Of 72 E. coli isolates, 76.4% were resistant to at least one of the 12 antibiotics tested and 47.2% isolates were resistant to three or four classes of antibiotics. Around 18% isolates were extended spectrum β- lactamase producing and of them 6 were positive for blaTEM specific gene, 4 for blaCTX-M gene, 1 for blaOXA gene and 2 for both blaTEM and blaCTX-M genes. Moreover, among 72 isolates, 4.2% carried virulence genes of enterotoxigenic E. coli; two isolates were positive for st and one was positive for both st and lt genes. In addition, 59.7% of the isolates contained plasmids (range 1.4 to 140 MDa) of which 19.5% isolates contained a single plasmid and 40.2% contained multiple plasmids. Conclusions: The presence of pathogenic, drug resistant E. coli in human sludge necessitates a regular surveillance before using as a biofertilizer.
Natural pterocarpan Medicarpin (Med) has been shown to have various beneficial biological roles, including inhibition of osteoclastogenesis, stimulation of bone regeneration and induction of apoptosis. However, the effect of the Med on lipolysis in adipocytes has not been reported. Here, we show the effect of Med on lipolysis in different mouse adipocytes and elucidate the underlying mechanism. We observed that Med treatment promoted release of glycerol in the media. Differentiated mouse brown adipose tissue cells were treated with Med. RNA-Seq analysis was performed to elucidate the effect of med and subsequently was confirmed by qRT-PCR and western blotting analyses. Med treatment increased both protein and gene expression levels of hormone-sensitive lipase (Hsl) and adipose triglyceride lipase (Atgl), which are two critical enzymes necessary for lipolysis. Mechanistic study showed that Med activates Protein Kinase A (PKA) and phosphorylates Hsl at PKA target position at Serine660. Silencing of PKA gene by short interfering RNA attenuated the Med-induced increase in glycerol release and Hsl phosphorylation. The results unveil that Med boosts lipolysis via a PKA-dependent pathway in adipocytes and may provide a possible avenue of further research of Med mediated reduction of body fat.
In this study, we investigated the effect of Biochanin A (BioA), an O‐methylated isoflavone on the brown‐fat phenotype formation and on the associated thermogenic program including mitochondrial biogenesis and lipolysis in C3H10T1/2 MSCs. Our data demonstrates that Treatment with BioA in an adipogenic differentiation cocktail induced formation of brown‐fat–like adipocytes from C3H10T1/2 MSCs without treatment with a known browning inducer (rosiglitazone or T3) at an early stage of differentiation. The formation of brown‐fat–like adipocytes by BioA treatment was evidenced by upregulation of key thermogenic markers: Ucp1, Pgc1α, Prdm16, and Pparγ. BioA also increased the expression of beige (Cd137 and Fgf21) and brown (Elovl3 and Zic1)‐specific markers. Additionally, BioA treatment promoted mitochondrial biogenesis, judging by the upregulation of genes; Cox8b, Cidea, Dio2, Sirt1, Opa1, and Fis1. BioA treatment increased the amount of mitochondrial DNA and its encoded proteins: oxidative phosphorylation complexes (I–V); this change was associated with high oxygen consumption by C3H10T1/2 MSCs. A small‐interfering‐RNA–induced gene knockdown and experiments with dorsomorphin‐driven competitive inhibition revealed that BioA exerts the thermogenic action via activation of AMPK signaling. Our study shows the mechanism of BioA‐induced promotion of a brown‐fat phenotype. Nonetheless, clinical research is necessary to validate BioA as a brown‐fat‐like signature inducer.
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