SummaryMutations in the NPHS1 gene, which encodes NEPHRIN, cause congenital nephrotic syndrome, resulting from impaired slit diaphragm (SD) formation in glomerular podocytes. However, methods for SD reconstitution have been unavailable, thereby limiting studies in the field. In the present study, we established human induced pluripotent stem cells (iPSCs) from a patient with an NPHS1 missense mutation, and reproduced the SD formation process using iPSC-derived kidney organoids. The mutant NEPHRIN failed to become localized on the cell surface for pre-SD domain formation in the induced podocytes. Upon transplantation, the mutant podocytes developed foot processes, but exhibited impaired SD formation. Genetic correction of the single amino acid mutation restored NEPHRIN localization and phosphorylation, colocalization of other SD-associated proteins, and SD formation. Thus, these kidney organoids from patient-derived iPSCs identified SD abnormalities in the podocytes at the initial phase of congenital nephrotic disease.

Shiga toxin-producing Escherichia coli (STEC) is one of the major causes of human gastrointestinal disease and has been implicated in sporadic cases and outbreaks of diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome worldwide. In this study, we determined the molecular characteristics and phylogenetic relationship of STEC isolates, and their genetic diversity was compared to that of other E. coli populations. Whole genome sequencing was performed on 132 clinical STEC isolates obtained from the faeces of 129 Dutch patients with gastrointestinal complaints. STEC isolates of this study belonged to 44 different sequence types (STs), 42 serogenotypes and 14 stx subtype combinations. Antibiotic resistance genes were more frequently present in stx1-positive isolates compared to stx2 and stx1 + stx2-positive isolates. The iha, mchB, mchC, mchF, subA, ireA, senB, saa and sigA genes were significantly more frequently present in eae-negative than in eae-positive STEC isolates. Presence of virulence genes encoding type III secretion proteins and adhesins was associated with isolates obtained from patients with bloody diarrhoea. Core genome phylogenetic analysis showed that isolates clustered according to their ST or serogenotypes irrespective of stx subtypes. Isolates obtained from patients with bloody diarrhoea were from diverse phylogenetic backgrounds. Some STEC isolates shared common ancestors with non-STEC isolates. Whole genome sequencing is a powerful tool for clinical microbiology, allowing high-resolution molecular typing, population structure analysis and detailed molecular characterization of strains. STEC isolates of a substantial genetic diversity and of distinct phylogenetic groups were observed in this study.

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Environmental variables influence Faecal Indicator Bacteria (FIB) in surface water. Understanding that influence is important, because presence of FIB, which are an indication of faecal contamination, means that harmful pathogens could be present that could also be influenced by environmental variables. Although some recent studies have focused on this topic, most of this work has been conducted in developed countries. Similar studies in developing countries and in a (sub)tropical climate are lacking. In this study we assess the influence of environmental variables on fluctuations in FIB concentrations of the Betna River in southwest Bangladesh that floods almost every year. Monthly water samples from five locations along Betna River were tested for FIB (E. coli and enterococci) in 2014-2015. A linear regression model was developed to assess the effect of the environmental variables on FIB concentrations. The study revealed increased FIB concentrations during wet weather conditions. Precipitation and water temperature were positively correlated with FIB concentrations. Water temperature was positively correlated, because the warm May to September period coincides with frequent precipitation. Precipitation increases manure release from land to surface water. The regression model explains nearly half of the variability in FIB concentrations (R 2 of 0.46 for E. coli and 0.48 for enterococci). This study indicates that increased precipitation combined with higher water temperature, as is expected in this region with climate change, likely increases FIB concentrations. Waterborne pathogens are expected to respond similarly to these environmental changes, indicating that disease outbreaks could well become more frequent and severe.

Trace metals accumulation in soil irrigated with polluted water and human health risk from vegetable consumption was assessed based on the data available in the literature on metals pollution of water, soil, sediment and vegetables from the cites of Bangladesh. The quantitative data on metal concentrations, their contamination levels and their pollution sources have not been systematically gathered and studied so far. The data on metal concentrations, sources, contamination levels, sample collection and analytical tools used were collected, compared and discussed. The USEPA-recommended method for health risk assessment was used to estimate human risk from vegetable consumption. Concentrations of metals in water were highly variable, and the mean concentrations of Cd, Cr, Cu and As in water were found to be higher than the FAO irrigation water quality standard. In most cases, mean concentrations of metals in soil were higher than the Bangladesh background value. Based on geoaccumulation index (I ) values, soils of Dhaka city are considered as highly contaminated. The I shows Cd, As, Cu, Ni, Pb and Cr contamination of agricultural soils and sediments of the cities all over the Bangladesh. Polluted water irrigation and agrochemicals are identified as dominant sources of metals in agricultural soils. Vegetable contamination by metals poses both non-carcinogenic and carcinogenic risks to the public. Based on the results of the pollution and health risk assessments, Cd, As, Cr, Cu, Pb and Ni are identified as the priority control metals and the Dhaka city is recommended as the priority control city. This study provides quantitative evidence demonstrating the critical need for strengthened wastewater discharge regulations in order to protect residents from heavy metal discharges into the environment.

This study was conducted to assess the bacteriological quality of alternative drinking water supply options in southwest coastal areas of Bangladesh. A total of 90 water samples were collected during both dry and wet seasons from household based rainwater harvesting systems (RWHSs), community based rain water harvesting systems (CRWHSs), pond-sand filters (PSFs) and ponds. The samples were evaluated for faecal coliform, Escherichia coli and Heterotrophic Plate Count, as well as Vibrio cholerae, Salmonella spp., Shigella spp. and Pseudomonas spp. Physico-chemical parameters (pH, electrical conductivity, and color) were also examined. In addition, sanitary inspections were conducted to identify faecal contamination sources. All options showed varying degrees of indicator bacterial contamination. The median E. coli concentrations measured for RWHSs, CRWHSs, PSFs, and ponds were 16, 7, 11, and 488 cfu/100 ml during the wet season, respectively. Vibrio cholerae O1/O139, Salmonella and Shigella spp. were not found in any samples. However, Vibrio cholerae Non-O1/Non-O139 and Pseudomonas spp. were isolated from 74.4% and 91.1% of the water samples collected during the wet season. A maximum pH of 10.4 was found in CRWHSs. Estimation of the disease burden for all options in disability adjusted life years (DALYs) showed an increased disease burden during the wet season. According to sanitary inspections, poor maintenance and unprotected ponds were responsible for rainwater and PSF water contamination, respectively.The findings of the present study suggest that alternative drinking water supply options available in southwest coastal Bangladesh pose a substantial risk to public health.

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