Vibrio cholerae is a leading waterborne pathogen worldwide. Continuous monitoring of V. cholerae contamination in aquatic products and identification of risk factors are crucial for assuring food safety. In this study, we determined the virulence, antimicrobial susceptibility, heavy metal tolerance, and genetic diversity of 400 V. cholerae isolates recovered from commonly consumed freshwater fish (Aristichthys nobilis, Carassius auratus, Ctenopharyngodon idellus, and Parabramis pekinensis) collected in July and August of 2017 in Shanghai, China. V. cholerae has not been previously detected in the half of these fish species. The results revealed an extremely low occurrence of pathogenic V. cholerae carrying the major virulence genes ctxAB (0.0%), tcpA (0.0%), ace (0.0%), and zot (0.0%). However, high incidence of virulence-associated genes was observed, including the RTX toxin gene cluster (rtxA-D) (83.0–97.0%), hlyA (87.8%), hapA (95.0%), and tlh (76.0%). Meanwhile, high percentages of resistance to antimicrobial agents streptomycin (65.3%), ampicillin (44.5%), and rifampicin (24.0%) were observed. Approximately 30.5% of the isolates displayed multidrug resistant (MDR) phenotypes with 42 resistance profiles, which were significantly different among the four fish species (MARI, P = 0.001). Additionally, tolerance of isolates to heavy metals Hg2+ (49.3%), Zn2+ (30.3%), and Pb2+ (12.0%) was observed. The enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR)-based fingerprinting of the 400 V. cholerae isolates revealed 328 ERIC-genotypes, which demonstrated a large degree of genomic variation among the isolates. Overall, the results of this study support the need for food safety risk assessment of aquatic products.Electronic supplementary materialThe online version of this article (10.1007/s11356-019-05287-8) contains supplementary material, which is available to authorized users.
Recent experimental results have shown that active enzymes can diffuse faster when they are in the presence of their substrates. Fluorescence correlation spectroscopy (FCS), which relies on analyzing the fluctuations in fluorescence intensity signal to measure the diffusion coefficient of particles, has typically been employed in most of the prior studies. However, flaws in the FCS method, due to its high sensitivity to the environment, have recently been evaluated, calling the prior diffusion results into question. It behooves us to adopt complementary and direct methods to measure the mobility of enzymes in solution. Herein, we use a novel technique of direct single-molecule imaging to observe the diffusion of single enzymes. This technique is less sensitive to intensity fluctuations and gives the diffusion coefficient directly based on the trajectory of the enzymes. Our measurements recapitulate that enzyme diffusion is enhanced in the presence of its substrate and find that the relative increase in diffusion of a single enzyme is even higher than those previously reported using FCS. We also use this complimentary method to test if the total enzyme concentration affects the relative increase in diffusion and if enzyme oligomerization state changes during catalytic turnover. We find that the diffusion increase is independent of the total background concentration of enzyme and the catalysis of substrate does not change the oligomerization state of enzymes.
Accumulated pieces of evidence have proved the beneficial effects of melatonin on myocardial ischemia/reperfusion (MI/R) injury, and these effects were largely dependent on melatonin membrane receptor activation. In humans and other mammals, there are two types of melatonin receptors, including the melatonin receptor 1 (MT1, melatonin receptor 1a or MTNR1A) and melatonin receptor 1 (MT2, melatonin receptor 1b or MTNR1B) receptor subtypes. However, which receptor mediates melatonin‐conferred cardioprotection remains unclear. In this study, we employed both loss‐of‐function and gain‐of‐function approaches to reveal the answer. Mice (wild‐type; MT1 or MT2 silencing by in vivo minicircle vector; and those overexpressing MT1 or MT2 by in vivo AAV9 vector) were exposed to MI/R injury. Both MT1 and MT2 were present in wild‐type myocardium. MT2, but not MT1, was essentially upregulated after MI/R Melatonin administration significantly reduced myocardial injury and improved cardiac function after MI/R Mechanistically, melatonin treatment suppressed MI/R‐initiated myocardial oxidative stress and nitrative stress, alleviated endoplasmic reticulum stress and mitochondrial injury, and inhibited myocardial apoptosis. These beneficial actions of melatonin were absent in MT2‐silenced heart, but not the MT1 subtype. Furthermore, AAV9‐mediated cardiomyocyte‐specific overexpression of MT2, but not MT1, mitigated MI/R injury and improved cardiac dysfunction, which was accompanied by significant amelioration of oxidative stress, endoplasmic reticulum stress, and mitochondrial dysfunction. Mechanistically, MT2 protected primary cardiomyocytes against hypoxia/reoxygenation injury via MT2/Notch1/Hes1/RORα signaling. Our study presents the first direct evidence that the MT2 subtype, but not MT1, is a novel endogenous cardiac protective receptor against MI/R injury. Medications specifically targeting MT2 may hold promise in fighting ischemic heart disease.
The aim of this study was to evaluate whether Schisandra chinensis extract (SCE) administration influences chronic unpredictable mild stress (CUMS)-induced depression and cognitive impairment, and explores underlying mechanisms. Sucrose preference test (SPT) and forced swimming test (FST) were used for assessing depressive symptoms, and Y-maze, Morris water maze were used for evaluating cognition processes. The results showed that CUMS (4 weeks) was effective in producing both depression and memory deficits in mice. Additionally, CUMS exposure significantly decreased brain derived neurotrophic factor (BDNF) levels in hippocampus as indicated by ELISA, immunohistochemistry and immunofluorescence assays, accompanied by down-regulated tyrosine kinase receptor B (TrkB)/cAMP-response element binding protein (CREB)/extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3 kinase (PI3K)/ protein kinase B (AKT)/ glycogen synthase kinase-3β (GSK-3β) signaling pathways. Chronic administration of SCE (600 or 1200 mg/kg, i.g.) significantly prevented all these CUMS-induced behavioral and biochemical alterations. It suggested that SCE could improve the depression-like emotional status and associated cognitive deficits in CUMS mice, which might be mediated by regulation of BDNF levels in hippocampus, as well as up-regulating of TrkB/CREB/ERK and PI3K/AKT/GSK-3β pathways.
Background: Individual clinical trials suggested that when treated with probiotic foods or supplements with Lactobacillus and Bifidobacterium, specific symptoms of metabolic syndrome (MetS) could be alleviated, but the results have been inconclusive. Aims: The objective of the present meta-analysis was to use anthropometric and biochemical as indicators to evaluate the efficacy of using these probiotic foods or supplements among individuals with MetS. Methods: PubMed, Cochrane Library, and CINAHL Plus were used to collect randomized controlled trials (RCTs) studies published from January 2000 to January 2018. Studies were included if they had at least one of the following outcome measurements: body mass index (BMI), waist circumference, hip circumference, waist-to-hip ratio, body fat mass (BFM), body fat percentage (BFP), systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting glucose, fasting insulin, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides, and/or hemoglobin A1c (HbA1c). Results: The 356 records were identified during the literature search, of which only 18 met the selection criteria. The 18 RCTs with a total of 1,544 participants were included in this analysis. This meta-analysis indicated that there were no significant differences of BMI, BFM, waist circumference, hip circumference, waist-to-hip ratio, SBP, DBP, fasting glucose, fasting insulin, TC, HDL-C, HbA1c, or triglycerides between the intervention and control groups. Significant standardized mean net differences were found in the BFP and LDL-C between the intervention and control groups. Conclusions: The results indicated that probiotic food and supplement with Lactobacillus and Bifidobacterium could be used as interventions to improve specific anthropometric and biochemical outcomes among individuals with MetS. However, probiotic treatment alone could not reduce overall health risks. In addition, there were methodological drawbacks among reviewed studies, and further research is needed.
Virulence-associated gene Type I: ctxAB + tcp + hapA + mshA + pilA + tlh + ; Type II: ctxAB − tcp − hapA + mshA + pilA − tlh + ; Type III: ctxAB − tcp − hapA + mshA − pilA + tlh + ; Type IV: ctxAB − tcp − hapA + mshA − pilA − tlh + ; Type V: ctxAB − tcp − hapA + mshA − pilA − tlh − ; Type VI: ctxAB − tcp − hapA − mshA − pilA − tlh − .
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