1,149 research outputs found

    Characterization of tetracycline modifying enzymes using a sensitive in vivo reporter system

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    Abstract Background Increasing our understanding of antibiotic resistance mechanisms is critical. To enable progress in this area, methods to rapidly identify and characterize antibiotic resistance conferring enzymes are required. Results We have constructed a sensitive reporter system in Escherichia coli that can be used to detect and characterize the activity of enzymes that act upon the antibiotic, tetracycline and its derivatives. In this system, expression of the lux operon is regulated by the tetracycline repressor, TetR, which is expressed from the same plasmid under the control of an arabinose-inducible promoter. Addition of very low concentrations of tetracycline derivatives, well below growth inhibitory concentrations, resulted in luminescence production as a result of expression of the lux genes carried by the reporter plasmid. Introduction of another plasmid into this system expressing TetX, a tetracycline-inactivating enzyme, caused a marked loss in luminescence due to enzyme-mediated reduction in the intracellular Tc concentration. Data generated for the TetX enzyme using the reporter system could be effectively fit with the known Km and kcat values, demonstrating the usefulness of this system for quantitative analyses. Conclusion Since members of the TetR family of repressors regulate enzymes and pumps acting upon almost every known antibiotic and a wide range of other small molecules, reporter systems with the same design as presented here, but employing heterologous TetR-related proteins, could be developed to measure enzymatic activities against a wide range of antibiotics and other compounds. Thus, the assay described here has far-reaching applicability and could be adapted for high-throughput applications

    EffectS of non-nutritive sWeetened beverages on appetITe during aCtive weigHt loss (SWITCH): Protocol for a randomized, controlled trial assessing the effects of non-nutritive sweetened beverages compared to water during a 12-week weight loss period and a follow up weight maintenance period

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    Background Acute and medium-term intervention studies suggest that non-nutritive sweeteners (NNS) are beneficial for weight loss, however there is limited human data on the long-term effects of consuming NNS on weight loss, maintenance, and appetite. Further research is therefore required to elucidate the prolonged impact of NNS consumption on these outcome measures. Methods/design A randomized parallel groups design will be used to assess whether regular NNS beverage intake is equivalent to a water control in promoting weight loss over 12-weeks (weekly weight loss sessions; Phase I), then supporting weight maintenance over 40-weeks (monthly sessions; Phase II) and subsequently independent weight maintenance over 52-weeks (Phase III) in 432 participants. A subset of these participants (n = 116) will complete laboratory-based appetite probe days (15 sessions; 3 sessions each at baseline, at the start of phase I and the end of each phase). A separate subset (n = 50) will complete body composition scans (DXA) at baseline and at the end of each phase. All participants will regularly be weighed and will complete questionnaires and cognitive tasks to assess changes in body weight and appetitive behaviours. Measures of physical activity and biochemical markers will also be taken. Discussion The trial will assess the efficacy of NNS beverages compared to water during a behavioural weight loss and maintenance programme. We aim to understand whether the impact of NNS on weight, dietary adherence and well-being are beneficial or transient and effects on prolonged successful weight loss and weight maintenance through sustained changes in appetite and eating behaviour. Trial registration: Clinical Trials: NCT02591134; registered: 23.10.201

    Invasion dynamics of Asian hornet, Vespa velutina (Hymenoptera: Vespidae) : a case study of a commune in south-west France

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    Asian hornet, Vespa velutina Lepeletier nests were discovered in 2007 in Andernos-les-Bains on the south-west coast of France, 3 years after the first reported sightings in France. The number of nests increased in the commune over the following 7 years, despite local authorities enacting a destruction policy. The nests existed in close proximity to one another leading to a high density of over 10 nests per square kilometre in urban areas. New information on the chosen habitat for nests is presented, and the differences between primary and secondary locations are evident, with primary nests mostly occupying buildings and man-made structures, while secondary nests were found on trees. Using Bayesian inference methods, we fit a basic model to the observational data, which allows us to estimate key demographic parameters. This model fit is highly informative for predicting V. velutina spread and colonisation of other at-risk regions, and suggests that local control has a limited impact on the spread of V. velutina once established within a region

    Historical development of accelerometry measures and methods for physical activity and sedentary behavior research worldwide: A scoping review of observational studies of adults

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    This scoping review identified observational studies of adults that utilized accelerometry to assess physical activity and sedentary behavior. Key elements on accelerometry data collection were abstracted to describe current practices and completeness of reporting. We searched three databases (PubMed, Web of Science, and SPORTDiscus) on June 1, 2021 for articles published up to that date. We included studies of non-institutionalized adults with an analytic sample size of at least 500. The search returned 5686 unique records. After reviewing 1027 full-text publications, we identified and abstracted accelerometry characteristics on 155 unique observational studies (154 cross-sectional/cohort studies and 1 case control study). The countries with the highest number of studies included the United States, the United Kingdom, and Japan. Fewer studies were identified from the continent of Africa. Five of these studies were distributed donor studies, where participants connected their devices to an application and voluntarily shared data with researchers. Data collection occurred between 1999 to 2019. Most studies used one accelerometer (94.2%), but 8 studies (5.2%) used 2 accelerometers and 1 study (0.6%) used 4 accelerometers. Accelerometers were more commonly worn on the hip (48.4%) as compared to the wrist (22.3%), thigh (5.4%), other locations (14.9%), or not reported (9.0%). Overall, 12.7% of the accelerometers collected raw accelerations and 44.6% were worn for 24 hours/day throughout the collection period. The review identified 155 observational studies of adults that collected accelerometry, utilizing a wide range of accelerometer data processing methods. Researchers inconsistently reported key aspects of the process from collection to analysis, which needs addressing to support accurate comparisons across studies

    The identification and neurochemical characterization of central neurons that target parasympathetic preganglionic neurons involved in the regulation of choroidal blood flow in the rat eye using pseudorabies virus, immunolabeling and conventional pathway tracing methods

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    The choroidal blood vessels of the eye provide the main vascular support to the outer retina. These blood vessels are under parasympathetic vasodilatory control via input from the pterygopalatine ganglion (PPG), which in turn receives its preganglionic input from the superior salivatory nucleus (SSN) of the hindbrain. The present study characterized the central neurons projecting to the SSN neurons innervating choroidal PPG neurons, using pathway tracing and immunolabeling. In the initial set of studies, minute injections of the Bartha strain of the retrograde transneuronal tracer pseudorabies virus (PRV) were made into choroid in rats in which the superior cervical ganglia had been excised (to prevent labeling of sympathetic circuitry). Diverse neuronal populations beyond the choroidal part of ipsilateral SSN showed transneuronal labeling, which notably included the parvocellular part of the paraventricular nucleus of the hypothalamus (PVN), the periaqueductal gray, the raphe magnus (RaM), the B3 region of the pons, A5, the nucleus of the solitary tract (NTS), the rostral ventrolateral medulla (RVLM), and the intermediate reticular nucleus of the medulla. The PRV+ neurons were located in the parts of these cell groups that are responsive to systemic blood pressure signals and involved in systemic blood pressure regulation by the sympathetic nervous system. In a second set of studies using PRV labeling, conventional pathway tracing, and immunolabeling, we found that PVN neurons projecting to SSN tended to be oxytocinergic and glutamatergic, RaM neurons projecting to SSN were serotonergic, and NTS neurons projecting to SSN were glutamatergic. Our results suggest that blood pressure and volume signals that drive sympathetic constriction of the systemic vasculature may also drive parasympathetic vasodilation of the choroidal vasculature, and may thereby contribute to choroidal baroregulation during low blood pressure
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