Caractérisation des mécanismes d'action de la kinase mTOR et des acides gras polyinsaturés sur la transcription du gène de la stéroyl CoA désaturase-1 au niveau hépatique

Une des enzymes clés de la lipogenèse hépatique, la stéaroyl-CoA désaturase 1 (SCD1), qui est impliquée dans la biosynthèse des acides gras monoinsaturés, semble jouer un rôle important dans le développement de l'obésité et de ses désordres métaboliques associés. En fait, une modification du ratio d'acides gras saturés versus acides gras mono-insaturés a été impliquée dans le développement de l'obésité, du diabète de type II et des maladies cardiovasculaires. L'expression de Scd1 est sous la dépendance de diverses hormones qui sont fortement modulées par la diète. L'insuline, dont la concentration augmente après un repas, permet d'activer la transcription du gène Scd1, tandis qu'une diète riche en acides gras polyinsaturés (PUFAs) permet d'inhiber son effet. La régulation de Scd1 est accomplie, entre autres, par la présence d'un élément de réponse à l'insuline sur le promoteur qui fixe les facteurs de transcription (TF) SREBP-1 et NF-Y et par l'activation de voies de signalisation spécifiques impliquant la voie PI3-K/mTOR. Ce projet vise à établir les mécanismes d'action des PUFAs et leurs liens potentiels avec PI3-K/mTOR au niveau hépatique. Grâce à des analyses de retard sur gel, nous avons démontré que l'insuline régulait la fixation de ces TFs sur le promoteur Scd1 et que cette fixation était modulée par les inhibiteurs de cette voie et par l'acide arachidonique. De plus, nous avons évalué par PCR en temps réel le rôle de l'insuline, et donc PI3-K/mTOR, ainsi que celui de l'acide arachidonique sur le niveau d'expression de Scdl et de ses TFs. Nos résultats démontrent au niveau hépatique que la voie PI3-K/mTOR cible des TFs spécifiques sur le promoteur Scd1 activant ainsi l'expression de ce gène en réponse à l'insuline et que les \ud PUFAs permettent de contrer ses effets. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Lipogenèse de nova, SCD1, Insuline, mTOR, PUFA, SREBP-1, NF-Y

Pseudomonas aeruginosa in premise plumbing of large buildings.

International audiencePseudomonas aeruginosa is an opportunistic bacterial pathogen that is widely occurring in the environment and is recognized for its capacity to form or join biofilms. The present review consolidates current knowledge on P. aeruginosa ecology and its implication in healthcare facilities premise plumbing. The adaptability of P. aeruginosa and its capacity to integrate the biofilm from the faucet and the drain highlight the role premise plumbing devices can play in promoting growth and persistence. A meta-analysis of P. aeruginosa prevalence in faucets (manual and electronic) and drains reveals the large variation in device positivity reported and suggest the high variability in the sampling approach and context as the main reason for this variation. The effects of the operating conditions that prevail within water distribution systems (disinfection, temperature, and hydraulic regime) on the persistence of P. aeruginosa are summarized. As a result from the review, recommendations for proactive control measures of water contamination by P. aeruginosa are presented. A better understanding of the ecology of P. aeruginosa and key influencing factors in premise plumbing are essential to identify culprit areas and implement effective control measure

Controlling Legionella pneumophila in Showerheads: Combination of Remedial Intervention and Preventative Flushing

RÉSUMÉ: Shock chlorination and remedial flushing are suggested to address Legionella pneumophila (Lp) contamination in buildings or during their (re)commissioning. However, data on general microbial measurements (adenosine tri-phosphate [ATP], total cell counts [TCC]), and the abundance of Lp are lacking to support their temporary implementation with variable water demands. In this study, the weekly short-term (3-week) impact of shock chlorination (20–25 mg/L free chlorine, 16 h) or remedial flushing (5-min flush) combined with distinct flushing regimes (daily, weekly, stagnant) was investigated in duplicates of showerheads in two shower systems. Results showed that the combination of stagnation and shock chlorination prompted biomass regrowth, with ATP and TCC in the first draws reaching large regrowth factors of 4.31–7.07-fold and 3.51–5.68-fold, respectively, from baseline values. Contrastingly, remedial flushing followed by stagnation generally resulted in complete or larger regrowth in Lp culturability and gene copies (gc). Irrespective of the intervention, daily flushed showerheads resulted in significantly (p < 0.05) lower ATP and TCC, as well as lower Lp concentrations than weekly flushes, in general. Nonetheless, Lp persisted at concentrations ranging from 11 to 223 as the most probable number per liter (MPN/L) and in the same order of magnitude (103–104 gc/L) than baseline values after remedial flushing, despite daily/weekly flushing, unlike shock chlorination which suppressed Lp culturability (down 3-log) for two weeks and gene copies by 1-log. This study provides insights on the most optimal short-term combination of remedial and preventative strategies that can be considered pending the implementation of suitable engineering controls or building-wide treatment

Pseudomonas aeruginosa in premise plumbing of large buildings

Pseudomonas aeruginosa is an opportunistic bacterial pathogen that is widely occurring in the environment and is recognized for its capacity to form or join biofilms. The present review consolidates current knowledge on P.aeruginosa ecology and its implication in healthcare facilities premise plumbing. The adaptability of P.aeruginosa and its capacity to integrate the biofilm from the faucet and the drain highlight the role premise plumbing devices can play in promoting growth and persistence. A meta-analysis of P.aeruginosa prevalence in faucets (manual and electronic) and drains reveals the large variation in device positivity reported and suggest the high variability in the sampling approach and context as the main reason for this variation. The effects of the operating conditions that prevail within water distribution systems (disinfection, temperature, and hydraulic regime) on the persistence of P.aeruginosa are summarized. As a result from the review, recommendations for proactive control measures of water contamination by P.aeruginosa are presented. A better understanding of the ecology of P.aeruginosa and key influencing factors in premise plumbing are essential to identify culprit areas and implement effective control measures

Impact of stagnation and sampling volume on water microbial quality monitoring in large buildings

Microbial drinking water quality can be altered in large buildings, especially after stagnation. In this study, bacterial profiles were generated according to the stagnation time and the volume of water collected at the tap. Successive volumes of cold and hot water were sampled after controlled stagnation periods. Bacterial profiles revealed an important decline (> 2 log) in culturable cells in the first 500 mL sampled from the hot and cold water systems, with a steep decline in the first 15 mL. The strong exponential correlation (R2 >/= 0.97) between the culturable cell counts in water and the pipe surface-to-volume ratio suggests the biofilm as the main contributor to the rapid increase in suspended culturable cells measured after a short stagnation of one-hour. Results evidence the contribution of the high surface-to-volume ratio at the point of use and the impact of short stagnation times on the increased bacterial load observed. Simple faucets with minimal internal surface area should be preferred to minimize surface area. Sampling protocol, including sampling volume and prior stagnation, was also shown to impact the resulting culturable cell concentration by more than 1000-fold. Sampling a smaller volume on first draw after stagnation will help maximize recovery of bacteria

Impact of recommissioning flushing on Legionella pneumophila in a large building during the COVID-19 pandemic

ABSTRACT: COVID-19 shutdowns drastically increased the frequency and duration of water stagnation events in building plumbing systems, urging local authorities to issue guidance for the safe reopening of buildings mostly by recommissioning flushing. The objectives of this study were to document the dynamic changes of bacterial indicators [adenosine triphosphate (ATP), total and intact cell counts (TCC, ICC)] and the prevalence of Legionella pneumophila (Lp) in 20–21 showerheads in a large building before (16-week building closure) and then shortly (24 h) and monthly (4-week of distal water stagnation) after targeted recommissioning flushing. Following the 16-week shutdown, the highest mean of ATP (10 pg ATP/mL), TCC (1.7 × 106 count/mL) and ICC (5.2 × 105 count/mL) were measured in first draw samples. This bacterial amplification was mostly attributable to detachment from biofilm present in the distal devices and immediate connecting piping. Culture-based (mean of 4 487 MPN/L) and quantitative polymerase chain reaction (qPCR; mean of 63 822 gu/L) concentrations of Lp were respectively measured in 81 and 90% of first draw samples. Individual flushing of showerheads for 5 min resulted in 1.2–278-fold decreases in ATP, whereas TCC and ICC were lowered by 1.1- and 0.7-log on average. A one-log reduction in culture-based and qPCR Lp was only achieved in 63 and 29% of paired water samples, resulting in less than one-log reduction in mean risk values per exposure, thus demonstrating the limited effects of fixture-flushing for risk reduction. Clear short-term (24 h) benefits of device recommissioning flushing included lowered values of all bacterial indicators and Lp levels systematically under the common alert threshold of 1 000 MPN/L in first draws. However, after a period of 1 month without water use, these benefits were mostly lost with considerable rebounds of concentrations to similar levels than those measured following the 16-week building closure. Results highlight the temporary benefits of device recommissioning flushing for the control of Lp in shower systems, especially in buildings colonized by Legionella

Experimental determination of the oral bioavailability and bioaccessibility of lead particles

In vivo estimations of Pb particle bioavailability are costly and variable, because of the nature of animal assays. The most feasible alternative for increasing the number of investigations carried out on Pb particle bioavailability is in vitro testing. This testing method requires calibration using in vivo data on an adapted animal model, so that the results will be valid for childhood exposure assessment. Also, the test results must be reproducible within and between laboratories. The Relative Bioaccessibility Leaching Procedure, which is calibrated with in vivo data on soils, presents the highest degree of validation and simplicity. This method could be applied to Pb particles, including those in paint and dust, and those in drinking water systems, which although relevant, have been poorly investigated up to now for childhood exposure assessment

Analysis of steroid hormones and their conjugated forms in water and urine by on-line solid-phase extraction coupled to liquid chromatography tandem mass spectrometry

Background: In recent years, endocrine disrupting compounds (EDCs) have been found in rivers that receive significant inputs of wastewater. Among EDCs, natural and synthetic steroid hormones are recognized for their potential to mimic or interfere with normal hormonal functions (development, growth and reproduction), even at ultratrace levels (ng L-1). Although conjugated hormones are less active than free hormones, they can be cleaved and release the unconjugated estrogens through microbial processes before or during the treatment of wastewater. Due to the need to identify and quantify these compounds, a new fully automated method was developed for the simultaneous determination of the two forms of several steroid hormones (free and conjugated) in different water matrixes and in urine.Results: The method is based on online solid phase extraction coupled with liquid chromatography and tandem mass spectrometry (SPE-LC-MS/MS). Several parameters were assessed in order to optimize the efficiency of the method, such as the type and flow rate of the mobile phase, the various SPE columns, chromatography as well as different sources and ionization modes for MS. The method demonstrated good linearity (R-2 &gt; 0.993) and precision with a coefficient of variance of less than 10 %. The quantification limits vary from a minimum of 3-15 ng L-1 for an injection volume of 1 and 5 mL, respectively, with the recovery values of the compounds varying from 72 to 117 %.Conclusion: The suggested method has been validated and successfully applied for the simultaneous analysis of several steroid hormones in different water matrixes and in urine

Comparative study of deterministic and probabilistic assessments of microbial risk associated with combined sewer overflows upstream of drinking water intakes

RÉSUMÉ: Combined sewer overflows (CSOs) are a source of microbial contamination of drinking water intakes located downstream from their discharge. To safeguard the quality of the source water, it is essential to evaluate the risk levels associated with these municipal structures. This study compares two risk assessment approaches to test their applicability for assessing the risk of CSOs to drinking water intakes in a highly urbanized watershed. The first approach was based on a deterministic equation that combines the characteristics of an overflow structure allowing the risk to be rated as very low, low, medium, high, or very high. The second probabilistic risk assessment approach yielded findings that are probabilistically distributed across the five levels of risk. This approach was developed by constructing a novel Bayesian network to probabilistically link the different factors defining the exposure of water intakes to the hazards of CSOs. The comparison between the results of these two approaches highlighted the importance of simultaneously considering many scenarios for assessing the risk of contamination of source waters. It was possible to use the Bayesian network rather than the deterministic equation, which only supports one scenario at a time. It was also shown that the deterministic approach often overestimated risk levels for CSO outfalls close to the water intake. This occurred because the assessment process emphasized the distance factor between the discharge point and the water intake, while neglecting other crucial characteristics of the overflow, such as duration and frequency. In particular, the deterministic approach tended to underestimate risk for CSOs associated with low overflow frequencies as it did not support scenarios of overflow duration, unlike the probabilistic approach. The validation and sensitivity analysis of the Bayesian model revealed that the population residing in the CSO's drainage basin, along with the frequency and duration of the overflows, exerted the greatest influence on the resulting risk levels. These factors outweighed other variables utilized in the risk assessment, including vulnerability of the drinking water intake, the type of overflow recorder, pipe diameter, and variables defining the exposure of the water intake to the discharge. In the context of implementing action plans, the Bayesian network is estimated as a cost-effective technique as it prioritized overflow structures needing special attention in a highly urbanized watershed, where the same CSOs were deterministically rated as having the same risk level. The results also demonstrated the effectiveness of the Bayesian model in addressing data gaps faced by water managers and stakeholders. The Bayesian model proved capable of assessing risks with uncertainties for CSOs, even with limited input data available. These findings can assist managers in identifying problematic structures by considering various scenarios, unlike the deterministic approach, which left almost half (n = 42) of the study site's overflow structures unassessed due to data limitations