Summary
Phospholipid homeostasis of the bacterial membrane is maintained by biochemical regulation of the synthesis enzymes depending on the environment. However, genes encoding phospholipid ...synthesis enzymes might also be regulated during stress responses, in order for the bacteria to adapt their growth to changing environments. While few studies have addressed this question, global analyses show that specific genes are activated by alternative Sigma factors, and that phospholipid synthesis genes are co‐ordinately regulated during stringent response. In Escherichia coli, the genes coding for glycerol‐3‐phosphate acyltransferase and diacylglycerol kinase (plsB and dgkA) are found next to each other in divergent orientations, suggesting a co‐ordinated regulation. We investigated their regulation and found that these two genes are inversely regulated by a diversity of stress responses. plsB activation by σE is concomitant with a reduced DgkA amount. A second proximal promoter for plsB expression is responsible for basal plsB expression and is inhibited during stringent response. Finally, dgkA is activated by the two‐component regulator BasR, linking dgkA function of phospholipid recycling to LPS modifications. In E. coli, PlsB and DgkA are key enzymes in the phospholipid synthesis pathway. Our results show that their expression is a crucial point of integration for different stress signals.
The Enterococcus faecalis leucine-rich protein ElrA promotes virulence by stimulating bacterial persistence in macrophages and production of the interleukin-6 (IL-6) cytokine. The ElrA protein is ...encoded within an operon that is poorly expressed under laboratory conditions but induced in vivo. In this study, we identify ef2687 (renamed elrR), which encodes a member of the Rgg (regulator gene for glucosyltransferase) family of putative regulatory proteins. Using quantitative reverse transcription-PCR, translational lacZ fusions, and electrophoretic mobility shift assays, we demonstrate that ElrR positively regulates expression of elrA. These results correlate with the attenuated virulence of the ΔelrR strain in a mouse peritonitis model. Virulence of simple and double elrR and elrA deletion mutants also suggests a remaining ElrR-independent expression of elrA in vivo and additional virulence-related genes controlled by ElrR.
A vacant unit, once used by a Portuguese Deli, was converted to a bar/music room in Toronto. The unit was divided into two spaces along its north-south axis. The western portion was designed as a ...music room that would provide a performance space from a solo artist to a Jazz combo to a small rock band. The eastern part was designed as a regular bar/dining area. The plan also called for a microbrewery unit at the back of the unit. The bar music can be loud, while the music room can be pianissimo to forte depending on the type of performance. The acoustical design aspects are critical for the music room. In addition, the acoustical separation between the two spaces is equally important. The music room/bar is currently in use. The design results are compared to actual field measurements. The results showed that the music venue performed satisfactorily. The acoustical separation between the music venue and the bar/restaurant was better than expected other than an installation deficiency of the south side sound lock doors. The background sound along the northern portion was NC-35 or less. However, the southern portion’s background sound exceeded NC-35 due to the hissing of the return air grille. The acoustical design and the performance results of the music venue-bar/restaurant are presented in this paper.
We propose an ecofriendly, efficient, stereoselective procedure for the two-carbon elongation of nonphosphorylated aldoses (C4–C6) to the corresponding C n+2 ketoses (C6–C8) in one step, using ...hydroxypyruvate (HPA) as a ketol donor substrate and an evolved thermostable transketolase from Geobacillus stearothermophilus (TKgst) as a biocatalyst. Simultaneous site saturation mutagenesis (SSM) at two or three key positions in the TKgst active site yielded efficient variants, L382F/F435Y, R521Y/S385/H462N, and R521V/S385D/H462S, with increased activity compared to wild-type TKgst for conversion of two tetroses (d-threose, l-erythrose), two pentoses (d-xylose, d-ribose), and two hexoses (d-allose, d-glucose), respectively. These six C n aldoses as acceptor and HPA as donor substrates were transformed by the TKgst variants at 60 °C with practically complete conversion. The corresponding C n+2 ketoses, including two hexuloses (d-tagatose, l-psicose), two heptuloses (d-altro-heptulose, d-ido-heptulose), and two octuloses (d-glycero- d-ido-octulose, d-glycero-d-altro-octulose) are naturally rare compounds with important biological functions, which were obtained with high diastereoselectivity.
The same gene was then reported in Europe (Denmark) among extended-spectrum β lactamase (ESBL) and AmpC-producing E coli isolates from chicken meat and human infections, but at a very low ...prevalence.2 We screened ESBL-positive E coli isolates collected in France for colistin resistance.
Despite the fact that noise-induced hearing loss remains the number one occupational disease in developed countries, individual noise exposure levels are still rarely known and infrequently tracked. ...Indeed, efforts to standardize noise exposure levels present disadvantages such as costly instrumentation and difficulties associated with on site implementation. Given their advanced technical capabilities and widespread daily usage, mobile phones could be used to measure noise levels and make noise monitoring more accessible. However, the use of mobile phones for measuring noise exposure is currently limited due to the lack of formal procedures for their calibration and challenges regarding the measurement procedure. Our research investigated the calibration of mobile phone-based solutions for measuring noise exposure using a mobile phone's built-in microphones and wearable external microphones. The proposed calibration approach integrated corrections that took into account microphone placement error. The corrections were of two types: frequency-dependent, using a digital filter and noise level-dependent, based on the difference between the C-weighted noise level minus A-weighted noise level of the noise measured by the phone. The electro-acoustical limitations and measurement calibration procedure of the mobile phone were investigated. The study also sought to quantify the effect of noise exposure characteristics on the accuracy of calibrated mobile phone measurements. Measurements were carried out in reverberant and semi-anechoic chambers with several mobiles phone units of the same model, two types of external devices (an earpiece and a headset with an in-line microphone) and an acoustical test fixture (ATF). The proposed calibration approach significantly improved the accuracy of the noise level measurements in diffuse and free fields, with better results in the diffuse field and with ATF positions causing little or no acoustic shadowing. Several sources of errors and uncertainties were identified including the errors associated with the inter-unit-variability, the presence of signal saturation and the microphone placement relative to the source and the wearer. The results of the investigations and validation measurements led to recommendations regarding the measurement procedure including the use of external microphones having lower sensitivity and provided the basis for a standardized and unique factory default calibration method intended for implementation in any mobile phone. A user-defined adjustment was proposed to minimize the errors associated with calibration and the acoustical field. Mobile phones implementing the proposed laboratory calibration and used with external microphones showed great potential as noise exposure instruments. Combined with their potential as training and prevention tools, the expansion of their use could significantly help reduce the risks of noise-induced hearing loss.
Mechanisms underlying the transition from commensalism to virulence in Enterococcus faecalis are not fully understood. We previously identified the enterococcal leucine-rich protein A (ElrA) as a ...virulence factor of E. faecalis. The elrA gene is part of an operon that comprises four other ORFs encoding putative surface proteins of unknown function.
In this work, we compared the susceptibility to phagocytosis of three E. faecalis strains, including a wild-type (WT), a ΔelrA strain, and a strain overexpressing the whole elr operon in order to understand the role of this operon in E. faecalis virulence. While both WT and ΔelrA strains were efficiently phagocytized by RAW 264.7 mouse macrophages, the elr operon-overexpressing strain showed a decreased capability to be internalized by the phagocytic cells. Consistently, the strain overexpressing elr operon was less adherent to macrophages than the WT strain, suggesting that overexpression of the elr operon could confer E. faecalis with additional anti-adhesion properties. In addition, increased virulence of the elr operon-overexpressing strain was shown in a mouse peritonitis model.
Altogether, our results indicate that overexpression of the elr operon facilitates the E. faecalis escape from host immune defenses.
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