Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage

Enterococci account for nearly 10% of all nosocomial infections and constitute a significant treatment challenge due to their multidrug resistance properties. One of the well-studied virulence factors of Enterococcus faecalis is a secreted bacterial protease, termed gelatinase, which has been shown to contribute to the process of biofilm formation. Gelatinase belongs to the M4 family of bacterial zinc metalloendopeptidases, typified by thermolysin. Gelatinase is synthesized as a preproenzyme consisting of a signal sequence, a putative propeptide, and then the mature enzyme. We determined that the molecular mass of the mature protein isolated from culture supernatant was 33,030 Da, which differed from the predicted molecular mass, 34,570 Da, by over 1,500 Da. Using N-terminal sequencing, we confirmed that the mature protein begins at the previously identified sequence VGSEV, thus suggesting that the 1,500-Da molecular mass difference resulted from a C-terminal processing event. By using mutants with site-directed mutations within a predicted C-terminal processing site and mutants with C-terminal deletions fused to a hexahistidine tag, we determined that the processing site is likely to be between residues D304 and 1305 and that it requires the Q306 residue. The results suggest that the E. faecalis gelatinase requires C-terminal processing for full activation of protease activity, making it a unique enzyme among the members of the M4 family of proteases of gram-positive bacteria.Instituto de Biotecnologia y Biologia Molecula

Continuous Gravitational Waves from Galactic Neutron Stars: Demography, Detectability, and Prospects

We study the prospects for the detection of continuous gravitational signals from normal Galactic neutron stars, i.e., nonrecycled stars. We use a synthetic population generated by evolving stellar remnants in time, according to several models. We consider the most recent constraints set by all-sky searches for continuous gravitational waves and use them for our detectability criteria. We discuss the detection prospects for the current and the next generation of gravitational-wave detectors. We find that neutron stars whose ellipticity is solely caused by magnetic deformations cannot produce any detectable signal, not even by third-generation detectors. The currently detectable sources all have B ≲ 1012 G and deformations that are not solely due to the magnetic field. For these, we find in fact that the larger the magnetic field, the higher the ellipticity required for the signal to be detectable, and this ellipticity is well above the value induced by the magnetic field. Third-generation detectors such as the Einstein Telescope and Cosmic Explorer will be able to detect up to ≈250 more sources than current detectors. We briefly treat the case of recycled neutron stars with a simplified model. We find that continuous gravitational waves from these objects will likely remain elusive to detection by current detectors, but should be detectable with the next generation of detectors

Les accidents de cyclomoteurs: mécanismes lésionnels et aspects anatomo-cliniques

Le but de notre étude est de décrire les mécanismes lésionnels et les aspects anatomo-cliniques des  traumatismes par accident de cyclomoteur. C'est une étude transversale menée au niveau du Centre  Hospitalier Régional de Kaffrine sur une période de 12 mois. Elle portait sur les patients admis au service d'accueil pour accident de la voie publique impliquant un cyclomoteur. Il s'agissait de 129 patients (112  hommes et de 17 femmes). L'âge moyen était de 30,5 ans. Soixante-treize patients étaient conducteurs de cyclomoteur, 31 piétons et 25 passagers arrière. Le mécanisme le plus fréquent était une chute de moto. Les lésions prédominaient au niveau des membres. Les accidents de cyclomoteur sont un problème de santé publique.Key words: Cyclomoteur, lésion, mécanisme, cliniqu

Cold preservation of the human colon and ileum with University of Wisconsin solution

The inclusion of the colon in the intestinal graft resulted in worsening patient and graft outcome and increased the incidence of infection and rejection. In this study, we examine the role of ischemia on the barrier function of the epithelium during cold ischemia. Samples were collected from 15 harvested and transplanted human donor grafts (colon, 10; ileum, 6), which were immersed in University of Wisconsin (UW) solution. Ischemia (6, 12, 24, and 45 h) and reoxygenation were performed to evaluate the mucosal electrical status using the Ussing chamber technique. The functions of enterocytes and crypt cells were tested by glucose and theophylline challenge. Modified Park's classification was applied to evaluate the severity of mucosal damage under light microscopy. The colon had higher levels of baseline potential difference, short-circuit current, and resistance than the ileum during 6-48 h of ischemia. Colonic epithelial cells responded well to theophylline stimulation at 24 h of ischemia, while there was no ileal response. The colonic mucosa was histopathologically well preserved in UW solution for 48 h, and mucosal damage induced by reoxygenation was less than in the ileum. In conclusion, electrophysiologically and histopathologically, the colon is less susceptible to cold preservation damage than the ileum during storage with UW solution

Toward a Real-Time Index of Pupillary Activity as an Indicator of Cognitive Load

The Low/High Index of Pupillary Activity (LHIPA), an eye-tracked measure of pupil diameter oscillation, is redesigned and implemented to function in real-time. The novel Real-time IPA (RIPA) is shown to discriminate cognitive load in re-streamed data from earlier experiments. Rationale for the RIPA is tied to the functioning of the human autonomic nervous system yielding a hybrid measure based on the ratio of Low/High frequencies of pupil oscillation. The paper\u27s contribution is drawn from provision of documentation of the calculation of the RIPA. As with the LHIPA, it is possible for researchers to apply this metric to their own experiments where a measure of cognitive load is of interest

Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage

Enterococci account for nearly 10% of all nosocomial infections and constitute a significant treatment challenge due to their multidrug resistance properties. One of the well-studied virulence factors of Enterococcus faecalis is a secreted bacterial protease, termed gelatinase, which has been shown to contribute to the process of biofilm formation. Gelatinase belongs to the M4 family of bacterial zinc metalloendopeptidases, typified by thermolysin. Gelatinase is synthesized as a preproenzyme consisting of a signal sequence, a putative propeptide, and then the mature enzyme. We determined that the molecular mass of the mature protein isolated from culture supernatant was 33,030 Da, which differed from the predicted molecular mass, 34,570 Da, by over 1,500 Da. Using N-terminal sequencing, we confirmed that the mature protein begins at the previously identified sequence VGSEV, thus suggesting that the 1,500-Da molecular mass difference resulted from a C-terminal processing event. By using mutants with site-directed mutations within a predicted C-terminal processing site and mutants with C-terminal deletions fused to a hexahistidine tag, we determined that the processing site is likely to be between residues D304 and 1305 and that it requires the Q306 residue. The results suggest that the E. faecalis gelatinase requires C-terminal processing for full activation of protease activity, making it a unique enzyme among the members of the M4 family of proteases of gram-positive bacteria.Instituto de Biotecnologia y Biologia Molecula

Activation of cardiac renin-angiotensin system in unstable angina

AbstractOBJECTIVESThe aim of this study was to investigate the activity of the cardiac renin-angiotensin system (RAS) in unstable angina (UA).BACKGROUNDAngiotensin (Ang) II locally produced by continuously operating cardiac RAS may affect the pathophysiology of UA.METHODSIn 35 patients with UA, 32 with stable effort angina (SA) and 21 with atypical chest pain (controls), cardiac RAS was investigated during coronary angiography after five days of Holter monitoring by combining the measurement of aorta-coronary sinus gradient for Ang I and Ang II with the kinetics study of 125I-Ang I. Messenger RNAs (mRNA) for all the components of RAS were also quantified with the reverse transcriptase-polymerase chain reaction (RT-PCR) and localized by in situ hybridization in myocardial biopsy specimens from patients who underwent aorta-coronary bypass surgery.RESULTSCardiac Ang II generation was higher in patients with UA than it was in patients with SA or in controls (p < 0.001) due to increased de novo cardiac Ang I formation and its enhanced fractional conversion rate to Ang II. Messenger RNA levels for angiotensinogen (AGTN), angiotensin-converting enzyme (ACE) and Ang II type 1 (AT1) subtype receptors were higher in patients with UA (p < 0.01) than they were in patients with SA or in control hearts. Messenger RNAs for AGTN and ACE were almost exclusively expressed on endothelial and interstitial cells. Angiotensin II formation was correlated with ischemia burden (p < 0.001). However, the amount of Ang II formed and the expression levels of mRNAs for AGTN, ACE and AT1 were not related to the time that had elapsed since the last anginal attack.CONCLUSIONSIn patients with UA, cardiac RAS is activated, resulting in increased Ang II formation. Myocardial ischemia is essential for RAS activation, but it is unlikely to be a direct and immediate cause of RAS activation