Dual Role for Pilus in Adherence to Epithelial Cells and Biofilm Formation in Streptococcus agalactiae

Streptococcus agalactiae is a common human commensal and a major life-threatening pathogen in neonates. Adherence to host epithelial cells is the first critical step of the infectious process. Pili have been observed on the surface of several gram-positive bacteria including S. agalactiae. We previously characterized the pilus-encoding operon gbs1479-1474 in strain NEM316. This pilus is composed of three structural subunit proteins: Gbs1478 (PilA), Gbs1477 (PilB), and Gbs1474 (PilC), and its assembly involves two class C sortases (SrtC3 and SrtC4). PilB, the bona fide pilin, is the major component; PilA, the pilus associated adhesin, and PilC, are both accessory proteins incorporated into the pilus backbone. We first addressed the role of the housekeeping sortase A in pilus biogenesis and showed that it is essential for the covalent anchoring of the pilus fiber to the peptidoglycan. We next aimed at understanding the role of the pilus fiber in bacterial adherence and at resolving the paradox of an adhesive but dispensable pilus. Combining immunoblotting and electron microscopy analyses, we showed that the PilB fiber is essential for efficient PilA display on the surface of the capsulated strain NEM316. We then demonstrated that pilus integrity becomes critical for adherence to respiratory epithelial cells under flow-conditions mimicking an in vivo situation and revealing the limitations of the commonly used static adherence model. Interestingly, PilA exhibits a von Willebrand adhesion domain (VWA) found in many extracellular eucaryotic proteins. We show here that the VWA domain of PilA is essential for its adhesive function, demonstrating for the first time the functionality of a prokaryotic VWA homolog. Furthermore, the auto aggregative phenotype of NEM316 observed in standing liquid culture was strongly reduced in all three individual pilus mutants. S. agalactiae strain NEM316 was able to form biofilm in microtiter plate and, strikingly, the PilA and PilB mutants were strongly impaired in biofilm formation. Surprisingly, the VWA domain involved in adherence to epithelial cells was not required for biofilm formation

Calculation of the orientational linear and nonlinear correlation factors of polar liquids from the rotational Dean-Kawasaki equation

International audienc

Sun backward gazing method for measuring optomechanical errors of solar concentrators: experimental results

International audienceThis paper presents the first experimental results obtained with the four cameras backward gazing method for measuring the opto-mechanical errors of solar concentrating surfaces in operation. It consists in combining the images simultaneously acquired by four cameras placed near the solar receiver in order to reconstruct the slopes errors of the mirrors digitally. New algorithms adapted to real acquired images are described. The experiment has been conducted on a sun-tracking heliostat of the experimental solar tower plant THEMIS with slopes error measurement accuracy estimated as 0.2 mrad. The capacity of characterizing shape distortions due to wind or gravity loads is also investigated. The main experimental error sources are analysed and possible ways of mitigation are proposed, giving reasonable hope in reaching an ultimate measurement accuracy of 0.1 mrad

Monte-Carlo and sensitivity transport models for domain deformation

International audienceWe address the question of evaluating shape derivatives of objective functions for radiative-transfer engineering involving semi-transparent media. After recalling the standard Monte-Carlo approach to sensitivity estimation and its current limitations, a new method is presented for the specific case of geometrical sensitivities. This method is then tested on configurations with multiple-scattering and absorbing (non-emitting) semi-transparent medium. A new geometrical sensitivity algorithm is presented with full details in order to extend, on several examples, its implementation in complex geometries