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Horizontal gene transfer is a principal source of evolution leading to change in the ecological character of bacterial species 1 . Bacterial conjugation 2 , which promotes the horizontal transfer of genetic material between donor and recipient cells by physical contact, is a phenomenon of fundamental evolutionary consequence 3 . Although conjugation has been studied primarily in liquid, most natural bacterial populations are found associated with environmental surfaces in complex multispecies communities called biofilms 4 . Biofilms are ideally suited to the exchange of genetic material of various origins, and it has been shown that bacterial conjugation occurs within biofilms 5 , 6 . Here I investigate the direct contribution of conjugative plasmids themselves to the capacity of the bacterial host to form a biofilm. Natural conjugative plasmids expressed factors that induced planktonic bacteria to form or enter biofilm communities, which favour the infectious transfer of the plasmid. This general connection between conjugation and biofilms suggests that medically relevant plasmid-bearing strains are more likely to form a biofilm. This may influence both the chances of biofilm-related infection risks and of conjugational spread of virulence factors.

Several pathogenic bacteria are able to use heme and hemoproteins as iron sources independent of siderophore production by mechanisms involving outer membrane heme-binding proteins and heme transport systems. Here we show that Serratia marcescens has such a property and we identify an extracellular heme-binding protein, HasA (for heme acquisition system), allowing the release of heme from hemoglobin. This protein is secreted by S. marcescens under conditions of iron depletion and is essential for heme acquisition.

mRNA profiling of pathogens during the course of human infections gives detailed information on the expression levels of relevant genes that drive pathogenicity and adaptation and at the same time allows for the delineation of phylogenetic relatedness of pathogens that cause specific diseases. In this study, we used mRNA sequencing to acquire information on the expression of Escherichia coli pathogenicity genes during urinary tract infections (UTI) in humans and to assign the UTI-associated E. coli isolates to different phylogenetic groups. Whereas the in vivo gene expression profiles of the majority of genes were conserved among 21 E. coli strains in the urine of elderly patients suffering from an acute UTI, the specific gene expression profiles of the flexible genomes was diverse and reflected phylogenetic relationships. Furthermore, genes transcribed in vivo relative to laboratory media included well-described virulence factors, small regulatory RNAs, as well as genes not previously linked to bacterial virulence. Knowledge on relevant transcriptional responses that drive pathogenicity and adaptation of isolates to the human host might lead to the introduction of a virulence typing strategy into clinical microbiology, potentially facilitating management and prevention of the disease. IMPORTANCE Urinary tract infections (UTI) are very common; at least half of all women experience UTI, most of which are caused by pathogenic Escherichia coli strains. In this study, we applied massive parallel cDNA sequencing (RNA-seq) to provide unbiased, deep, and accurate insight into the nature and the dimension of the uropathogenic E. coli gene expression profile during an acute UTI within the human host. This work was undertaken to identify key players in physiological adaptation processes and, hence, potential targets for new infection prevention and therapy interventions specifically aimed at sabotaging bacterial adaptation to the human host. Urinary tract infections (UTI) are very common; at least half of all women experience UTI, most of which are caused by pathogenic Escherichia coli strains. In this study, we applied massive parallel cDNA sequencing (RNA-seq) to provide unbiased, deep, and accurate insight into the nature and the dimension of the uropathogenic E. coli gene expression profile during an acute UTI within the human host. This work was undertaken to identify key players in physiological adaptation processes and, hence, potential targets for new infection prevention and therapy interventions specifically aimed at sabotaging bacterial adaptation to the human host.

One of the strategies used by Gram-negative bacteria to secrete proteins across the two membranes which delimit the cells, issec independent and dedicated to proteins lacking an N-terminal signal peptide. It depends on ABC protein-mediated exporters, which consist of three cell envelope proteins: two inner membrane proteins: an ATPase (the ABC protein), a membrane fusion protein (MFP) and an outer membrane polypeptide.Erwinia chrysanthemi metalloproteinases B and C, andSerratia marcescens hemoprotein HasA are secreted by such homologous pathways and interact with the ABC protein. Interaction between the ABC protein and its substrate has also been evidenced by studies on proteinase and HasA hybrid transporters obtained by combining components from each system. Association between hemoprotein HasA and the three exporter/secretion proteins was demonstrated by affinity chromatography on hemin agarose on which the substrate remained bound with the three secretion proteins. The three component association was ordered and substrate binding was required for the formation of this multiprotein complex.

The Beam Expander Testing X-ray facility (BEaTriX) is a unique X-ray apparatus now operated at the Istituto Nazionale di Astrofisica (INAF), Osservatorio Astronomico di Brera (OAB), in Merate, Italy. It has been specifically designed to measure the point spread function (PSF) and the effective area (EA) of the X-ray mirror modules (MMs) of the Advanced Telescope for High-ENergy Astrophysics (ATHENA), based on silicon pore optics (SPO) technology, for verification before integration into the mirror assembly. To this end, BEaTriX generates a broad, uniform, monochromatic, and collimated X-ray beam at 4.51 keV. [...] In BEaTriX, a micro-focus X-ray source with a titanium anode is placed in the focus of a paraboloidal mirror, which generates a parallel beam. A crystal monochromator selects the 4.51 keV line, which is expanded to the final size by a crystal asymmetrically cut with respect to the crystalline planes. [...] After characterization, the BEaTriX beam has the nominal dimensions of 170 mm x 60 mm, with a vertical divergence of 1.65 arcsec and a horizontal divergence varying between 2.7 and 3.45 arcsec, depending on the monochromator setting: either high collimation or high intensity. The flux per area unit varies from 10 to 50 photons/s/cm2 from one configuration to the other. The BEaTriX beam performance was tested using an SPO MM, whose entrance pupil was fully illuminated by the expanded beam, and its focus was directly imaged onto the camera. The first light test returned a PSF and an EA in full agreement with expectations. As of today, the 4.51 keV beamline of BEaTriX is operational and can characterize modular X-ray optics, measuring their PSF and EA with a typical exposure of 30 minutes. [...] We expect BEaTriX to be a crucial facility for the functional test of modular X-ray optics, such as the SPO MMs for ATHENA.

Because of its nicely chromatic behavior, Calcium Fluoride (CaF2) is a nice choice for an optical designer as it can easily solve a number of issues, giving the right extra degree of freedom in the optical design tuning. However, switching from tablet screens to real life, the scarcity of information -and sometimes the bad reputation in term of fragility- about this material makes an overall test much more than a \"display determination\" experiment. We describe the extensive tests performed in ambient temperature and in thermo-vacuum of a prototype, consistent with flight CTEs, of a 200mm class camera envisaged for the PLATO (PLAnetary Transit and Oscillations of Stars) mission. We show how the CaF2 lens uneventfully succeeded to all the tests and handling procedures, and discuss the main results of the very intensive test campaign of the PLATO Telescope Optical Unit prototype.

The BEaTriX (Beam Expander Testing X-ray facility) project is an X-ray apparatus under construction at INAF/OAB to generate a broad (200 x 60 mm2), uniform and low-divergent X-ray beam within a small lab (6 x 15 m2). BEaTriX will consist of an X-ray source in the focus a grazing incidence paraboloidal mirror to obtain a parallel beam, followed by a crystal monochromation system and by an asymmetrically-cut diffracting crystal to perform the beam expansion to the desired size. Once completed, BEaTriX will be used to directly perform the quality control of focusing modules of large X-ray optics such as those for the ATHENA X-ray observatory, based on either Silicon Pore Optics (baseline) or Slumped Glass Optics (alternative), and will thereby enable a direct quality control of angular resolution and effective area on a number of mirror modules in a short time, in full X-ray illumination and without being affected by the finite distance of the X-ray source. However, since the individual mirror modules for ATHENA will have an optical quality of 3-4 arcsec HEW or better, BEaTriX is required to produce a broad beam with divergence below 1-2 arcsec, and sufficient flux to quickly characterize the PSF of the module without being significantly affected by statistical uncertainties. Therefore, the optical components of BEaTriX have to be selected and/or manufactured with excellent optical properties in order to guarantee the final performance of the system. In this paper we report the final design of the facility and a detailed performance simulation.

Thin slumped glass foils are considered good candidates for the realization of future X-ray telescopes with large effective area and high spatial resolution. However, the hot slumping process affects the glass strength, and this can be an issue during the launch of the satellite because of the high kinematical and static loads occurring during that phase. In the present work we have investigated the possible use of Gorilla glass (produced by Corning), a chemical tempered glass that, thanks to its strength characteristics, would be ideal. The un-tempered glass foils were curved by means of an innovative hot slumping technique and subsequently chemically tempered. In this paper we show that the chemical tempering process applied to Gorilla glass foils does not affect the surface micro-roughness of the mirrors. On the other end, the stress introduced by the tempering process causes a reduction in the amplitude of the longitudinal profile errors with a lateral size close to the mirror length. The effect of the overall shape changes in the final resolution performance of the glass mirrors was studied by simulating the glass foils integration with our innovative approach based on glass reinforcing ribs. The preliminary tests performed so far suggest that this approach has the potential to be applied to the X-ray telescopes of the next generation.

The optics of a number of future X-ray telescopes will have very long focal lengths (10 - 20 m), and will consist of a number of nested/stacked thin, grazing-incidence mirrors. The optical quality characterization of a real mirror can be obtained via profile metrology, and the Point Spread Function of the mirror can be derived via one of the standard computation methods. However, in practical cases it can be difficult to access the optical surfaces of densely stacked mirror shells, after they have been assembled, using the widespread metrological tools. For this reason, the assessment of the imaging resolution of a system of mirrors is better obtained via a direct, full-illumination test in X-rays. If the focus cannot be reached, an intra-focus test can be performed, and the image can be compared with the simulation results based on the metrology, if available. However, until today no quantitative information was extracted from a full-illumination, intra-focal exposure. In this work we show that, if the detector is located at an optimal distance from the mirror, the intensity variations of the intra-focal, full-illumination image in single reflection can be used to reconstruct the profile of the mirror surface, without the need of a wavefront sensor. The Point Spread Function can be subsequently computed from the reconstructed mirror shape. We show the application of this method to an intra-focal (8 m distance from mirror) test performed at PANTER on an optical module prototype made of hot-slumped glass foils with a 20 m focal length, from which we could derive an expected imaging quality near 16 arcsec HEW.