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Sophorolipids are bioderived glycolipids displaying interesting antimicrobial properties. We show that they can be used to develop biocidal monolayers against Listeria ivanovii, a Gram-positive bacterium. The present work points out the dependence between the surface density and the antibacterial activity of grafted sophorolipids. It also emphasizes the broad spectrum of activity of these coatings, demonstrating their potential against both Gram-positive strains ( Enteroccocus faecalis, Staphylococcus epidermidis, Streptococcus pyogenes) and Gram-negative strains ( Escherichia coli, Pseudomonas aeruginosa and Salmonella typhymurium ). After exposure to sophorolipids grafted onto gold, all these bacterial strains show a significant reduction in viability resulting from membrane damage as evidenced by fluorescent labelling and SEM-FEG analysis. [ABSTRACT FROM AUTHOR]

The development of solid-state chemistry at the end of the 19th century offered a variety of routes to colour a glass matrix. Eight enamelled glass objects made by Philippe-Joseph Brocard and two representative objects made by Emile Gallé have been analysed using a mobile Raman instrument at the Musée des arts décoratifs (Paris) in order to compare their colouration technology. White, blue, yellow, green, orange, red, brown and black pigments have been identified. If most of the pigment palette is common to both craftsmen and typical of the second half of the 19th century, innovative uses are recognized for Gallé (wollastonite as an opacifier, manganese oxides in black mixtures) and Brocard (specific black and grey, pigment mixture, shade modification by small addition of white and red pigments). This preliminary work confirms the potential of Raman spectroscopy, not as a simple analytical method but as a way to document the ancient technology of fine art objects and to discriminate between different genuine productions and/or copies. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The zoonotic pathogen Brucella abortus is part of the Rhizobiales, which are alpha‐proteobacteria displaying unipolar growth. Here, we show that this bacterium exhibits heterogeneity in its outer membrane composition, with clusters of rough lipopolysaccharide co‐localizing with the essential outer membrane porin Omp2b, which is proposed to allow facilitated diffusion of solutes through the porin. We also show that the major outer membrane protein Omp25 and peptidoglycan are incorporated at the new pole and the division site, the expected growth sites. Interestingly, lipopolysaccharide is also inserted at the same growth sites. The absence of long‐range diffusion of main components of the outer membrane could explain the apparent immobility of the Omp2b clusters, as well as unipolar and mid‐cell localizations of newly incorporated outer membrane proteins and lipopolysaccharide. Unipolar growth and limited mobility of surface structures also suggest that new surface variants could arise in a few generations without the need of diluting pre‐existing surface antigens. Synopsis: The outer membrane of Gram‐negative bacteria is an asymmetric bilayer. Single cell analysis reveals that cell envelope proteins and saccharides of Brucella abortus are targeted to active membrane growth sites and exhibit low mobility increasing surface antigen variability. The outer membrane composition of B. abortus is heterogeneous.The essential outer membrane porin Omp2b co‐localizes with LPS.Omp25 and peptidoglycan are selectively incorporated at the new growth pole and the division site.The cell envelope layers LPS, peptidoglycan and outer membrane proteins display limited mobility. Cell envelope proteins and saccharides of Brucella abortus are targeted to active membrane growth sites and exhibit low mobility increasing surface antigen variability. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a leading cause of endovascular infections. This bacterial pathogen uses a diverse array of surface adhesins to clump in blood and adhere to vessel walls, leading to endothelial damage, development of intravascular vegetations and secondary infectious foci, and overall disease progression. In this work, we describe a novel strategy used by S. aureus to control adhesion and clumping through activity of the ArlRS two-component regulatory system, and its downstream effector MgrA. Utilizing a combination of in vitro cellular assays, and single-cell atomic force microscopy, we demonstrated that inactivation of this ArlRS—MgrA cascade inhibits S. aureus adhesion to a vast array of relevant host molecules (fibrinogen, fibronectin, von Willebrand factor, collagen), its clumping with fibrinogen, and its attachment to human endothelial cells and vascular structures. This impact on S. aureus adhesion was apparent in low shear environments, and in physiological levels of shear stress, as well as in vivo in mouse models. These effects were likely mediated by the de-repression of giant surface proteins Ebh, SraP, and SasG, caused by inactivation of the ArlRS—MgrA cascade. In our in vitro assays, these giant proteins collectively shielded the function of other surface adhesins and impaired their binding to cognate ligands. Finally, we demonstrated that the ArlRS—MgrA regulatory cascade is a druggable target through the identification of a small-molecule inhibitor of ArlRS signaling. Our findings suggest a novel approach for the pharmacological treatment and prevention of S. aureus endovascular infections through targeting the ArlRS—MgrA regulatory system. [ABSTRACT FROM AUTHOR]

Physical contact is prevalent in the animal kingdom to recognize suitable mates by decoding information about sex, species, and maturity. Although chemical cues for mate recognition have been extensively studied, the role of mechanical cues remains elusive. Here, we show that C. elegans males recognize conspecific and reproductive mates through short-range cues, and that the attractiveness of potential mates depends on the sex and developmental stages of the hypodermis. We find that a particular group of cuticular collagens is required for mate attractiveness. These collagens maintain body stiffness to sustain mate attractiveness but do not affect the surface properties that evoke the initial step of mate recognition, suggesting that males utilize multiple sensory mechanisms to recognize suitable mates. Manipulations of body stiffness via physical interventions, chemical treatments, and 3D-printed bionic worms indicate that body stiffness is a mechanical property for mate recognition and increases mating efficiency. Our study thus extends the repertoire of sensory cues of mate recognition in C. elegans and provides a paradigm to study the important roles of mechanosensory cues in social behaviors. [Display omitted] • C. elegans males recognize species, sex, and reproductive stages of their mates • Cuticular cues on adult hermaphrodites evoke transient contact response of males • Proper body stiffness is a ubiquitous mechanical factor for mates to retain males • Cuticular cues and body stiffness are inseparable for mate recognition by contact Weng et al. identify two inseparable sensory cues by which C. elegans males identify suitable mating partners. A cuticular signal evokes contact behaviors of males, and then body stiffness maintained by collagens ensures that males stay in contact with their partners. This study reveals a vital role of the mechanical signal for social communications. [ABSTRACT FROM AUTHOR]

Antibodies against the carboxy-terminal section of the membrane-proximal external region (C-MPER) of the HIV-1 envelope glycoprotein (Env) are considered as nearly pan-neutralizing. Development of vaccines capable of producing analogous broadly neutralizing antibodies requires deep understanding of the mechanism that underlies C-MPER recognition in membranes. Here, we use the archetypic 10E8 antibody and a variety of biophysical techniques including single-molecule approaches to study the molecular recognition of C-MPER in membrane mimetics. In contrast to the assumption that an interfacial MPER helix embodies the entire C-MPER epitope recognized by 10E8, our data indicate that transmembrane domain (TMD) residues contribute to binding affinity and specificity. Moreover, anchoring to membrane the helical C-MPER epitope through the TMD augments antibody binding affinity and relieves the effects exerted by the interfacial MPER helix on the mechanical stability of the lipid bilayer. These observations support that addition of TMD residues may result in more efficient and stable anti-MPER vaccines. The molecular recognition of the carboxy-terminal section of the membrane-proximal external region (C-MPER) of the HIV-1 envelope glycoprotein in membrane mimetics is characterised to help shape continued development of more efficient and stable anti-MPER vaccines. [ABSTRACT FROM AUTHOR]

The micellar structure of sophorolipids, a glycolipid bolaamphiphile, is analyzed using a combination of small-angle X-ray scattering (SAXS), small-angle neutron scattering (SANS), and molecular dynamics (MD) simulations. Numerical modeling of SAXS curves shows that micellar morphology in the noncharged system (pH< 5) is made of prolate ellipsoids of revolution with core–shell morphology. Opposed to most surfactant systems, the hydrophilic shell has a nonhomogeneous distribution of matter: the shell thickness in the axial direction of the ellipsoid is found to be practically zero, while it measures about 12 Å at its cross-section, thus forming a “coffee bean”-like shape. The use of a contrast-matching SANS experiment shows that the hydrophobic component of sophorolipids is actually distributed in a narrow spheroidal region in the micellar core. These data seem to indicate a complex distribution of sophorolipids within the micelle, divided into at least three domains: a pure hydrophobic core, a hydrophilic shell, and a region of less defined composition in the axial direction of the ellipsoid. To account for these results, we make the hypothesis that sophorolipid molecules acquire various configurations within the micelle including bent and linear, crossing the micellar core. These results are confirmed by MD simulations which do show the presence of multiple sophorolipid configurations when passing from spherical to ellipsoidal aggregates. Finally, we also used Rb+ and Sr2+ counterions in combination with anomalous SAXS experiments to probe the distribution of the COO– group of sophorolipids upon small pH increase (5 < pH < 7), where repulsive intermicellar interactions become important. The poor ASAXS signal shows that the COO– groups are rather diffused in the broad hydrophilic shell rather than at the outer micellar/water interface. [ABSTRACT FROM AUTHOR]

Raw sophorolipids, a variety of bioderived glycolipid biosurfactants produced by the yeast Candida bombicola, generally present themselves as a mixture of acetylated acidic and lactonic forms mixed with residual fatty acids. To recover the acidic form alone in large amounts, e.g., tens of grams, which is interesting for various applications in cleaning technologies, material science, medicine, one must either purify the mixture through column chromatography, thus tremendously reducing the final yield, or remove the fatty acids via hot hexane extraction followed by alkaline hydrolysis and a liquid-liquid extraction process; in the latter, pentanol, used as extraction medium, is a high boiling point solvent and, consequently, difficult to remove under standard vacuum extraction conditions. Here, we compare, quantify, and draw the positive and negative aspects of the typical hexane/pentanol extraction route compared to two additional ones, never reported for this system: a cold extraction from a pentanol/alkane mixture and a consecutive filtration on inverse C18-modified silica phase and silica gel in, respectively, water/acetonitrile and methanol/dichloromethane mixtures. The first one generates highly pure products but quantitative extraction is quite long (up to several days) while the second is an extremely handy and rapid (few hours) way but less quantitative as it provides a product with not less than 6% impurities. [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidis and Staphylococcus aureus are two important nosocomial pathogens that form biofilms on indwelling medical devices. Biofilm infections are difficult to fight as cells within the biofilm show increased resistance to antibiotics. Our understanding of the molecular interactions driving bacterial adhesion, the first stage of biofilm formation, has long been hampered by the paucity of appropriate force-measuring techniques. In this minireview, we discuss how atomic force microscopy techniques have enabled to shed light on the molecular forces at play during staphylococcal adhesion. Specific highlights include the study of the binding mechanisms of adhesion molecules by means of single-molecule force spectroscopy, the measurement of the forces involved in whole cell interactions using single-cell force spectroscopy, and the probing of the nanobiophysical properties of living bacteria via multiparametric imaging. Collectively, these findings emphasize the notion that force and function are tightly connected in staphylococcal adhesion. [ABSTRACT FROM AUTHOR]