In vitro characterization of PlyE146, a novel phage lysin that targets Gram-negative bacteria.

The recent rise of multidrug-resistant Gram-negative bacteria represents a serious threat to public health and makes the search for novel effective alternatives to antibiotics a compelling need. Bacteriophage (Phage) lysins are enzymes that hydrolyze the cell wall of bacteria and represent a promising alternative to tackle this ever-increasing problem. Despite their use is believed to be restricted to Gram-positive bacteria, recent findings have shown that they can also be used against Gram-negative bacteria. By using a phage genome-based screening approach, we identified and characterized a novel lysin, PlyE146, encoded by an Escherichia coli prophage and with a predicted molecular mass of ca. 17 kDa. PlyE146 is composed of a C-terminal cationic peptide and a N-terminal N-acetylmuramidase domain. Histidine-tagged PlyE146 was overexpressed from a plasmid in Lactococcus lactis NZ9000 and purified by NI-NTA chromatography. PlyE146 exhibited in vitro optimal bactericidal activity against E. coli K12 (3.6 log10 CFU/mL decrease) after 2 h of incubation at 37°C at a concentration of 400 μg/mL in the absence of NaCl and at pH 6.0. Under these conditions, PlyE146 displayed antimicrobial activity towards several other E. coli, Pseudomonas aeruginosa (3 to 3.8-log10 CFU/mL decrease) and Acinetobacter baumannii (4.9 to >5-log10 CFU/mL decrease) strains. Therefore, PlyE146 represents a promising therapeutic agent against E. coli, P. aeruginosa and A. baumannii infections. However, further studies are required to improve the efficacy of PlyE146 under physiological conditions

Engineered Liposomes Protect Immortalized Immune Cells from Cytolysins Secreted by Group A and Group G Streptococci.

The increasing antibiotic resistance of bacterial pathogens fosters the development of alternative, non-antibiotic treatments. Antivirulence therapy, which is neither bacteriostatic nor bactericidal, acts by depriving bacterial pathogens of their virulence factors. To establish a successful infection, many bacterial pathogens secrete exotoxins/cytolysins that perforate the host cell plasma membrane. Recently developed liposomal nanotraps, mimicking the outer layer of the targeted cell membranes, serve as decoys for exotoxins, thus diverting them from attacking host cells. In this study, we develop a liposomal nanotrap formulation that is capable of protecting immortalized immune cells from the whole palette of cytolysins secreted by Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis-important human pathogens that can cause life-threatening bacteremia. We show that the mixture of cholesterol-containing liposomes with liposomes composed exclusively of phospholipids is protective against the combined action of all streptococcal exotoxins. Our findings pave the way for further development of liposomal antivirulence therapy in order to provide more efficient treatment of bacterial infections, including those caused by antibiotic resistant pathogens

Host-Derived Microvesicles Carrying Bacterial Pore-Forming Toxins Deliver Signals to Macrophages: A Novel Mechanism of Shaping Immune Responses

Bacterial infectious diseases are a leading cause of death. Pore-forming toxins (PFTs) are important virulence factors of Gram-positive pathogens, which disrupt the plasma membrane of host cells and can lead to cell death. Yet, host defense and cell membrane repair mechanisms have been identified: i.e., PFTs can be eliminated from membranes as microvesicles, thus limiting the extent of cell damage. Released into an inflammatory environment, these host-derived PFTs-carrying microvesicles encounter innate immune cells as first-line defenders. This study investigated the impact of microvesicle- or liposome-sequestered PFTs on human macrophage polarization in vitro. We show that microvesicle-sequestered PFTs are phagocytosed by macrophages and induce their polarization into a novel CD14+MHCIIlowCD86low phenotype. Macrophages polarized in this way exhibit an enhanced response to Gram-positive bacterial ligands and a blunted response to Gram-negative ligands. Liposomes, which were recently shown to sequester PFTs and so protect mice from lethal bacterial infections, show the same effect on macrophage polarization in analogy to host-derived microvesicles. This novel type of polarized macrophage exhibits an enhanced response to Gram-positive bacterial ligands. The specific recognition of their cargo might be of advantage in the efficiency of targeted bacterial clearance

Biodiversity recovery of Neotropical secondary forests

Old-growth tropical forests harbor an immense diversity of tree species but are rapidly being cleared, while secondary forests that regrow on abandoned agricultural lands increase in extent. We assess how tree species richness and composition recover during secondary succession across gradients in environmental conditions and anthropogenic disturbance in an unprecedented multisite analysis for the Neotropics. Secondary forests recover remarkably fast in species richness but slowly in species composition. Secondary forests take a median time of five decades to recover the species richness of old-growth forest (80% recovery after 20 years) based on rarefaction analysis. Full recovery of species composition takes centuries (only 34% recovery after 20 years). A dual strategy that maintains both old-growth forests and species-rich secondary forests is therefore crucial for biodiversity conservation in human-modified tropical landscapes. Copyright © 2019 The Authors, some rights reserved

Which tool to distinguish transient alumina from alpha alumina in thermally grown alumina scales

International audienceAlumina scales constitute excellent protective barriers when they form on alumina-forming steels. If they keep tightly adherent to the underlying substrate, they isolate it from the surrounding aggressive atmosphere at high temperature. The protectiveness of the alumina scale is highly dependant upon its growth mechanism. The nucleation and transformation of transient alumina (mainly g-Al2O3 and y-Al2O3) is known to play an important role on alumina scale formation. It is therefore fundamental to characterise these transient alumina especially during the early stages of the oxidation process. The morphology of the transient alumina was observed by scanning electron microscopy (SEM), their crystallographic phases determined by X-ray diffraction (XRD) and transmission electron microscopy (TEM). X-ray photoelectron spectrometry (XPS) analyses were performed on reference samples and then compared to the alloys oxidised 5 and 30 minutes at 850, 900 and 950°

Influence of a reactive element oxide coating on the high temperature oxidation of chromia-former alloys

MOCVD technique was used to deposit Nd2O3 coatings on two chromia-formers alloys. Oxidation experiments were performed at 1000 ○C under air at atmospheric pressure on uncoated alloys and Nd2O3-coated alloys. The beneficial effects generally ascribed to the reactive elements were confirmed: the oxidation rates were decreased and the adherence of the oxide scales was drastically improved. The reactive element was located at the top of the oxide scale after oxidation. X-ray diffraction measurements identified NdCrO3 as the phase containing the reactive element. This work is compared to a previous study [1] concerning the influence of Nd2O3 on the high temperature oxidation behaviour of a Ti-containing chromia-former alloy in the oxide scale of which a complex phase close to CeTi21O38 was characterized

High-temperature oxidation of Fe3Al and Fe3AlZr intermetallics

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High temperature corrosion of FeCr and FeCrAl alloys

The preparation of model alloys with controlled levels of minor elements, as C or S, is very useful to study the high temperature formation of chromia and alumina scales. The absence of any parasitic influence of classical alloying elements (Si, Mn, Ti, ...) allows to carefully analyse the mechanisms of oxide scale growth. High purity Fe-30Cr and Fe-20Cr-5Al alloys were synthesised and oxidised in air under atmospheric pressure in the temperature range from 900 to 1050°C for the chromia-forming steel and from 1000 to 1200°C for the aluminaforming steel. Oxide scale formation was studied using successive oxidation steps under 16O2 and l8O2. The location of 18O isotope within the pre-formed Cr2l6O3 or A1216O3 scales enabled an interpretation of the oxide scale growth mechanisms. The introduction of Y2O3 or Y in Fe-30Cr and Fe-20Cr-5Al alloys respectively, decreased the oxidation rate and changed the oxide scale growth mechanisms in connection with different oxide scale morphologies on the “doped” materials, giving better protective properties to chromia and alumina scales