Thermo-mechanical FE model with memory effect for 304L austenitic stainless steel presenting microstructure gradient

The main purpose of this study is to determine, via a three dimensions Finite Element analysis (FE), the stress and strain fields at the inner surface of a tubular specimen submitted to thermo-mechanical fatigue. To investigate the surface finish effect on fatigue behaviour at this inner surface, mechanical tests were carried out on real size tubular specimens under various thermal loadings. X ray measurements, Transmission Electron Microscopy observations and micro-hardness tests performed at and under the inner surface of the specimen before testing, revealed residual internal stresses and a large dislocation microstructure gradient in correlation with hardening gradients due to machining. A memory effect, bound to the pre-hardening gradient, was introduced into an elasto-visco-plastic model in order to determine the stress and strain fields at the inner surface. The temperature evolution on the inner surface of the tubular specimen was first computed via a thermo-elastic model and then used for our thermo-mechanical simulations. Identification of the thermo-mechanical model parameters was based on the experimental stabilized cyclic tension-compression tests performed at 20^{\circ}C and 300^{\circ}C. A good agreement was obtained between numerical stabilized traction-compression cycle curves (with and without pre-straining) and experimental ones. This 3 dimensional simulation gave access to the evolution of the axial and tangential internal stresses and local strains during the tests. Numerical results showed: a decreasing of the tangential stress and stabilization after 40 cycles, whereas the axial stress showed weaker decreasing with the number of cycles. The results also pointed out a ratcheting and a slightly non proportional loading at the inner surface. The computed mean stress and strain values of the stabilized cycle being far from the initial ones, they could be used to get the safety margins of standard design related to fatigue, as well as to get accurate loading conditions needed for the use of more advanced fatigue analysis and criteria

Arbidol: a broad-spectrum antiviral that inhibits acute and chronic HCV infection

Arbidol (ARB) is an antiviral compound that was originally proven effective for treatment of influenza and several other respiratory viral infections. The broad spectrum of ARB anti-viral activity led us to evaluate its effect on hepatitis C virus (HCV) infection and replication in cell culture. Long-term ARB treatment of Huh7 cells chronically replicating a genomic length genotype 1b replicon resulted in sustained reduction of viral RNA and protein expression, and eventually cured HCV infected cells. Pre-treatment of human hepatoma Huh7.5.1 cells with 15 μM ARB for 24 to 48 hours inhibited acute infection with JFH-1 virus by up to 1000-fold. The inhibitory effect of ARB on HCV was not due to generalized cytotoxicity, nor to augmentation of IFN antiviral signaling pathways, but involved impaired virus-mediated membrane fusion. ARB's affinity for membranes may inhibit several aspects of the HCV lifecycle that are membrane-dependent

Reconstitution of Membrane Proteins into Giant Unilamellar Vesicles via Peptide-Induced Fusion

In this work, we present a protocol to reconstitute membrane proteins into giant unilamellar vesicles (GUV) via peptide-induced fusion. In principle, GUV provide a well-defined lipid matrix, resembling a close-to-native state for biophysical studies, including optical microspectroscopy, of transmembrane proteins at the molecular level. Furthermore, reconstitution in this manner would also eliminate potential artifacts arising from secondary interactions of proteins, when reconstituted in planar membranes supported on solid surfaces. However, assembly procedures of GUV preclude direct reconstitution. Here, for the first time, a method is described that allows the controlled incorporation of membrane proteins into GUV. We demonstrate that large unilamellar vesicles (LUV, diameter 0.1 µm), to which the small fusogenic peptide WAE has been covalently attached, readily fuse with GUV, as revealed by monitoring lipid and contents mixing by fluorescence microscopy. To monitor contents mixing, a new fluorescence-based enzymatic assay was devised. Fusion does not introduce changes in the membrane morphology, as shown by fluorescence correlation spectroscopy. Analysis of fluorescence confocal imaging intensity revealed that ~6 to 10 LUV fused per µm2 of GUV surface. As a model protein, bacteriorhodopsin (BR) was reconstituted into GUV, using LUV into which BR was incorporated via detergent dyalisis. BR did not affect GUV-LUV fusion and the protein was stably inserted into the GUV and functionally active. Fluorescence correlation spectroscopy experiments show that BR inserted into GUV undergoes unrestricted Brownian motion with a diffusion coefficient of 1.2 µm2/s. The current procedure offers new opportunities to address issues related to membrane-protein structure and dynamics in a close-to-native state

Multiple effects of silymarin on the hepatitis C virus lifecycle

Silymarin, an extract from milk thistle (Silybum marianum), and its purified flavonolignans have been recently shown to inhibit hepatitis C virus (HCV) infection, both in vitro and in vivo. In the current study, we further characterized silymarin's antiviral actions. Silymarin had antiviral effects against hepatitis C virus cell culture (HCVcc) infection that included inhibition of virus entry, RNA and protein expression, and infectious virus production. Silymarin did not block HCVcc binding to cells but inhibited the entry of several viral pseudoparticles (pp), and fusion of HCVpp with liposomes. Silymarin but not silibinin inhibited genotype 2a NS5B RNA-dependent RNA polymerase (RdRp) activity at concentrations 5 to 10 times higher than required for anti-HCVcc effects. Furthermore, silymarin had inefficient activity on the genotype 1b BK and four 1b RDRPs derived from HCV-infected patients. Moreover, silymarin did not inhibit HCV replication in five independent genotype 1a, 1b, and 2a replicon cell lines that did not produce infectious virus. Silymarin inhibited microsomal triglyceride transfer protein activity, apolipoprotein B secretion, and infectious virion production into culture supernatants. Silymarin also blocked cell-to-cell spread of virus. CONCLUSION: Although inhibition of in vitro NS5B polymerase activity is demonstrable, the mechanisms of silymarin's antiviral action appear to include blocking of virus entry and transmission, possibly by targeting the host cell

The influence of ecological infrastructures adjacent to crops on their carabid assemblages in intensive agroecosystems

peer reviewedBackground. Conserving biodiversity and enhancing ecosystem services of interest in intensive agroecosystems is a major challenge. Perennial ecological infrastructures (EIs), such as hedges and grassy strips, and annual EI under Agri-Environment Schemes appear to be good candidates to promote both. Our study focused on carabids, an indicator group responding both at the species and functional trait level to disturbances and supporting pest control and weed seed consumption services. Methods. We compared carabid assemblages at the species and functional traits levels, sampled via pitfall trapping, in three types of EIs (hedges, grassy strips and annual flower strips) and crops. We also tested via GLMs the effect of (1) the type of EI at the crops' border and (2) the distance from the crops' border (two meters or 30 meters) on carabid assemblages of crops. Tested variables comprised: Activity-density, species richness, functional dispersion metrics (FDis) and proportions of carabids by functional categories (Diet: Generalist predators/specialist predators/seed-eaters; Size: Small/medium/large/very large; Breeding period: Spring/autumn). Results and Discussion. Carabid assemblages on the Principal Coordinate Analysis split in two groups: Crops and EIs. Assemblages from all sampled EIs were dominated by mobile generalist predator species from open-land, reproducing in spring. Assemblages of hedges were poor in activity-density and species richness, contrarily to grassy and annual flower strips. Differences in carabid assemblages in crops were mainly driven by the presence of hedges. The presence of hedges diminished the Community Weighted Mean size of carabids in crops, due to an increased proportion of small (<5 mm) individuals, while distance from crops' border favoured large (between 10-15 mm) carabids. Moreover, even if they were attracted by EIs, granivorous carabid species were rare in crops. Our results underlie the importance of local heterogeneity when adapting crops' borders to enhance carabid diversity and question the relevance of hedge implantation in intensive agrolandscapes, disconnected from any coherent ecological network. Moreover, this study emphasizes the difficulty to modify functional assemblages of crops for purposes of ecosystem services development, especially for weed seed consumption, as well as the role of distance from the crops' border in the shaping of crop carabid assemblages. © 2020 Pecheur et al

Material characterization and finite element modelling of cyclic plasticity behavior for 304 stainless steel using a crystal plasticity model

Low cycle fatigue tests were carried out for a 304 stainless steel at room temperature. A series of experimental characterisations, including SEM, TEM, and XRD were conducted for the 304 stainless steel to facilitate the understanding of the mechanical responses and microstructural behaviour of the material under cyclic loading including nanostructure, crystal structure and the fractured surface. The crystal plasticity finite element method (CPFEM) is a powerful tool for studying the microstructure influence on the cyclic plasticity behaviour. This method was incorporated into the commercially available software ABAQUS by coding a UMAT user subroutine. Based on the results of fatigue tests and material characterisation, the full set of material constants for the crystal plasticity model was determined. The CPFEM framework used in this paper can be used to predict the crack initiation sites based on the local accumulated plastic deformation and local plastic dissipation energy criterion, but with limitation in predicting the crack initiation caused by precipitates

Mechanism of Inhibition of Enveloped Virus Membrane Fusion by the Antiviral Drug Arbidol

The broad-spectrum antiviral arbidol (Arb) inhibits cell entry of enveloped viruses by blocking viral fusion with host cell membrane. To better understand Arb mechanism of action, we investigated its interactions with phospholipids and membrane peptides. We demonstrate that Arb associates with phospholipids in the micromolar range. NMR reveals that Arb interacts with the polar head-group of phospholipid at the membrane interface. Fluorescence studies of interactions between Arb and either tryptophan derivatives or membrane peptides reconstituted into liposomes show that Arb interacts with tryptophan in the micromolar range. Interestingly, apparent binding affinities between lipids and tryptophan residues are comparable with those of Arb IC50 of the hepatitis C virus (HCV) membrane fusion. Since tryptophan residues of membrane proteins are known to bind preferentially at the membrane interface, these data suggest that Arb could increase the strength of virus glycoprotein's interactions with the membrane, due to a dual binding mode involving aromatic residues and phospholipids. The resulting complexation would inhibit the expected viral glycoprotein conformational changes required during the fusion process. Our findings pave the way towards the design of new drugs exhibiting Arb-like interfacial membrane binding properties to inhibit early steps of virus entry, i.e., attractive targets to combat viral infection

Differential In Vitro Effects of Intravenous versus Oral Formulations of Silibinin on the HCV Life Cycle and Inflammation

Silymarin prevents liver disease in many experimental rodent models, and is the most popular botanical medicine consumed by patients with hepatitis C. Silibinin is a major component of silymarin, consisting of the flavonolignans silybin A and silybin B, which are insoluble in aqueous solution. A chemically modified and soluble version of silibinin, SIL, has been shown to potently reduce hepatitis C virus (HCV) RNA levels in vivo when administered intravenously. Silymarin and silibinin inhibit HCV infection in cell culture by targeting multiple steps in the virus lifecycle. We tested the hepatoprotective profiles of SIL and silibinin in assays that measure antiviral and anti-inflammatory functions. Both mixtures inhibited fusion of HCV pseudoparticles (HCVpp) with fluorescent liposomes in a dose-dependent fashion. SIL inhibited 5 clinical genotype 1b isolates of NS5B RNA dependent RNA polymerase (RdRp) activity better than silibinin, with IC50 values of 40–85 µM. The enhanced activity of SIL may have been in part due to inhibition of NS5B binding to RNA templates. However, inhibition of the RdRps by both mixtures plateaued at 43–73%, suggesting that the products are poor overall inhibitors of RdRp. Silibinin did not inhibit HCV replication in subgenomic genotype 1b or 2a replicon cell lines, but it did inhibit JFH-1 infection. In contrast, SIL inhibited 1b but not 2a subgenomic replicons and also inhibited JFH-1 infection. Both mixtures inhibited production of progeny virus particles. Silibinin but not SIL inhibited NF-κB- and IFN-B-dependent transcription in Huh7 cells. However, both mixtures inhibited T cell proliferation to similar degrees. These data underscore the differences and similarities between the intravenous and oral formulations of silibinin, which could influence the clinical effects of this mixture on patients with chronic liver diseases