Efficacy of laser shock processing of biodegradable Mg and Mg-1Zn alloy on their in vitro corrosion and bacterial response

Laser shock processing (LSP) is increasingly applied as an effective technology for improving the properties of different metallic components. This is done principally to enhance their corrosion and fatigue life behaviour, stress corrosion cracking resistance, etc. In this paper, LSP has been applied to a commercially pure Mg and a Mg-1Zn alloy (wt%) which is aimed to be used as a biodegradable material for biomedical applications. The rational for microalloying with Zn is not only influencing the bacterial response, but also enhancing corrosion resistance and mechanical strength of Mg without causing any toxic effect. The present work is focussed on the examination of the effects of the LSP treatment on the relevant surface related properties of the samples and their correlation with the surface and subsurface induced modifications such as residual stress state, microstructural, roughness, hardness, etc. Central to this investigation is the study of the corrosion response and antibacterial properties against Staphylococcus epidermidis of the different samples as a function of material and LSP parameters. The results show that the application of LSP introduces compressive residual stresses up to 1 mm deep. This occurs together with a significant improvement in corrosion resistance, and less bacterial colonization.Peer reviewe

Enhanced Astrocytic Nitric Oxide Production and Neuronal Modifications in the Neocortex of a NOS2 Mutant Mouse

BACKGROUND: It has been well accepted that glial cells in the central nervous system (CNS) produce nitric oxide (NO) through the induction of a nitric oxide synthase isoform (NOS2) only in response to various insults. Recently we described rapid astroglial, NOS2-dependent, NO production in the neocortex of healthy mice on a time scale relevant to neuronal activity. To explore a possible role for astroglial NOS2 in normal brain function we investigated a NOS2 knockout mouse (B6;129P2-Nos2(tm1Lau)/J, Jackson Laboratory). Previous studies of this mouse strain revealed mainly altered immune responses, but no compensatory pathways and no CNS abnormalities have been reported. METHODOLOGY/PRINCIPAL FINDINGS: To our surprise, using NO imaging in brain slices in combination with biochemical methods we uncovered robust NO production by neocortical astrocytes of the NOS2 mutant. These findings indicate the existence of an alternative pathway that increases basal NOS activity. In addition, the astroglial mutation instigated modifications of neuronal attributes, shown by changes in the membrane properties of pyramidal neurons, and revealed in distinct behavioral abnormalities characterized by an increase in stress-related parameters. CONCLUSIONS/SIGNIFICANCE: The results strongly indicate the involvement of astrocytic-derived NO in modifying the activity of neuronal networks. In addition, the findings corroborate data linking NO signaling with stress-related behavior, and highlight the potential use of this genetic model for studies of stress-susceptibility. Lastly, our results beg re-examination of previous studies that used this mouse strain to examine the pathophysiology of brain insults, assuming lack of astrocytic nitrosative reaction

Contribution of DNA adenine methylation to gene expression heterogeneity in Salmonella enterica

Expression of Salmonella enterica loci harboring undermethylated GATC sites at promoters or regulatory regions was monitored by single cell analysis. Cell-to-cell differences in expression were detected in ten such loci (carA, dgoR, holA, nanA, ssaN, STM1290, STM3276, STM5308, gtr and opvAB), with concomitant formation of ON and OFF subpopulations. The ON and OFF subpopulation sizes varied depending on the growth conditions, suggesting that the population structure can be modulated by environmental control. All the loci under study except STM5308 displayed altered patterns of expression in strains lacking or overproducing Dam methylase, thereby confirming control by Dam methylation. Bioinformatic analysis identified potential binding sites for transcription factors OxyR, CRP and Fur, and analysis of expression in mutant backgrounds confirmed transcriptional control by one or more of such factors. Surveys of gene expression in pairwise combinations of Dam methylation-dependent loci revealed independent switching, thus predicting the formation of a high number of cell variants. This study expands the list of S. enterica loci under transcriptional control by Dam methylation, and underscores the relevance of the DNA adenine methylome as a source of phenotypic heterogeneity.Ministerio de Ciencia e Innovación, Spain [BIO2016-75235-P]; European Regional Fund. Funding for open access charge: Ministerio de Ciencia e Innovación [BIO2016-75235-P]

Contribution of DNA adenine methylation to gene expression heterogeneity in Salmonella enterica

Expression of Salmonella enterica loci harboring undermethylated GATC sites at promoters or regulatory regions was monitored by single cell analysis. Cell-to-cell differences in expression were detected in ten such loci (carA, dgoR, holA, nanA, ssaN, STM1290, STM3276, STM5308, gtr and opvAB), with concomitant formation of ON and OFF subpopulations. The ON and OFF subpopulation sizes varied depending on the growth conditions, suggesting that the population structure can be modulated by environmental control. All the loci under study except STM5308 displayed altered patterns of expression in strains lacking or overproducing Dam methylase, thereby confirming control by Dam methylation. Bioinformatic analysis identified potential binding sites for transcription factors OxyR, CRP and Fur, and analysis of expression in mutant backgrounds confirmed transcriptional control by one or more of such factors. Surveys of gene expression in pairwise combinations of Dam methylation-dependent loci revealed independent switching, thus predicting the formation of a high number of cell variants. This study expands the list of S. enterica loci under transcriptional control by Dam methylation, and underscores the relevance of the DNA adenine methylome as a source of phenotypic heterogeneity.Ministerio de Ciencia e Innovación, Spain [BIO2016-75235-P]; European Regional Fund. Funding for open access charge: Ministerio de Ciencia e Innovación [BIO2016-75235-P]

Epigenetic biosensors for bacteriophage detection and phage receptor discrimination

International audienceEnvironmental monitoring of bacteria using phage-based biosensors has been widely developed for many different species. However, there are only a few available methods to detect specific bacteriophages in raw environmental samples. In this work, we developed a simple and efficient assay to rapidly monitor the phage content of a given sample. The assay is based on the bistable expression of the Salmonella enterica opvAB operon. Under regular growth conditions, opvAB is only expressed by a small fraction of the bacterial sub-population. In the OpvAB ON subpopulation, synthesis of the OpvA and OpvB products shortens the O-antigen and confers resistance to phages that use LPS as a receptor. As a consequence, the OpvAB ON sub-population is selected in the presence of such phages. Using an opvAB::gfp fusion, we could monitor LPS-binding phages in various media, including raw water samples. To enlarge our phage-biosensor panoply, we also developed biosensors able to detect LPS, as well as protein-binding coliphages. Moreover, the combination of these tools allowed to identify the bacterial receptor triggering phage infection. The epigenetic opvAB::gfp biosensor thus comes in different flavours to detect a wide range of bacteriophages and identify the type of receptor they recognize

A portable epigenetic switch for bistable gene expression in bacteria

We describe a portable epigenetic switch based on opvAB, a Salmonella enterica operon that undergoes bistable expression under DNA methylation control. A DNA fragment containing the opvAB promoter and the opvAB upstream regulatory region confers bistability to heterologous genes, yielding OFF and ON subpopulations. Bistable expression under opvAB control is reproducible in Escherichia coli, showing that the opvAB switch can be functional in a heterologous host. Subpopulations of different sizes can be produced at will using engineered opvAB variants. Controlled formation of antibiotic-resistant and antibiotic-susceptible subpopulations may allow use of the opvAB switch in the study of bacterial heteroresistance to antibiotics

Understanding Everything NPCs Can Do: Metrics for Action Similarity in Non-Player Characters

Non-Player Characters (NPCs) have actions that allow them to reason about what they can do in a game and how they can do it. The background information about what they can do are the components of the action, and how they can do it is the form or shape of an action, which may be built up from several sub- actions. The components and shape of an action must be fully defined in a game, which can be tedious when several similar actions are needed. Furthermore, as the number of nuanced actions grows, more pressure is placed on an already constrained reasoning system. By discovering the similarities between actions an NPC can do, a given action set can be intelligently organized with similar components being generalized using an action taxonomy. To understand the similarity between actions we have developed measures of action similarity based on their constituent components and form. From this, wediscover a metric to determine the generalization ability of an organization strategy for an NPC action set. We examine the use of our measures on a previously developed action set to show the nuances between those actions. Lastly, we find that intelligently organizing actions has a positive effect on virtual character reasoning abilities.Green Open Access added to TU Delft Institutional Repository ‘You share, we take care!’ – Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public.Comp Graphics & Visualisatio