Contribution of SecDF to Staphylococcus aureus resistance and expression of virulence factors

Background SecDF is an accessory factor of the conserved Sec protein translocation machinery and belongs to the resistance-nodulation-cell division (RND) family of multidrug exporters. SecDF has been shown in Escherichia coli and Bacillus subtilis to be involved in the export of proteins. RND proteins can mediate resistance against various substances and might be of relevance in antimicrobial therapy. The role of RND proteins in Staphylococcus aureus has not yet been determined. Results Markerless deletion mutants were constructed to analyze the impact of the so far uncharacterized RND proteins in S. aureus. While the lack of Sa2056 and Sa2339 caused no phenotype regarding growth and resistance, the secDF mutant resulted in a pleiotropic phenotype. The secDF mutant was cold sensitive, but grew normally in rich medium at 37°C. Resistance to beta-lactams, glycopeptides and the RND substrates acriflavine, ethidium bromide and sodium dodecyl sulfate was reduced. The secDF mutant showed an aberrant cell separation and increased spontaneous and Triton X-100 induced autolysis, although the amounts of penicillin-binding proteins in the membrane were unchanged. The impact of secDF deletion on transcription and expression of specific virulence determinants varied: While coagulase transcription and activity were reduced, the opposite was observed for the autolysin Atl. A reduction of the transcription of the cell wall anchored protein A (spa) was also found. The accumulation of SpA in the membrane and lowered amounts in the cell wall pointed to an impaired translocation. Conclusions The combination of different effects of secDF deletion on transcription, regulation and translocation lead to impaired cell division, reduced resistance and altered expression of virulence determinants suggesting SecDF to be of major relevance in S. aureus. Thus SecDF could be a potential target for the control and eradication of S. aureus in the future

The PURPLE mystery: Semantic meaning of three purple terms in French speakers from Algeria, France, and Switzerland

Studies on the colour category PURPLE yielded inconsistent category boundaries, focal colours, and colour-emotion associations. In French, there are at least three colour terms referring to the shades of purple, potentially weighing on these inconsistencies. Thus, we tested the semantic breadth and richness in semantic meaning of violet (basic term), lilas (non-basic), and pourpre (non-basic). We collected free associations in 274 French speakers from Algeria, France, and Switzerland, yielding 2,079 responses, of which 436 were discrete and 275 were unique. Frequency analyses and semantic coding supported the basicness status of violet in French, within a hierarchically structured semantic system. Moreover, the meaning of the three terms was not synonymous. Violet had the most abstract meaning. Lilas had the narrowest meaning, mainly referring to Natural Entities. Pourpre seemed close to RED. We found no differences between the countries. Future studies should extend this approach to other languages and other colour terms

Première évaluation du risque toxique lié aux cyanobactéries d'eau douce en France : le programme " EFFLOCYA "

Une enquête menée à l'aide d'un questionnaire couvrant l'ensemble du territoire français ainsi que l'étude spécifique d'écosystèmes aquatiques de type lacs et réservoirs ont permis de montrer que les proliférations de cyanobactéries toxiques pouvaient a priori affecter n'importe quel plan d'eau de l'Héxagone. Les genres rencontrés responsables de la production de toxines, le plus souvent hépatiques, sont Microcystis, Planktothrix, Anabœna et Cylindrospermopsis.Les efflorescences peuvent se produire tout au long de l'année car certaines espèces sont adaptées aux eaux froides et elles ne sont pas liées de manière univoque à un état eutrophe de l'écosystème.La production toxinique est la plus élevée lorsque les populations cyanobactériennes sont peu ou pas diversifiées et à la lumière de travaux récents, cette production serait favorisée, en ce qui concerne les microcystines, par un milieu riche en nitrate, pauvre en ammonium et fortement carencé en fer.La suite à donner à ce travail pourrait être la création d'un observatoire national des efflorescences toxiques.An inquiry covering the whole French territory together with specific studies of natural and artificial reservoirs has been made to assess the human health risk related to the prolifération of toxin producing blue green algae (cyanobacteria). The conclusions show that any lentic ecosystem can be affected like it has been shown already in other countries all over the world and more specifically within the EU. The main genus concemed are: Microcystis, Planktothrix, Anabœna and Cylindrospermopsis.Blooms can occur juring the whole year since some species are well adapted to cold water and are not clearly linked with the trophic status of the aquatic ecosystem.The toxinic production is the largest when the cyanobacteria populations contain only a few number of species. The results of recent works are well correlated with our findings which show that the production of toxic heptapeptides (microcystins) could be correlated with high levels of nitrate and depletions of ammonium and iron.The follow up of this work should now include the set up of a national observatory of toxic algal blooms

Chord distribution functions of three-dimensional random media: Approximate first-passage times of Gaussian processes

The main result of this paper is a semi-analytic approximation for the chord distribution functions of three-dimensional models of microstructure derived from Gaussian random fields. In the simplest case the chord functions are equivalent to a standard first-passage time problem, i.e., the probability density governing the time taken by a Gaussian random process to first exceed a threshold. We obtain an approximation based on the assumption that successive chords are independent. The result is a generalization of the independent interval approximation recently used to determine the exponent of persistence time decay in coarsening. The approximation is easily extended to more general models based on the intersection and union sets of models generated from the iso-surfaces of random fields. The chord distribution functions play an important role in the characterization of random composite and porous materials. Our results are compared with experimental data obtained from a three-dimensional image of a porous Fontainebleau sandstone and a two-dimensional image of a tungsten-silver composite alloy.Comment: 12 pages, 11 figures. Submitted to Phys. Rev.

Structure-property correlations in model composite materials

We investigate the effective properties (conductivity, diffusivity and elastic moduli) of model random composite media derived from Gaussian random fields and overlapping hollow spheres. The morphologies generated in the models exhibit low percolation thresholds and give a realistic representation of the complex microstructure observed in many classes of composites. The statistical correlation functions of the models are derived and used to evaluate rigorous bounds on each property. Simulation of the effective conductivity is used to demonstrate the applicability of the bounds. The key morphological features which effect composite properties are discussed

Molecular approaches for monitoring potentially toxic marine and freshwater phytoplankton species

International audienceHarmful phytoplankton species are a growing problem in freshwater and marine ecosystems, because of their ability to synthesize toxins that threaten both animal and human health. The monitoring of these microorganisms has so far been based on conventional methods, mainly involving the microscopic counting and identification of cells, and using analytical and bioanalytical methods to identify and quantify the toxins. However, the increasing number of microbial sequences in the GeneBank database and the development of new tools in the last 15 years nowadays enables the use of molecular methods for detection and quantification of harmful phytoplankton species and their toxins. These methods provide species-level identification of the microorganisms of interest, and their early detection in the environment by PCR techniques. Moreover, real time PCR can be used to quantify the cells of interest, and in some cases to evaluate the proportion of toxin-producing and non-toxin-producing genotypes in a population. Recently, microarray technologies have also been used to achieve simultaneous detection and semiquantification of harmful species in environmental samples. These methods look very promising, but so far their use remains limited to research. The need for validation for routine use and the cost of these methods still hamper their use in monitoring programs

Phytoplankton distribution as determinated by HPLC pigment analysis

International audienc

Population-based inference of aminoglycoside resistance mechanisms in Escherichia coli

BACKGROUND Interpretative reading of antimicrobial susceptibility test (AST) results allows inferring biochemical resistance mechanisms from resistance phenotypes. For aminoglycosides, however, correlations between resistance pathways inferred on the basis of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) clinical breakpoints and expert rules versus genotypes are generally poor. This study aimed at developing and validating a decision tree based on resistance phenotypes determined by disc diffusion and based on epidemiological cut-offs (ECOFFs) to infer the corresponding resistance mechanisms in Escherichia coli. METHODS Phenotypic antibiotic susceptibility of thirty wild-type and 458 aminoglycoside-resistant E. coli clinical isolates was determined by disc diffusion and the genomes were sequenced. Based on well-defined cut-offs, we developed a phenotype-based algorithm (Aminoglycoside Resistance Mechanism Inference Algorithm - ARMIA) to infer the biochemical mechanisms responsible for the corresponding aminoglycoside resistance phenotypes. The mechanisms inferred from susceptibility to kanamycin, tobramycin and gentamicin were analysed using ARMIA- or EUCAST-based AST interpretation and validated by whole genome sequencing (WGS) of the host bacteria. FINDINGS ARMIA-based inference of resistance mechanisms and WGS data were congruent in 441/458 isolates (96·3%). In contrast, there was a poor correlation between resistance mechanisms inferred using EUCAST CBPs/expert rules and WGS data (418/488, 85·6%). Based on the assumption that resistance mechanisms can result in therapeutic failure, EUCAST produced 63 (12·9%) very major errors (vME), compared to only 2 (0·4%) vME with ARMIA. When used for detection and identification of resistance mechanisms, ARMIA resolved >95% vMEs generated by EUCAST-based AST interpretation. INTERPRETATION This study demonstrates that ECOFF-based analysis of AST data of only four aminoglycosides provides accurate information on the resistance mechanisms in E. coli. Since aminoglycoside resistance mechanisms, despite having in certain cases a minimal effect on the minimal inhibitory concentration, may compromise the bactericidal activity of aminoglycosides, prompt detection of resistance mechanisms is crucial for therapy. Using ARMIA as an interpretative rule set for editing AST results allows for better predictions of in vivo activity of this drug class