Whole Genome Sequencing and Complete Genetic Analysis Reveals Novel Pathways to Glycopeptide Resistance in Staphylococcus aureus

The precise mechanisms leading to the emergence of low-level glycopeptide resistance in Staphylococcus aureus are poorly understood. In this study, we used whole genome deep sequencing to detect differences between two isogenic strains: a parental strain and a stable derivative selected stepwise for survival on 4 µg/ml teicoplanin, but which grows at higher drug concentrations (MIC 8 µg/ml). We uncovered only three single nucleotide changes in the selected strain. Nonsense mutations occurred in stp1, encoding a serine/threonine phosphatase, and in yjbH, encoding a post-transcriptional negative regulator of the redox/thiol stress sensor and global transcriptional regulator, Spx. A missense mutation (G45R) occurred in the histidine kinase sensor of cell wall stress, VraS. Using genetic methods, all single, pairwise combinations, and a fully reconstructed triple mutant were evaluated for their contribution to low-level glycopeptide resistance. We found a synergistic cooperation between dual phospho-signalling systems and a subtle contribution from YjbH, suggesting the activation of oxidative stress defences via Spx. To our knowledge, this is the first genetic demonstration of multiple sensor and stress pathways contributing simultaneously to glycopeptide resistance development. The multifactorial nature of glycopeptide resistance in this strain suggests a complex reprogramming of cell physiology to survive in the face of drug challenge

Solid-state reactivity explored in situ by synchrotron radiation on single crystals: from SrFeO 2.5 to SrFeO 3 via electrochemical oxygen intercalation

In this study we demonstrate the feasibility of following up a chemical reaction by single crystal x-ray (synchrotron) diffraction under operando conditions, carried out in a specially designed electrochemical cell mounted on the BM01A at the European Synchrotron Radiation Facility (ESRF). We investigated in detail the electrochemical oxidation of SrFeO2.5 to SrFeO3 on a spherical single crystal of 70 μm diameter by in situ diffraction at an ambient temperature. Complete data sets were obtained by scanning the whole reciprocal space using a 2M Pilatus detector, resulting in 3600 frames with a resolution of 0.1° per data set, each obtained in 18 min. The crystal was mounted in a specially designed electrochemical cell with 1N KOH used as the electrolyte. During the electrochemical oxidation, the reaction proceeds following the phase sequence SrFeO2.5/SrFeO2.75/SrFeO2.875/SrFeO3, structurally accompanied by establishing a complex series of long-range oxygen vacancy ordering, which gets instantly organized at ambient temperature. The topotactic reaction pathway is discussed in terms of the evolution of the twin domain structure. The formation of SrFeO2.875 is accompanied by the formation of diffuse streaks along the [1 0 0]-direction of the perovskite cell, reaching high d-spacings. The diffuse streaks are discussed and are thought to originate from a modified twin structure induced by the SrFeO2.75 to SrFeO2.875 transition, and the associated changes in the domain structure, developed during the oxygen intercalation. We equally analysed and discussed in detail the twin structure of all the title compounds. We confirm the ground state of SrFeO2.5 is able to adopt the Imma space group symmetry, showing stacking faults of the tetrahedral layers along the stacking axis of the brownmillerite unit cell, indicated by the 1D diffuse rods. We showed that in situ single crystal diffraction has huge potential in the study of nonstoichiometric compounds under operando conditions, in order to obtain structural information i.e. about diffuse scattering, and microstructural information related to domain effects such as twinning—information far beyond that which powder diffraction methods allow us to obtain

Results of the Marine Ice Sheet Model Intercomparison Project, MISMIP

Predictions of marine ice-sheet behaviour require models that are able to robustly simulate grounding line migration. We present results of an intercomparison exercise for marine ice-sheet models. Verification is effected by comparison with approximate analytical solutions for flux across the grounding line using simplified geometrical configurations (no lateral variations, no effects of lateral buttressing). Unique steady state grounding line positions exist for ice sheets on a downward sloping bed, while hysteresis occurs across an overdeepened bed, and stable steady state grounding line positions only occur on the downward-sloping sections. Models based on the shallow ice approximation, which does not resolve extensional stresses, do not reproduce the approximate analytical results unless appropriate parameterizations for ice flux are imposed at the grounding line. For extensional-stress resolving “shelfy stream” models, differences between model results were mainly due to the choice of spatial discretization. Moving grid methods were found to be the most accurate at capturing grounding line evolution, since they track the grounding line explicitly. Adaptive mesh refinement can further improve accuracy, including fixed grid models that generally perform poorly at coarse resolution. Fixed grid models, with nested grid representations of the grounding line, are able to generate accurate steady state positions, but can be inaccurate over transients. Only one full-Stokes model was included in the intercomparison, and consequently the accuracy of shelfy stream models as approximations of full-Stokes models remains to be determined in detail, especially during transients

Electrical transport in porous silicon from improved complex impedance analysis

AbstractAn improved analysis of the electrical transport parameters in meso-porous silicon is presented. Our approach is based on a separate contribution of the crystallites and their interconnections to the total impedance of meso-porous silicon layers. Meso-porous silicon morphology exhibits a columnar structure without quantum confinement. The electrical conduction is thus, partially due to the bulk conductivity within continuous paths of crystallites. The samples were realized on 0.02ω-cm p-type Si substrates. Porous silicon layers of 100µm of thickness and 50% of porosity were inserted in Al/SiO2/porous-Si/Si structures. Their electronic transport parameters were determined using complex impedance measurements. A frequency range of 102 - 107 Hz was used allowing an accurate determination of the impedance components. Combined with thermal stimulation, theses measurements provide a powerful tool for the interpretation of basic properties such as the carriers density in the crystallites and the trapping mechanisms. Our results were interpreted in terms of free carriers conduction in partially compensated crystallites prevailing at low frequencies. At high frequencies (above 10 kHz), the electrical conductivity is mainly controlled by hopping transport on localized states in the chaotic porous structure. Finally, the free carriers mobility, evaluated from SCLC measurement is discussed.</jats:p

INTERETHNIC POLYMORPHISM OF THE BETA-GLOBIN GENE LOCUS-CONTROL REGION (LCR) IN SICKLE-CELL-ANEMIA PATIENTS

Sequence polymorphisms within the 5'HS2 segment of human locus control region is described among sickle cell anemia patients. Distinct polymorphic patterns of a simple sequence repeat are observed in strong linkage disequilibrium with each of the five major betas haplotypes. Potential functional relevance of this polymorphic region in globin gene expression is discussed