Regulatory Requirements for Staphylococcus aureus Nitric Oxide Resistance

ABSTRACT The ability of Staphylococcus aureus to resist host innate immunity augments the severity and pervasiveness of its pathogenesis. Nitric oxide (NO˙) is an innate immune radical that is critical for the efficient clearance of a wide range of microbial pathogens. Exposure of microbes to NO˙ typically results in growth inhibition and induction of stress regulons. S. aureus , however, induces a metabolic state in response to NO˙ that allows for continued replication and precludes stress regulon induction. The regulatory factors mediating this distinctive response remain largely undefined. Here, we employ a targeted transposon screen and transcriptomics to identify and characterize five regulons essential for NO˙ resistance in S. aureus : three virulence regulons not formerly associated with NO˙ resistance, SarA, CodY, and Rot, as well as two regulons with established roles, Fur and SrrAB. We provide new insights into the contributions of Fur and SrrAB during NO˙ stress and show that the S. aureus Δ sarA mutant, the most sensitive of the newly identified mutants, exhibits metabolic dysfunction and widespread transcriptional dysregulation following NO˙ exposure. Altogether, our results broadly characterize the regulatory requirements for NO˙ resistance in S. aureus and suggest an intriguing overlap between the regulation of NO˙ resistance and virulence in this well-adapted human pathogen. IMPORTANCE The prolific human pathogen Staphylococcus aureus is uniquely capable of resisting the antimicrobial radical nitric oxide (NO˙), a crucial component of the innate immune response. However, a complete understanding of how S. aureus regulates an effective response to NO˙ is lacking. Here, we implicate three central virulence regulators, SarA, CodY, and Rot, as major players in the S. aureus NO˙ response. Additionally, we elaborate on the contribution of two regulators, SrrAB and Fur, already known to play a crucial role in S. aureus NO˙ resistance. Our study sheds light on a unique facet of S. aureus pathogenicity and demonstrates that the transcriptional response of S. aureus to NO˙ is highly pleiotropic and intrinsically tied to metabolism and virulence regulation

Biochemical Variation Among Normal Equine Carpal and Tarsocrural Joint Fluids are Detected by Infrared Spectral Characteristics and A Modified Approach to Linear Discriminant Analysis

Research into osteoarthritis diagnostics has evolved from traditional methods that are only useful in more advanced clinical disease, towards the discovery of biomarkers that are predictive or reflective of preclinical joint disease. The potential of Fourier transform infrared spectroscopy (FTIR) coupled with chemometrics has been demonstrated as useful for the assessment of biomolecular responses to disease. Joint fluid was collected from 105 clinically normal antebrachiocarpal (AC), midcarpal (MC) and tarsocrural (TCRL) joints. Thin films were prepared and FTIR absorbance spectra in the mid-infrared region recorded. Interferograms were signal averaged and Fourier transformed to generate spectra with a nominal resolution of 4 cm-1. Comparisons among joints were made using a novel modified method similar to linear discriminant analysis, which maximized the difference of between-group variance minus within-group variance, followed by permutation testing. Differences within animal between contralateral pairs of joints were minimal. Significant differences among AC, MC and TCRL joint fluid spectra were found. The range of biomolecular differences among these normal joints as characterized by FTIR indicates that interarticular variation within the horse needs to be considered for ongoing research, especially when utilizing within-horse joints as controls

The thermodynamics of prediction

A system responding to a stochastic driving signal can be interpreted as computing, by means of its dynamics, an implicit model of the environmental variables. The system's state retains information about past environmental fluctuations, and a fraction of this information is predictive of future ones. The remaining nonpredictive information reflects model complexity that does not improve predictive power, and thus represents the ineffectiveness of the model. We expose the fundamental equivalence between this model inefficiency and thermodynamic inefficiency, measured by dissipation. Our results hold arbitrarily far from thermodynamic equilibrium and are applicable to a wide range of systems, including biomolecular machines. They highlight a profound connection between the effective use of information and efficient thermodynamic operation: any system constructed to keep memory about its environment and to operate with maximal energetic efficiency has to be predictive.Comment: 5 pages, 1 figur

Electrochemical investigation of the kinetics of chloride substitution upon reduction of [Ru(porphyrin)(NO)Cl] complexes in THF.

The electrochemistry of several ruthenium porphyrin nitrosyl chloride complexes [Ru(por)(NO)Cl] have been examined in tetrahydrofuran. The complexes undergo 1-electron irreversible reductions which result in the diffusion-limited substitutions of the chloride ligands for THF. This chloride metathesis is reversible in the presence of added NBu4Cl, and equilibrium constants and rate constants for chloride loss have been estimated. These parameters correlate with the NO stretching frequencies of the parent complexes, with more electron-donating porphyrin ligands favouring chloride loss from the reduced complexes. The [Ru(por)(NO)(THF)] products of the reductions can be detected by IR, EPR and visible spectroscopies. These species undergo three further reductions, with good reversibility at scan rates \u3e0.40 V s-1. The [Ru(por)(NO)(THF)]+/0 couples have also been determined, and the rate constants and equilibrium constants for recombination with chloride have been estimated. One-electron reductions of the [Ru(por)(NO)Cl] complexes result in ~1018 enhancement of the rates of chloride loss

Halogen Bonding with Phosphine: Evidence for Mulliken Inner Complexes and the Importance of Relaxation Energy

Intermolecular halogen bonding in complexes of phosphine and dihalogens has been theoretically investigated using explicitly correlated coupled cluster methods and symmetry adapted perturbation theory. The complexes H3P· · · ClF, H3P· · · BrF and H3P· · ·IF are demonstrated to possess unusually strong interactions that are accompanied by an increase in the induction component of the interaction energy and significant elongation of the X–Y halogen distance on complex formation. The combination of these factors is indicative of Mulliken inner complexes and criteria for identifying this classification are further developed. The importance of choosing an electronic structure method that describes both dispersion and longer range interactions is demonstrated, along with the need to account for the change in geometry on complexation formation via relaxation energy and overall stabilisation energies

A comparison of methods for estimating Reynolds stress from ADCP measurements in wavy environments

Author Posting. © American Meteorological Society, 2011. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Atmospheric and Oceanic Technology 28 (2011): 1539–1553, doi:10.1175/JTECH-D-11-00001.1.Turbulent Reynolds stresses are now routinely estimated from acoustic Doppler current profiler (ADCP) measurements in estuaries and tidal channels using the variance method, yet biases due to surface gravity waves limit its use in the coastal ocean. Recent modifications to this method, including spatially filtering velocities to isolate the turbulence from wave velocities and fitting a cospectral model to the below-wave band cospectra, have been used to remove this bias. Individually, each modification performed well for the published test datasets, but a comparative analysis over the range of conditions in the coastal ocean has not yet been performed. This work uses ADCP velocity measurements from five previously published coastal ocean and estuarine datasets, which span a range of wave and current conditions as well as instrument configurations, to directly compare methods for estimating stresses in the presence of waves. The computed stresses from each were compared to bottom stress estimates from a quadratic drag law and, where available, estimates of wind stress. These comparisons, along with an analysis of the cospectra, indicated that spectral fitting performs well when the wave climate is wide-banded and/or multidirectional as well as when instrument noise is high. In contrast, spatial filtering performs better when waves are narrow-banded, low frequency, and when wave orbital velocities are strong relative to currents. However, as spatial filtering uses vertically separated velocity bins to remove the wave bias, spectral fitting is able to resolve stresses over a larger fraction of the water column.J. Rosman acknowledges funding from the National Science Foundation (OCE-1061108)

Functional responses can unify invasion ecology.

We contend that invasion ecology requires a universal, measurable trait of species and their interactions with resources that predicts key elements of invasibility and ecological impact; here, we advocate that functional responses can help achieve this across taxonomic and trophic groups, among habitats and contexts, and can hence help unify disparate research interests in invasion ecology

Precision Determination of the Neutron Spin Structure Function g1n

We report on a precision measurement of the neutron spin structure function $g^n_1$ using deep inelastic scattering of polarized electrons by polarized ^3He. For the kinematic range 0.014<x<0.7 and 1 (GeV/c)^2< Q^2< 17 (GeV/c)^2, we obtain $\int^{0.7}_{0.014} g^n_1(x)dx = -0.036 \pm 0.004 (stat) \pm 0.005 (syst)$ at an average $Q^2=5 (GeV/c)^2$. We find relatively large negative values for $g^n_1$ at low $x$. The results call into question the usual Regge theory method for extrapolating to x=0 to find the full neutron integral $\int^1_0 g^n_1(x)dx$, needed for testing quark-parton model and QCD sum rules.Comment: 5 pages, 3 figures To be published in Phys. Rev. Let

Calculating Reynolds stresses from ADCP measurements in the presence of surface gravity waves using the cospectra-fit method

Author Posting. © American Meteorological Society, 2010. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Atmospheric and Oceanic Technology 27 (2010): 889-907, doi:10.1175/2009JTECHO682.1.Recently, the velocity observations of acoustic Doppler current profilers (ADCPs) have been successfully used to estimate turbulent Reynolds stresses in estuaries and tidal channels. However, the presence of surface gravity waves can significantly bias stress estimates, limiting application of the technique in the coastal ocean. This work describes a new approach to estimate Reynolds stresses from ADCP velocities obtained in the presence of waves. The method fits an established semiempirical model of boundary layer turbulence to the measured turbulent cospectra at frequencies below those of surface gravity waves to estimate the stress. Applied to ADCP observations made in weakly stratified waters and variable significant wave heights, estimated near-bottom and near-surface stresses using this method compared well with independent estimates of the boundary stresses in contrast to previous methods. Additionally, the vertical structure of tidal stress estimated using the new approach matched that inferred from a linear momentum balance at stress levels below the estimated stress uncertainties. Because the method makes an estimate of the horizontal turbulent length scales present as part of the model fit, these results can also enable a direct correction for the mean bias errors resulting from instrument tilt, if these scales are long relative to the beam separation.AK acknowledges support from the WHOI Coastal Ocean Institute, and SL acknowledges support from NSF Ocean Science Grant OCE-0548961

HIV-1 Clade D Is Associated with Increased Rates of CD4 Decline in a Kenyan Cohort

HIV-1 is grouped phylogenetically into clades, which may impact rates of HIV-1 disease progression. Clade D infection in particular has been shown to be more pathogenic. Here we confirm in a Nairobi-based prospective female sex worker cohort (1985-2004) that Clade D (n = 54) is associated with a more rapid CD4 decline than clade A1 (n = 150, 20.6% vs 13.4% decline per year, 1.53-fold increase, p = 0.015). This was independent of "protective" HLA and country of origin (p = 0.053), which in turn were also independent predictors of the rate of CD4 decline (p = 0.026 and 0.005, respectively). These data confirm that clade D is more pathogenic than clade A1. The precise reason for this difference is currently unclear, and requires further study. This is first study to demonstrate difference in HIV-1 disease progression between clades while controlling for protective HLA alleles