57 research outputs found

    Absence of the KhpA and KhpB (JAG/EloR) RNA-Binding Proteins Suppresses the Requirement for PBP2b by Overproduction of FtsA in Streptococcus pneumoniae D39

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    Suppressor mutations were isolated that obviate the requirement for essential PBP2b in peripheral elongation of peptidoglycan from the midcells of dividing Streptococcus pneumoniae D39 background cells. One suppressor was in a gene encoding a single KH-domain protein (KhpA). ΔkhpA suppresses deletions in most, but not all (mltG), genes involved in peripheral PG synthesis and in the gpsB regulatory gene. ΔkhpA mutations reduce growth rate, decrease cell size, minimally affect shape, and induce expression of the WalRK cell-wall stress regulon. Reciprocal co-immunoprecipitations show that KhpA forms a complex in cells with another KH-domain protein (KhpB/JAG/EloR). ΔkhpA and ΔkhpB mutants phenocopy each other exactly, consistent with a direct interaction. RNA-immunoprecipitation showed that KhpA/KhpB bind an overlapping set of RNAs in cells. Phosphorylation of KhpB reported previously does not affect KhpB function in the D39 progenitor background. A chromosome duplication implicated FtsA overproduction in Δpbp2b suppression. We show that cellular FtsA concentration is negatively regulated by KhpA/B at the post-transcriptional level and that FtsA overproduction is necessary and sufficient for suppression of Δpbp2b. However, increased FtsA only partially accounts for the phenotypes of ΔkhpA mutants. Together, these results suggest that multimeric KhpA/B may function as a pleiotropic RNA chaperone controlling pneumococcal cell division

    Characterize the assembly of dark matter halos with protohalo size histories: I. Redshift evolution, relation to descendant halos, and halo assembly bias

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    We propose a novel method to quantify the assembly histories of dark matter halos with the redshift evolution of the mass-weighted spatial variance of their progenitor halos, i.e. the protohalo size history. We find that the protohalo size history for each individual halo at z~0 can be described by a double power-law function. The amplitude of the fitting function strongly correlates to the central-to-total stellar mass ratios of descendant halos. The variation of the amplitude of the protohalo size history can induce a strong halo assembly bias effect for massive halos. This effect is detectable in observation using the central-to-total stellar mass ratio as a proxy of the protohalo size. The correlation to the descendant central-to-total stellar mass ratio and the halo assembly bias effect seen in the protohalo size are much stronger than that seen in the commonly adopted half-mass formation time derived from the mass accretion history. This indicates that the information loss caused by the compression of halo merger trees to mass accretion histories can be captured by the protohalo size history. Protohalo size thus provides a useful quantity to connect protoclusters across cosmic time and to link protoclusters with their descendant clusters in observations.Comment: 19 pages, 12 + 8 figures, comments are welcome

    Deep Learning of Dark Energy Spectroscopic Instrument Mock Spectra to Find Damped Ly alpha Systems

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    We have updated and applied a convolutional neural network (CNN) machine-learning model to discover and characterize damped Lyα systems (DLAs) based on Dark Energy Spectroscopic Instrument (DESI) mock spectra. We have optimized the training process and constructed a CNN model that yields a DLA classification accuracy above 99% for spectra that have signal-to-noise ratios (S/N) above 5 per pixel. The classification accuracy is the rate of correct classifications. This accuracy remains above 97% for lower S/N ≈1 spectra. This CNN model provides estimations for redshift and H i column density with standard deviations of 0.002 and 0.17 dex for spectra with S/N above 3 pixel-1. Also, this DLA finder is able to identify overlapping DLAs and sub-DLAs. Further, the impact of different DLA catalogs on the measurement of baryon acoustic oscillations (BAO) is investigated. The cosmological fitting parameter result for BAO has less than 0.61% difference compared to analysis of the mock results with perfect knowledge of DLAs. This difference is lower than the statistical error for the first year estimated from the mock spectra: above 1.7%. We also compared the performances of the CNN and Gaussian Process (GP) models. Our improved CNN model has moderately 14% higher purity and 7% higher completeness than an older version of the GP code, for S/N > 3. Both codes provide good DLA redshift estimates, but the GP produces a better column density estimate by 24% less standard deviation. A credible DLA catalog for the DESI main survey can be provided by combining these two algorithms

    SalK/SalR, a Two-Component Signal Transduction System, Is Essential for Full Virulence of Highly Invasive Streptococcus suis Serotype 2

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    BACKGROUND: Streptococcus suis serotype 2 (S. suis 2, SS2) has evolved into a highly infectious entity, which caused the two recent large-scale outbreaks of human SS2 epidemic in China, and is characterized by a toxic shock-like syndrome. However, the molecular pathogenesis of this new emerging pathogen is still poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: 89K is a newly predicted pathogenicity island (PAI) which is specific to Chinese epidemic strains isolated from these two SS2 outbreaks. Further bioinformatics analysis revealed a unique two-component signal transduction system (TCSTS) located in the candidate 89K PAI, which is orthologous to the SalK/SalR regulatory system of Streptococcus salivarius. Knockout of salKR eliminated the lethality of SS2 in experimental infection of piglets. Functional complementation of salKR into the isogenic mutant DeltasalKR restored its soaring pathogenicity. Colonization experiments showed that the DeltasalKR mutant could not colonize any susceptible tissue of piglets when administered alone. Bactericidal assays demonstrated that resistance of the mutant to polymorphonuclear leukocyte (PMN)-mediated killing was greatly decreased. Expression microarray analysis exhibited a transcription profile alteration of 26 various genes down-regulated in the DeltasalKR mutant. CONCLUSIONS/SIGNIFICANCE: These findings suggest that SalK/SalR is requisite for the full virulence of ethnic Chinese isolates of highly pathogenic SS2, thus providing experimental evidence for the validity of this bioinformatically predicted PAI

    Robust estimation of bacterial cell count from optical density

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    Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

    Case Study: Comparative Analysis of Hydrologic Simulations with Areal-Averaging of Moving Rainfall

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    The goal of this investigation is to compare the hydrologic simulations caused by the areal-averaging of dynamic moving rainfall. Two types of synthetic rainfall are developed: spatially varied rainfall (SVR) is the typical input to a distributed model while temporally varied rainfall (TVR) emulates SVR but is spread uniformly over the entire watershed as in the case of a lumped model. This study demonstrates a direct comparison of peak discharge and peak timing generated by synthetic moving storms over idealized rectangular basins and a real watershed. It is found that the difference between the hydrologic responses from SVR and TVR reflects the impact from the areal-averaging of rainfall; the areal-averaging of rainfall for the movement from upstream to downstream over a lumped model can result in underestimated and delayed peak values in comparison to those from a distributed model; the flood peaks from SVR and TVR are found similar when the storm moves from downstream to upstream. The findings of the study suggest that extra cautions are needed for practitioners when evaluating simulated results from distributed and lumped modeling approaches even using the same rainfall information
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