Comparative Genomic Analysis of the Human Pathogen Wohlfahrtiimonas Chitiniclastica Provides Insight Into the Identification of Antimicrobial Resistance Genotypes and Potential Virulence Traits

Recent studies suggest that Wohlfahrtiimonas chitiniclastica may be the cause of several diseases in humans including sepsis and bacteremia making the bacterium as a previously underappreciated human pathogen. However, very little is known about the pathogenicity and genetic potential of W. chitiniclastica ; therefore, it is necessary to conduct systematic studies to gain a deeper understanding of its virulence characteristics and treatment options. In this study, the entire genetic repertoire of all publicly available W. chitiniclastica genomes was examined including in silico characterization of bacteriophage content, antibiotic resistome, and putative virulence profile. The pan-genome of W. chitiniclastica comprises 3819 genes with 1622 core genes (43%) indicating a putative metabolic conserved species. Furthermore, in silico analysis indicated presumed resistome expansion as defined by the presence of genome-encoded transposons and bacteriophages. While macrolide resistance genes macA and macB are located within the core genome, additional antimicrobial resistance genotypes for tetracycline ( tetH, tetB , and tetD ), aminoglycosides ( ant(2’’)-Ia, aac(6’)-Ia , aph(3’’)-Ib , aph(3’)-Ia , and aph(6)-Id )), sulfonamide ( sul2 ), streptomycin ( strA ), chloramphenicol ( cat3 ), and beta-lactamase ( blaVEB ) are distributed among the accessory genome. Notably, our data indicate that the type strain DSM 18708 T does not encode any additional clinically relevant antibiotic resistance genes, whereas drug resistance is increasing within the W. chitiniclastica clade. This trend should be monitored with caution. To the best of our knowledge, this is the first comprehensive genome analysis of this species, providing new insights into the genome of this opportunistic human pathogen

Sample deposition onto cryo-EM grids: from sprays to jets and back

Despite the great strides made in the field of single-particle cryogenic electron microscopy (cryo-EM) in microscope design, direct electron detectors and new processing suites, the area of sample preparation is still far from ideal. Traditionally, sample preparation involves blotting, which has been used to achieve high resolution, particularly for well behaved samples such as apoferritin. However, this approach is flawed since the blotting process can have adverse effects on some proteins and protein complexes, and the long blot time increases exposure to the damaging air-water interface. To overcome these problems, new blotless approaches have been designed for the direct deposition of the sample on the grid. Here, different methods of producing droplets for sample deposition are compared. Using gas dynamic virtual nozzles, small and high-velocity droplets were deposited on cryo-EM grids, which spread sufficiently for high-resolution cryo-EM imaging. For those wishing to pursue a similar approach, an overview is given of the current use of spray technology for cryo-EM grid preparation and areas for enhancement are pointed out. It is further shown how the broad aspects of sprayer design and operation conditions can be utilized to improve grid quality reproducibly

Isotopic evidence (⁸⁷Sr/⁸⁶Sr, δ⁷Li) for alteration of the oceanic crust at deep-rooted mud volcanoes in the Gulf of Cadiz, NE Atlantic Ocean [(Sr-87/Sr-86, delta Li-7) ]

The chemical and isotopic composition of pore fluids is presented for five deep-rooted mud volcanoes aligned on a transect across the Gulf of Cadiz continental margin at water depths between 350 and 3860 m. Generally decreasing interstitial Li concentrations and Sr-87/Sr-86 ratios with increasing distance from shore are attributed to systematically changing fluid sources across the continental margin. Although highest Li concentrations at the near-shore mud volcanoes coincide with high salinities derived from dissolution of halite and late-stage evaporites, clayey, terrigenous sediments are identified as the ultimate Li source to all pore fluids investigated. Light delta Li-7 values, partly close to those of hydrothermal vent fluids (delta Li-7: +11.9 parts per thousand), indicate that Li has been mobilized during high-temperature fluid/sediment or fluid/rock interactions in the deep sub-surface. Intense leaching of terrigenous clay has led to radiogenic Sr-87/Sr-86 ratios (similar to 0.7106) in pore fluids of the near-shore mud volcanoes. In contrast, non-radiogenic Sr-87/Sr-86 ratios (similar to 0.7075) at the distal locations are attributed to admixing of a basement-derived fluid component, carrying an isotopic signature from interaction with the basaltic crust. This inference is substantiated by temperature constraints from Li isotope equilibrium calculations suggesting exchange processes at particularly high temperatures (>200 degrees C) for the least radiogenic pore fluids of the most distal location.Advective pore fluids in the off-shore reaches of the Gulf of Cadiz are influenced by successive exchange processes with both oceanic crust and terrigenous, fine-grained sediments, resulting in a chemical and isotopic signature similar to that of fluids in near-shore ridge flank hydrothermal systems. This suggests that deep-rooted mud volcanoes in the Gulf of Cadiz represent a fluid pathway intermediate between mid-ocean ridge hydrothermal vent and shallow, marginal cold seep. Due to the thicker sediment coverage and slower fluid advection rates, the overall geochemical signature is shifted towards the sediment-diagenetic signal compared to ridge flank hydrothermal environments. (C) 2009 Elsevier Ltd. All rights reserved

psychotree - Recursive partitioning based on psychometric models: Version 0.12-1

Recursive partitioning based on psychometric models,employing the general MOB algo- rithm (from package party) to obtain Bradley-Terry trees and Rasch trees

Demolition and recycling of carbon reinforced concrete

Carbon reinforced concrete is an artificially produced composite construction material consisting of two components: concrete and continuous carbon filaments formed as roving fabric or bars. The research project C³-V1.5 “Demolition, Dismantling and Recycling” aims to answer the key questions regarding the LCA. Under the leadership of the “Institute of Construction Technology” (TECHNISCHE UNIVERSITÄT DRESDEN), 5 companies and 4 research institutes are involved in the project. The main goal of the research project is to keep carbon concrete materials in the life cycle and to avoid down-cycling. Furthermore, investigations concerning health and safety will be carried out to make sure that no hazardous emissions are released while working with the material. To reach this goal, laboratory tests will be conducted to set the basis for large-scale experiments. Within these experiments relevant concrete processing and recycling technologies will be assessed. First results of the laboratory tests indicate that the wear on the machines while processing carbon concrete is significantly lower than processing steel reinforced concrete. Furthermore, carbon concrete allows the use of much smaller machines due to the lack of steel reinforcement. Concerning the measurements of emissions, first results show that no dangerous fibre particles (WHO fibres) were released

siRNA screen of early poxvirus genes identifies the AAA+ ATPase D5 as the virus genome-uncoating factor

SummaryPoxvirus genome uncoating is a two-step process. First, cytoplasmic viral cores are activated and early viral genes are expressed. Next, cores are disassembled and the genomes released. This second step depends on an early viral factor(s) that has eluded identification for over 40 years. We used a large-scale, high-throughput RNAi screen directed against vaccinia virus (VACV) to identify the VACV AAA+ ATPase D5 as the poxvirus uncoating factor. We show that the ATPase activity of D5 is required for uncoating. Superresolution microscopy suggests that D5 acts directly at viral cores for genome release. Thus, the putative helicase D5 is a multifunctional protein required for genome uncoating and replication. Additionally, in vivo delivery of anti-D5 siRNAs reduced virus production in a mouse model of VACV infection. These results demonstrate the use of virus-targeting RNAi libraries to investigate viral gene function and suggest therapeutic avenues

Immunological Predictors of Dimethyl Fumarate-Induced Lymphopenia

Treatment with dimethyl fumarate (DMF) leads to lymphopenia and infectious complications in a subset of patients with multiple sclerosis (MS). Here, we aimed to reveal immune markers of DMF-associated lymphopenia. This prospective observational study longitudinally assessed 31 individuals with MS by single-cell mass cytometry before and after 12 and 48 weeks of DMF therapy. Employing a neural network-based representation learning approach, we identified a CCR4-expressing T helper cell population negatively associated with relevant lymphopenia. CCR4-expressing T helper cells represent a candidate prognostic biomarker for the development of relevant lymphopenia in patients undergoing DMF treatment. ANN NEUROL 2022ISSN:0364-5134ISSN:1531-824