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Arthur Aronson
The spore coat of Bacilli is a relatively complex structure comprised of about 70 species of proteins in 2 or 3 layers. While some are involved in assembly or protection, the regulation of many are not well defined so lacZ transcriptional fusions were constructed to six Bacillus anthracis spore coat genes in order to gain insight into their possible functions. The genes were selected on the basis of the location of the encoded proteins within the coat and distribution among spore forming species. Conditions tested were temperature and media either as solid or liquid...
March 17, 2018: FEMS Microbiology Letters
Roger D Plaut, Andrea B Staab, Mark A Munson, Joan S Gebhardt, Christopher P Klimko, Avery V Quirk, Christopher K Cote, Tony L Buhr, Rebecca D Rossmaier, Robert C Bernhards, Courtney E Love, Kimberly L Berk, Teresa G Abshire, David A Rozak, Linda C Beck, Scott Stibitz, Bruce G Goodwin, Michael A Smith, Shanmuga Sozhamannan
The revelation in May 2015 of the shipment of γ irradiation-inactivated wild-type Bacillus anthracis spore preparations containing a small number of live spores raised concern about the safety and security of these materials. The finding also raised doubts about the validity of the protocols and procedures used to prepare them. Such inactivated reference materials were used as positive controls in assays to detect suspected B. anthracis in samples because live agent cannot be shipped for use in field settings, in improvement of currently deployed detection methods or development of new methods, or for quality assurance and training activities...
April 2018: Emerging Infectious Diseases
Vatika Gupta, Kanika Jain, Rajni Garg, Anshu Malik, Pooja Gulati, Rakesh Bhatnagar
Microbial colonization is an outcome of appropriate sensing and regulation of its gene expression. Bacillus anthracis adapts and thrives in its environment through complex regulatory mechanisms, among them, the two component systems (TCS). Many bacteria respond to the oxygen fluctuations via TCS. In the present work, a previously uncharacterized TCS, Bas1213-1214, of B. anthracis with a probable role in oxygen sensing has been characterized as a functional TCS. A substantial increase in the expression of Bas1213 was observed during the stationary growth phase, in presence of bicarbonate ions, and under oxidative stress thereby speculating the role of Bas1213 in toxin production and adaptive responses...
March 14, 2018: Journal of Cellular Biochemistry
Alanson W Girton, Narcis I Popescu, Ravi S Keshari, Tarea Burgett, Florea Lupu, K Mark Coggeshall
We showed that human IgG supported the response by human innate immune cells to peptidoglycan (PGN) from Bacillus anthracis and PGN-induced complement activation. However, other serum constituents have been shown to interact with peptidoglycan, including the IgG-like soluble pattern recognition receptor serum amyloid P (SAP). Here, we compared the ability of SAP and of IgG to support monocyte and complement responses to PGN. Utilizing in vitro methods, we demonstrate that SAP is superior to IgG in supporting monocyte production of cytokines in response PGN...
March 12, 2018: Infection and Immunity
Abbas Shali, Sadegh Hasannia, Fatemeh Gashtasbi, S Masoud Abdous, S Shirin Shahangian, Shirin Jalili
Since anthrax is an acute infectious disease, detection and neutralization of the toxins of pathogenic Bacillus anthracis are of great importance. The critical role of protective antigen (PA) component of tripartite anthrax toxin in toxin entry into the host cell cytosol provided a great deal of effort to generate monoclonal antibodies against this constitute. Regarding the importance of anthrax detection/neutralization and unique physicochemical and pharmacological features of VHHs as single domain antibodies, the present study aimed to generate VHHs against the receptor binding domain of PA, termed PAD4...
March 7, 2018: International Journal of Biological Macromolecules
David Sychantha, Robert Chapman, Natalie C Bamford, Geert-Jan Boons, P Lynne Howell, Anthony John Clarke
Bacterial surface (S)-layers are paracrystalline arrays of protein assembled on the bacterial cell wall which serve as protective barriers and scaffolds for housekeeping enzymes and virulence factors. The attachment of S-layer proteins to the cell walls of the Bacillus cereus sensu lato, which includes the pathogen Bacillus anthracis, occurs through non-covalent interactions between their S-layer homology domains and secondary cell wall polysaccharides. To promote recognition for these interactions, it is presumed that the terminal N-acetylmannosamine (ManNAc) residues of the secondary cell wall polysaccharides must be ketal-pyruvylated...
March 9, 2018: Biochemistry
Viktoria I Pauker, Bryan R Thoma, Gregor Grass, Pauline Bleichert, Matthias Hanczaruk, Lothar Zöller, Sabine Zange
Discrimination of highly pathogenic bacteria, such as Bacillus anthracis , from closely related species based on molecular biological methods is challenging. We applied Matrix Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry (MALDI-TOF MS) to a collection of B. anthracis strains and close relatives in order to significantly improve the statistical confidence of identification results for this group of bacteria. Protein mass spectra of 189 verified and diverse Bacillus strains of the Bacillus cereus sensu lato group were generated using MALDI-TOF MS and subsequently analyzed with supervised and unsupervised statistical methods, such as shrinkage discriminant analysis (SDA) and principle component analysis (PCA)...
March 7, 2018: Journal of Clinical Microbiology
Guillaume Méric, Leonardos Mageiros, Ben Pascoe, Dan J Woodcock, Evangelos Mourkas, Sarah Lamble, Rory Bowden, Keith A Jolley, Ben Raymond, Samuel K Sheppard
Bacterial plasmids can vary from small selfish genetic elements to large autonomous replicons that constitute a significant proportion of total cellular DNA. By conferring novel function to the cell, plasmids may facilitate evolution but their mobility may be opposed by co-evolutionary relationships with chromosomes or encouraged via the infectious sharing of genes encoding public goods. Here, we explore these hypotheses through large-scale examination of the association between plasmids and genomes in the phenotypically diverse Bacillus cereus group...
March 6, 2018: Molecular Ecology
Miloslava Duracova, Jana Klimentova, Alena Fucikova, Jiri Dresler
Biological toxins are a heterogeneous group of compounds that share commonalities with biological and chemical agents. Among them, protein toxins represent a considerable, diverse set. They cover a broad range of molecular weights from less than 1000 Da to more than 150 kDa. This review aims to compare conventional detection methods of protein toxins such as in vitro bioassays with proteomic methods, including immunoassays and mass spectrometry-based techniques and their combination. Special emphasis is given to toxins falling into a group of selected agents, according to the Centers for Disease Control and Prevention, such as Staphylococcal enterotoxins , Bacillus anthracis toxins, Clostridium botulinum toxins, Clostridium perfringens epsilon toxin, ricin from Ricinus communis , Abrin from Abrus precatorius or control of trade in dual-use items in the European Union, including lesser known protein toxins such as Viscumin from Viscum album ...
February 28, 2018: Toxins
Natosha L Finley
Dissecting how bacterial pathogens escape immune destruction and cause respiratory infections in humans is a work in progress. One tactic employed by microbes is to use bacterial adenylate cyclase toxins (ACTs) to disarm immune cells and disrupt cellular signaling in host cells, which facilitates the infection process. Several clinically significant pathogens, such as Bacillus anthracis and Bordetella pertussis, have ACTs that are stimulated by an activator protein in human cells. Research has shown that these bacterial ACTs have evolved distinct ways of controlling their activities, but our understanding of how the B...
February 27, 2018: PLoS Biology
Abdurahman H Hirad, Javed Ahmad, Abdulaziz A Alkhedhairy, Ali H Bahkali, Shams T Khan
Customary consumption of unpasteurized milk by the population in the central Najed region of Saudi Arabia may pose a health risk. Therefore, 80 camel milk samples were collected aseptically from seven different stations of Riyadh region. The biochemical and microbiological properties of these milk samples were determined. Nutrient agar and brain heart infusion agar were used to determine mesophilic aerobic counts (MACs). The MAC in each mL of milk varied from 60 to 16 × 104  CFU/mL on nutrient agar. Based on the colony morphology, 176 colonies were collected from different samples, and these isolates were de-replicated into 80 unique isolates using rep-PCR analysis...
March 2018: APMIS: Acta Pathologica, Microbiologica, et Immunologica Scandinavica
Marybeth Langer, Alanson W Girton, Narcis I Popescu, Tarea Burgett, Jordan P Metcalf, K Mark Coggeshall
Peptidoglycan (PGN), a major component of bacterial cell walls, is a pathogen-associated molecular pattern (PAMP) that causes innate immune cells to produce inflammatory cytokines that escalate the host response during infection. In order to better understand the role of PGN in infection, we wanted to gain insight into the cellular receptor for PGN. Although the receptor was initially identified as Toll-like receptor 2 (TLR2), this receptor has remained controversial and other PGN receptors have been reported...
2018: PloS One
Kevin M Claunch, Madeline Bush, Christopher R Evans, Jacob A Malmquist, Matthew C Hale, Shauna M McGillivray
ClpX functions as either an independent chaperone or a component of the ClpXP protease, a conserved intracellular protease that acts as a global regulator in the bacterial cell by degrading regulatory proteins, stress response proteins and rate-limiting enzymes. Previously, we found that loss of clpX in Bacillus anthracis Sterne leads to increased susceptibility to antimicrobial agents that target the cell envelope. The aim of this study was to identify genes within the regulatory network of clpX that contribute to antimicrobial resistance...
February 23, 2018: Microbiology
Tiantian Wang, Dongshu Wang, Yufei Lyu, Erling Feng, Zhu Li, Chunjie Liu, Yanchun Wang, Xiankai Liu, Hengliang Wang
To investigate gene function in Bacillus anthracis, a high-efficiency cloning system is required with an increased rate of allelic exchange. Golden Gate cloning is a molecular cloning strategy allowing researchers to simultaneously and directionally assemble multiple DNA fragments to construct target plasmids using type IIs restriction enzymes and T4 DNA ligase in the same reaction system. Here, a B. anthracis S-layer protein EA1 allelic exchange vector was successfully constructed using the Golden Gate method...
February 10, 2018: Journal of Biotechnology
Mesut Ortatatli, Kadir Canitez, Sermet Sezigen, Ruşen Koray Eyison, Levent Kenar
Decontamination of suspected packages, such as sealed envelopes, liquids and tools that are likely contaminated with biological agents is of great importance. In this study, we aimed to determine the gamma radiation dose required for the decontamination of paper, fabric and liquid materials without causing any damage to the structure of these materials. Each study group included 11 pieces of paper, fabric and sterile saline contaminated with 0.8 × 105 virulent Bacillus anthracis (B. anthracis) spores. These specimens were exposed to doses of 5...
March 2018: Indian Journal of Microbiology
Aparna Banerjee, Shalini Gaur Rudra, Koushik Mazumder, Vinod Nigam, Rajib Bandopadhyay
Exopolysaccharide produced by a unique avirulent Bacillus anthracis strain PFAB2 of hot spring origin has been characterized and its functional properties are investigated which is a first report. Maximum yield of EPS is 7.66 g/l with 2% glucose and 1% peptone as optimum carbon and nitrogen source respectively. The EPS is found to be a homopolymer consisting of only glucose as principle monosaccharide component. Through 1H NMR study, different dextran-like proton peaks are observed. Molecular weight of the EPS resembles low molecular weight bacterial origin polysaccharides...
March 2018: Indian Journal of Microbiology
Joel W Francis, Christopher J Royer, Paul D Cook
Bacillithiol is a low-molecular weight thiol produced by many gram-positive organisms, including Staphylococcus aureus and Bacillus anthracis. It is the major thiol responsible for maintaining redox homeostasis and cellular detoxification, including inactivation of the antibiotic fosfomycin. The metal-dependent bacillithiol transferase BstA is likely involved in these sorts of detoxification processes, but the exact substrates and enzyme mechanism have not been identified. Here we report the 1.34 Å resolution X-ray crystallographic structure of BstA from S...
February 8, 2018: Protein Science: a Publication of the Protein Society
Janet Z Liu, Syed R Ali, Ethan Bier, Victor Nizet
Bacillus anthracis, the causative agent of anthrax, has been a focus of study in host-pathogen dynamics since the nineteenth century. While the interaction between anthrax and host macrophages has been extensively modeled, comparatively little is known about the effect of anthrax on the immune function of neutrophils, a key frontline effector of innate immune defense. Here we showed that depletion of neutrophils significantly enhanced mortality in a systemic model of anthrax infection in mice. Ex vivo, we found that freshly isolated human neutrophils can rapidly kill anthrax, with specific inhibitor studies showing that phagocytosis and reactive oxygen species (ROS) generation contribute to this efficient bacterial clearance...
2018: Frontiers in Cellular and Infection Microbiology
Pieter Johan Steenkamp, Henriette van Heerden, Ockert Louis van Schalkwyk
The spores of the soil-borne bacterium, Bacillus anthracis, which causes anthrax are highly resistant to adverse environmental conditions. Under ideal conditions, anthrax spores can survive for many years in the soil. Anthrax is known to be endemic in the northern part of Kruger National Park (KNP) in South Africa (SA), with occasional epidemics spreading southward. The aim of this study was to identify and map areas that are ecologically suitable for the harboring of B. anthracis spores within the KNP. Anthrax surveillance data and selected environmental variables were used as inputs to the maximum entropy (Maxent) species distribution modeling method...
2018: PloS One
Janine R Hutchison, Greg F Piepel, Brett G Amidan, Becky M Hess, Michael A Sydor, Brooke L Deatherage Kaiser
AIMS: We evaluated the effects of Bacillus anthracis surrogates, low surface concentrations, surface materials, and assay methods on false-negative rate (FNR) and limit of detection (LOD95 ) for recovering Bacillus spores using a macrofoam-swab sampling procedure. METHODS AND RESULTS: Bacillus anthracis Sterne or Bacillus atrophaeus Nakamura spores were deposited over a range of low target concentrations (2 - 500 coupon-1 ) onto glass, stainless steel, vinyl tile, and plastic...
January 21, 2018: Journal of Applied Microbiology
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