DOCSLIB.ORG

Functional Similarity of PRD-Containing Virulence Regulators in Bacillus Anthracis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Functional Similarity of PRD-Containing Virulence Regulators in Bacillus Anthracis The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 5-2018 Functional Similarity of PRD-Containing Virulence Regulators in Bacillus anthracis Malik Raynor Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Bacteriology Commons, Biochemistry Commons, Molecular Biology Commons, Molecular Genetics Commons, and the Pathogenic Microbiology Commons Recommended Citation Raynor, Malik, "Functional Similarity of PRD-Containing Virulence Regulators in Bacillus anthracis" (2018). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 857. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/857 This Dissertation (PhD) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. FUNCTIONAL SIMILARITY OF PRD-CONTAINING VIRULENCE REGULATORS IN BACILLUS ANTHRACIS by Malik Jamaal Raynor, B.S. APPROVED: ______________________________ Theresa M. Koehler, Ph.D. Advisory Professor ______________________________ Peter J. Christie, Ph.D. ______________________________ Kevin A. Morano, Ph.D. ______________________________ Hung Ton-That, Ph.D. ______________________________ Eric J. Wagner, Ph.D. APPROVED: ____________________________ Dean, The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences FUNCTIONAL SIMILARITY OF PRD-CONTAINING VIRULENCE REGULATORS IN BACILLUS ANTHRACIS A DISSERTATION Presented to the Faculty of The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY by Malik Jamaal Raynor, B.S. Houston, Texas Date of Graduation (May, 2018) FUNCTIONAL SIMILARITY OF PRD-CONTAINING VIRULENCE REGULATORS IN BACILLUS ANTHRACIS Malik Jamaal Raynor, B.S. Advisory Professor: Theresa M. Koehler, Ph.D. Bacillus anthracis produces three regulators, AtxA, AcpA, and AcpB, that control virulence gene expression and are members of an emerging class of regulators termed “PCVRs” (Phosphoenolpyruvate-dependent phosphotransferase regulation Domain-Containing Virulence Regulators). AtxA controls expression of the toxin genes; lef, cya, and pag, and is the master virulence regulator and archetype PCVR. AcpA and AcpB are less well studied. AcpA and AcpB independently positively control transcription of the capsule biosynthetic operon capBCADE, and culture conditions that enhance AtxA activity result in capBCADE transcription in strains lacking acpA and acpB. RNA-Seq was used to assess the regulons of the paralogs in strains producing individual PCVRs at native levels. Plasmid- and chromosome-borne genes were PCVR-controlled, with AtxA, AcpA, and AcpB having a ≥4-fold effect on transcript levels of 145, 130, and 49 genes respectively. Several genes were co-regulated by two or three PCVRs. Results from transcriptional reporters of PCVR-regulated promoters fused to promoterless lacZ genes largely mirrored RNA-Seq data showing AtxA alone had activity on Plef-lacZ, and AcpA and AcpB had more activity than AtxA on PcapB-lacZ. Studies to test the effect of AtxA levels on virulence and sporulation used atxA mutants. A mutant that overexpressed atxA and exhibited elevated AtxA and toxin levels in vitro, was not increased for virulence in a murine anthrax infection model. AtxA levels also affected sporulation efficiency. Culture of B. anthracis in medium containing bicarbonate and elevated carbon dioxide increased PCVR activity compared to culture in ambient air in medium lacking bicarbonate. However, neither the solubility nor stability of the regulators was affected by carbon dioxide concentration. AcpA and AcpB form homomultimers and multimerization was dependent on the iii EIIB-like domains, as shown previously for AtxA. Heteromultimers of AtxA-AcpA were detected and in co-expression experiments, AcpA activity was reduced by increased levels of AtxA. An AtxA orthologue in Bacillus cereus, AtxA2, had less activity than AtxA from B. anthracis potentially due to reduced dimer formation. The results provided in this dissertation increase our knowledge of virulence gene expression in B. anthracis, while advancing our understanding of this newly-discovered class of transcriptional regulators. iv Table of Contents Approval Sheet..............................................................................................................................i Title Page......................................................................................................................................ii Abstract........................................................................................................................................iii Table of Contents.........................................................................................................................v List of Figures.............................................................................................................................viii List of Tables................................................................................................................................x Chapter I: Introduction...............................................................................................................1 1.1 Gene expression modulated by environmental conditions.........................................2 1.2 PRD-containing virulence regulators..........................................................................5 1.3 Bacillus anthracis and anthrax disease......................................................................7 1.4 Virulence factors in anthrax pathogenesis.................................................................9 1.5 AtxA: the most well-characterized PCVR.................................................................11 1.6 AcpA and AcpB: regulators of capsule expression...................................................14 1.7 AtxA2, an AtxA homologue in Bacillus cereus strain G9241....................................14 1.8 Gaps in knowledge and significance of this work.....................................................15 Chapter II: Materials and Methods..........................................................................................17 2.1 Culture conditions.....................................................................................................18 2.2 Strain construction....................................................................................................18 2.3 Native and IPTG-induced PCVR expression............................................................21 2.4 RNA isolation for RNA-seq.......................................................................................21 2.5 Creation of Next Generation Sequencing (NGS) libraries for RNA-seq and sequencing..................................................................................................22 2.6 RNA-seq and bioinformatic analysis.........................................................................23 2.7 Western blotting........................................................................................................24 2.8 India ink exclusion assay..........................................................................................25 v 2.9 Co-affinity purification...............................................................................................25 2.10 AcpA-His and AcpB-His purification.......................................................................26 2.11 Bis-maleimidohexane crosslinking.........................................................................26 2.12 Mouse infections.....................................................................................................27 2.13 Heat-resistant CFU determination..........................................................................27 2.14 Real-time quantitative PCR (RT-qPCR).................................................................28 Chapter III: Regulons of PRD-containing Bacillus anthracis virulence regulators reveal overlapping but distinct functions..........................................................35 3.1 Introduction...............................................................................................................36 3.2 Results......................................................................................................................38 3.2.1 Amino acid sequence and predicted domain similarity of the B. anthracis PCVRs...................................................................................38 3.2.2 Native PCVR protein and transcript levels.................................................41 3.2.3 Overlapping regulons of AtxA, AcpA, and AcpB........................................45 3.2.4 Loci of PCVR-regulated genes..................................................................49
Load more

Recommended publications
  • Developing Drugs for Prophylaxis of Inhalational Anthrax Guidance for Industry
    Anthrax: Developing Drugs for Prophylaxis of Inhalational Anthrax Guidance for Industry U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) May 2018 Clinical/Antimicrobial Anthrax: Developing Drugs for Prophylaxis of Inhalational Anthrax Guidance for Industry Additional copies are available from: Office of Communications, Division of Drug Information Center for Drug Evaluation and Research Food and Drug Administration 10001 New Hampshire Ave., Hillandale Bldg., 4th Floor Silver Spring, MD 20993-0002 Phone: 855-543-3784 or 301-796-3400; Fax: 301-431-6353; Email: [email protected] https://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/default.htm U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) May 2018 Clinical/Antimicrobial TABLE OF CONTENTS I. INTRODUCTION............................................................................................................. 1 II. BACKGROUND ............................................................................................................... 2 A. Historical Background................................................................................................................... 2 B. Indication for Prophylaxis of Inhalational Anthrax ................................................................... 2 III. DEVELOPMENT PROGRAM ....................................................................................... 3 A. General
    [Show full text]
  • Amerithrax Investigative Summary
    The United States Department of Justice AMERITHRAX INVESTIGATIVE SUMMARY Released Pursuant to the Freedom of Information Act Friday, February 19, 2010 TABLE OF CONTENTS I. THE ANTHRAX LETTER ATTACKS . .1 II. EXECUTIVE SUMMARY . 4 A. Overview of the Amerithrax Investigation . .4 B. The Elimination of Dr. Steven J. Hatfill as a Suspect . .6 C. Summary of the Investigation of Dr. Bruce E. Ivins . 6 D. Summary of Evidence from the Investigation Implicating Dr. Ivins . .8 III. THE AMERITHRAX INVESTIGATION . 11 A. Introduction . .11 B. The Investigation Prior to the Scientific Conclusions in 2007 . 12 1. Early investigation of the letters and envelopes . .12 2. Preliminary scientific testing of the Bacillus anthracis spore powder . .13 3. Early scientific findings and conclusions . .14 4. Continuing investigative efforts . 16 5. Assessing individual suspects . .17 6. Dr. Steven J. Hatfill . .19 7. Simultaneous investigative initiatives . .21 C. The Genetic Analysis . .23 IV. THE EVIDENCE AGAINST DR. BRUCE E. IVINS . 25 A. Introduction . .25 B. Background of Dr. Ivins . .25 C. Opportunity, Access and Ability . 26 1. The creation of RMR-1029 – Dr. Ivins’s flask . .26 2. RMR-1029 is the source of the murder weapon . 28 3. Dr. Ivins’s suspicious lab hours just before each mailing . .29 4. Others with access to RMR-1029 have been ruled out . .33 5. Dr. Ivins’s considerable skill and familiarity with the necessary equipment . 36 D. Motive . .38 1. Dr. Ivins’s life’s work appeared destined for failure, absent an unexpected event . .39 2. Dr. Ivins was being subjected to increasing public criticism for his work .
    [Show full text]
  • Molecular Subtyping of Bacillus Anthracis and the 2001 Bioterrorism-Associated Anthrax Outbreak, United States Alex R
    BIOTERRORISM-RELATED ANTHRAX Molecular Subtyping of Bacillus anthracis and the 2001 Bioterrorism-Associated Anthrax Outbreak, United States Alex R. Hoffmaster,* Collette C. Fitzgerald,* Efrain Ribot,* Leonard W. Mayer,* and Tanja Popovic* Molecular subtyping of Bacillus anthracis played an important role in differentiating and identifying strains during the 2001 bioterrorism-associated outbreak. Because B. anthracis has a low level of genetic variabil- ity, only a few subtyping methods, with varying reliability, exist. We initially used multiple-locus variable- number tandem repeat analysis (MLVA) to subtype 135 B. anthracis isolates associated with the outbreak. All isolates were determined to be of genotype 62, the same as the Ames strain used in laboratories. We sequenced the protective antigen gene (pagA) from 42 representative outbreak isolates and determined they all had a pagA sequence indistinguishable from the Ames strain (PA genotype I). MLVA and pagA sequencing were also used on DNA from clinical specimens, making subtyping B. anthracis possible with- out an isolate. Use of high-resolution molecular subtyping determined that all outbreak isolates were indis- tinguishable by the methods used and probably originated from a single source. In addition, subtyping rapidly identified laboratory contaminants and nonoutbreak–related isolates. he recent bioterrorism-associated anthrax outbreak dem- subtype 26 diverse B. anthracis isolates into six PA genotypes T onstrated the need for rapid molecular subtyping of Bacil- (8). Although sequencing of pagA results in limited numbers lus anthracis isolates. Numerous methods, including multiple- of subtypes, it does have the added benefit of determining if locus enzyme electrophoresis (MEE) and multiple-locus the pagA gene has been altered or engineered.
    [Show full text]
  • Bioterrorism & Biodefense
    Hugh-Jones et al. J Bioterr Biodef 2011, S3 Bioterrorism & Biodefense http://dx.doi.org/10.4172/2157-2526.S3-001 Review Article Open Access The 2001 Attack Anthrax: Key Observations Martin E Hugh-Jones1*, Barbara Hatch Rosenberg2 and Stuart Jacobsen3 1Professor Emeritus, Louisiana State University; Anthrax Moderator, ProMED-mail, USA 2Sloan-Kettering Institute for Cancer Research and State Univ. of NY-Purchase (retired); Scientists Working Group on CBW, Center for Arms Control and Non-Proliferation, USA 3Technical Consultant Silicon Materials, Dallas, TX,USA Abstract Unresolved scientificquestions, remaining ten years after the anthrax attacks, three years after the FBI accused a dead man of perpetrating the 2001 anthrax attacks singlehandedly, and more than a year since they closed the case without further investigation, indictment or trial, are perpetuating serious concerns that the FBI may have accused the wrong person of carrying out the anthrax attacks. The FBI has not produced concrete evidence on key questions: • Where and how were the anthrax spores in the attack letters prepared? There is no material evidence of where the attack anthrax was made, and no direct evidence that any specific individual made the anthrax, or mailed it. On the basis of a number` of assumptions, the FBI has not scrutinized the most likely laboratories. • How and why did the spore powders acquire the high levels of silicon and tin found in them? The FBI has repeatedly insisted that the powders in the letters contained no additives, but they also claim that they have not been able to reproduce the high silicon content in the powders, and there has been little public mention of the extraordinary presence of tin.
    [Show full text]
  • Vaccination of Rhesus Macaques with the Anthrax Vaccine Adsorbed
    CLINICAL AND VACCINE IMMUNOLOGY, Nov. 2010, p. 1753–1762 Vol. 17, No. 11 1556-6811/10/$12.00 doi:10.1128/CVI.00174-10 Copyright © 2010, American Society for Microbiology. All Rights Reserved. Vaccination of Rhesus Macaques with the Anthrax Vaccine Adsorbed Vaccine Produces a Serum Antibody Response That Effectively Neutralizes Receptor-Bound Protective Antigen In Vitroᰔ Kristin H. Clement,1* Thomas L. Rudge, Jr.,1 Heather J. Mayfield,1 Lena A. Carlton,1 Arelis Hester,1 Nancy A. Niemuth,1 Carol L. Sabourin,1 April M. Brys,1 and Conrad P. Quinn2 Battelle Memorial Institute, 505 King Avenue, Columbus, Ohio 43201,1 and Meningitis and Vaccine Preventable Diseases Branch, Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 303332 Received 29 April 2010/Returned for modification 17 June 2010/Accepted 19 August 2010 Anthrax toxin (ATx) is composed of the binary exotoxins lethal toxin (LTx) and edema toxin (ETx). They have separate effector proteins (edema factor and lethal factor) but have the same binding protein, protective antigen (PA). PA is the primary immunogen in the current licensed vaccine anthrax vaccine adsorbed (AVA [BioThrax]). AVA confers protective immunity by stimulating production of ATx-neutralizing antibodies, which could block the intoxication process at several steps (binding of PA to the target cell surface, furin cleavage, toxin complex formation, and binding/translocation of ATx into the cell). To evaluate ATx neutral- ization by anti-AVA antibodies, we developed two low-temperature LTx neutralization activity (TNA) assays that distinguish antibody blocking before and after binding of PA to target cells (noncomplexed [NC] and receptor-bound [RB] TNA assays).
    [Show full text]
  • Gao-15-80, Anthrax
    United States Government Accountability Office Report to Congressional Requesters December 2014 ANTHRAX Agency Approaches to Validation and Statistical Analyses Could Be Improved GAO-15-80 December 2014 ANTHRAX Agency Approaches to Validation and Statistical Analyses Could Be Improved Highlights of GAO-15-80, a report to congressional requesters Why GAO Did This Study What GAO Found In 2001, the FBI investigated an After the 2001 Anthrax attacks, the genetic tests that were conducted by the intentional release of B. anthracis, a Federal Bureau of Investigation’s (FBI) four contractors were generally bacterium that causes anthrax, which scientifically verified and validated, and met the FBI’s criteria. However, GAO was identified as the Ames strain. found that the FBI lacked a comprehensive approach—or framework—that could Subsequently, FBI contractors have ensured standardization of the testing process. As a result, each of the developed and validated several contractors developed their tests differently, and one contractor did not conduct genetic tests to analyze B. anthracis verification testing, a key step in determining whether a test will meet a user’s samples for the presence of certain requirements, such as for sensitivity or accuracy. Also, GAO found that the genetic mutations. The FBI had contractors did not develop the level of statistical confidence for interpreting the previously collected and maintained testing results for the validation tests they performed. Responses to future these samples in a repository. incidents could be improved by using a standardized framework for achieving GAO was asked to review the FBI’s minimum performance standards during verification and validation, and by genetic test development process and incorporating statistical analyses when interpreting validation testing results.
    [Show full text]
  • Plague Fact Sheet
    SPARVAX™ - RECOMBINANT PROTECTIVE ANTIGEN (rPA) ANTHRAX VACCINE – NOVEL SECOND GENERATION VACCINE TECHNOLOGY Bacillus anthracis (Anthrax) Infection Bacillus anthracis is a spore forming, gram positive bacterium that has potential to be used as a weapon of bioterror when delivered in an aerosolized form. Following germination of the spores, the bacteria replicates and produces three toxins. Anthrax Protective Antigen (PA) initiates the onset of the illness by attaching to cells in the infected person where it then facilitates entry of the two additional destructive toxins - Lethal Factor (LF) and Edema Factor (EF) into the cell. Current Standard of Care Antibiotics are the first line of defense against anthrax infection. However, early identification and treatment are critical for successful outcome. Even with aggressive antibiotic therapy, five of the eleven victims of the 2001 anthrax postal attacks died, underscoring the need for improved vaccines and anti-toxins for civilian protection. The current FDA licensed anthrax vaccine (BioThrax® Anthrax Vaccine Adsorbed) is approved for the prevention of anthrax infection, but requires six doses over a period of eighteen months to achieve protective immunity. AVA is a first generation anthrax vaccine made from cell free filtrates of whole bacterial cultures of Bacillus anthracis. This vaccine was FDA licensed in 1970. SparVax™ Key Characteristics SparVax™ is a novel second generation recombinant protective (rPA) anthrax vaccine being developed for administration by intramuscular injection. Phase I and Phase II clinical trials involving more than 700 healthy human subjects have been completed and showed that SparVax™ appears to be well tolerated and induces an immune response in humans. These studies suggest that three doses of SparVax™, administered several weeks apart, should be sufficient to induce protective immunity.
    [Show full text]
  • Clpx Contributes to Innate Defense Peptide Resistance and Virulence Phenotypes of Bacillus Anthracis
    Research Article Journal of Innate J Innate Immun 2009;1:494–506 Received: March 3, 2009 Immunity DOI: 10.1159/000225955 Accepted after revision: April 7, 2009 Published online: June 18, 2009 ClpX Contributes to Innate Defense Peptide Resistance and Virulence Phenotypes of Bacillus anthracis a a f b Shauna M. McGillivray Celia M. Ebrahimi Nathan Fisher Mojgan Sabet a g c d Dawn X. Zhang Yahua Chen Nina M. Haste Raffi V. Aroian b b f Richard L. Gallo Donald G. Guiney Arthur M. Friedlander g a, c, e Theresa M. Koehler Victor Nizet a b c Departments of Pediatrics and Medicine, Skaggs School of Pharmacy and Pharmaceutical Sciences, and d e Division of Biological Sciences, University of California San Diego, La Jolla, Calif., Rady Children’s Hospital, f San Diego, Calif. , United States Army Medical Research Institute of Infectious Diseases, Frederick, Md. , and g Department of Microbiology and Molecular Genetics, University of Texas Houston Health Science Center Medical School, Houston, Tex. , USA Key Words tion was linked to degradation of cathelicidin antimicrobial -Antimicrobial peptides ؒ Bacillus anthracis ؒ Bacterial peptides, a front-line effector of innate host defense. B. an infection ؒ Cathelicidins ؒ Hemolysis ؒ Innate immunity ؒ thracis lacking ClpX were rapidly killed by cathelicidin and Protease ؒ Transposon mutagenesis ؒ Virulence factor ␣ -defensin antimicrobial peptides and lysozyme in vitro. In turn, mice lacking cathelicidin proved hyper-susceptible to lethal infection with wild-type B. anthracis Sterne, confirm- Abstract ing cathelicidin to be a critical element of innate defense Bacillus anthracis is a National Institute of Allergy and Infec- against the pathogen.
    [Show full text]
  • Anti-B. Anthracis Polyclonal Antibody (DPAB0093) This Product Is for Research Use Only and Is Not Intended for Diagnostic Use
    Anti-B. anthracis polyclonal antibody (DPAB0093) This product is for research use only and is not intended for diagnostic use. PRODUCT INFORMATION Product Overview Goat Antibody to Anthrax (Multi Strain) Specificity Detects Ames, Sterne and Vollum strains of Bacillus anthracis. B. anthracis is a rod-shaped gram positive bacterium which causes anthrax, an acute infectious disease resulting in macrophage infection and immune suppression. Anthrax mainly affects ruminants, but can also affect humans. Immunogen Gamma inactivated, purified spore preparation of Bacillus anthracis using a mixture of Ames, Sterne and Vollum strains Source/Host Goat Species Reactivity B. anthracis Purification Ion exchange chromatography Conjugate Unconjugated Applications Suitable for use in ELISA. Each laboratory should determine an optimum working titer for use in its particular application. Other applications have not been tested but use in such assays should not necessarily be excluded. Format Purified, Liquid Concentration 1mg/ml (OD280nm) Buffer PBS Preservative 0.09% Sodium Azide Storage Short-term (up to 2 weeks) store at 2–8°C. Long term, store at -20°C. Avoid multiple freeze/thaw cycles. Warnings This product contains sodium azide, which has been classified as Xn (Harmful), in European Directive 67/548/EEC in the concentration range of 0.1–1.0%. When disposing of this reagent through lead or copper plumbing, flush with copious volumes of water to prevent azide build-up in drains. 45-1 Ramsey Road, Shirley, NY 11967, USA Email: [email protected] Tel: 1-631-624-4882 Fax: 1-631-938-8221 1 © Creative Diagnostics All Rights Reserved BACKGROUND Introduction Bacillus anthracis is a rod shaped gram positive bacterium which causes anthrax, an acute infectious disease resulting in macrophage infection and immune suppression.
    [Show full text]
  • The Anthrax Vaccine Debate: a Medical Review for Commanders
    The Anthrax Vaccine Debate: A Medical Review For Commanders Colonel Richard A. Hersack, USAF US Air Force Counterproliferation Center 10 Future Warfare Series No. 10 THE ANTHRAX VACCINE DEBATE: A MEDICAL REVIEW FOR COMMANDERS by Richard A. Hersack, Col, USAF, MC, CFS The Counterproliferation Papers Future Warfare Series No. 10 USAF Counterproliferation Center Air War College Air University Maxwell Air Force Base, Alabama The Anthrax Vaccine Debate: A Medical Review for Commanders Richard A. Hersack, Col, USAF, MC, CFS April 2001 The Counterproliferation Papers Series was established by the USAF Counterproliferation Center to provide information and analysis to assist the understanding of the U.S. national security policy-makers and USAF officers to help them better prepare to counter the threat from weapons of mass destruction. Copies of No. 10 and previous papers in this series are available from the USAF Counterproliferation Center, 325 Chennault Circle, Maxwell AFB AL 36112-6427. The fax number is (334) 953- 7530; phone (334) 953-7538. Counterproliferation Paper No. 10 USAF Counterproliferation Center Air War College Air University Maxwell Air Force Base, Alabama 36112-6427 The internet address for the USAF Counterproliferation Center is: http://www.au.af.mil/au/awc/awcgate/awc-cps.htm Executive Summary There are two distinct yet related aspects to the debate over the safety and efficacy of the anthrax vaccine. - An assessment of the clinical safety and efficacy of the anthrax vaccine. - The policy level decision to vaccinate military personnel based on intelligence reports and assessments. - The policy decision to vaccinate is based on an assessment of relative risk.
    [Show full text]
  • Anthrax As a Biological Weapon, 2002 Updated Recommendations for Management
    CONSENSUS STATEMENT Anthrax as a Biological Weapon, 2002 Updated Recommendations for Management Thomas V. Inglesby, MD Objective To review and update consensus-based recommendations for medical Tara O’Toole, MD, MPH and public health professionals following a Bacillus anthracis attack against a civilian population. Donald A. Henderson, MD, MPH Participants The working group included 23 experts from academic medical cen- John G. Bartlett, MD ters, research organizations, and governmental, military, public health, and emer- Michael S. Ascher, MD gency management institutions and agencies. Edward Eitzen, MD, MPH Evidence MEDLINE databases were searched from January 1966 to January 2002, using the Medical Subject Headings anthrax, Bacillus anthracis, biological weapon, Arthur M. Friedlander, MD biological terrorism, biological warfare, and biowarfare. Reference review identified Julie Gerberding, MD, MPH work published before 1966. Participants identified unpublished sources. Jerome Hauer, MPH Consensus Process The first draft synthesized the gathered information. Written comments were incorporated into subsequent drafts. The final statement incorpo- James Hughes, MD rated all relevant evidence from the search along with consensus recommendations. Joseph McDade, PhD Conclusions Specific recommendations include diagnosis of anthrax infection, in- Michael T. Osterholm, PhD, MPH dications for vaccination, therapy, postexposure prophylaxis, decontamination of the environment, and suggested research. This revised consensus statement presents new Gerald Parker, PhD, DVM information based on the analysis of the anthrax attacks of 2001, including develop- Trish M. Perl, MD, MSc ments in the investigation of the anthrax attacks of 2001; important symptoms, signs, and laboratory studies; new diagnostic clues that may help future recognition of this Philip K. Russell, MD disease; current anthrax vaccine information; updated antibiotic therapeutic consid- Kevin Tonat, DrPH, MPH erations; and judgments about environmental surveillance and decontamination.
    [Show full text]
  • Anthrax Lethal Factor Inhibition
    Anthrax lethal factor inhibition W. L. Shoop*†, Y. Xiong*, J. Wiltsie*, A. Woods*, J. Guo*, J. V. Pivnichny*, T. Felcetto*, B. F. Michael*, A. Bansal*, R. T. Cummings*, B. R. Cunningham*, A. M. Friedlander‡, C. M. Douglas*, S. B. Patel*, D. Wisniewski*, G. Scapin*, S. P. Salowe*, D. M. Zaller*, K. T. Chapman*, E. M. Scolnick§, D. M. Schmatz*, K. Bartizal*, M. MacCoss*, and J. D. Hermes* *Merck Research Laboratories, Rahway, NJ 07065; ‡United States Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702; and §Department of Biology, Massachusetts institute of Technology, Cambridge, MA 02139 Communicated by William C. Campbell, Drew University, Madison, NJ, April 12, 2005 (received for review November 5, 2004) The primary virulence factor of Bacillus anthracis is a secreted phylactically if intentional release of anthrax were suspected) or, zinc-dependent metalloprotease toxin known as lethal factor (LF) more probably, an LFI would be used to block late stage effects that is lethal to the host through disruption of signaling pathways, of LF during an active infection and increase the probability of cell destruction, and circulatory shock. Inhibition of this proteolyt- host survival. This latter aspect would unquestionably be used in ic-based LF toxemia could be expected to provide therapeutic value adjunct therapy with an antibiotic. Herein, we reveal the crystal in combination with an antibiotic during and immediately after an structure of a hydroxamate LFI and its intimate interaction with active anthrax infection. Herein is shown the crystal structure of an LF and present a sequence of in vitro and in vivo studies, intimate complex between a hydroxamate, (2R)-2-[(4-fluoro-3- including those with active B.
    [Show full text]