Diphtheria Toxin Post Exposure Guidance Revised: 1/14/2019 Tent/EHS/Homepage/Diphtheriaprintandgo.Pdf
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Pertussis Toxin
Pertussis Toxin Publication Number MAN0004270 Revision Date 09 May 2011 Catalog Number: PHZ1174 Quantity: 50 μg Lot Number: See product label. Appearance: Lyophilized solid. Origin: Bordetella pertussis. Purity: >99%. This preparation migrates as five distinct bands when analyzed by SDS–Urea PAGE. The five bands correspond to one A protomer subunit, designated S1 (Mr=26.2 kDa), and four B oligomer subunits, designated S2- S5 (Mr’s= 21.9, 21.9, 12.1, and 10.9 kDa, respectively). Summary: Islet-activating protein Pertussis toxin consists of an A protomer subunit (S1) which possesses both NAD+ glycohydrolase and ADP–ribosyltransferase activities, and B oligomer subunits (S2, S3, S4, and S5) which are responsible for attachment of the native toxin to eukaryotic cell surfaces. Pertussis toxin uncouples G proteins from receptors by ADP ribosylating a cysteine residue near the carboxyl terminus of the α subunit. Biological Activity: The lowest concentration which produces a clustered growth pattern with CHO cells is 0.03 ng/mL. The adenylate cyclase activity of this preparation is 20.1 picomoles/minute/μg in the presence of 1 μg calmodulin. Reconstitution Reconstitute the contents of this vial with 500 μL sterile, distilled water. The composition of the solution will be Recommendation: 50 μg Pertussis toxin, 10 mM sodium phosphate, pH 7.0, 50 mM sodium chloride. Because Pertussis toxin is relatively insoluble, the resulting suspension should be made uniform by gentle mixing prior to withdrawing aliquots. It is important to note that this suspension should not be sterile-filtered. This preparation is not activated. For use with intact cells or extracts, activation is not necessary. -
Biological Toxins Fact Sheet
Work with FACT SHEET Biological Toxins The University of Utah Institutional Biosafety Committee (IBC) reviews registrations for work with, possession of, use of, and transfer of acute biological toxins (mammalian LD50 <100 µg/kg body weight) or toxins that fall under the Federal Select Agent Guidelines, as well as the organisms, both natural and recombinant, which produce these toxins Toxins Requiring IBC Registration Laboratory Practices Guidelines for working with biological toxins can be found The following toxins require registration with the IBC. The list in Appendix I of the Biosafety in Microbiological and is not comprehensive. Any toxin with an LD50 greater than 100 µg/kg body weight, or on the select agent list requires Biomedical Laboratories registration. Principal investigators should confirm whether or (http://www.cdc.gov/biosafety/publications/bmbl5/i not the toxins they propose to work with require IBC ndex.htm). These are summarized below. registration by contacting the OEHS Biosafety Officer at [email protected] or 801-581-6590. Routine operations with dilute toxin solutions are Abrin conducted using Biosafety Level 2 (BSL2) practices and Aflatoxin these must be detailed in the IBC protocol and will be Bacillus anthracis edema factor verified during the inspection by OEHS staff prior to IBC Bacillus anthracis lethal toxin Botulinum neurotoxins approval. BSL2 Inspection checklists can be found here Brevetoxin (http://oehs.utah.edu/research-safety/biosafety/ Cholera toxin biosafety-laboratory-audits). All personnel working with Clostridium difficile toxin biological toxins or accessing a toxin laboratory must be Clostridium perfringens toxins Conotoxins trained in the theory and practice of the toxins to be used, Dendrotoxin (DTX) with special emphasis on the nature of the hazards Diacetoxyscirpenol (DAS) associated with laboratory operations and should be Diphtheria toxin familiar with the signs and symptoms of toxin exposure. -
Pertussis Toxin
PLEASE POST THIS PAGE IN AREAS WHERE PERTUSSIS TOXIN IS USED IN RESEARCH LABORATORIES UNIVERSITY OF CALIFORNIA, SAN FRANCISCO ENVIRONMENT, HEALTH AND SAFETY/BIOSAFETY PERTUSSIS TOXIN EXPOSURE/INJURY RESPONSE PROTOCOL Organism or Agent: Pertussis Toxin Exposure Risk; Multiple Endocrine/Metabolic Effects Exposure Hotline Pager: 415/353-7842 (353-STIC) (Available 24 hours) Office of Environment, Health & Safety: 415/476-1300 (Available during work hours) 415/476-1414 or 9-911 (In case of emergency, available 24 hours) EH&S Biosafety Officer 415/514-2824 EH&S Public Health Officer: 415/514-3531 UCSF Occupational Health Services: 415/885-7580 (Available during work hours) California Poison Control: 800/222-1222 SFDPH Emergency Number: 415/554-2830 CDC Emergency Operations: 770/488-7100 _________________________________________________________________________ PROTOCOL SUMMARY In the event of an accidental exposure or injury, the protocol is as follows: 1. Modes of Exposure: a. Skin puncture or injection b. Ingestion c. Contact with mucous membranes (eyes, nose, mouth) d. Contact with non-intact skin e. Exposure to aerosols f. Respiratory exposure from inhalation of toxin 2. First Aid: a. Skin Exposure, immediately go to the sink and thoroughly wash the skin with soap and water. If working with pertussis, decontaminate any exposed skin with an antiseptic scrub solution. b. Skin Wound, immediately go to the sink and thoroughly wash the wound with soap and water and pat dry. c. Splash to Eye(s), Nose or Mouth, immediately flush the area with running water for at least 5- 10 minutes. d. Splash Affecting Garments, remove garments that may have become soiled or contaminated and place them in a double red plastic bag. -
Targeted Neuronal Cell Ablation in the Drosophila Embryo: Pathfinding by Follower Growth Cones in the Absence of Pioneers
Neuron, Vol. 14, 707-715, April, 1995, Copyright© 1995 by Cell Press Targeted Neuronal Cell Ablation in the Drosophila Embryo: Pathfinding by Follower Growth Cones in the Absence of Pioneers David M. Lin,* Vanessa J. Auld,*t reach their targets in the CNS (Wigglesworth, 1953). The and Corey S. Goodman ability of the initial axons to guide later growth cones has Howard Hughes Medical Institute led to the suggestion that pioneering growth cones might Division of Neurobiology be endowed with special pathfinding abilities. Do subse- Department of Molecular and Cell Biology quent growth cones possess the same pathfinding abili- Life Science Addition, Room 519 ties, or are they different from the pioneers? University of California, Berkeley Ablation experiments in a variety of organisms thus far Berkeley, California 94720 have provided a range of conflicting answers to this ques- tion. In the grasshopper embryo CNS, the G growth cone turns anteriorly along the P axons in the NP fascicle, a Summary pathway formed by the three descending P and the two ascending A axons. Although the A and P axons normally We developed a rapid method that uses diphtheria fasciculate and follow one another, either can pioneer the toxin, the flp recognition target sequences, and the complete pathway on its own. Furthermore, any one of GAL4-UAS activation system, to ablate specific neu- the P axons will suffice, since when any two of them are rons in the Drosophila embryo and to examine the con- ablated the remaining P pioneers the pathway. When the sequences in large numbers of embryos at many time three P axons are ablated, the G growth cone stalls, and points. -
Human Peptides -Defensin-1 and -5 Inhibit Pertussis Toxin
toxins Article Human Peptides α-Defensin-1 and -5 Inhibit Pertussis Toxin Carolin Kling 1, Arto T. Pulliainen 2, Holger Barth 1 and Katharina Ernst 1,* 1 Institute of Pharmacology and Toxicology, Ulm University Medical Center, 89081 Ulm, Germany; [email protected] (C.K.); [email protected] (H.B.) 2 Institute of Biomedicine, Research Unit for Infection and Immunity, University of Turku, FI-20520 Turku, Finland; arto.pulliainen@utu.fi * Correspondence: [email protected] Abstract: Bordetella pertussis causes the severe childhood disease whooping cough, by releasing several toxins, including pertussis toxin (PT) as a major virulence factor. PT is an AB5-type toxin, and consists of the enzymatic A-subunit PTS1 and five B-subunits, which facilitate binding to cells and transport of PTS1 into the cytosol. PTS1 ADP-ribosylates α-subunits of inhibitory G-proteins (Gαi) in the cytosol, which leads to disturbed cAMP signaling. Since PT is crucial for causing severe courses of disease, our aim is to identify new inhibitors against PT, to provide starting points for novel therapeutic approaches. Here, we investigated the effect of human antimicrobial peptides of the defensin family on PT. We demonstrated that PTS1 enzyme activity in vitro was inhibited by α-defensin-1 and -5, but not β-defensin-1. The amount of ADP-ribosylated Gαi was significantly reduced in PT-treated cells, in the presence of α-defensin-1 and -5. Moreover, both α-defensins decreased PT-mediated effects on cAMP signaling in the living cell-based interference in the Gαi- mediated signal transduction (iGIST) assay. -
Release of Pertussis Toxin and Its Interaction with Outer-Membrane Antigens
Journal of General Microbiology (1987), 133, 2427-2435. Printed in Great Britain 2427 Release of Pertussis Toxin and Its Interaction With Outer-membrane Antigens ByV. Y. PERERA, A. C. WARDLAW AND J. H. FREER* Department of Microbiology, University of Glasgow, Alexander Stone Building, Garscube Estate, Bearsden, Glasgow G61 IQH, UK (Received 16 December 1986 ;revised 22 April 1987) The absence of subunit S3 in cell-associated pertussis toxin (PT) from a mutant of Bordetella pertussis which failed to produce cell-free toxin suggested that this subunit was involved in the release of PT into the culture medium. The addition of methylated P-cyclodextrin (MCD) to the culture medium caused a small but consistent increase in the release of lipopolysaccharide (LPS) by four wild-type strains of B. pertussis. Since previous studies have shown that MCD also enhances the levels of PT in culture supernates, it seemed probable that the increased shedding of outer-membrane vesicles (OMV) may explain the increased levels of both cell-free PT and LPS. Release of PT was inhibited in media buffered with HEPES but was unaffected in Tris/HCl buffer. This suggested that in addition to shedding of the outer membrane, increased permeability and greater destabilization of the outer membrane, as caused by Tris/HCl buffer, may be important in the release of PT. Our data do not support the idea that PT is packaged into OMV because only an insignificant proportion (0.01 %) of the total cell-free PT was associated with LPS. The association of PT with small micelles derived from outer-membrane amphiphiles may be more important since the LPS content of PT purified from culture supernates (containing no large OMV) was nearly 18% by weight. -
Safe Handling of Acutely Toxic Chemicals Safe Handling of Acutely
Safe Handling of Acutely Toxic Chemicals , Mutagens, Teratogens and Reproductive Toxins October 12, 2011 BSBy Sco ttBthlltt Batcheller R&D Manager Milwaukee WI Hazards Classes for Chemicals Flammables • Risk of ignition in air when in contact with common energy sources Corrosives • Generally destructive to materials and tissues Energetic and Reactive Materials • Sudden release of destructive energy possible (e.g. fire, heat, pressure) Toxic Substances • Interaction with cells and organs may lead to tissue damage • EfftEffects are t tilltypically not general ltllti to all tissues, bttbut target tdted to specifi c ones • Examples: – Cancers – Organ diseases – Inflammation, skin rashes – Debilitation from long-term Poison Acute Cancer, health or accumulation with delayed (ingestion) risk reproductive risk emergence 2 Toxic Substances Are All Around Us Pollutants Natural toxins • Cigare tte smo ke • V(kidbt)Venoms (snakes, spiders, bees, etc.) • Automotive exhaust • Poison ivy Common Chemicals • Botulinum toxin • Pesticides • Ricin • Fluorescent lights (mercury) • Radon gas • Asbestos insulation • Arsenic and heavy metals • BPA ((pBisphenol A used in some in ground water plastics) 3 Application at UNL Chemicals in Chemistry Labs Toxin-producing Microorganisms • Chloro form • FiFungi • Formaldehyde • Staphylococcus species • Acetonitrile • Shiga-toxin from E. coli • Benzene Select Agent Toxins (see register) • Sodium azide • Botulinum neurotoxins • Osmium/arsenic/cadmium salts • T-2 toxin Chemicals in Biology Labs • Tetrodotoxin • Phenol -
Diphtheria Toxin
Duke University / Duke University Health System Policy on working with: Diphtheria Toxin I. Revision Page II. Background III. Risk Assessment IV. Policy Statement V. Emergency Information VI. Appendix I: EOHW Exposure/Injury Response Protocol VII. Appendix II: OESO SOP Template VIII. References This policy was first approved on December, 2018. Occupational and Environmental Safety Office (OESO) Employee Occupational Health and Wellness (EOHW) Page | 1 I. Revision Page Date Revision(s) / Comment(s) 12/06/2018 New policy document Page | 2 II. Background Diphtheria toxin (DT) is a biological toxin and is secreted by the bacterium Corynebacterium diphtheriae. DT is useful in biomedical research using mice because it can be used to selectively target and kill cells or organs without requiring surgery. Wild type mice do not have DT receptors, so they are relatively resistant to DT. The median lethal dose (LD50) for mice has been estimated at 1.6 mg/kg by subcutaneous injection compared to an estimated human LD50 of less than 100 ng/kg by IM injection. The gene for DT receptors can be inserted into a mouse genome so that the transgenic mice will express DT receptors only on specific cells. For example, a transgenic mouse engineered to express DT receptors only on hepatocytes can be injected with DT, which will only kill the hepatocytes, creating a nonsurgical mouse model without a functional liver. Diphtheria toxin doses used in transgenic mice range from 0.5 µg/kg to 50 µg/kg depending on the scientific goal (10 to 1000 ng for a 20-gram mouse).[Saito et al, 2001; Cha et al, 2003; Cerpa et al, 2014] A common dose is 100 ng per injection, sometimes administered in repeated doses to achieve a cumulative effect. -
Cellular Activity of Salmonella Typhimurium Artab Toxin and Its Receptor-Binding Subunit
toxins Article Cellular Activity of Salmonella Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit Elise Overgaard 1, Brad Morris 2, Omid Mohammad Mousa 2 , Emily Price 3, Adriana Rodriguez 2, Leyla Cufurovic 2, Richard S. Beard 4 and Juliette K. Tinker 1,2,* 1 Biomolecular Sciences Graduate Program, Boise State University, Boise, ID 83725, USA; [email protected] 2 Department of Biology, Boise State University, Boise, ID 83725, USA; [email protected] (B.M.); [email protected] (O.M.M.); [email protected] (A.R.); [email protected] (L.C.) 3 Idaho Veterans Research and Education Foundation, Infectious Diseases Section, Boise, ID 83702, USA; [email protected] 4 Biomolecular Research Center, Boise State University, Boise, ID 83725, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-208-426-5472 Abstract: Salmonellosis is among the most reported foodborne illnesses in the United States. The Salmonella enterica Typhimurium DT104 phage type, which is associated with multidrug-resistant disease in humans and animals, possesses an ADP-ribosylating toxin called ArtAB. Full-length artAB has been found on a number of broad-host-range non-typhoidal Salmonella species and serovars. ArtAB is also homologous to many AB5 toxins from diverse Gram-negative pathogens, including cholera toxin (CT) and pertussis toxin (PT), and may be involved in Salmonella pathogenesis, however, in vitro cellular toxicity of ArtAB has not been characterized. artAB was cloned into E. coli and initially isolated using a histidine tag (ArtABHIS) and nickel chromatography. ArtABHIS was found to bind to African green monkey kidney epithelial (Vero) cells using confocal microscopy and to Citation: Overgaard, E.; Morris, B.; interact with glycans present on fetuin and monosialotetrahexosylganglioside (GM1) using ELISA. -
Cloning and Sequencing of the Pertussis Toxin Genes
Proc. Nati. Acad. Sci. USA Vol. 83, pp. 4631-4635, July 1986 Biochemistry Cloning and sequencing of the pertussis toxin genes: Operon structure and gene duplication (bacterial toxins/Bordetella peflussis/ADP-ribosylation/trnshlaonal signals/secreted proteins) ALFREDO NICOSIA*, MARIA PERUGINI*, CARLO FRANZINI*, MARIA CRISTINA CASAGLI*, MARIA GIUSEPPINA BORRI*, GUIDO ANTONI*, MARCO ALMONIt, PAOLO NERIt, GIULIO RATTI*, AND RINO RAPPUOLI*t *Sclavo Research Center, Via Fiorentina, 1, 53100 Siena, and tCRISMA, University of Siena "Le Scotte," 53100 Siena, Italy Communicated by A. M. Pappenheimer, Jr., February 26, 1986 ABSTRACT Pertussis toxin, a protein composed of five A B different subunits (S1, S2, S3, S4, and S5), is the major virulence factor of Bordetela pertussis. We have cloned and * sequenced a DNA fragment of 4.7 kilobases that contains the SiP.~ genes coding for the 'five subunits. The genes are clustered S3C. , within 3.2 kilobases in the following order: S1, S2, SA, S5, and S3. A sequence closely resembling Escherichia coli promoters is found only before the S1 gene, and a possible termination S5S 0 S1~~~~~~~~~~~~ S4 signal is present at the end of the S3 gene, which suggests that S4 -- - S5 the pertussis toxin genes are organized in a single operon. A possible Shine-Dalgarno sequence is present before the S1 gene but not before the other four genes that 8-12 nucleotides FIG. 1. NaDodSO4/polyacrylamide gel electrophoresis of affin- upstream from the ATG codon show a new consensus sequence, ity-purified PT. The toxin in lane A has been treated with a reducing 5'TCC(T)GG3', possibly involved in the regulation'of trans- agent before loading the gel. -
Intracerebroventricular Antisense Knockdown of Gαi2 Results in Ciliary
BMC Neuroscience BioMed Central Research article Open Access α Intracerebroventricular antisense knockdown of G i2 results in ciliary stasis and ventricular dilatation in the rat Kati S Mönkkönen*1, Juhana M Hakumäki2, Robert A Hirst3, Riitta A Miettinen4, Christopher O'Callaghan3, Pekka T Männistö5 and Jarmo T Laitinen6 Address: 1Department of Pharmacology & Toxicology, University of Kuopio, Kuopio, FIN-70211, Finland, 2Department of Biomedical NMR, National Bio-NMR Facility, A.I. Virtanen Institute for Molecular Sciences, University of Kuopio, Kuopio, FIN-70211, Finland, 3Department of Infection, Immunity and Inflammation, University of Leicester, Leicester LE2 7LX, UK, 4Department of Neuroscience and Neurology, University of Kuopio, Finland and Department of Neurology, Kuopio University Hospital, Kuopio, FIN-70211, Finland, 5Division of Pharmacology & Toxicology, University of Helsinki, Helsinki, FIN-00014, Finland and 6Institute of Biomedicine, University of Kuopio, Kuopio, FIN-70211, Finland Email: Kati S Mönkkönen* - [email protected]; Juhana M Hakumäki - [email protected]; Robert A Hirst - [email protected]; Riitta A Miettinen - [email protected]; Christopher O'Callaghan - [email protected]; Pekka T Männistö - [email protected]; Jarmo T Laitinen - [email protected] * Corresponding author Published: 12 April 2007 Received: 7 September 2006 Accepted: 12 April 2007 BMC Neuroscience 2007, 8:26 doi:10.1186/1471-2202-8-26 This article is available from: http://www.biomedcentral.com/1471-2202/8/26 © 2007 Mönkkönen et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. -
Diphtheria. In: Epidemiology and Prevention of Vaccine
Diphtheria Anna M. Acosta, MD; Pedro L. Moro, MD, MPH; Susan Hariri, PhD; and Tejpratap S.P. Tiwari, MD Diphtheria is an acute, bacterial disease caused by toxin- producing strains of Corynebacterium diphtheriae. The name Diphtheria of the disease is derived from the Greek diphthera, meaning ● Described by Hippocrates in ‘leather hide.’ The disease was described in the 5th century 5th century BCE BCE by Hippocrates, and epidemics were described in the ● Epidemics described in 6th century AD by Aetius. The bacterium was first observed 6th century in diphtheritic membranes by Edwin Klebs in 1883 and cultivated by Friedrich Löffler in 1884. Beginning in the early ● Bacterium first observed in 1900s, prophylaxis was attempted with combinations of toxin 1883 and cultivated in 1884 and antitoxin. Diphtheria toxoid was developed in the early ● Diphtheria toxoid developed 7 1920s but was not widely used until the early 1930s. It was in 1920s incorporated with tetanus toxoid and pertussis vaccine and became routinely used in the 1940s. Corynebacterium diphtheria Corynebacterium diphtheriae ● Aerobic gram-positive bacillus C. diphtheriae is an aerobic, gram-positive bacillus. ● Toxin production occurs Toxin production (toxigenicity) occurs only when the when bacillus is infected bacillus is itself infected (lysogenized) by specific viruses by corynebacteriophages (corynebacteriophages) carrying the genetic information for carrying tox gene the toxin (tox gene). Diphtheria toxin causes the local and systemic manifestations of diphtheria. ● Four biotypes: gravis, intermedius, mitis, and belfanti C. diphtheriae has four biotypes: gravis, intermedius, mitis, ● All isolates should be tested and belfanti. All biotypes can become toxigenic and cause for toxigenicity severe disease.