3 edition of Construction and screening of a combinatorial shiga toxin library found in the catalog.
Construction and screening of a combinatorial shiga toxin library
Thesis (M.Sc.) -- University of Toronto, 1998.
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Shiga toxin (Stx)-producing Escherichia coli (STEC) bacteria are causative pathogens of diarrhea, hemorrhagic colitis (HC), and hemolytic uremic syndrome (HUS), and these bacteria are also the most common cause of acute renal failure in children [ 1–4].Human STEC isolates are also designated enterohemorrhagic E coli, and serotype OH7 is one of the predominant serotypes responsible for. STFRP: Shiga toxins (also known as Shiga-like toxins, Vero toxins, or Vero-like toxins) are encoded by some strains of Escherichia coli, most notably OH7. Shiga toxin can also be produced by other serogroups of enterohemorrhagic E coli (EHEC), as well as Shigella dysenteriae type 1. Generally, Shiga toxin-producing organisms cause bloody diarrhea, although this is not universal.
Qualitative EIA detection of Shiga toxins 1 (ST-1)and 2 (ST-2) produced by some strains of Escherichia coli is included in all stool culture requests _____ _____ October Diagnostic Laboratory Services Inc. Iwaiwa Street Aiea, HI () Clinical Background Shiga toxin--producing E. coli (STEC) cause. Shiga-like toxins (verotoxins) are a class of AB5 holotoxins that are primarily responsible for the virulence associated with Shiga-like toxin producing Escherichia coli (STEC) infections. The holotoxins are composed of a pentamer of identical subunits (B subunit) responsible for delivering the catalytic subunit (A subunit) to a host cell and facilitating endocytosis of the toxin into the cell.
Enterohemorrhagic Escherichia coli (EHEC) OH11, a serotype within Shiga toxin-producing E. coli (STEC) that causes severe human disease, has been considered to have evolved from attaching and effacing E. coli (AEEC) OH11 through the acquisition of a Shiga toxin-encoding gene. Targeted amplicon sequencing using next-generation sequencing technology of 48 phylogenetically informative. The AIMS-5 library was constructed as described structure of the pSCCAE8d vector used to construct this library is shown in Fig. 1A. The variable region of heavy chain (V H) and the full-length light chain (V L –C L) genes were joined using a flexible peptide linker sequence to generate an scFv-C L segment. The 3′ end of this segment was then fused with a gene .
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ABSTRACT Shiga-like toxin I (SLTI) is a bacterial toxin that is able to penetrate and kill mammalian this projecf a library of SLTI mutants with altered receptor specificity was constructed using a synthetic oligonucleotide that varied 9 amino acids in the specificity-determining region of the B : Wai-May Lim.
Shiga-like toxin 1 was used as a molecular template in developing cytotoxic agents. The receptor binding portion of the toxin template could be altered, and the retained cytotoxic activity of each toxin variant could be exploited as a molecular search engine in designing, constructing, and screening combinatorial SLT protein by: The resulting focused library also can be screened for different phenotypes and in different cell lines.
To illustrate this strategy, we determined resistance phenotypes for ricin in K, HeLa, and Raji cells and for Shiga toxin in HeLa and Raji cells. (K cells are not Shiga toxin-sensitive because they lack the cell-surface receptor CD)Cited by: Introduction.
Ever since the first description of an outbreak of hemorrhagic colitis (HC) associated with Shiga toxin-producing Escherichia coli (STEC) in the United States inSTEC have been identified worldwide as an important cause of human disease, mostly foodborne .In addition of uncomplicated to severe bloody diarrhea, STEC infections can evolve into the development of the life Cited by: 8.
Construction of SLT-1 libraries the library were conducted to identify cytotoxic mutants Shiga-like toxin 1 (SLT-1) is a ribosome-inactivating pro- able to kill these two cell lines.
A collection of single tein composed of two functional domains: a catalytic A clones was selected from the library. Briefly, individual. The Retro-1 molecule was identified in a high-throughput screening as an inhibitor of ricin and Shiga toxins by diminishing their intracellular trafficking via the retrograde route, from early endosomes to the Golgi apparatus.
In order to improve the activity of Retro-1, a SAR study was undertaken yielding an analog that possesses roughly fold better EC50 against Shiga toxin cytotoxicity.
In essence, a combinatorial peptide library (numbering – different molecules) is chemically synthesized and subjected to screening for biological activity. Methodology for scheduling pre-board screening staff at the Vancouver International Airport Book.
Jan ; Math Gaz; Donald Gross Construction and screening of a combinatorial shiga toxin. The resulting focused library also can be screened for different phenotypes and in different cell lines.
To illustrate this strategy, we determined resistance phenotypes for ricin in K, HeLa, and Raji cells and for Shiga toxin in HeLa and Raji cells. (K cells are not Shiga toxin-sensitive because they lack the cell-surface receptor CD).
Shiga toxins have an AB 5 structure with one enzymatically active A subunit non-covalently linked to a pentamer of B subunits responsible for Gb3 binding. After binding of Stx to lipid raft-associated Gb3, Stx is internalised into early endosomes and then travels further to the trans-Golgi network (Figure 1).
This application relates to libraries of ABx toxin mutants, in which a peptide insert is introduced into the protease- sensitive loop of the A-chain sequence to alter the type of cells to which toxic species are delivered. Said libraries are used in the development of therapeutics targeted against specific cell types.
Shiga toxin (Stx)-producing Escherichia coli (STEC) causes diarrhea and colitis in humans that can develop into a life-threatening hemolytic uremic syndrome (HUS).
Developing efficient means of controlling STEC diseases, for which no drugs or vaccines are currently available, remains a high priority.
We report here the construction and development of chitosan conjugates bearing the Stx ligand. Integrated platform for genome-wide screening and construction of high-density genetic interaction maps in mammalian cells Martin Kampmann a,b, Michael C.
Bassik, and Jonathan S. Weissmana,b,1 aDepartment of Cellular and Molecular Pharmacology, California Institute for Quantitative Biomedical Research and bHoward Hughes Medical Institute, University of California, San Francisco, CA Identification of SLT-1A IYSNKLM: a human melanoma-specific scRIP toxin variant.
SLT-1 is a bacterial type II ribosome-inactivating protein produced by enteropathogenic E. coli strains such as OH7. Previously, the full-length SLT-1 (70 kDa; AB 5) was used by our group as a scaffold to construct a combinatorial peptide elements were embedded within its receptor-binding B.
Shiga toxin‐producing Escherichia coli (STEC) strains are food‐borne pathogens of public health concern. Despite ruminants are the most important reservoir, STEC human infections have also been attributed to pigs.
We examined for the presence of STEC in samples of swine caecal content collected during the year at Italian abattoirs in the framework of the harmonized monitoring of. INTRODUCTION. Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen frequently identified as causative agent of acute diarrheal disease in humans.
The outcomes of STEC infections may range from asymptomatic carriage and mild diarrhea to severe disease, such as hemorrhagic colitis (HC) and hemolytic-uremic syndrome (HUS) (1,– 3).Based on pathogenic properties, a subgroup of.
Discovery of Inhibitors of Shiga Toxin Type 2 by On-Plate Generation and Screening of a Focused Compound Library. Angewandte Chemie(6), DOI: /ange Shiga toxin positive, unknown organism Overview (1,2,3) Shiga toxin-producing Escherichia coli (STEC) is the term used to refer to a group of E.
coli bacteria that produce powerful toxins, which can cause severe illness. Most cases in North America are caused by E. coli OH7, but other serotypes of E.
coli can also express Shiga toxins. The. Shiga toxin-producing Escherichia coli (STEC) causes outbreaks and sporadic cases of gastroenteritis. STEC OH7 is the most clinically relevant serotype in the world.
The major virulence determinants of STEC OH7 are the Shiga toxins and the locus of enterocyte effacement. However, several accessory virulence factors, mainly outer membrane proteins (OMPs) that interact with the host.
The subtilase cytotoxin (SubAB) is an AB 5 toxin described in certain Shiga toxin (Stx)-producing Escherichia coli (STEC) strains that usually lack the locus for enterocyte effacement (LEE).
We report for the first time the production of SubAB by two Stx-negative E. coli strains, isolated from unrelated cases of childhood diarrhea. The characterization of the SubAB-coding genes showed a 90%.
Shiga toxin-producing E. coli (STEC) Screen EIA: Back to Test Directory: Test Description: Contact: Enteric Bacteriology () Test Request Form: Bacteriology SHL OpenELIS Code: Test detects the Shiga toxins, sensitive markers of toxigenic Escherichia coli but does not determine specific strains of E.
coli. (e.g., E. coli OH7). While genomes are being sequenced at an accelerating pace, the elucidation of the function of human genes and their interactions remains a formidable challenge.
Genetic interaction maps, which report on how genes work together, provide a powerful tool for systematically defining gene function and pathways. Here we present the complete quantitative framework underlying our recently .Aims: To develop and evaluate a real‐time PCR assay incorporating an internal amplification control (IAC) suitable for the screening of Shiga toxin (Stx)‐producing Escherichia coli (STEC) in foods.
Methods and Results: A competitive IAC was constructed and included in an stx‐specific real‐time PCR d to 18‐h enrichment and automated DNA extraction, the assay could reliably.