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[PMID]:28883040
[Au] Autor:Selyunin AS; Iles LR; Bartholomeusz G; Mukhopadhyay S
[Ad] Endereço:Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas at Austin, Austin, TX.
[Ti] Título:Genome-wide siRNA screen identifies UNC50 as a regulator of Shiga toxin 2 trafficking.
[So] Source:J Cell Biol;216(10):3249-3262, 2017 Oct 02.
[Is] ISSN:1540-8140
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Shiga toxins 1 and 2 (STx1 and STx2) undergo retrograde trafficking to reach the cytosol. Early endosome-to-Golgi transport allows the toxins to evade degradation in lysosomes. Targeting this trafficking step has therapeutic promise, but the mechanism of trafficking for the more potent toxin STx2 is unclear. To identify host factors required for early endosome-to-Golgi trafficking of STx2, we performed a viability-based genome-wide siRNA screen in HeLa cells. 564, 535, and 196 genes were found to be required for toxicity induced by STx1 only, STx2 only, and both toxins, respectively. We focused on validating endosome/Golgi-localized hits specific for STx2 and found that depletion of UNC50 blocked early endosome-to-Golgi trafficking and induced lysosomal degradation of STx2. UNC50 acted by recruiting GBF1, an ADP ribosylation factor-guanine nucleotide exchange factor (ARF-GEF), to the Golgi. These results provide new information about STx2 trafficking mechanisms and may advance efforts to generate therapeutically viable toxin-trafficking inhibitors.
[Mh] Termos MeSH primário: Estudo de Associação Genômica Ampla
Proteínas de Membrana
RNA Interferente Pequeno
Proteínas de Ligação a RNA
Toxina Shiga II/metabolismo
[Mh] Termos MeSH secundário: Endossomos/genética
Endossomos/metabolismo
Complexo de Golgi/genética
Complexo de Golgi/metabolismo
Células HeLa
Seres Humanos
Lisossomos/genética
Lisossomos/metabolismo
Proteínas de Membrana/genética
Transporte Proteico/genética
Proteólise
Proteínas de Ligação a RNA/genética
Toxina Shiga I/genética
Toxina Shiga I/metabolismo
Toxina Shiga II/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Membrane Proteins); 0 (RNA, Small Interfering); 0 (RNA-Binding Proteins); 0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 0 (UNC50 protein, human)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171007
[Lr] Data última revisão:
171007
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170909
[St] Status:MEDLINE
[do] DOI:10.1083/jcb.201704015


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[PMID]:28493685
[Au] Autor:Li CH; Bai YL; Selvaprakash K; Mong KT; Chen YC
[Ad] Endereço:Department of Applied Chemistry, National Chiao Tung University , Hsinchu 300, Taiwan.
[Ti] Título:Selective Detection of Shiga-like Toxin 1 from Complex Samples Using Pigeon Ovalbumin Functionalized Gold Nanoparticles as Affinity Probes.
[So] Source:J Agric Food Chem;65(21):4359-4365, 2017 May 31.
[Is] ISSN:1520-5118
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Escherichia coli O157:H7 is a foodborne pathogen. This bacterial strain can generate Shiga-like toxins (SLTs), which can cause serious sickness and even death. Thus, it is important to develop effective and sensitive methods that can be used to rapidly identify the presence of SLTs from complex samples. Pigeon egg white (PEW) contains abundant glycoproteins, including pigeon ovalbumin (POA) (∼60%). POA possesses Gal-α(1→4)-Gal-ß(1→4)-GlcNAc termini, which can recognize the B subunits in SLT type 1 (SLT-1B). Thus, POA is a suitable probe for trapping SLT-1B. In this work, we used PEW proteins as starting materials to react with aqueous tetrachloroauric acid for generation of PEW-protein-immobilized gold nanoparticles (AuNPs@PEW) via one-pot reactions. We demonstrated that the generated AuNPs@PEW were mainly dominated by POA-immobilized Au NPs. The as-prepared AuNPs@PEW were used as affinity probes to selectively probe SLT-1B from complex cell lysates derived from E. coli O157:H7. The selective trapping step can be completed within ∼90 s under microwave heating (power = 450 W) to enrich sufficient SLT-1B for matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analysis. Furthermore, this approach can be used to detect SLT-1B at a concentration as low as ∼40 pM. The feasibility of using the proposed method to selectively detect SLT-1B from ham contaminated by E. coli O157:H7 was also demonstrated.
[Mh] Termos MeSH primário: Técnicas Biossensoriais/métodos
Escherichia coli O157/metabolismo
Ouro/química
Produtos da Carne/microbiologia
Nanopartículas/química
Ovalbumina/química
Toxina Shiga I/análise
Toxina Shiga II/análise
[Mh] Termos MeSH secundário: Animais
Técnicas Biossensoriais/instrumentação
Columbidae
Escherichia coli O157/isolamento & purificação
Contaminação de Alimentos/análise
Produtos da Carne/análise
Toxina Shiga I/metabolismo
Toxina Shiga II/metabolismo
Suínos
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 7440-57-5 (Gold); 9006-59-1 (Ovalbumin)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170608
[Lr] Data última revisão:
170608
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170512
[St] Status:MEDLINE
[do] DOI:10.1021/acs.jafc.7b00863


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[PMID]:28434982
[Au] Autor:He X; Patfield S; Rasooly R; Mavrici D
[Ad] Endereço:Western Regional Research Center, U.S. Department of Agriculture, Agricultural Research Service, 800 Buchanan Street, Albany, CA 94710, United States. Electronic address: Xiaohua.he@ars.usda.gov.
[Ti] Título:Novel monoclonal antibodies against Stx1d and 1e and their use for improving immunoassays.
[So] Source:J Immunol Methods;447:52-56, 2017 Aug.
[Is] ISSN:1872-7905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Shiga toxins (Stxs) are major causative agents for bloody diarrhea and hemolytic uremic syndrome, a life-threatening disease in humans. No effective treatment is available. Early detection of Stxs in clinical samples is critical for disease management. As bacteria evolve, new Stxs are produced; therefore, methods used to identify them need to be improved as well. In this study, new monoclonal antibodies (mAbs) against Stx1d and 1e were developed and used to improve a commercial Stx1 kit. Incorporation of the new mAbs into the Abraxis Stx1 kit not only increased the assay sensitivity to Stx1d, but the assay was conferred the ability to detect Stx1e, a newly identified subtype of Stx1 produced by an atypical Stx-producing bacterial strain, Enterobacter cloacae M12X01451, isolated from a clinical specimen. This toxin was not detectable using existing commercial kits. The signal to noise ratio (s/n) of the new assay was increased 3-fold for Stx1d, and 44-fold for Stx1e at toxin concentration of 10ng/mL. The limit of detection (LOD) was 10pg/mL for Stx1a, and 100pg/mL for Stx1c, 1d and 1e. When used for bacterial strains, the sensitivity of the new assay was improved 2.5- to 60-fold depending on subtypes produced. In summary, high affinity mAbs against Stx1d and 1e were developed and incorporation of these mAbs into the Stx1 kit significantly enhanced the assay sensitivity and broadened the subtype-specificity. This improvement should be useful for reducing product recalls and disease mistreatment due to failures of detecting less common but clinically important subtypes of Stxs.
[Mh] Termos MeSH primário: Anticorpos Monoclonais/imunologia
Infecções por Enterobacteriaceae/diagnóstico
Ensaio de Imunoadsorção Enzimática/métodos
Toxina Shiga I/análise
Toxina Shiga I/imunologia
[Mh] Termos MeSH secundário: Anticorpos Monoclonais/isolamento & purificação
Afinidade de Anticorpos
Enterobacter cloacae/metabolismo
Infecções por Enterobacteriaceae/microbiologia
Seres Humanos
Limite de Detecção
Kit de Reagentes para Diagnóstico
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Monoclonal); 0 (Reagent Kits, Diagnostic); 0 (Shiga Toxin 1)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170915
[Lr] Data última revisão:
170915
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170425
[St] Status:MEDLINE


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[PMID]:28403141
[Au] Autor:Gordon DE; Chia J; Jayawardena K; Antrobus R; Bard F; Peden AA
[Ad] Endereço:University of California San Francisco, Department of Cellular and Molecular Pharmacology, San Francisco, CA, United States of America.
[Ti] Título:VAMP3/Syb and YKT6 are required for the fusion of constitutive secretory carriers with the plasma membrane.
[So] Source:PLoS Genet;13(4):e1006698, 2017 Apr.
[Is] ISSN:1553-7404
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The cellular machinery required for the fusion of constitutive secretory vesicles with the plasma membrane in metazoans remains poorly defined. To address this problem we have developed a powerful, quantitative assay for measuring secretion and used it in combination with combinatorial gene depletion studies in Drosophila cells. This has allowed us to identify at least three SNARE complexes mediating Golgi to PM transport (STX1, SNAP24/29 and Syb; STX1, SNAP24/29 and YKT6; STX4, SNAP24 and Syb). RNAi mediated depletion of YKT6 and VAMP3 in mammalian cells also blocks constitutive secretion suggesting that YKT6 has an evolutionarily conserved role in this process. The unexpected role of YKT6 in plasma membrane fusion may in part explain why RNAi and gene disruption studies have failed to produce the expected phenotypes in higher eukaryotes.
[Mh] Termos MeSH primário: Membrana Celular/genética
Proteínas de Drosophila/genética
Proteínas R-SNARE/genética
Proteínas SNARE/genética
Proteína 3 Associada à Membrana da Vesícula/genética
[Mh] Termos MeSH secundário: Animais
Membrana Celular/metabolismo
Proteínas de Drosophila/metabolismo
Drosophila melanogaster/genética
Drosophila melanogaster/metabolismo
Complexo de Golgi/genética
Complexo de Golgi/metabolismo
Heterozigoto
Seres Humanos
Fusão de Membrana/genética
Transporte Proteico/genética
Proteínas R-SNARE/metabolismo
Interferência de RNA
Proteínas SNARE/metabolismo
Toxina Shiga I/genética
Toxina Shiga I/metabolismo
Proteínas de Ligação a Fator Solúvel Sensível a N-Etilmaleimida/genética
Proteínas de Ligação a Fator Solúvel Sensível a N-Etilmaleimida/metabolismo
Proteína 3 Associada à Membrana da Vesícula/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Drosophila Proteins); 0 (R-SNARE Proteins); 0 (SNARE Proteins); 0 (Shiga Toxin 1); 0 (Snap24 protein, Drosophila); 0 (Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins); 0 (Vesicle-Associated Membrane Protein 3); 0 (synaptobrevin protein, Drosophila)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170530
[Lr] Data última revisão:
170530
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170414
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pgen.1006698


  5 / 1481 MEDLINE  
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[PMID]:28398250
[Au] Autor:Li XP; Tumer NE
[Ad] Endereço:Department of Plant Biology, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901, USA. xpli@aesop.rutgers.edu.
[Ti] Título:Differences in Ribosome Binding and Sarcin/Ricin Loop Depurination by Shiga and Ricin Holotoxins.
[So] Source:Toxins (Basel);9(4), 2017 Apr 11.
[Is] ISSN:2072-6651
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:Both ricin and Shiga holotoxins display no ribosomal activity in their native forms and need to be activated to inhibit translation in a cell-free translation inhibition assay. This is because the ribosome binding site of the ricin A chain (RTA) is blocked by the B subunit in ricin holotoxin. However, it is not clear why Shiga toxin 1 (Stx1) or Shiga toxin 2 (Stx2) holotoxin is not active in a cell-free system. Here, we compare the ribosome binding and depurination activity of Stx1 and Stx2 holotoxins with the A1 subunits of Stx1 and Stx2 using either the ribosome or a 10-mer RNA mimic of the sarcin/ricin loop as substrates. Our results demonstrate that the active sites of Stx1 and Stx2 holotoxins are blocked by the A2 chain and the B subunit, while the ribosome binding sites are exposed to the solvent. Unlike ricin, which is enzymatically active, but cannot interact with the ribosome, Stx1 and Stx2 holotoxins are enzymatically inactive but can interact with the ribosome.
[Mh] Termos MeSH primário: Ribossomos/metabolismo
Ricina/metabolismo
Toxina Shiga I/metabolismo
Toxina Shiga II/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 9009-86-3 (Ricin)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171109
[Lr] Data última revisão:
171109
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170412
[St] Status:MEDLINE


  6 / 1481 MEDLINE  
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[PMID]:28167125
[Au] Autor:Yan M; Xu L; Jiang H; Zhou Z; Zhou S; Zhang L
[Ad] Endereço:Department of Haematology, Guangzhou Women and Children's Medical Centre, Guangzhou Medical University, Guangzhou 510623, PR China. Electronic address: robinbobo@qq.com.
[Ti] Título:PMA-LAMP for rapid detection of Escherichia coli and shiga toxins from viable but non-culturable state.
[So] Source:Microb Pathog;105:245-250, 2017 Apr.
[Is] ISSN:1096-1208
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:In exposure to outer pressure, microorganisms are capable of entry into the Viable But Non-Culturable (VBNC) state, and thus survive under various elimination processing. The survival microorganisms may yield negative results on culturing, and cause false negative for this golden standard methodology. In this study, a novel PMA-LAMP assay on the detection of Enterohemorrhage E. coli and shiga toxins has been developed and evaluated, with further application on a number of food borne E. coli strains. LAMP primers were designed on the target of rfbe for Enterohemorrhage E. coli and stx1with stx2 for shiga toxins. Via specific penetration through the damaged cell membrane of dead cells and intercalating into DNA, PMA could prevent DNA amplification of dead bacteria from LAMP, which enabled the differentiation of bacteria between VBNC state and dead state. The established PMA-LAMP showed significant advantage in rapidity, sensitivity and specificity, compared with regular PCR assay. The applicability had also been verified, demonstrating the PMA-LAMP was capable of detection on Enterohemorrhage E. coli and shiga toxins.
[Mh] Termos MeSH primário: Técnicas Bacteriológicas/métodos
Escherichia coli/isolamento & purificação
Microbiologia de Alimentos/métodos
Toxinas Shiga/isolamento & purificação
[Mh] Termos MeSH secundário: Carboidratos Epimerases/genética
Primers do DNA
DNA Bacteriano/genética
Escherichia coli/genética
Escherichia coli O157/genética
Escherichia coli O157/isolamento & purificação
Inocuidade dos Alimentos/métodos
Doenças Transmitidas por Alimentos/microbiologia
Testes de Sensibilidade Microbiana
Viabilidade Microbiana
Técnicas de Amplificação de Ácido Nucleico/métodos
Reação em Cadeia da Polimerase
Toxina Shiga I/genética
Toxina Shiga II/genética
Toxinas Shiga/genética
Transaminases/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Primers); 0 (DNA, Bacterial); 0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 0 (Shiga Toxins); EC 2.6.1.- (Transaminases); EC 5.1.3.- (Carbohydrate Epimerases); EC 5.1.3.- (perosamine synthetase)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170505
[Lr] Data última revisão:
170505
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170208
[St] Status:MEDLINE


  7 / 1481 MEDLINE  
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[PMID]:28011696
[Au] Autor:Picozzi C; Antoniani D; Vigentini I; Foschino R; Kneifel W
[Ti] Título:Genotypic characterization and biofilm formation of Shiga toxin-producing Escherichia coli.
[So] Source:FEMS Microbiol Lett;364(2), 2017 Jan 01.
[Is] ISSN:1574-6968
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Shiga toxin-producing Escherichia coli (STEC) are recognized as one of the most dangerous foodborne pathogens. The production of Shiga toxins together with intimin protein is among the main virulence factors. However, the ability to form biofilm can protect bacteria against environmental factors (i.e. desiccation, exposure to UV rays, predation, etc.) and sanitization procedures (cleaning, rinsing, chlorination), increasing their survival on food products and in manufacturing plants. Forty-five isolates collected from food and fecal samples were genotyped by pulsed field gel electrophoresis analysis with XbaI restriction enzyme and investigated by searching for toxins (stx1, stx2) and intimin (eae) genes and serogroup (O157, O26, O145, O111, O103 and O104). Afterward, the ability to develop biofilm in microtiter assay and the production of adhesive curli fimbriae and cellulose on agar plates were tested. Our study demonstrated that biofilm formation has a great variability among STEC strains and cannot be related to a specific pulsotype nor even to serogroup or presence of virulence genes.
[Mh] Termos MeSH primário: Biofilmes/crescimento & desenvolvimento
Genótipo
Escherichia coli Shiga Toxigênica/genética
Escherichia coli Shiga Toxigênica/fisiologia
[Mh] Termos MeSH secundário: Adesinas Bacterianas/genética
Proteínas de Bactérias/metabolismo
Celulose/metabolismo
Eletroforese em Gel de Campo Pulsado
Proteínas de Escherichia coli/genética
Fezes/microbiologia
Microbiologia de Alimentos
Seres Humanos
Tipagem Molecular
Sorogrupo
Toxina Shiga I/genética
Toxina Shiga II/genética
Escherichia coli Shiga Toxigênica/classificação
Escherichia coli Shiga Toxigênica/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Adhesins, Bacterial); 0 (Bacterial Proteins); 0 (Escherichia coli Proteins); 0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 147094-99-3 (eaeA protein, E coli); 148349-72-8 (Crl protein, Bacteria); 9004-34-6 (Cellulose)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170508
[Lr] Data última revisão:
170508
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161225
[St] Status:MEDLINE
[do] DOI:10.1093/femsle/fnw291


  8 / 1481 MEDLINE  
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[PMID]:28221953
[Au] Autor:Carrillo CD; Koziol AG; Mathews A; Goji N; Lambert D; Huszczynski G; Gauthier M; Amoako K; Blais BW
[Ad] Endereço:Food Microbiology Research Team, Canadian Food Inspection Agency, 960 Carling Avenue, Building 22, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6.
[Ti] Título:Comparative Evaluation of Genomic and Laboratory Approaches for Determination of Shiga Toxin Subtypes in Escherichia coli.
[So] Source:J Food Prot;79(12):2078-2085, 2016 Dec.
[Is] ISSN:1944-9097
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The determination of Shiga toxin (ST) subtypes can be an important element in the risk characterization of foodborne ST-producing Escherichia coli (STEC) isolates for making risk management decisions. ST subtyping methods include PCR techniques based on electrophoretic or pyrosequencing analysis of amplicons and in silico techniques based on whole genome sequence analysis using algorithms that can be readily incorporated into bioinformatics analysis pipelines for characterization of isolates by their genetic composition. The choice of technique will depend on the performance characteristics of the method and an individual laboratory's access to specialized equipment or personnel. We developed two whole genome sequence-based ST subtyping tools: (i) an in silico PCR algorithm requiring genome assembly to replicate a reference PCR-based method developed by the Statens Serum Institut (SSI) and (ii) an assembly-independent routine in which raw sequencing results are mapped to a database of known ST subtype sequence variants (V-Typer). These tools were evaluated alongside the SSI reference PCR method and a recently described PCR-based pyrosequencing technique. The V-Typer method results corresponded closely with the reference method in the analysis of 67 STEC cultures obtained from a World Health Organization National Reference Laboratory. In contrast, the in silico PCR method failed to detect ST subtypes in several cases, a result which we attribute to assembly-induced errors typically encountered with repetitive gene sequences. The V-Typer can be readily integrated into bioinformatics protocols used in the identification and characterization of foodborne STEC isolates.
[Mh] Termos MeSH primário: Infecções por Escherichia coli/diagnóstico
Toxina Shiga/genética
[Mh] Termos MeSH secundário: Proteínas de Escherichia coli/genética
Genômica
Seres Humanos
Toxina Shiga I/genética
Toxina Shiga II/genética
Escherichia coli Shiga Toxigênica/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Escherichia coli Proteins); 0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 75757-64-1 (Shiga Toxin)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170605
[Lr] Data última revisão:
170605
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170222
[St] Status:MEDLINE
[do] DOI:10.4315/0362-028X.JFP-16-228


  9 / 1481 MEDLINE  
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[PMID]:27792838
[Au] Autor:Goto T; Tsuji M; Kanemaru K; Yokoigawa K
[Ad] Endereço:Graduate School of Integrated Arts and Sciences, Tokushima Univ, 1-1 Minamijosanjima-cho, Tokushima, 770-8502, Japan.
[Ti] Título:Adsorption of Shiga Toxin to Poly-γ-Glutamate Precipitated.
[So] Source:J Food Sci;81(12):M2977-M2981, 2016 Dec.
[Is] ISSN:1750-3841
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We screened foods containing indigestible ingredients in the ability to adsorb Shiga toxin (Stx). When 5 mg of foods and dietary fibers such as dry vegetables and inulin were mixed and incubated with 0.5 mL of Stx solution (100 ng/mL) containing 0.5% bovine serum albumin, both Stx1 and Stx2 seemed to be adsorbed by only a fermented food, natto (a traditional Japanese food prepared from steamed soybeans by the biological action of Bacillus subtilis). We purified the Stx-adsorbing substance from natto by extraction with H O, acid treatment, Proteinase K treatment, and an ion exchange chromatography. The purified substance showed an average molecular mass of about 600 kDa. We identified it as poly-γ-glutamate (PGA) by amino acid analysis of its hydrolysate and peptide analysis after its treatment with Proteinase K. Purified PGA (MW: molecular weight = about 600 kDa) was considered to adsorb both Stx1 and Stx2 when we separated adsorbed and unadsorbed Stxs (MW = about 72 kDa) by an ultrafiltration method with a centrifugal filter unit (MWCO: molecular weight cut-off = 100 K). However, PGA with the ability to adsorb Stx was an insoluble form precipitated in the filter unit during centrifugation. PGA precipitated beyond the saturated density was also confirmed to well adsorb both Stx1 and Stx2 by an equilibrated dialysis method. To the best of our knowledge, this is the 1st report on food-adsorbing Stx.
[Mh] Termos MeSH primário: Ácido Poliglutâmico/análogos & derivados
Toxina Shiga I/química
Toxina Shiga II/química
[Mh] Termos MeSH secundário: Adsorção
Bacillus subtilis/metabolismo
Cromatografia por Troca Iônica
Fibras na Dieta/análise
Endopeptidase K/metabolismo
Escherichia coli O157/metabolismo
Contaminação de Alimentos
Peso Molecular
Ácido Poliglutâmico/química
Ultrafiltração
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Dietary Fiber); 0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 0 (poly(gamma-glutamic acid)); 25513-46-6 (Polyglutamic Acid); EC 3.4.21.64 (Endopeptidase K)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161030
[St] Status:MEDLINE
[do] DOI:10.1111/1750-3841.13540


  10 / 1481 MEDLINE  
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[PMID]:27733635
[Au] Autor:Boone JT; Campbell DE; Dandro AS; Chen L; Herbein JF
[Ad] Endereço:TechLab, Inc., Blacksburg, Virginia, USA jeboone@techlab.com.
[Ti] Título:A Rapid Immunoassay for Detection of Shiga Toxin-Producing Escherichia coli Directly from Human Fecal Samples and Its Performance in Detection of Toxin Subtypes.
[So] Source:J Clin Microbiol;54(12):3056-3063, 2016 Dec.
[Is] ISSN:1098-660X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Fecal samples (n = 531) submitted to a regional clinical laboratory during a 6-month period were tested for the presence of Shiga toxin using both a Vero cell cytotoxicity assay and the Shiga Toxin Quik Chek test (STQC), a rapid membrane immunoassay. Testing the samples directly (without culture), 9 positives were identified by the Vero cell assay, all of which were also detected by the STQC. The correlation between the two assays was 100%. Not all of the identified positive samples were detected when fecal broth cultures were tested. By testing broth cultures of characterized isolates representing all described Shiga toxin subtypes, the STQC detected all subtypes. Levels of induction of toxin production by ciprofloxacin differed among the strains tested, with more toxin induction seen in strains harboring Stx2 phages than in those harboring Stx1 phages.
[Mh] Termos MeSH primário: Ciprofloxacino/farmacologia
Infecções por Escherichia coli/diagnóstico
Escherichia coli O157/isolamento & purificação
Fezes/microbiologia
Toxina Shiga I/biossíntese
Toxina Shiga II/biossíntese
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Cercopithecus aethiops
Infecções por Escherichia coli/microbiologia
Seres Humanos
Imunoensaio/métodos
Células Vero
[Pt] Tipo de publicação:COMPARATIVE STUDY; EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Shiga Toxin 1); 0 (Shiga Toxin 2); 5E8K9I0O4U (Ciprofloxacin)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170803
[Lr] Data última revisão:
170803
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161014
[St] Status:MEDLINE



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