Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Plasmid Preparation, Mutagenesis, Transformation Assay, Clone Assay

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: LPS analysis of an E. coli O157:H7 strain and derivatives of this strain. (A) LPS from E. coli O157:H7 strain ATCC 43895 (wild type), the O-antigen-negative mutant (Δper), and the complemented mutant (compl). LPS samples from equal numbers of bacterial cells were loaded in the lanes, separated by Tricine-SDS-PAGE on a 12% acrylamide gel, and visualized by silver staining. (B) Immunoblot of bacterial LPS. Components separated by Tricine-SDS-PAGE were transferred to Immobilon-P and probed with antibody to the O157 antigen.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Mutagenesis, SDS Page, Acrylamide Gel Assay, Silver Staining, Western Blot

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Secreted and membrane proteins of E. coli O157:H7 strain ATCC 43895 (wild type) and the O-antigen-negative mutant (Δper). Proteins were separated by 10% SDS-PAGE. (A) Secreted proteins from culture supernatants visualized by silver staining. (B) Membrane proteins visualized by Coomassie blue staining. The positions of molecular mass markers (in kilodaltons) are indicated on the left. The proteins in similar gels were transferred to PVDF membranes for immunoblotting and probed with (C) anti-EspA and anti-EspB or (D) anti-intimin, reacted with secondary antibodies conjugated with HRP, and visualized by autoradiography. Δeae, E. coli O157:H7 with the intimin gene deleted.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Membrane, Mutagenesis, SDS Page, Silver Staining, Staining, Western Blot, Autoradiography

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Autoaggregation of O-antigen-negative mutants. Wild-type E. coli O157:H7 strain ATCC 43895 (43895) and strain 8624 (8624) and the corresponding O-antigen mutants (43895Δper and 8624 mutant) were grown overnight with shaking in LB broth. Cells taken directly from the shaker were observed microscopically by phase-contrast microscopy (magnification, ×1,000) to examine clumping (A), or cultures were monitored visually for sedimentation after they stood undisturbed for 3 h at room temperature (B). Cell aggregation is indicated by arrows.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Mutagenesis, Microscopy, Sedimentation

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Motility and flagellin expression in E. coli O157:H7 strain ATCC 43895 (wild type), the O-antigen-negative mutant (Δper), and the complemented strain (compl). Bacteria were stabbed into 0.3% motility agar and incubated at 37°C for 18 h (A) or 40 h (B). White areas represent bacterial growth (motility halo). Flagellin expression was examined using cells from the agar plates by immunoblotting (C) after 18 h of growth or 40 h of growth (Δper 40 h). ΔfliC, E. coli O157:H7 strain ATCC 43894 with the flagellin gene deleted.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Expressing, Mutagenesis, Bacteria, Incubation, Western Blot

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Transmission electron microscopy. Bacterial cells grown overnight on LB agar were visualized by electron microscopy. Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown. Among the O-antigen-negative mutant (Δper) cells, cells without flagella were observed, and the cells were closer to each other than wild-type cells and electron-dense material apparently linked some cells together (indicated by arrows). Total magnification, ×20,000.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Transmission Assay, Electron Microscopy, Mutagenesis

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Fecal shedding of E. coli O157:H7 by mice. Three groups of six mice received a single oral dose of 108 CFU E. coli O157:H7 strain ATCC 43895 (wild type), the O-antigen-negative mutant (Δper), or the complemented strain (compl). The geometric means of log-transformed viable counts (CFU/fecal pellet) are shown for each strain cultured from fecal samples on the indicated days. E+, fecal cultures positive for E. coli O157:H7 only as determined by using enrichment cultures and containing <10 E. coli O157 CFU/pellet; Neg, feces cultures that were negative as determined by both direct and enrichment culture procedures. The error bars indicate standard errors of the means.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Mutagenesis, Transformation Assay, Cell Culture

Journal:

Article Title: Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa ▿

doi: 10.1128/AEM.00743-08

Figure Lengend Snippet: Bacterial colonization of the bovine rectal-anal junction mucosa. The numbers of E. coli O157:H7 strain ATCC 43895 (wild-type) or O-antigen-negative mutant (Δper) bacteria in RAMS samples from experimentally inoculated steers were determined. Two groups of seven steers received a single rectal application of 1010 CFU E. coli O157:H7 strain ATCC 43895 or the Δper mutant. The geometric means of log-transformed bacterial numbers (CFU/swab) are shown for each strain isolated from RAMS cultures on the indicated days. E+, RAMS cultures positive for E. coli O157:H7 as determined only by an enrichment procedure and containing <30 E. coli O157 CFU/swab; Neg, RAMS cultures that were negative as determined by both direct and enrichment culture procedures.

Article Snippet: Typical E. coli O157:H7 strain ATCC 43895 (wild-type) cells with flagella (indicated by arrowheads) are shown.

Techniques: Mutagenesis, Bacteria, Transformation Assay, Isolation

Journal: BMC Nephrology

Article Title: Early graft loss due to acute thrombotic microangiopathy accompanied by complement gene variants in living-related kidney transplantation: case series report

doi: 10.1186/s12882-022-02868-7

Figure Lengend Snippet: Biopsy findings of three cases

Article Snippet: Light microscopy (equipment, Nikon ECLIPSE 80i; software, NIS-Elementary TS BR 3.2) on transplant biopsy revealed glomerulitis, acute tubulointerstitial nephritis (ATIN), and typical TMA.

Techniques: Biomarker Discovery

Journal: BMC Nephrology

Article Title: Early graft loss due to acute thrombotic microangiopathy accompanied by complement gene variants in living-related kidney transplantation: case series report

doi: 10.1186/s12882-022-02868-7

Figure Lengend Snippet: Hypothesis based on our case series: both recipients and donors carried the complement genetic abnormality, contributing to the continuous injury of endothelial cells and leading to early graft loss after activation of TMA by triggers in our case series. Advice for these patients: (1) before transplantation, recipients with kidney disease of unknown etiology may consider complement genetic testing. A deceased donor kidney transplant should be recommended if testing reveals complement genetic variant in both donor and recipient; (2) after transplantation, intensive monitoring and timely treatment of triggers are critical. Besides, surveillance allograft biopsy, more testing, and exome sequence may help to diagnosis; (3) since diagnosis of post-transplant TMA, early anti-complement treatment is necessary. LKT, living-relative kidney disease; DD, deceased donor; ABMR, antibody-mediated rejection; CNIs, calcineurin inhibitors; TMA, thrombotic microangiopathy; LDH, lactic dehydrogenase; PE, plasma exchange

Article Snippet: Light microscopy (equipment, Nikon ECLIPSE 80i; software, NIS-Elementary TS BR 3.2) on transplant biopsy revealed glomerulitis, acute tubulointerstitial nephritis (ATIN), and typical TMA.

Techniques: Activation Assay, Transplantation Assay, Variant Assay, Sequencing, Biomarker Discovery, Clinical Proteomics