Journal: PLoS ONE

Article Title: Evidence for Conserved Function of γ–Glutamyltranspeptidase in Helicobacter Genus

doi: 10.1371/journal.pone.0030543

Figure Lengend Snippet: Helicobacter species used in this study.

Article Snippet: To determine the frequency of bgh1 and bgh2 in H. bilis , 33 H. bilis strains from our collection , including H. bilis type strain ATCC 51630 T , H. bilis ATCC 49314 and H. bilis ATCC 49320, were subjected to a specific BclI RFLP-PCR for bgh1 and a specific PCR for bgh2 .

Techniques:

Journal: Pathogens

Article Title: Infection with Helicobacter pylori Induces Epithelial to Mesenchymal Transition in Human Cholangiocytes

doi: 10.3390/pathogens9110971

Figure Lengend Snippet: Exposure to CagA + Helicobacter pylori strain NCTC 11637 (ATCC 43504) induced morphological alteration in cholangiocytes that included cellular elongation, terminal thread-like filopodia and diminished cell-to-cell contacts, indicative of epidermal to mesenchymal transition. Panels A – C : photomicrographs documenting morphology of H69 cells exposed to H. pylori at 0, 10, and 100 bacilli per cholangiocyte, respectively (left to right). The cellular appearance changed from an epithelial phenotype to a mesenchymal phenotype as evidenced by the loss of cell-cell contact, an elongated and spindle-shaped morphology, along with growth as individual cells by 24 h following exposure to H. pylori , in a dose-dependent manner. Scale bars, 5 μm (right), 20× magnification. The length-to-width ratio of single, isolated cells was determined to document elongation and scattering of the cell population ( D , E ). The number of elongated cells increased in a dose-dependent fashion in response to H. pylori ( F ). By contrast, the number of isolated, individual cholangiocytes, indicative of cell scattering, was also significantly increased in dose-dependent fashion ( G ). Data are presented as the mean ± standard error of three biological replicates. Means were compared using a one-way ANOVA. Asterisks indicate levels of statistical significance of experimental compared to control groups at 24 h; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

Article Snippet: Helicobacter pylori NCTC 11637 = CCUG 17874 = ATCC 43504 (Epsilon-proteobacteria), infraspecific name, strain NCTC 11637, from human gastric antrum [ ], and Helicobacter bilis (ATCC ® 49314™), originally from aborted sheep fetus, liver, and fluids, from Brookings, South Dakota [ , , ], were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Isolation

Journal: Pathogens

Article Title: Infection with Helicobacter pylori Induces Epithelial to Mesenchymal Transition in Human Cholangiocytes

doi: 10.3390/pathogens9110971

Figure Lengend Snippet: Differential transcript fold change of EMT-related and cancer stem cell marker genes after exposure to Helicobacter pylori . Messenger RNA expression of six EMT-related genes and two cancer stem cell markers were determined after 24 h of infection. Expression of Snail, Slug, vimentin, JAM1, MMP7, and CD44 increased in a dose-dependent fashion, whereas CD24 transcription did not change significantly. Expression of the regulatory transcriptional factor, Snail, was notably up-regulated by 6.27-fold ± 1.02-fold, 9.05-fold ± 1.28-fold, and 12.25-fold ± 0.78-fold at an MOI 10, 50, and 100, respectively. MMP7 expression was markedly up-regulated by 3.3- to 5.3-fold at each MOI. Expression of each of Slug, ZEB1, vimentin, and JAM1 was also up-regulated in a dose-dependent fashion. Transcription of the cancer stem cell marker CD44 was significantly up-regulated by 2.02-fold ± 0.88-fold at an MOI 50 and by 3.75-fold ± 0.60-fold at MO of 100, whereas significant change was not evident with CD24. Three biological replicates were carried out. The qPCR findings were normalized to the expression levels of GAPDH in each sample, with the mean ± S.D. values shown for the seven genes at an MOI of 10, 50, and 100, and compared using a two-way ANOVA multiple comparison with a 95% confidence interval of difference. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001.

Article Snippet: Helicobacter pylori NCTC 11637 = CCUG 17874 = ATCC 43504 (Epsilon-proteobacteria), infraspecific name, strain NCTC 11637, from human gastric antrum [ ], and Helicobacter bilis (ATCC ® 49314™), originally from aborted sheep fetus, liver, and fluids, from Brookings, South Dakota [ , , ], were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Marker, RNA Expression, Infection, Expressing

Journal: Pathogens

Article Title: Infection with Helicobacter pylori Induces Epithelial to Mesenchymal Transition in Human Cholangiocytes

doi: 10.3390/pathogens9110971

Figure Lengend Snippet: Anchorage-independent cell growth in soft agar revealed cellular transformation of cholangiocytes following exposure to H. pylori . Representative micrographs revealing the appearance of colonies of H69 cholangiocytes at 30 days following exposure to H. pylori and to H. bilis , as indicated ( A ). At an MOI of 10, the number of H69 cell colonies increased significantly (**, p ≤ 0.01), whereas they did not at an MOI of 50. By contrast, there was a significant decrease in colony numbers at an MOI of 100 when compared with the non-infected control cells (***, p ≤ 0.001) ( B ). Exposure of H69 cells to H. bilis resulted in markedly reduced numbers of colonies, in a dose-dependent fashion, indicating an inhibitory effect of H. bilis on anchorage-independent cell growth and/or cellular transformation of cholangiocytes ( C ). Three biological replicates were performed; mean ± S.E; (****, p ≤ 0.0001).

Article Snippet: Helicobacter pylori NCTC 11637 = CCUG 17874 = ATCC 43504 (Epsilon-proteobacteria), infraspecific name, strain NCTC 11637, from human gastric antrum [ ], and Helicobacter bilis (ATCC ® 49314™), originally from aborted sheep fetus, liver, and fluids, from Brookings, South Dakota [ , , ], were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Transformation Assay, Infection