Journal: Chemical Communications

Article Title: Highly efficient chemoenzymatic synthesis of β1–4-linked galactosides with promiscuous bacterial β1–4-galactosyltransferases

doi: 10.1039/c0cc01381a

Figure Lengend Snippet: Scheme 1 One-pot multienzyme synthesis of b1–4-linked galactosides with in situ generation of UDP-Gal. GalU, glucose-1-phosphate uridylyltransferase (or UDP-glucose pyrophosphorylase); GalE, UDP-galactose C4-epimerase; PpA, Pasteurella multocida inorganic pyrophosphatase. NmLgtB, Neiserria meningitidis b1–4-galactosylatrans- ferase; Hp1–4GalT, Helicobacter pylori b1–4-galactosylatransferase.

Article Snippet: Here, we report the cloning of two b1–4GalTs, from Neisseria meniningitidis serogroup B strain MC58 genomic DNA (ATCC BAA-335D) (GenBank accession number AE002098) and Helicobacter pylori strain J99 genomic DNA (ATCC 700824D) (GenBank accession number AB035971)

Techniques: In Situ

Journal: Gut

Article Title: Helicobacter pylori potentiates epithelial:mesenchymal transition in gastric cancer: links to soluble HB-EGF, gastrin and matrix metalloproteinase-7

doi: 10.1136/gut.2009.199794

Figure Lengend Snippet: Epithelial:mesenchymal transition gene expression following co-culture of gastric epithelial cells with Helicobacter pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. (a) Snail gene expression: *significantly higher gene expression level compared with untreated control cells (strain 60190 treated: AGS p<0.001, MGLVA1 p<0.05, ST16 p<0.005; strain Tx30a treated: AGS p<0.005, MGLVA1 p<0.05, ST16 p<0.02); #significantly higher gene expression level compared with Tx30a treated cells (AGS p<0.05, ST16 p<0.05). (b) Slug gene expression: *significantly higher gene expression level compared with untreated control cells (strain 60190 treated: p<0.001 in all cell lines; strain Tx30a treated: AGS p<0.001, MGLVA1 p<0.005, ST16 p<0.005); #significantly higher gene expression level compared with Tx30a treated cells (AGS p<0.04, MGLVA1 p<0.05, ST16 p<0.002). (c) Vimentin gene expression: *significantly higher gene expression level compared with untreated control cells (strain 60190 treated: AGS p<0.05, MGLVA1 p<0.001, ST16 p<0.005; strain Tx30a treated: MGLVA1 p<0.05); #significantly higher gene expression level compared with Tx30a treated cells (MGLVA1 p<0.04). (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence).

Article Snippet: H pylori strains 60190 (American Type Culture Collection (ATCC) 49503) and Tx30a (ATCC 51932) were used in this study.

Techniques: Gene Expression, Co-Culture Assay, Control

Journal: Gut

Article Title: Helicobacter pylori potentiates epithelial:mesenchymal transition in gastric cancer: links to soluble HB-EGF, gastrin and matrix metalloproteinase-7

doi: 10.1136/gut.2009.199794

Figure Lengend Snippet: Effect of co-culture of gastric epithelial cells with Helicobacter pylori strain 60190 on heparin-binding epidermal growth factor (HB-EGF) expression. (a) HB-EGF gene expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains; *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (b) Cell-associated HB-EGF expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. Cells were stained for HB-EGF (red fluorescence) and nuclei were counterstained with Hoechst (blue fluorescence); (i) AGS, (ii) AGS/ H pylori 60190, (iii) AGS/ H pylori Tx30a. Magnification ×20. (c) HB-EGF protein expression level quantified by image analysis following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains; *significantly higher HB-EGF protein expression level compared with untreated control cells (p<0.03). (d) HB-EGF shedding level following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains; *significantly higher HB-EGF shedding level compared with untreated control cells (strain 60190 treated: p<0.01 in all cell lines; strain Tx30a treated: p<0.05 in AGS and ST16); #significantly higher HB-EGF shedding level compared with Tx30a treated (p<0.05 in AGS and ST16). (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence).

Article Snippet: H pylori strains 60190 (American Type Culture Collection (ATCC) 49503) and Tx30a (ATCC 51932) were used in this study.

Techniques: Co-Culture Assay, Binding Assay, Expressing, Gene Expression, Staining, Fluorescence, Control

Journal: Gut

Article Title: Helicobacter pylori potentiates epithelial:mesenchymal transition in gastric cancer: links to soluble HB-EGF, gastrin and matrix metalloproteinase-7

doi: 10.1136/gut.2009.199794

Figure Lengend Snippet: Effect of co-culture of gastric epithelial cells with Helicobacter pylori strain 60190 on matrix metalloproteinase 7 (MMP-7) expression. (a) MMP-7 gene expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (b) MMP-7 protein expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. Cells were stained for MMP-7 (green fluorescence) and nuclei were counterstained with Hoechst (blue fluorescence). (i) AGS, (ii) AGS/ H pylori 60190, (iii) AGS H pylori Tx30a. Magnification ×20. (c) MMP-7 protein expression level quantified by image analysis following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains; *significantly higher MMP-7 protein expression level compared with untreated control cells (p<0.001). (d) MMP-7 and heparin-binding epidermal growth factor (HB-EGF) co-localisation: MMP-7 expression was stained by green fluorescence (i), HB-EGF by red fluorescence (ii), nuclei were stained with Hoechst blue fluorescence (iii). Merged image shows co-localisation of MMP-7 and HB-EGF (iv). Magnification ×20. (e) MMP-7 gene expression in MMP-7 siRNA treated gastric cell lines. MMP-7 siRNA significantly reduced MMP-7 mRNA expression in all three gastric cell lines tested. *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (f) HB-EGF gene expression in MMP-7 siRNA treated gastric cell lines exposed to H pylori 60190. MMP-7 siRNA significantly reduced HB-EGF shedding in all three gastric cell lines tested. *p<0.02, **p<0.03 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (g) HB-EGF gene expression in MMP-7 neutralising antibody treated gastric cell lines exposed to H pylori 60190. *p<0.03 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (h) HB-EGF shedding levels in gastric cell lines exposed to H pylori 60190 treated with an MMP-7 neutralising antibody. *p<0.04, **p<0.03 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (i) EMT gene expression in MMP-7 siRNA treated gastric cell lines exposed to H pylori strain 60190: Snail, p<0.01; Slug, p<0.005; vimentin, p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence).

Article Snippet: H pylori strains 60190 (American Type Culture Collection (ATCC) 49503) and Tx30a (ATCC 51932) were used in this study.

Techniques: Co-Culture Assay, Expressing, Gene Expression, Staining, Fluorescence, Control, Binding Assay

Journal: Gut

Article Title: Helicobacter pylori potentiates epithelial:mesenchymal transition in gastric cancer: links to soluble HB-EGF, gastrin and matrix metalloproteinase-7

doi: 10.1136/gut.2009.199794

Figure Lengend Snippet: Effect of co-culture of gastric epithelial cells with Helicobacter pylori strain 60190 on gastrin expression and the effect of gastrin siRNA on matrix metalloproteinase 7 (MMP-7) and epithelial:mesenchymal transition (EMT). (a) Gastrin gene expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (b) Gastrin gene expression in gastrin siRNA treated gastric cell lines. Gastrin siRNA significantly reduced gastrin mRNA expression in all three gastric cell lines tested. *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (c) MMP-7 expression in gastric cells exposed to H pylori and gastrin siRNA.*p<0.02 and **p<0.05 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (d) MMP-7 protein expression in gastrin siRNA transfected gastric cell lines exposed to H pylori . Cells were stained for MMP-7 (green fluorescence) and nuclei were counterstained with Hoescht (blue fluorescence). Magnification ×20. (e) MMP-7 protein expression level quantified by image analysis in gastrin siRNA transfected gastric cell lines exposed to H pylori ; *significantly higher MMP-7 protein expression level compared with untreated control cells (p<0.001). (f) Gastrin siRNA reverses H pylori induced EMT in gastric cell lines.*p<0.01 (n=3 replicates per condition and a representative graph is shown. Error bars indicate 95% confidence).

Article Snippet: H pylori strains 60190 (American Type Culture Collection (ATCC) 49503) and Tx30a (ATCC 51932) were used in this study.

Techniques: Co-Culture Assay, Expressing, Gene Expression, Transfection, Staining, Fluorescence, Control