Expression of carbohydrate antigens ^a in control, <i>Helicobacter pylori</i> infected and <i>Pteridium aquilinum</i> treated with or without infection mice.

a<p>Staining was graded in three categories: (-) negative, (+) positive, and (+ +) strongly positive.</p>b<p>Fisher’s exact test (p-value); n.a. – not applicable.</p

Glycosylation alterations in mice gastric mucosa upon <i>Pteridium aquilinum</i> treatment and concomitant <i>Helicobacter pylori</i> infection.

<p><b>Panel A –</b> Heatmap of the Glyco-gene Chip array analysis obtained when comparing group 1 (control) with group 4 (treated with <i>Pteridium aquilinum</i> and <i>Helicobacter pylori</i> infected). Red indicates increased and blue indicates decreased expression relative to the mean transcript expression value. The transcripts identified as differentially expressed were those with adjusted p-value <0.1 and fold change >1.3. <b>Panel B</b> – Schematic representation of the biosynthesis of sialylated terminal structures. The enzymes or family of enzymes responsible for each step of the biosynthesis are indicated. The enzymes identified as showing different expression levels in <i>Pteridium aquilinum</i> treated and <i>Helicobacter pylori</i> infected mice (Group 4) are highlighted in red (increased expression) and blue (decreased expression). <b>Panel C</b> – Immunohistochemistry for evaluation of expression of Sialyl-Lewis a (SLe^a) antigen (a–d) and for Sialyl-Lewis x (SLe^x) (e–h). Additional immunofluorescence evaluation performed for SLe^x (i–l). Bar = 20 µm.</p

Specificity and dilution of monoclonal antibodies.

a<p>Sialyl-T antigen was detected using the anti-T antibody after neuraminidase treatment as described in Material and Methods.</p

Histochemical characterization of mice gastric mucosa treated with <i>Pteridium aquilinum</i> and/or infected with <i>Helicobacter pylori.</i>

<p>Histochemical staining with hematoxylin and eosin (<b>Panel A</b>) and Alcian Blue (pH = 2,5) with nuclear red staining (<b>Panel B</b>) of mice gastric mucosa of: Control (Group 1) (a,b,i,j), <i>Pteridium aquilinum</i> treated (Group 2) (c,d,k,l), <i>Helicobacter pylori</i> infected (Group 3) (e,f,m,n) and both <i>Pteridium aquilinum</i> treated and <i>Helicobacter pylori</i> infected (Group 4) (g,h,o,p). Since the results observed at 4 and 7 weeks time points were identical, only a representative picture is included for each experimental condition. Bar = 40 µm (a,c,e,g and i-p) and bar = 20 µm (b,d,f,h). Arrows indicate inflammatory cells.</p

Alterations in simple mucin-type carbohydrate antigens in gastric mucosa from mice exposed to <i>Pteridium aquilinum.</i> Panel A –

<p>Schematic representation of the biosynthesis of simple mucin-type carbohydrate antigens. The enzymes or family of enzymes responsible for each step of the biosynthesis are indicated. The enzymes identified as showing different expression levels in <i>Pteridium aquilinum</i> treated mice (Group 2) are highlighted in red (increased expression) and blue (decreased expression). <b>Panel B</b> – Immunohistochemistry of simple mucin-type carbohydrate antigens, Tn (a,b), STn (c,d), T (e,f) and ST (g,h) in gastric mucosa of control (Group 1) (a,c,e,g) and <i>Pteridium aquilinum</i> treated (Group 2) (b,d,f,h) mice. Bar = 20 µm.</p

Primer sequences and Real-time PCR conditions for the analysis of gene expression in mice gastric mucosa.

<p>Primer sequences and Real-time PCR conditions for the analysis of gene expression in mice gastric mucosa.</p

Glycosyltransferases expression analysis of gastric mucosa from <i>Pteridium aquilinum</i> exposed mice. Panel A –

<p>Heatmap of the Glyco-gene Chip array analysis obtained when comparing group 1 (control mice) with group 2 (mice treated with <i>Pteridium aquilinum</i>). Red and blue indicate increased and decreased expression relative to the mean transcript expression value, respectively. The transcripts identified as differentially expressed were those with adjusted p-value <0.1 and fold change >1.3. <b>Panel B –</b> Real-time PCR analysis for expression of <i>St6galnac6</i>, <i>Galntl4</i>, <i>St3gal2</i> and <i>C1galt1</i> in gastric mucosa from control (Group 1), <i>Pteridium aquilinum</i> treated (Group 2) and concomitantly <i>Pteridium aquilinum</i> treated and <i>Helicobacter pylori</i> infected (Group 4) mice. The gene used as reference was <i>Hprt1</i>. Significance was evaluated using the Shapiro-Wilk test. n.s. not significant p<0.05(*); p<0.01(**); p<0.005 (***).</p

Characterization of cystically dilated gland of gastric mucosa of mouse exposed to <i>Pteridium aquilinum.</i>

<p><i> Pteridium aquilinum</i> treated mice (Group 2) displaying a cystically dilated gland stained with hematoxylin and eosin (a), Alcian Blue (pH = 2,5) (b), and immunostained for Sialyl-Lewis x antigen (c). Bar = 20 µm.</p