13 research outputs found

    Effect of glucose and lactate on cellular ATP content in IPEC-1 cells after 1 h treatment.

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    <p>IPEC-1 cells were grown to confluence (7 d) and treated for 1 h with HBSS-based solutions. Supernatant was removed and ATP content was measured by a luminescence based assay. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p<0.05) differences.</p

    Bray-Curtis OUT’s level principal component analysis (PCA).

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    <p>(A) Individual fecal samples of sow, piglets at one week and four weeks of age. (B) Individual sow fecal samples in the control group and YD group. (C) Individual sow fecal samples based on the sow colostrum yield, high (≥ 3500 g) and low (< 3500 g).</p

    Microbiota profiles showing normalized square root transformed abundances of genera in two groups of sow.

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    <p>Genera are colored according to their phyla. <i>Firmicutes</i> (blue), <i>Bacteroides</i> (orange), and <i>Proteobacteria</i> (green). *<i>P</i><0.05, **<i>P</i><0.01.</p

    Quantification of cytosol / nucleus NADH ratio in IPEC-1 cells after 1 h treatment with glucose and lactate without further challenge.

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    <p>Cellular autofluorescence of IPEC-1 cells was monitored at ex: 340 nm / em: 510 nm. <b>(A)</b> Change in spatial distribution of autofluorescence in response to Anti A (100 μM). <b>(B)</b> Cytosolic autofluorescence in response to complex III inhibitor Anti A (100 μM) and uncoupler FCCP (1 μM). <b>(C)</b> IPEC-1 cells were treated as indicated for 1 h. Glucose (gluc) concentration was 1 g/L if not otherwise indicated. Cells were treated with lactate (lac) 25 mM or increased gluc 4.5 g/L. Autofluorescence measured after 1 h and ratio of cytosolic (F<sub>cytosol</sub>) and nuclei (F<sub>nucleus</sub>) was calculated (ex: 340 nm / em: 510 nm) from individual cells without further stimulation. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p<0.05) differences.</p

    Attachment and structural effects of lactobacilli on IPEC-1 cells.

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    <p><b>(A)</b> Three dimensional reconstruction of lactobacilli attached to IPEC-1 cells. <i>Lactobacillus amlylovorus</i> strain GRL1110 (arrows) was applied for 6 h on confluent IPEC-1 cells. The bacterial suspension was then carefully removed; cells and attached bacteria were fixed and stained with DAPI (nuclei, blue) and CM-Dil (plasma membrane, red). Confocal sections were reconstructed to obtain a 3D view. Note the thin, barely visible cytosolic layer above nucleus (*). (<b>B-H</b>) Actin structure of IPEC-1 cells treated with lactate and <i>Lactobacillus amylovorus</i> supernatants. IPEC-1 cells were cultured on glass, incubated as indicated and stained for F-actin (Phalloidin-PromoFluor 546) and nuclei (DAPI). (<b>B</b>) control buffer pH7, (<b>C</b>) Na-DL-lactate (25 mM, pH 7), (<b>D</b>) control buffer pH 4, (<b>E</b>) lactic acid pH 4. (<b>F-H</b>) supernatants of <i>Lactobacillus amylovorus</i> strains. (<b>F</b>) GRL1110, (<b>G</b>) GRL1112, (<b>H</b>) GRL1115. Predominantly basal F-actin fibre structure was reduced by lactic acid and bacterial supernatants (pH 4), (<b>I</b>) F-actin staining of freeze-section of porcine jejunal epithel. Note positive F-actin signal indicating the apical terminal web (arrowhead), the low F-actin signal at the lateral side and the faint signal in the basal region of the enterocytes (arrow). Bar: 5 μm.</p

    Effect of weaning on intestinal lactate concentration and lactobacilli load of piglets.

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    <p><b>(A)</b> Lactate concentration <b>(B)</b> <i>Lactobacillus</i> load and <b>(C)</b> pH in chyme of suckling and weaned piglets along the intestinal axis was measured. Statistical analysis was performed with the procedure “MIXED” (SAS, version 8) and comparison between pre- and postweaning using the t-test (p<0.01).</p

    Normalised Anti A triggered ROS generation in IPEC cells after lactate and glucose treatment.

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    <p><b>(A)</b> IPEC-1 cells were cultured on glass (7 d), incubated for 1 h in buffer (HBSS), placed on microscopic stage and challenged with Anti A in the presence of DHE. Mean fluorescence of nuclei (regions of interest, white circles) was monitored over time. <b>(B)</b> Quantification of nucleic fluorescence of ethidium cation (ethidium cation is the oxidation product of DHE) binding to nuclei. Anti A (10 / 100 μM) or control buffer was applied in presence of DHE 1.5 min after starting the measurement. <b>(C)</b> Anti A (100 μM) triggered superoxide generation rate (DHE oxidation) of IPEC-1 and IPEC-J2 cells after 1 h treatment (39°C). Cells were treated with lactate (lac, 25 mM); lac + inhibitor of monocarboxylate transporter (MCT inhibitor, ARC155858, 10 μM); increased glucose (gluc, 4.5 g/L) or glycolysis inhibitor 2-Deoxyglucose (2DG, 10 mM). Glucose concentration was 1 g/L if not otherwise indicated. Slope of fluorescence increase of individual cell nuclei was calculated and normalised to the control. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p<0.05) differences.</p
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