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, 2007). Segments were incubated in oxygenated (95% O2; 5% CO2) bicarbonate buffer at 37��C. Tissue was equilibrated for 15 min, thereafter a 3 mA current pulse was applied across the intestinal wall every 6 s for 30 min. The trans-epithelial potential was measured and recorded by the Acquire and Analyze software, and the change in potential induced by the current pulse was used to calculate trans-epithelial resistance (TER) according to Ohm's Law. Higher values of TER represent lower gut permeability. Immunohistochemical staining Four micro miter-thick cross sections of Selleckchem JAK inhibitor formalin-fixed, paraffin-embedded ileum and colon tissue were cut, deparaffinized, and subjected to a heat-induced epitope retrieval step. Slides were rinsed in cool running water, washed in Tris-buffered saline (pH7.4) before incubation with primary polyclonal antibody against mouse Muc2 (USCN, USA, dilution 1:100) overnight at 4��C. For detection, rabbit anti-rat secondary antibody was used followed by application of the peroxidase kit (USCN, USA). Alkaline phosphatase was revealed by Fast Red as chromogen and peroxidase was developed with a highly sensitive diaminobenzidine (DBA) chromogenic substrate for approximately 10 min. Negative controls were performed by omitting the primary antibody. Statistical analysis The relative position and intensity of DNA bands on DGGE profiles were used for principal component analysis (PCA) by SPSS 16.0 statistical package (SPSS Inc. IL, USA). Statistical analyses were performed using a two-tailed Student t-test, with the assistance from GraphPad Prism Program (version 5.01, GraphPad Software Inc., USA). All values were expressed as means �� standard deviation (SD) and the sample sizes. Significance was accepted at p *p