The effects of variations in dose and method of administration on glucagon like peptide-2 activity in the rat

The influence of nutrients, biliary-pancreatic secretions, and systemic trophic hormones on intestinal adaptation in a Roux-en-Y bypass model

“…After rinsing in saline, the length and the weight of each were measured, as described in previous work [38,39].…”

“…Tissues were fixed in 10% formalin. The remaining mucosa was pinned open, and mucosal scrapings of at least 5 cm of mucosa were taken from each limb immediately adjacent to the site of the histologic sample and frozen for future analysis (−80°C) [12,39]. For histologic examination, 6-μm paraffin sections of the formalin-fixed tissues were cut and stained with H&E. An observer blinded to the origin of the tissue assessed the morphology.…”

“…Crypt cell proliferation rates were quantified using BrdU as a marker of cell division, as previously described [39,40]. In brief, BrdU staining was done using deparaffinized sections treated with 2N HCl for 1 hour at 60°C, incubated overnight in a 1:100 dilution of BrdU primary antibody (Seralab, Sacramento, Calif), and detected with a Cy3-conjugated antirat IgG secondary antibody (Jackson Immunoresearch Laboratories, West Grove, Pa).…”

“…In brief, BrdU staining was done using deparaffinized sections treated with 2N HCl for 1 hour at 60°C, incubated overnight in a 1:100 dilution of BrdU primary antibody (Seralab, Sacramento, Calif), and detected with a Cy3-conjugated antirat IgG secondary antibody (Jackson Immunoresearch Laboratories, West Grove, Pa). Ten consecutive well-oriented crypts per slide were reviewed by an observer blinded as to tissue source and reported as the ratio of BrdU staining nuclei to total cells per crypt [39]. Similarly, apoptosis was quantified using immunohistologic detection of activated caspase-3 as a marker for apoptosis [39,40].…”

“…Ten consecutive well-oriented crypts per slide were reviewed by an observer blinded as to tissue source and reported as the ratio of BrdU staining nuclei to total cells per crypt [39]. Similarly, apoptosis was quantified using immunohistologic detection of activated caspase-3 as a marker for apoptosis [39,40]. In brief, 6-μm paraffin sections were deparaffinized and blocked with 3% hydrogen peroxide for 30 minutes.…”