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The wheat-grain proteome was investigated, as a basis for devising more efficient methods of cultivar identification or discrimination. Australian wheats (Halberd, Cranbrook, CD87 and Katepwa) were used as the basis of this study. These cultivars were selected on the basis of differences in the quality types represented, in terms of dough-processing attributes that can suit one cultivar better than another for specific types of industrial utilisation. Total wheat endosperm (flour) protein extracts were prepared from mature wheat for two-dimensional electrophoresis, across both acidic (pH 4-7) and basic (pH 6-11) pH ranges. Three particular regions of the proteome maps were chosen for close comparison, involving two sets of gluten proteins and a nongluten protein region (involving small heat shock proteins), based on previous protein characterisation. Differences in the nongluten protein regions (heat shock proteins and other unidentified polypeptides) are of particular interest as being possible targets for use in developing new approaches to cultivar discrimination, such as the development of simple immunoassays.
BACKGROUND - Cytochrome P450 (CYP) 3A is constitutively expressed in human intestinal villi and may account for significant "first-pass" prehepatic metabolism of a number of drugs in the intestine. Celiac disease results in small intestinal atrophy. We hypothesized that this would result in a loss of CYP3A.
METHODS - Formalin-fixed jejunal biopsy specimens taken from nine patients with celiac disease at variable times before and after treatment with a gluten-free diet were immunoperoxidase stained after incubation with anti-CYP3A4 rabbit antisera (1:2000 dilution). The amount of immunoreactive CYP3A was determined by two observers who were blinded to the treatment states of the patients. Staining intensity was graded on a numerical scale from 1 to 4+ on the basis of intensity of staining in individual enterocytes, as well as the total number of enterocytes stained.
RESULTS - Slides of biopsy specimens from all nine untreated patients with celiac disease were graded 1. Treatment with a gluten-free diet was associated with a significant increase in CYP3A immunoreactive protein in small bowel biopsy specimens (p < 0.05, Wilcoxon signed-rank test).
CONCLUSIONS - We conclude that patients with celiac disease have low intestinal CYP3A immunoreactivity and that treatment with a gluten-free diet is associated with an increase in intestinal CYP3A immunoreactive protein. Our findings suggest that intestinal disease and variability in intestinal CYP3A activity might be an unexamined variable that may contribute to interindividual variability in drug disposition.