Expression of NOD2 in Paneth cells: a possible link to Crohn's ileitis.

BACKGROUND AND AIMS: Genetic variation in NOD2 has been associated with susceptibility to Crohn's disease (CD) and specifically with ileal involvement. The reason for the unique association of NOD2 mutations with ileal disease is unclear. To identify a possible link, we tested expression of NOD2 in intestinal tissue of CD patients and controls. PATIENTS AND METHODS: Fifty five specimens of ileum or colon from 21 CD patients, seven ulcerative colitis (UC) patients, and five controls with pathology other than CD or UC were stained for NOD2 using an immunoperoxidase method. RESULTS: Using a monoclonal antibody against NOD2 developed in our laboratory, we detected uniform expression of NOD2 in terminal ileum Paneth cells from controls and patients as well as in metaplastic Paneth cells in the colon. Mechanical purification showed enriched expression of NOD2 mRNA in ileal crypts. In Paneth cells, NOD2 was located in the cytosol in close proximity to the granules that contain antimicrobial peptides. We detected minimal NOD2 in the villous epithelium of the ileum or in the colonic epithelium from both CD patients and controls. CONCLUSIONS: These results suggest a role for NOD2 in the regulation of Paneth cell mediated responses against intestinal bacteria and a plausible mechanism to explain the selective association of NOD2 mutations with ileal disease. The impaired capacity of CD associated mutations to sense luminal bacteria may result in increased susceptibility to certain gut microbes.

Endogenous and exogenous ARC in serum withdrawal mediated PC12 cell apoptosis: a new pro-apoptotic role for ARC.

The role of ARC (Apoptosis Repressor with Caspase Recruitment Domain) in PC12 cell serum withdrawal driven apoptosis was studied. A progressive and massive increase in ARC protein occurs during serum withdrawal that correlates with declining survival and processing of caspase-2, previously shown to associate with ARC. This accumulation of ARC occurs in a transcriptional and translational independent manner. Additionally, ARC is localized exclusively in the nucleus of PC12 cells. Furthermore, transfection of PC12 cells with hARC-Flag promotes death and fails to protect the cells from apoptosis by serum withdrawal. Therefore, ARC functions in a pro-apoptotic manner in PC12 cell serum withdrawal induced apoptosis.

ANG II stimulates endothelial nitric oxide synthase expression in bovine pulmonary artery endothelium.

Although angiotensin II (ANG II) is a known pulmonary vasoconstrictor, the purpose of this study was to examine the effect of ANG II on pulmonary artery endothelial cell nitric oxide synthase (ecNOS) mRNA and protein expression. Cultured bovine pulmonary artery endothelial (BPAE; passages 5-8) cells were incubated for 0-12 h with 10(-6) M ANG II. Total RNA was extracted, and ecNOS expression was assessed by Northern blot analysis. In BPAE cells, ecNOS mRNA was significantly increased 2.4 +/- 0.3-fold (P < 0.05 vs. basal; n = 5) 6 h after the addition of ANG II over basal levels. In & similar time course, it was found that ecNOS protein concentrations are increased 247 +/- 62% (P < 0.05 vs. basal; n = 8) over basal levels 4 h after ANG II addition. There is a second protein peak 8 h after ANG II addition in which ecNOS was increased 333 +/- 145% over basal (P < 0.05, n = 3). These data suggest that ANG II stimulates ecNOS mRNA expression and are followed by increased levels of ecNOS protein in cultured BPAE cells, consistent with an observed increase in nitrite production. Both the increase in ecNOS protein and mRNA expression could be inhibited with the ANG II receptor antagonist saralasin. Additionally, actinomycin D, an inhibitor of transcription, prevented the rise in mRNA at 6 h while cycloheximide inhibited the initial protein peak. The effects of ANG II on ecNOS were specific for the pulmonary artery endothelium. Addition of ANG II did not increase ecNOS protein or mRNA expression in parallel studies in bovine coronary artery endothelium. The stimulation of ecNOS by ANG II may act to protect the lung and maintain low pulmonary artery pressures in the renin-angiotensin model of systemic hypertension.

Development and validation of the Stirling Eating Disorder Scales.

The development and reliability/validity check of an 80-item, 8-scale measure for use with eating disorder patients is presented. The Stirling Eating Disorder Scales (SEDS) assess anorexic dietary behavior, anorexic dietary cognitions, bulimic dietary behavior, bulimic dietary cognitions, high perceived external control, low assertiveness, low self-esteem, and self-directed hostility. The SEDS were administered to 82 eating disorder patients and 85 controls. Results indicate that the SEDS are acceptable in terms of internal consistency, reliability, group validity, and concurrent validity.

Comparison of eating disorders and other dietary/weight groups on measures of perceived control, assertiveness, self-esteem, and self-directed hostility.

Anorexic and bulimic patients were compared to obese dieters, nonobese dieters, and normal controls on measures of perceived control, assertiveness, self-esteem, self-directed hostility, and psychiatric caseness. The anorexic and bulimic groups both scored significantly differently in the expected direction from the other three groups on all measures. There were no significant differences between the anorexic and bulimic groups and in turn, no significant differences among the obese, nonobese dieters, and normal controls. Results are in keeping with the notion that perceived control, low assertiveness, low self-esteem, and self-directed hostility are characteristics of eating disorder patients that differentiate them from individuals who display dietary/weight features, as well as from normal controls.