Bile acid metabolism was studied by means of the fractional turnover rate or orally ingested 14C-labeled taurocholic acid and by gas chromatographic determination of fecal excretion of the bile acids cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), and lithocholic acid (LCA). Thirty patients with Crohn's disease (CD) of the small bowel, of whom 19 had been operated on with limited ileal resections, were studied and compared with 11 healthy volunteers. The unoperated group of CD patients did not show significant increase in bile acid excretion in the stools in contrast to the CD patients with ileal resection. The fecal excretion consisted mostly of primary bile acids, and a significant correlation between length of resection and bile acid excretion was found (rs = 0.81, p less than 0.01). The fractional turnover rate of CA + DCA was significantly increased in both unoperated (0.21 l/day) and operated (0.44 l/day) patients compared with normal controls (0.06 l/day). The bile acid pool of CA + DCA, however, was normal in patients with ileal resections, indicating a compensatory increase in bile acid synthesis. In unoperated patients the bile acid pool of CA + DCA was slightly decreased (3.1 mmol) compared with operated patients (6.2 mmol) and normal controls (4.8 mmol). The pool size was not significantly correlated to mean transit time of dietary residue, feces excretion, loss of weight, or amount of fat in feces. The mean transit time of dietary residue was decreased in both operated and unoperated CD patients.
We report a simple method to establish culture of human colonic epithelial cells from endoscopically obtained biopsy specimens, producing sufficient viable cells to perform metabolic studies pertinent to the pathogenesis of IBD and related human disorders.
of protein kinase C during interferon-y-and phorbol ester-stimulated immunocytochemical expression of ICAM-1 in human renal carcinoma cells. APMIS 101: 437448, 1993.Incubation of the human renal carcinoma cell line CaKi-1 with interferon-y (IFN-y) or the phorbol ester, phorbol-12-myristate 13-acetate (PMA) strongly stimulated the immunocytochemical expression of the intercellular adhesion molecule-I (ICAM-I) in a dose-dependent manner. Since PMA is capable of activating the Ca2+ /phospholipid-dependent protein kinase C (PKC), we investigated the role of this kinase during IFN-y signal transduction. Calcium ionophore A231 87 significantly enhanced IFN-y-and PMA-induced ICAM-1 staining. While staurosporine, H7 and sphingosine, three known PKC inhibitors, blocked the PMA effect, only staurosporine abrogated the action of IFN-y. Finally, 24 h of PMA pretreatment with subsequent IFN-y stimulation enhanced ICAM-1 staining above values from cultures where IFN-y was omitted. This occurred despite the fact that 24 h of PMA pretreatment abolished the effect of IFN-y on PKC activation, as determined by acetylated myelin basic protein 4 1 4 phosphorylation. In conclusion, these results suggest that additional events other than PKC activation are required for complete regulation of ICAM-1 antigen by IFN--y in the whole cell population. Hence, other Ca2+-dependent signalling pathway(s) mediated by IFN-y receptors must act. Further studies are needed to elucidate these specific pathway(s) activated during IFN-y stimulation in our model.
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