Upregulation of mucosal soluble fas ligand and interferon-gamma may be involved in ulcerogenesis in patients with Helicobacter pylori-positive gastric ulcer.

BACKGROUND: Excessive upregulation of gastric epithelial cell apoptosis is speculated to be associated with ulcerogenesis in Helicobacter pylori-positive peptic ulcer disease. H. pylori may have an ulcerogenic effect through induction of gastric epithelial cell apoptosis mediated by infiltrating T cells and their soluble products. METHODS: The contents of soluble Fas ligand (sFasL) and interferon-gamma (IFN-gamma) in organ cultures and the degree of apoptosis and the expression of apoptosis-related proteins in the gastric epithelium were examined using the mucosal tissues obtained from the antrum and the ulcer site in patients with H. pylori-positive gastric ulcer (GU). The molecular mechanisms of gastric epithelial cell apoptosis induced by sFasL and IFN-gamma were analyzed using epithelial cell lines, MKN 45 and KATO III. RESULTS: The mucosal tissues of the ulcer site had substantially higher contents of sFasL and IFN-gamma in organ cultures regardless of its healing stage in association with increased numbers of apoptotic cells and enhanced expression of proapoptotic proteins Bak and Bax in the surface foveolar epithelium as compared with the antral tissues in patients with H. pylori-positive GU. The addition of sFasL caused increases in cytotoxic cell death and caspase-3 activation in MKN 45 and KATO III cells in which IFN-gamma treated cells had more prominent effects than untreated cells. The expression of Bak in MKN 45 cells increased when they were treated with IFN-gamma. CONCLUSIONS: Upregulation of mucosal sFasL and IFN-gamma may be involved in ulcerogenesis in patients with H. pylori-positive GU through induction of gastric epithelial cell apoptosis.

Resistance of Crohn's disease T cells to multiple apoptotic signals is associated with a Bcl-2/Bax mucosal imbalance.

Crohn's disease (CD) is a condition characterized by excessive numbers of activated T cells in the mucosa. We investigated whether a defect in apoptosis could prolong T cell survival and contribute to their accumulation in the mucosa. Apoptotic, Bcl-2+, and Bax+ cells in tissue sections were detected by the TUNEL method and immunohistochemistry. T cell apoptosis was induced by IL-2 deprivation, Fas Ag ligation, and exposure to TNF-alpha and nitric oxide. TUNEL+ leukocytes were few in control, CD, and ulcerative colitis (UC) mucosa, with occasional CD68+ and myeloperoxidase+, but no CD45RO+, apoptotic cells. Compared with control and UC, CD T cells grew remarkably more in response to IL-2 and were significantly more resistant to IL-2 deprivation-induced apoptosis. CD T cells were also more resistant to Fas- and nitric oxide-mediated apoptosis, whereas TNF-alpha failed to induce cell death in all groups. Compared with control, CD mucosa contained similar numbers of Bcl-2+, but fewer Bax+, cells, while UC mucosa contained fewer Bcl-2+, but more Bax+, cells. Hence, the Bcl-2/Bax ratio was significantly higher in CD and lower in UC. These results indicate that CD may represent a disorder where the rate of T cell proliferation exceeds that of cell death. Insufficient T cell apoptosis may interfere with clonal deletion and maintenance of tolerance, and result in inappropriate T cell accumulation contributing to chronic inflammation.

Increased mucosal production of granulocyte colony-stimulating factor is related to a delay in neutrophil apoptosis in Inflammatory Bowel disease.

Tissue accumulation of polymorphonuclear neutrophils (PMN) in Inflammatory Bowel disease (IBD) might be, in part, due to a delay in apoptotic processes associated with the effects of their specific growth factors and inflammatory cytokines. We addressed this hypothesis by examining the activity of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage CSF (GM-CSF) in the organ culture supernatants of colonic mucosal specimens and their regulatory effects on PMN apoptosis in patients with IBD. The contents of G-CSF and GM-CSF in the supernatants were measured by the enzyme-linked immunosorbent assays and PMN apoptosis was evaluated by acridine orange/ethidium bromide staining, respectively. Mucosal specimens obtained from patients with active IBD exhibited higher levels of G-CSF and GM-CSF activity than controls. Notably, the levels of G-CSF activity were approximately 1000-fold higher than those of GM-CSF activity. Freshly isolated PMN showed a time-related increase in the proportion of cells with characteristic features of apoptosis when they were incubated with the culture medium alone and exposure of PMN to recombinant G-CSF and GM-CSF caused a concentration-dependent inhibition of apoptosis. Incubation of PMN with the supernatants from patients with active IBD induced an inhibitory effect on PMN apoptosis; this effect was abrogated to a significant degree by pre-incubation of the supernatants with anti-G-CSF serum. This study suggests that PMN apoptosis may be delayed under the influence of soluble mediators, especially G-CSF, in the microenvironment of IBD-affected mucosa, thus providing possible mechanisms for tissue accumulation of PMN in IBD.

Expression of Bcl-2 and p53 correlates with the morphology of gastric neoplasia.

The growth of a tumour can be determined by an interplay between cell proliferation and loss. The expression of apoptosis-related proteins (Bcl-2 and p53), cell proliferation (Ki-67), and apoptotic cell death were investigated using immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling in gastric neoplams, to evaluate whether they correlate with the morphology of the tumour. The materials included ten cases of gastric adenoma and 40 cases of early gastric carcinoma consisting of differentiated adenocarcinomas (n = 20) and undifferentiated carcinomas (n = 20). All cases of adenoma and eight cases of differentiated adenocarcinoma were of the elevated type, while 12 differentiated adenocarcinomas and all of the undifferentiated carcinomas were of the depressed type. The diffuse expression of Bcl-2 was observed in all cases of adenoma and seven out of eight (88 per cent) of elevated-type differentiated adenocarcinoma. In contrast, Bcl-2 expression was absent or focal in the depressed type of carcinoma. Overexpression of p53 was found exclusively in the depressed type of carcinoma. Thus, Bcl-2 and p53 expression was associated with tumour morphology. It seemed unlikely that Bcl-2 and p53 expression was involved in the morphogenesis of the gastric tumours through inhibiting apoptotic cell death, since the degree of apoptosis in Bcl-2-positive gastric tumours was rather higher than that in Bcl-2-negative ones and it did not differ significantly between p53-positive and p53-negative tumours. Instead, the diffuse distribution of Bcl-2 correlated with the superficial distribution of Ki-67-positive proliferating cells, and the overexpression of p53 had a tendency to correlate with the diffuse distribution of proliferating cells. These results suggest that diffuse Bcl-2 expression and a superficial distribution of proliferating cells may contribute to the elevated configuration, and that overexpression of p53 and a diffuse distribution of proliferating cells may result in the depressed configuration in the relatively early stages of gastric tumourigenesis.

Successful treatment of primary sclerosing cholangitis with cyclosporine and corticosteroid.

A case of primary sclerosing cholangitis (PSC) successfully treated with cyclosporine is described. A 65-year-old man who presented with jaundice and anemia was diagnosed as having PSC, accompanied by pancreatic duct abnormalities. Cholangiography and pancreatography showed marked improvements following combined therapy with cyclosporine and methylprednisolone. Cyclosporine seems to be a promising drug for the treatment of patients with PSC. This report reasons that the same disease process affects both the pancreatic ducts and the bile ducts, and that stagnation of pancreatic juice may have a role to play in the pancreatic duct abnormalities.

[Treatment of nausea and vomiting induced by a 24-hour i.v. infusion of cisplatin].

Nine advanced gastric cancer patients were given 17 courses of cisplatin administrations by means of a 24-hour intravenous infusion at a dose of 100 mg/m2. For an anti-emetic, 40 mg of metoclopramide was administered 5 times at 6-hour intervals, along with a 500 mg hydrocortisone administration at the start of the cisplatin infusion. Despite this preventative treatment, nausea and/or vomiting occurred in over one-third of all the courses. Thus, to combat this nausea and/or vomiting, a combination of lorazepam (1.5 mg/day, divided into 3 p.o.), dexamethasone (20, 10 and 10 mg by i.v. at 3, 8, and 24 hours, respectively, after start of the cisplatin infusion), and a 60 mg intravenous administration of metoclopramide (5 times at 6-hour intervals) was given, and it was found that this new method (Method IV) prevented both nausea and vomiting.

Human saliva inactivates mutagenicity of carcinogens.

Mutagenicities of AF-2, MNNG, 4NQO, aflatoxin B1, benzo [a] pyrene and Trp-P-1, with or without metabolic activation, were inactivated by treatment with human saliva to a great extent in the Ames test with Salmonella typhimurium test strains TA98 and TA100. Mutagenic activities of quercetin, pyrolysates of beef, salmon and sodium glutamate, and condensate of cigarette smoke were also decreased to some extent by saliva treatment, but no significant effect was found on the activity of MMS and pyrolysate of polypeptone. These effects showed individual variations. The inhibition of AF-2 mutagenicity by saliva varied with temperature in TA100 but not in TA98 cultures. Boiled saliva inactivated AF-2 mutagenicity in TA98 to some extent but not in TA100 cultures. Inactivation of AF-2 mutagenicity by saliva treatment was completed within 30 sec. Complex mechanisms may be involved in the inactivation of mutagenicity of carcinogens by saliva, including biochemical reactions with enzymes, vitamins, etc. and/or adsorption with high molecular weight substances in saliva such as proteins, bacterial cells, mucous materials, etc.