Temporal and spatial distribution of corticosteroidogenic enzymes immunoreactivity in developing human adrenal.

Steroid hormones secreted by fetal adrenocortical cells are considered to be a requirement for a fetus to maintain intrauterine life, but, to date, the regulation of steroid hormone secretion has not been studied in detail in the human fetal adrenal gland. In this study, we examined the immunolocalization of steroidogenic enzymes and their local regulation, including adrenal 4-binding protein (Ad4BP or NR5A1), steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc or CYP11A1), P450 17alpha-hydroxylase/17,20-lyase (P450c17 or CYP17), 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD), P450 21 hydroxylase (P450c21 or CYP21), dehydroepiandrosterone sulfotransferase (DHEA-ST), P450 oxidoreductase and cytochrome b5, in the human fetal adrenal gland (n=31) obtained from fetuses ranging in ages from 14 to 40 weeks of gestation. Ad4BP immunoreactivity was detected in all adrenocortical zones throughout gestation, suggesting that this nuclear protein is likely to be essential in the development of the human adrenal. Immunoreactivity for StAR, P450scc, P450c21, P450 oxidoreductase and cytochrome b5 was detected only in fetal and transitional zone between 14 and 22 weeks of gestation, but was detected in all three zones after 23 gestational weeks. 3beta-HSD immunoreactivity was not detected in any of the three cortical zones prior to 22 weeks of gestation, but became discernible in the transitional zone and definitive zone after 23 weeks. Immunoreactivity for P450c17 and DHEA-ST was detected in the transitional and fetal zones throughout gestation, but not in the definitive zone. These results suggest that the human adrenal cortex may produce dehydroepiandrosterone (DHEA) in the transitional and fetal zones throughout gestation, and cortisol in the transitional zone after the 23rd week of gestation.

Chicken ovalbumin upstream promoter transcription factor II in the human adrenal cortex and its disorders.

Chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is an orphan member of the steroid/thyroid hormone receptor superfamily. COUP-TFII has been demonstrated to negatively regulate the transcriptional activity of adrenal 4-binding protein, a steroidogenic cell-specific transcription factor that activates the transcription of various steroidogenic P450 genes. We therefore examined immunolocalization of COUP-TFII in the human adrenal cortex and its disorders, including functioning and nonfunctioning cortical tumors, to study its possible correlation with adrenocortical steroidogenesis. In nonpathological adrenal cortex, COUP-TFII immunoreactivity was marked in the nuclei of adrenocortical cells in definitive and fetal zones from 16 gestational weeks to 2 months after birth. Immunoreactivity for COUP-TFII was marked in the zona glomerulosa and weak in the zonae fasciculata and reticularis from 7 months to 8 yr of age, but thereafter markedly decreased in these zones (P < 0.05, between age 7 months to 8 yr and 24-62 yr of age, respectively). In adrenocortical tumors, COUP-TFII immunoreactivity was marked in the nuclei of tumor cells of aldosteroma (H score, 134 +/- 15.9; P < 0.001 vs. Cushing's adenoma and P < 0.05 vs. nonfunctioning adenoma and carcinoma), modest in nonfunctioning adenoma (82.7 +/- 19.8) and adrenocortical carcinoma (79.6 +/- 56.3), and low in Cushing's adenoma (38.2 +/- 24.5). Results from immunoblotting performed in seven cases of adenomas were consistent with those of immunohistochemistry. In the attached nonneoplastic adrenal cortex of the adenomas, immunoreactivity for COUP-TFII was markedly increased compared to that in nonpathological adrenal cortex in adults and was especially marked in the zona glomerulosa in the attached adrenal of aldosteroma (P < 0.001) and the zona fasciculata in that of Cushing's adenoma (P < 0.05). COUP-TFII immunoreactivity was universally detected in stromal cells of the adrenal glands. These results suggest that COUP-TFII plays an important role in the regulation of steroidogenesis in human adrenal cortex and its disorders.

Non-Helicobacter pylori ulcer disease in rheumatoid arthritis patients receiving long-term NSAID therapy.

Helicobacter pylori and nonsteroidal antiinflammatory drugs (NSAIDs) are considered the major causes of peptic ulcer. If ulcers are not attributable to H. pylori, most are thought to be attributable to NSAIDs. We have previously reported that rheumatoid arthritis (RA) patients taking NSAIDs long term (NSAIDs group) are more likely to develop gastric ulcers, which commonly occur in the gastric antrum. In addition, the morphology of gastric ulcers in the NSAIDs group differs from that in the non-NSAIDs group (control group), in whom NSAIDs are not involved in the occurrence of gastric ulcers. In this study, we compared gastric ulcers in the NSAIDs group with those in the control group in terms of H. pylori infection. The positive rate of H. pylori in gastric ulcers was significantly lower in the NSAIDs group than in control group (53.6% vs 91.5%). At the ulcer site they were seen significantly less often in the antrum than in either the angle or body of the stomach (35% vs 100%) in the NSAIDs group. On the other hand, the H. pylori-positive rate for ulcers in the antrum did not differ significantly from that in the angle and body of the stomach (81.8% vs 93.8%) in the control group. These findings suggest that H. pylori plays little role in antral ulcers in those taking NSAIDs.

17Beta-hydroxysteroid dehydrogenase type 2 and dehydroepiandrosterone sulfotransferase in the human liver.

The liver plays important roles in the clearance and metabolism of sex steroids. Its dysfunction is considered to influence the metabolic pathways of sex steroids, and to result in gynecomastia and other abnormalities of sex steroids. However, the details of its mechanism have not been well-characterized. We therefore examined the enzymes involved in the hepatic clearance and/or metabolism of sex steroids in human liver and its disorders using immunohistochemistry to determine whether there are any abnormalities of expression of these enzymes in human liver disorders. These enzymes are 17beta-hydroxysteroid dehydrogenase (17beta-HSD) type 2, an enzyme that catalyzes the biologically active estrogen, estradiol (E2), to inactive estrogen, estrone (El), and dehydroepiandrosterone sulfotransferase (DHEA-ST), which catalyzes sulfonation of dehydroepiandrosterone (DHEA) to form biologically inactive DHEA-S. A total of 162 cases including normal liver (n=31), chronic hepatitis (n=41), liver cirrhosis (n = 21), hepatocellular carcinoma (n = 47), cholangiocellular carcinoma (n = 22) and fetal liver (n = 4) were examined by immunohistochemistry. Both enzymes were expressed in the hepatocytes around portal area and central vein in normal liver. Immunopositive area for DHEA-ST was significantly larger in chronic hepatitis than in normal liver, but that of 17beta-HSD type 2 in chronic hepatitis was not different from normal liver. There were no significant differences in the immunopositive area for both enzymes between liver cirrhosis and normal liver. In hepatocellular carcinoma, immunoreactivity for both enzymes were categorized into Group A, or low positive group, and Group B, or high positive group. The latter tended to be poorly differentiated carcinoma. In cholangiocellular carcinoma, immunopositive areas of both enzymes were significantly smaller than those of normal liver. These findings indicate that the amount of expression of the enzymes involved in metabolism and/or clearance of sex steroids per hepatocyte did not decrease in liver cirrhosis. Therefore, sex steroids' abnormalities may be due to the decreased quantity of hepatocytes associated with liver cirrhosis. In hepatocellular carcinoma, some poorly differentiated cases were associated with increased expression of 17beta-HSD type 2 but its biological significance needs to be determined by further studies.

Studies on the fibercolonoscopy: with special reference to the latest procedures.

The Machida type fibercolonoscope, or FCS, devised by us is an anterior direct vision type of fiberscope. The most important feature of this new fiberscope consists of a tip of 5 cm size which is capable of being directed by flection up to 120 degrees in any direction and the tip up to 30 cm possessing a fishing-rod-like flexibility and elasticity by means of a special spiral spring. This angle-regulating mechanism makes it possible to insert this instrument along the course of the intestine under direct visual control. The fishing-rod-like construction where the maximal flexibility is greatest at the tip makes it considerably easier to pass sites of marked flexure in the colon. At examination the insertion of the instrument is guided by direct observation and by roentgenoscopy. The results in 200 cases in 1970 and thereafter, when operative techniques No. I, II and III devised previously by us were used, showed that successful insertion into the cecum was possible in 139 out of 173 cases (80%) except 27 cases of pathological stenosis due to cancer and other reasons. Thus fibercolonoscopy now enables the direct observation of the colon beyond the reach of the signoidoscope, providing a useful diagnostic procedure. Furthermore, a medium-long fibercolonoscope or FCS-M has been produced by us for examination of the lower portion of the large intestine. This is used in the inverted position without roentgenoscopic guidance as in conventional sigmoidoscopy. When this method, fibersigmoidoscopy, was used in 120 cases, it was possible to examine up to the transverse colon in 19 cases, the splenic flexure in 22 cases and the descending colon in 39 cases without the use of special techniques I, II and III. Thus in the majority of cases it is possible to carry out the examination of the left colon in this manner. The usefulness of fibersigmoidoscopy in the practical clinic is, therefore, rather larger than that of fibercolonoscopy.

Statistical analysis of diverticulosis of the colon.

Forty-four cases of diverticulosis of the colon were detected among 2,662 cases examined by barium enema in Tohoku University. This corresponds to 1.7% in frequency. On the other hand, 21 cases were found among 1,511 cases examined in Hirosaki University, giving 1.4% in frequency. It was more frequent in the male. The frequency, as calculated on the basis of the numbers of cases examined by barium enema in separate age groups, showed the gradual increase with age except for above the eighth decade. The location where the diverticulum was most frequently seen was the right colon; namely, cases in which the diverticulum occurred from the cecum to the ascending colon and those in which it was found only in the ascending colon occupied 61.4 and 71.4% of the total cases, respectively. Diverticulitis as a complication was rare, while coincidence of polyp or carcinoma was sometimes observed. Furthermore, the value of fiberscopy for the diagnosis of diverticulosis of the colon was discussed, and its usefulness for the detection of complication was emphasized.

Fibercolonoscopy for the diagnosis of granulomatous colitis.

In order to clarify the significance of fibercolonoscopy in diagnosis of granulomatous colitis, the pathological, radiological, endoscopic and clinical differences between granulomatous colitis and ulcerative colitis were investigated on 12 cases of granulomatous colitis and 23 cases of ulcerative colitis, which underwent colonic resections between 1954 and 1973. The comparative studies have revealed considerable differences in their characteristic features besides the overlapping spectrum. As a consequence of assessments made of the various diagnostic techniques as to usefulness in differential diagnosis between the two colitis conditions, it was noted that fibercolonoscopy and biopsy under direct vision, when applied in conjunction with the conventional procedures, readily facilitate the differential diagnosis without any surgical intervention in all aspects, excepting the depth of inflammatory involvement and the presence of lymphangiectasia. However, the depth of the inflammed lesion as well as lymphangiectasia can also be estimated indirectly. Therefore, it seems that it is possible to make the differential diagnosis between the two colitis conditions even prior to colectomy, with the exception of atypical forms which represent overlapping findings.