Saburo Ohshiba

Immunochemical detection of human lactoferrin in feces as a new marker for inflammatory gastrointestinal disorders and colon cancer

We have developed a new immunochemical test for fecal lactoferrin (LF) utilizing an enzyme-linked immunosorbent assay (ELISA). The ELISA had a sensitivity of about 10 micrograms/L of lactoferrin and the measurable range was 10.0-1000.0 micrograms/L (1.0-100.0 micrograms LF/g feces). The stability of lactoferrin in feces was greater than that of myeloperoxidase and leucocyte elastase. The fecal concentration of lactoferrin (mean +/- SD) in 35 normal subjects was 0.75 +/- 0.83 microgram/g feces, whereas that in 24 patients with colon cancer was 74.4 +/- 88.3 micrograms/g feces. The fecal lactoferrin concentration of 38 patient with active ulcerative colitis was 307.4 +/- 233.9 micrograms/g feces, and that in 36 patients with active Crohns disease was 191.7 +/- 231.1 micrograms/g feces. The ELISA for human fecal lactoferrin might be useful in the diagnosis of colon disease.

Intestinal Protein Loss and Bleeding Assessed by Fecal Hemoglobin, Transferrin, Albumin, and Alpha-1-Antitrypsin Levels in Patients with Colorectal Diseases

Four fecal proteins (hemoglobin, transferrin, albumin, and alpha 1-antitrypsin) were measured by enzyme-linked immunosorbent assay (ELISA) in patients with colorectal diseases. Levels of all 4 proteins were significantly increased in patients with colonic cancer and ulcerative colitis (UC) compared to levels in control subjects, while fecal alpha 1-antitrypsin was particularly elevated in colonic Crohns disease (CD). That is, the fecal protein pattern of CD was distinct from those of colonic polyps, colonic cancer, and UC. To investigate whether levels of these fecal proteins reflect disease activity in UC and CD, comparative evaluation of fecal proteins in the active and inactive phases was performed. In UC, differences in the fecal concentrations of all 4 proteins were significant between the active and inactive phases of the disease. In CD, however, the difference in alpha 1-antitrypsin concentration was significant. Our results suggest that measurements of these 4 fecal proteins would be useful in the screening of colorectal diseases. In addition, these markers can also be used as indicators of disease activity in inflammatory bowel diseases.

Immunochemical detection of human blood in feces

We have developed a new immunochemical test for fecal occult blood utilizing enzyme-linked immunosorbent assay (ELISA) of human hemoglobin (HbAo) and transferrin (Tf) simultaneously. The ELISA had a sensitivity of about 15 ng/ml Hb, and the measurable range was 1.5-750 micrograms Hb per g feces. The stability of Tf in feces was greater than that of Hb. In 17 out of 18 patients with colon cancer, 8 out of 15 patients with colon polyps, and 11 out of 20 patients with upper-gastrointestinal disorders. The Hb and Tf values were more than 10 micrograms/g feces, in terms of Hb concentration. The ELISA for human fecal HbAo and Tf might be useful for the diagnosis of gastrointestinal disorders.

Clinicopathological Study on Flat and Depressed Minute Colorecta Carcinomas

Abstract: Over the last two years, a total colonoscopy was performed on 3,006 patients and 232 colorectal carcinomas were detected in these patients. Seventy one (30.6%) of the carcinomas were found to be early cancers. Eighteen (25.4%) of these 71 early cancers were minute (>5 mm) colorectal carcinomas of the flat or depressed type and they were resected endoscopically. More than half of them were accompanied by colonic polyps.

Endoscopic evaluation of the therapeutic effect of sclerotherapy for esophageal varices

The rebleeding rate after endoscopic injection sclerotherapy was studied in 237 patients with esophageal varices, and the optimal outcome of treatment was determined. Two new categories, RC(2-) and F0, were added to the classification scheme of endoscopic findings of varices in Japan. RC(2-) represents a state in which no veins, not even small vessels, are observed by endoscopy after endoscopic injection sclerotherapy, and F0 represents a state in which no localized venous dilations in the esophagus exist. The criteria for defining the other categories were not altered, and the R-C sign was expressed as RC(2-), RC(-), or RC(+); the degree of dilation was classified as F0 to F3. By combining the R-C sign and the F number, endoscopic findings were classified 1 to 4 weeks after endoscopic injection sclerotherapy into groups designated RC(2-)F0, RC(-)F0, RC(-)F1, and RC(+)F1-RC(+)F2. The four groups were observed to determine the incidence of esophageal stricture and rebleeding. In the RC(2-)F0 group, the incidence of stricture was high, but the rebleeding rate was low. In the RC(-)F0 group, both the incidence of stricture and rebleeding rate were low. We conclude that the optimal outcome of endoscopic injection sclerotherapy is RC(-)F0.

Clinical study of a new fecal occult blood test using a combination assay of hemoglobin and transferrin

SummaryThe applicability of a new immunological fecal occult blood test in which hemoglobin (Hb) and transferrin (Tf) are simultaneously assayed was evaluated. The mean absorbance and standard deviation (510/630 nm) obtained by this test was 0.840±0.805 in 51 fecal samples from patients with colon cancer, 0.248±0.305 in 95 samples from patients with colon polyps, and 0.104±0.053 in 110 samples from control patients; these values differed significantly (P<0.005). Hb and Tf concentrations were separately determined in the same fecal sampels, and qualitative evaluation was performed with a cutoff value of 5.1µg/g feces for Hb and 0.4µg/g feces for Tf. Hb or Tf was positive in 41 of the 51 samples in the colon cancer group, 33 of the 95 in the colon polyp group, and 3 of the 110 in the control group. Qualitative analysis of the values obtained by the combination assay of Hb and Tf with a cutoff value of 0.200 revealed positive rates of 41/51 in the colon cancer gorup, 33/95 in the colon polyp group, and 4/110 in the control group. These results sugest the usefulness of a combination assay of Hb and Tf as a fecal occult blood test.

Immunohistochemical study of carbohydrate antigen expression in gastric carcinoma

SummaryThe expression of carbohydrate antigens in malignant and non-malignant gastric mucosa was studied immunohistochemically using the monoclonal antibody AH6 directed to Ley antigen, FH2 directed to Lex antigen, and FH6 directed to sialyl-Lex antigen. Formalin-fixed gastric tissue resected from 54 patients with gastric cancer and 20 patients with gastric ulcer were tested. The incidence of positive cases in gastric cancer patients with each antibody was as follows: AH6;85%, FH2;74%, FH6;74%. The Lex antigen was expressed in 81.5% of cases histologically classified as undifferentiated type, and 66.7% of cases classified as differentiated type. It was expressed in a higher incidence in early stage cancer (93.3%) than in advanced stage cancer (66.7%). Sialyl-Lex antigen was detected in more cases of differentiated type (88.9%) than in those of undifferentiated type (59.3%), whereas none of 8 early cancers of undifferentiated type expressed the antigen. The incidence of the expression of Ley antigen did not differ in relation to histological type or invasiveness. Lex and Ley antigens were detected in noncancerous gastric epithelium. Sialy-Lex antigen was not detected in the normal fundic gland region. These results demonstrate that Lex antigen may be a differentiation-associated antigen, and sialyl-Lex antigen might be useful as a marker of differentiated cancer and an indication for invasion of undifferentiated cancer.

The effect of acid secretagogues on mucin synthesis using primary monolayer culture of the guinea pig gastric mucous cells

SummaryMucin plays a principal role in protecting the gastric mucosa against injury. We investigated the effect of acid secretagogues on mucin synthesis using a primary gastric mucous cell monolayer cultue system of guinea pig. Significant increases in mucin synthesis were observed in response to the secretagogues pentagastrin (10-8M, 10-7M) and carbachol (10-4M, 10-3M), but not to histamine. After pretreatment with indomethacin (10-5M), 10-8M pentagastrin significantly increased mucin synthesis to 125.6±3.9%, but carbachol did not. Prostaglandin E2 release into the culture medium was signifiantly increased by 10-4M carbachol to 118.0±5.9%, but there was no change after application of pentagastrin. These findings suggest that pentagastrin and carbachol may act directly on mucous cells, and that part of the mucin synthesis-promoting action of carbachol is mediated by prostaglandins. There were no changes in intracellular cAMP concentration after the addition of these acid secretagogues. However, calcium ionophore (A23187) produced an increase in mucin synthesis, suggesting a Ca++ involvement in mucin synthesis. No differences were found in the sugar side chain structures of newly synthesized mucin glycoprotein as a result of exposure to acid secretagogues by histochemical or biochemical methods using lectins.

Kimura's disease (angiolymphoid hyperplasia with eosinophilia) associated with ulcerative colitis.

SummaryWe treated a 35-year-old Japanese woman who had Kimura’s disease associated with ulcerative colitis. Kimura’s disease is an uncommon chronic inflammatory disease considered to be of allergic origin based on the presence of eosinophilia and IgE hyperimmunoglobulinemia, and may be a manifestation of a systemic immunologic disturbance. Some immunological complications such as nephrotic syndrome have been reported in association with Kimura’s disease, but the present case is the first associated with ulcerative colitis.

Effects of 16, 16-dimethyl prostaglandin E2 on lysosomal membrane stability in rat stomach

The lysosomal membrane encloses numerous hydrolytic enzymes and prevents the cytoplasm from being damaged by these enzymes. It is possible that the fragility of this membrane may be implicated in the pathogenesis of gastric mucosal damage. We investigated the effects of 16,16-dimethyl prostaglandin E2 (dmPGE2), which is known to protect the gastric mucosa from various noxious agents, on lysosomal membrane stability in the rat stomach. Sodium taurocholate (TC) was used as the damaging agent. To assess lysosomal membrane stability in the gastric mucosa, we assayed acid phosphatase released from lysosomes isolated from a gastric mucosal homogenate. To assess lysosomal membrane stability in gastric surface epithelial cells, we used laser scanning confocal microscopy to observe the fading of red fluorescence in living cells vitally stained with acridine orange. Exogenous dmPGE2 enhanced lysosomal membrane stability in the gastric mucosa, whereas TC decreased it. In gastric surface epithelial cells, exogenous dmPGE2 protected the cells against TC-induced damage and prevented TC-induced decreased lysosmal membrane stability. It was concluded that a decrease in lysosomal membrane stability seemed to be closely involved in the pathogenesis of gastric mucosal damage. Moreover, it appears that stabilization of the lysosomal membrane by exogenous dmPGE2 may contribute to its protective effect in the gastric mucosa, both at the level of gastric surface epithelial cells and in regard to the entire gastric mucosa.