Kazuro Itoh

Propionibacterium freudenreichii component 1.4‐dihydroxy‐2‐naphthoic acid (DHNA) attenuates dextran sodium sulphate induced colitis by modulation of bacterial flora and lymphocyte homing

Background and aim: 1.4-Dihydroxy-2-naphthoic acid (DHNA), a bifidogenic growth stimulator from Propionibacterium freudenreichii, is thought to have a beneficial effect as a prebiotic; however, its in vivo effect on intestinal inflammation remains unknown. The aim of this study was to determine whether oral administration of DHNA can ameliorate dextran sodium sulphate (DSS) induced colitis and to determine the possible underlying mechanisms. Method: Colitis was induced in mice by treatment with 2.0% DSS for seven days. DHNA (0.6 or 2.0 mg/kg) was given in drinking water prior to (preventive study) or after (therapeutic study) DSS administration. Colonic damage was histologically scored, and mucosal addressin cell adhesion molecule 1 (MAdCAM-1) expression and β7 positive cell infiltration were determined by immunohistochemistry. mRNA levels of proinflammatory cytokines (interleukin (IL)-1β, IL-6 and tumour necrosis factor α (TNF-α)) were determined by quantitative real time polymerase chain reaction. In addition, bacterial flora in the caecum, concentrations of short chain acids, and luminal pH were examined. Results: DHNA improved survival rate and histological damage score in mice administered DSS in both the preventive and therapeutic studies. DHNA significantly attenuated the enhanced expression of MAdCAM-1, the increased β7 positive cell number, and the increased mRNA levels of IL-1β, IL-6, and TNF-α in DSS treated colon. In addition, the decreased number of Lactobacillus and Enterobacteriaceae induced by DSS was recovered by DHNA. Preventive effects on decrease in butyrate concentration and decrease in pH level in mice administered DSS were also observed in the DHNA preventive study. Conclusion: DHNA, a novel type of prebiotic, attenuates colonic inflammation not only by balancing intestinal bacterial flora but also by suppressing lymphocyte infiltration through reduction of MAdCAM-1.

Expression of PD-1, PD-L1, and PD-L2 in the liver in autoimmune liver diseases

OBJECTIVES:PD-L1 (also B7-H1) and PD-L2 (also B7-DC) are ligands for programmed death-1 (PD-1), which is a member of the CD28/B7 superfamily of costimulatory molecules and plays an inhibitory role on the periphery. Impaired regulation of this system may cause disruption to self-tolerance leading to autoimmunity; however, the role of these molecules in the liver is unknown. Therefore, we examined the expression of PD-1, PD-L1, and PD-L2 in the liver in autoimmune liver diseases.METHODS:We examined the liver expression of these molecules in autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) with no previous medical treatment using immunohistochemical staining and real-time PCR, and compared with chronic hepatitis type C (CHC) as a control.RESULTS:Although PD-1, PD-L1, and PD-L2 were expressed in the liver in AIH, PBC, as well as CHC, the expressions were relatively lower in PBC. In AIH, despite more severe inflammation than in CHC, the expression of these molecules was not greater than in CHC, and when compared with the relative expression of PD-L1, PD-L2 was lower in AIH. PD-L1 and PD-L2 expressions were well correlated with the level of IFN-γ; however, relatively decreased induction for PD-L1 and PD-L2 by IFN-γ was observed in AIH or PBC than in CHC.CONCLUSION:Modulation of PD-1/PD-L1 and PD-L2 systems may play a role in the development of autoimmune liver diseases.

SQUAMOUS CELL CARCINOMA OF GALLBLADDER

Two cases of a well‐differentiated keratinizing squamous cell carcinoma of the gallbladder were reported. Pathologic analysis of this rare neoplasm was made in conjunction with cases of the gallbladder carcinoma of a squamous cell variety reported in literatures. The squamous cell carcinoma is characterized by a well‐localized growth and a rarity or lack of metastasis. These characteristics make a good contrast with an adenosquamous carcinoma of the gallbladder which usually infiltrates rather extensively and metastasizes widely. Thus the adenosquamous carcinoma should be sequestered from group of squamous cell carcinoma. Radical operative procedures may well be encouraged on selected cases of squamous cell carcinoma.

Cysteamine-induced duodenal ulcer and the hepatoduodenal branch of the vagus nerve

SummaryWe investigated the role of the autonomic nervous system in gastric acid secretion, somatostatin concentration and PAS-positive mucus production in Brunner’s glands in cysteamine-induced duodenal ulcer. Vagotomized rats were used. No ulcers occurred in the groups with vagotomies of the hepatoduodenal, truncal or gastric branches after cysteamine administration. However, in the hepatoduodenal branch vagotomized group, there was an increases in gastric acid secretion after cysteamine administration. A similar increase was observed in the control group, but the decreases in somatostatin concentration and PAS-positive mucus seen in the control group were not found in the hepatoduodenal vagotomized group. These results suggest that the hepatoduodenal branch of the vagus nerve might play an important role in the ulcerogenic process of cysteamine-induced duodenal ulcer.

An experimental study of gastric mucosal blood flow in endotoxemia of the rat, with special reference to the vagus nerve and EDRF

Gastric mucosal lesions are an important complication in endotoxemia. In order to define the role played by the vagus nerve and endothelial-derived relaxing factor (EDRF) in gastric mucosal blood flow, an investigation was carried out on four groups of rats: a control group; a group given lipopolysaccharide (LPS, 5 mg/kg); a group given gossypol-acetic acid (gossypol), which has an injurious effect on the vascular endothelial cell; and a group given L-NG-monomethyl arginine (LNMMA). Following the administration of acetylcholine and papaverine hydrochloride (via the splenic artery) and vagus nerve stimulation in all four groups of rats, the effects of vagus nerve stimulation and EDRF on the gastric mucosal blood flow were determined with a laser Doppler rheometer. In the LPS group, the gastric mucosal blood flow was decreased after acetylcholine administration and vagus nerve stimulation. This was also the case in the gossypol group. These findings suggest that inhibition of EDRF release may be responsible for the reduced gastric mucosal blood flow observed in endotoxemia.

Biological Significance of Phospholipids in the Rat Stomach–From the Viewpoint of Electron Microscopy

The structures of gastric phospholipids (PLs) were further confirmed by employing an improved fixative and fixation method. PLs were detected on the free surface of superficial mucous epithelial cells and epithelial cells of the foveolar crypts. PLs were also detected inside superficial mucous cells and epithelial cells of the foveolar crypts. The gastric PLs were secreted through a merocrine-like mechanism. Apocrine-like secretion of PLs was also observed in some electron microscopic images. PLs were observed in the intercellular and intracellular secretory canaliculi of parietal cells and also in the rest of the parietal cytoplasm. The above findings indicated the localization of PLs on the surface of the gastric lumen and in the gastric mucosa, and suggested that, similar to gastric glycoproteins, gastric PLs play an important role in the gastric mucous barrier and mucosal barrier.

Effect of 15(R)-15-methyl PGE2 (arbaprostil) on duodenal bicarbonate secretion in rat

SummaryThe present study was designed to observe the effect of 15(R)-15-methyl PGE2 (arbaprostil) on duodenal bicarbonate secretion in male Wistar rats anesthetized with urethane. A proximal duodenal loop (1.5cm) was made and perfused with saline (pH4.5) in an air tight circle system. The pH and Pco2 of the perfusing fluid were measured continuously and the quantity of bicarbonate was calculated according to the Henderson-Hasselbalch equation. Vehicle or arbaprostil was injected, intravenously as a bolus injection. The total output of bicarbonate in 30 min was as follows; control: 0.449 ± 0.093 μEq (mean±SE); 1,10 and 100 μ/kg of arbaprostil: 0.752±0.218,2.75±0.430 and 5.958±0.578 μEq respectively. Arbaprostil (10 and 100 γ/kg) increased the total output of bicarbonate significantly (p< 0.001). The rate of secretion was also increased by arbaprostil.

S1233 Evaluation of Long Pentraxin Ptx3 for the Inflammation Marker in the Patients with Ulcerative Colitis

Backgrounds & Aims: CRP, short pentraxin C-reactive protein, is often used in the evaluation of inflammatory status in ulcerative colitis (UC). However, usually serum CRP level does not increase in active UC because the short pentraxin CRP is produced mainly by the liver in response to interleukin (IL)-6, as well as IL-6 level is not so high in patients with UC. Pentraxin-3 (PTX3) is a prototypic long pentraxin mainly by dendritic cells, macrophages and endothelial cells in response to primary inflammatory stimuli. We reported increased plasma PTX3 concentration in active inflammatory bowel diseases (Kato S et al. Dig Dis Sci 2007 in press). The aims of this study were to investigate the usefulness of plasma PTX3 concentration as an diagnositic and therapeutic marker for patients with UC. Materials & Methods: Plasma PTX3 levels were examined in 46 UC patients by sandwitch ELISA. Mayo Scoring system was used to evaluate disease activity in UC patients. Plasma PTX3 concentrations were evaluated for disease type and clinical course. Sensitivity, specificity and accuracywere evaluated Results: Plasma PTX3 concentration was decreased after treatment for UC. For disease type, plasma PTX3 concentration of total colitis , left side colitis and proctitis type were 8.05+-4.03, 9.22+-5.35 and 3.55+-1.91(concentration of of total colitis , left side colitis vs proctitis type, p<0.05). Serum CRP concentration of of total colitis , left side colitis and proctitis type were 2.01+-3.39, 1.1+-2.71 and 0.11+-0.1. For clinical course, plasma PTX3 concentration of one attack only , relapse-remitting and chronic continuous type were 3.79+-2.52, 8.06+-5.02 and 6.82+-2.93. Serum CRP concentration of one attack only , relapse-remitting and chronic continuous type were 0.12+-0.084, 1.83+-3.43 and 0.57+1.00. Sensitivity, specificity and accuracy of plasma PTX3 concentration against disease activity were 74%, 92% and 83% in UC patients. However, sensitivity, specificity and accuracy of serum CRP concentration were 39%, 100% and 70%. Conclusions: Plasma PTX3 concentration is a good diagnostic and therapeutic marker in patients with UC.

M1211 Anti-Inflammatory Effect of Novel Lactobacillus Isolated from Japanese “Funazushi” On DSS-Induced Colitis

BACKGROUND: Genus Lactobacillus is classified broadly into two categories. One is isolated from dairy products or gut and another is vegetable -derived Lactobacillus (VDL) isolated from fermented vegetable food such as pickles or a soy-sauce. Genus Lactobacillus isolated from dairy products or gut have been used for clinical trials in inflammatory bowel disease (IBD) as probiotics, however, VDL was poorly investigated. In this study, we newly isolated two VDL from Funazushi (Japanese old-style sushi; salted and fermented with a rice and a crusian carp) and examined its anti-inflammatory effect on dextran sodium sulfate (DSS)induced colitis. MATERIALS & METHODS: The surrounding rice material of Funazushi, was homogenized and plated on MRS medium for selection of Genus Lactobacillus. A single colony was picked and sorted out by its ability for suppression of proinflammatory cytokines at mRNA level in RAW264.7 cells after LPS stimulation (In Vitro screening). Intragastric administration of Lactobacillus isolated from Funazushi or L.gasseri (ATCC 33323) was performed in 8-week olds-female C57BL/6 mice every day during administration of 6% DSS in water for 7 days. The degree of colonic inflammation and mRNA levels of expression of cytokines in colonic mucosa were determined. Results: Among 500 screened colonies, two different anti-inflammatory types of Lactobacillus were isolated from Funazushi (s193 and s292). Thus, in addition to each Lactobacillus alone administration, we performed the combinatory administration with equivalent amount of s193 and s292 (dual administration). Although each Lactobacillus from Funazushi showed an attenuating effect on inflammation, dual administration significantly improved survival rate, body weight loss and histologic damage score in mice compared with either DSS-treated control, L.gasseri, s193 or s292 alone group. mRNA level of IL-6 was attenuated by s193 alone, and IL-12p40 or IL-23p19 level was decreased by s292 alone. mRNA levels of IL-1β and TNF-α were also remarkably inhibited by dual administration. Conclusions: Two strains of newly established Lactobacillus isolated from Funazushi showed a different anti-inflamattory effect on DSS-induced colitis and that effect was enhanced by simultaneous dual administration. These data suggested that VDL isolated from Funazushi may be useful for treatment for IBD with stronger antiinflammatory effects than Lactobacillus isolated from dairy products or gut.

Abstracts of selected papers presented at the 77th General Meeting of the Japanese Society of Gastroenterology

M E E T I N G OF T H E J A P A N E S E S O C I E T Y OF G A S T R O E N T E R O L O G Y March 28-30, 1991-Tokyo, Japan Chairman: Yutaka MATSUO, M.D.