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Biomedical subjects

M Tsuchiya

Publications and source records attributed to M Tsuchiya.

At least 127 records · Page 7Linked to original sources
PubMed1995

Spatial heterogeneity of mucosal blood flow during ischemia-reperfusion injury of rat stomach investigated by laser Doppler perfusion imaging.

Spatial alterations in blood flow during the development of mucosal injury induced by ischemia-reperfusion in rats were determined with a two-dimensional laser Doppler tissue perfusion imager. The rats were anesthetized with pentobarbital, and the stomach was exteriorized on a stage; the mucosa was then sequentially scanned. The mucosa was constantly superfused with 0.1 N HCl in physiological saline. Systemic arterial pressure was continuously monitored and blood was stepwisely withdrawn from the femoral artery by 20-mmHg stage and then maintained at 20 mmHg for 20 min. The shed blood was reinfused and the stomach was removed 30 min later. Under control conditions, the average perfusion of the forestomach was usually greater than that in the glandular stomach. When systemic blood pressure was stepwisely decreased, the extent of decrease in the mucosal blood perfusion unit was always greater than that in systemic blood pressure, but mucosal perfusion appeared to be uniformly decreased throughout the stomach. Ten min after reperfusion, a hypoperfused area began to appear in the corpus near the greater curvature, and this area subsequently increased. The area of ulcer formation corresponded with the hypoperfused area in the gastric mucosa 30 min after reperfusion. Pretreatment with CV-6209, a platelet-activating factor antagonist, significantly attenuated the hypoperfusion induced by reperfusion and also prevented gastric mucosal damage. Our results suggest that hypoperfusion in the mucosal microcirculation is indeed an important factor contributing to the localized occurrence of gastric mucosal lesions and that the laser Doppler perfusion imager is useful for the detection of local hypoperfused areas in the gastric mucosa.

Analysis of Variance↗
PubMed1995

Wound healing of acetic acid-induced gastric ulcer in rats and the effects of cimetidine and calcitonin, with special reference to prolylhydroxylase and collagenase enzyme activity.

The healing of acetic acid-induced gastric ulcer in rats and the effects of cimetidine and calcitonin were investigated with reference to the enzyme activity of both prolylhydroxylase and collagenase as related to histological findings. The rats were observed by endoscopy on the 3rd day after the subserosal injection of acetic acid; rats with ulcers were divided into three groups: non-treated, and cimetidine- and calcitonin-treated. The latter two groups were treated for 7 days. Prolylhydroxylase activity in active ulcers in the non-treated group was slightly higher on the 3rd day and significantly higher on the 10th day than the activity in control rats that had received subserosal injections of physiological saline solution on the respective days. In non-treated rats, the healed ulcer on the 10th day showed lower prolylhydroxylase activity than that in the active ulcer on the same day. Cimetidine did not affect prolylhydroxylase activity, but, with calcitonin, there was higher prolylhydroxylase activity in the healed than in the active ulcer, although the difference was not significant. Interstitial collagenase showed the highest activity on the 3rd day and decreased on the 10th day in non-treated rats. Collagenase activity was higher in the cimetidine-treated group, than that in the non-treated group, and numerous peroxidase-positive granulocytes were seen in the mucosa and submucosa. Calcitonin did not affect collagenase activity. The participation of both enzymes is indispensable in the healing process and the effects of anti-ulcer agents on these enzymes must be considered.

Acetates↗
PubMed1995

Antibody-dependent cell-mediated cytotoxicity against cell lines generated by liver-specific idiotype-bearing antibody.

We produced a murine monoclonal antibody (mAb), designated H2-mAb, against a fractionated soluble phase of human liver homogenate which antibody reacted with human liver cells. A human antibody possessing the same idiotype as the H2-mAb, designated LSIA (liver-specific idiotype-bearing antibody), can be measured by a sandwich enzyme-linked immunosorbent assay, using the anti-H2 idiotype antibody. The serum level of LSIA in patients with histologically proven chronic hepatitis (CH) was significantly higher than that in healthy subjects and it was also higher than that in subjects with other diseases, including systemic lupus erythematosus. In a comparison between patients with CH type B and those with CH type C, there was no significant difference in serum levels of LSIA. It was possible to purify LSIA from the sera of patients with CH. The purified LSIA bound to the human cell lines Chang and HCC-M, derived from liver cells and a hepatoma respectively, but not to HeLa cells, a uterine carcinoma derivative. The reactivity of this mAb to HCC-M was weaker than that to Change. Moreover, the presence of LSIA caused an antibody-dependent cell-mediated cytotoxic challenge against Change cells in vitro.

Adult↗
PubMed1995

Oxidative stress in gastric mucosal injury: role of platelet-activating factor-activated granulocytes.

Temporal and spatial changes due to oxidative stress in the rat gastric mucosa were visualized and quantified during the process of mucosal hemorrhagic change. The fluorescence associated with dichlorofluorescein (DCF), a hydroperoxide-sensitive fluorochrome, increased 30 min after repeated electrical stimuli to the gastric artery. The increase in the fluorescence was enhanced in the area between two adjacent collecting venules. The content of platelet-activating factor (PAF), the activity of myeloperoxidase (MPO) in the gastric mucosa, the area of mucosal lesions, and the luminol-dependent chemiluminescence activity in zymosan-treated blood samples, obtained from the gastric vein, were measured and found to increase significantly 30 min after the stimuli. The intravenous injection of CV-6209, a PAF antagonist, 5 min prior to the stimuli significantly inhibited the DCF activation, the increases in PAF level and MPO activity, the mucosal hemorrhagic change, and the elevation in chemiluminescence activity. In addition, continuous infusion of superoxide dismutase also inhibited all these changes, except for chemiluminescence activity. These results suggest that oxygen radicals derived from PAF-activated granulocytes induce oxidative stress, and that oxidative changes are actually implicated in the pathogenesis of gastric mucosal injury.

Animals↗
PubMed1995

Intravital demonstration of sequential migration process of lymphocyte subpopulations in rat Peyer's patches.

BACKGROUND & AIMS: Although recirculation of lymphocytes through Peyer's patches is important for specific immune defense, the intraorgan migration of lymphocyte subpopulations has not been clearly understood. The aim of this study was to compare the spatial distributions of labeled lymphocytes among various subpopulations in rat Peyer's patches. METHODS: Lymphocytes collected from intestinal lymph were separated into CD4+, CD8+, and T and B cells, labeled with a fluorochrome carboxyfluorescein diacetate succinimidyl ester, and injected into the jugular vein. Peyer's patches of recipient rats were observed by intravital fluorescence microscopy. RESULTS: No significant difference was found in the percentage of lymphocytes in transit or in the rolling velocity among different subpopulations. Lymphocytes sticking to the venules increased in number at 10-20 minutes, with preferential adherence of CD4+ cells to venules of 25-50 microns and preferential adherence of B cells to the venules of a wider size range. After 30 minutes, extravasated lymphocytes moved into the interstitium. B cells migrated from venules more quickly than CD4+ cells. CD8+ cells showed an intermediate pattern between CD4+ and B cells in sticking and migratory behaviors. Subsequently, CD4+ and CD8 cells preferentially appeared in parafollicular microlymphatics. CONCLUSIONS: Significant differences were observed among lymphocyte subpopulations in terms of spatial distribution of lymphocytes sticking to venules, migration into the interstitium, and their lymphatic transport.

Animals↗
PubMed1995

Humanization of mouse ONS-M21 antibody with the aid of hybrid variable regions.

Mouse monoclonal antibody, ONS-M21, directed against human medulloblastoma cells, has been humanized by complementarity determining region (CDR) grafting. A humanized ONS-M21 VH region, comparable to the original mouse ONS-M21 VH region, was easily constructed based on framework regions (FRs) 1, 2 and 3 from human EU antibody and on FR4 from human ND antibody. Five alterations in the FRs were made at amino acids 27, 28, 29, 30 and 94 which are all part of the canonical structure for CDR1 (H1). The humanized ONS-M21 VL regions were constructed based on the FRs from human REI antibody. We first identified five amino acid residues in the FRs at positions 20, 21, 71, 73 and 87 as having a possible adverse influences on antigen binding. None of the versions with a variety of combinations at these five positions showed any bindings to antigen. In order to identify the mouse residues that must be retained in the human FRs, hybrid VL regions were constructed by joining the mouse ONS-M21 VL region and the first humanized version within CDR2. The hybrid VL regions revealed that residues in FR1 and/or FR2 were critical in creating a functional antigen binding site. Redesigning several versions with alterations in FR1 and FR2 revealed that the Pro-46 residue was the only critical residue for creating an antigen binding site. This approach should be helpful in identifying key residues in difficult cases of antibody humanization.

Amino Acid Sequence↗
PubMed1995

Relationship between hepatitis C virus subtypes and clinical features of liver disease seen in alcoholics.

The influence of hepatitis C virus and its subtypes on the clinical course of liver disease in alcoholics was assessed. Hepatitis C virus infection was confirmed by a reverse transcription and polymerase chain reaction method for the hepatitis C virus NS-5 region in the sera of alcoholics with various stages of histologically proven liver disease. The frequency of hepatitis C virus was significantly higher in alcoholics with chronic hepatitis (73%) than in those with liver fibrosis (18%), alcoholic hepatitis (17%), and fatty liver (0%). Hepatitis C virus subtypes, namely K1 and K2, were determined by dot-blot hybridization analysis of the polymerase chain reaction products with specific probes, and their frequencies were 68% and 32%, respectively. The proportion of patients whose serum transaminase levels returned to normal following 4 weeks of abstinence in hospital was significantly lower in alcoholics with hepatitis C virus viremia (glutamic oxaloacetic transaminase: 53.8%; glutamic pyruvic transaminase: 42.3%) than in those without viremia (glutamic oxaloacetic transaminase: 86.2%, p < 0.01; glutamic pyruvic transaminase: 89.7%, p < 0.01). When alcoholics with the K1 and K2 subtypes of hepatitis C virus were compared, normalization of transaminase levels was less frequent in alcoholics with K1 (glutamic oxaloacetic transaminase: 42.8%; glutamic pyruvic transaminase: 28.6%) than in those with K2 (glutamic oxaloacetic transaminase: 88.9%, p < 0.05; glutamic pyruvic transaminase: 77.8%, P < 0.05). These data indicate that hepatitis C virus infection is associated with a reduced rate of recovery of serum transminase levels following abstinence in subjects with alcoholic liver disease, more so in the K1 subtype than in the K2 subtype.

Alcohol Drinking↗
PubMed1995

Substance P induces degranulation of mast cells and leukocyte adhesion to venular endothelium.

Substance P (SP), one of the established neurotransmitters, evokes an immunoinflammatory response involving leukocyte adhesion to venular endothelium and the degranulation of mast cells. The pathogenetic relationship between these responses, however, remains unresolved. In this study, we propose to examine the changes associated with the activation of mast cells, as well as leukocyte adhesion to venular endothelium by in vivo observation of the rat mesentery. The use of an in vitro assay for intracellular Ca2+ mobilization and the degranulation of mast cells demonstrated the significant upper shift of concentration response to SP (10(-4)-10(-5) M). In vivo experiments on the mesenteric microcirculation also showed that SP induced a significant increase in the number of degranulated mast cells as well as in the number of leukocytes adherent to the venular wall. Tranilast, a mast cell stabilizer, as well as SP antagonist (CP-96,345) significantly attenuated the extent of mast cell degranulation and leukocyte adhesion elicited by SP. Although an immunoneutralization against CD18 by WT-3 significantly attenuated the leukocyte adhesion, it had no influence on the mast cell degranulation after SP superfusion. These separate in vivo observations show that SP induces leukocyte adhesion to the venular endothelium, possibly through the degranulation of mast cells.

Animals↗
PubMed1995

Effects of aluminum adjuvant on systemic reactions of lipopolysaccharides in swine.

In vivo effects of aluminum adjuvant on systemic reaction of bacterial lipopolysaccharide (LPS) in piglets were investigated. Intramuscular injection of 0.1 mg kg-1 of LPS added to aluminum hydroxide gel (LPS(+)AL) mitigated the leukopenia, trembling and serum levels of TNF-alpha and cortisol compared with the injection of LPS suspended in LPS-free saline (LPS(+)SALINE). The serum endotoxin levels were reduced remarkably but relatively long-lasting in the LPS(+)AL. The lethality in mice injected with LPS added to aluminum hydroxide gel was significantly reduced. Likewise, the Limulus activity of a test LPS was reduced by the addition of aluminum hydroxide gel or aluminum chloride.

Adjuvants, Immunologic↗
PubMed1995

Formation of bioorganic compounds in planetary atmospheres by cosmic radiation.

Simulated planetary atmospheres (mixtures of simple gases) were irradiated with high energy particles to simulate an action of cosmic rays. When a mixture of carbon monoxide, nitrogen and water was irradiated with 2.8-40 MeV protons, a wide variety of bioorganic compounds including amino acids, imidazole, and uracil were identified in the products. The amount of amino acids was proportional to the energy deposit to the system. Various kinds of simulated planetary atmospheres, such as "Titan type" and "Jovian type", were also irradiated with high energy protons, and gave amino acids in the hydrolyzed products. Since cosmic rays are a universal energy source in space, it was suggested that formation of bioorganic compounds in planetary atmospheres is inevitable in the course of cosmic evolution.

Amino Acids↗
PubMed1995

Autoradiographic demonstration of lansoprazole uptake sites in rat antrum and colon.

To investigate possible effects of lansoprazole other than its acid suppression through the parietal cell, uptake sites for lansoprazole were studied by autoradiography in rat antral and colon mucosa. Autoradiography was performed using two different methods, i.e., routine fixation and in vitro autoradiography on cryostat sections. Our results showed that the uptake sites for lansoprazole in the gastric antrum were located in the interstitial cells, including unmyelinated nerve fibers, whereas those in the colon mucosa corresponded to epithelial cells and some of the interstitial cells. With electron microscopy, the uptake sites for lansoprazole in the colon epithelial cells were seen exclusively near the apical plasma membrane. On the basis of these findings, lansoprazole may have effects on autonomic nerve and colon epithelial function as well as on acid secretion.

2-Pyridinylmethylsulfinylbenzimidazoles↗
PubMed1995

Lansoprazole inhibits oxygen-derived free radical production from neutrophils activated by Helicobacter pylori.

We studied the influence of lansoprazole on Helicobacter pylori-elicited neutrophil activation, including the oxidative burst and infiltration of gastric mucosa, and confirmed whether radiolabeled lansoprazole is actually detected in gastric neutrophils. The oxidative burst of purified human neutrophils was measured by luminol-dependent chemiluminescence (ChL). [3H]Lansoprazole uptake sites in human gastric mucosa were observed by autoradiography. The magnitude of neutrophil infiltration of gastric mucosa was assessed by tissue myeloperoxidase (MPO) content. ChL assay indicated that oxygen-derived free radical production was increased twofold by adding H. pylori water extract, which was significantly inhibited by lansoprazole (10(-4) M). Gastric biopsy samples were obtained endoscopically from patients with H. pylori-positive gastritis. Autoradiographic examination revealed that the [3H]lansoprazole binding site was present in the cytoplasmic granules of infiltrated neutrophils. Tissue MPO content was significantly decreased after treatment with lansoprazole. These data suggest that lansoprazole binds directly to neutrophils, subsequently inhibiting neutrophil accumulation and release of toxic metabolites.

2-Pyridinylmethylsulfinylbenzimidazoles↗
PubMed1995

Effects of ventilation and pleural effusion on measurements of airway thermal volume and blood flow in dog lungs.

We studied the effects of ventilation and pleural effusion on measurements of airway thermal volume (ATV) and pulmonary blood flow (PBF) by using the airway gas thermometry method of V. B. Serikov, M. S. Rumm, K. Kambara, M. I. Bootomo, A. R. Osmack, and N. C. Staub (J. Appl. Physiol. 72: 944-953, 1992) in 39 anesthetized dogs with or without lung edema or pleural effusion. To examine the differential effects of increased-pressure and increased-permeability lung edema on accuracy and sensitivity of ATV and PBF, two models of lung edema were induced by intravenous infusion of a Dextran 70 solution and alloxan monohydrate, respectively. Dogs were hyperventilated for 3 min by using a wide range of minute ventilation (VE) to produce two steady-state conditions of airway temperature. Higher levels of VE increased an estimated amount of ATV. The ATV produced by hyperventilation at VE values of 559, 158, and 72 ml.min-1.kg-1 was consistent with the gravimetric total lung mass, the blood-free wet lung weight, and the extravascular lung water volume, respectively. The coefficient of lung thermal conductivity, a practical index of the rate of heat conduction through tissue from lung vessels, was related to the ratio of the decrease in expired air temperature to VE, and estimated PBF was consistent with the thermodilution cardiac output. Pleural effusion had little effect on measurements of ATV and PBF. However, ATV and PBF showed increased variation in dogs with dextran-induced lung edema.

Air Pressure↗
PubMed1995

Interleukin 7 is produced by human intestinal epithelial cells and regulates the proliferation of intestinal mucosal lymphocytes.

The interaction of mucosal lymphocytes and intestinal epithelial cells is thought to be important in regulating immune response in the intestinal mucosa, but conclusive evidence is limited. Here we demonstrate the expression of IL-7 mRNA in human intestinal mucosa by combined reverse transcription PCR and Southern blot hybridization. Immunohistochemistry and in situ hybridization confirm the presence of IL-7 in intestinal epithelial cells, especially in epithelial goblet cells. Moreover, IL-7 receptor expression in mucosal lymphocytes is demonstrated by immunohistochemistry and in situ hybridization, as well as by Southern blot and flow cytometric analysis of freshly isolated lamina propria lymphocytes. In contrast, IL-7 receptor could not be detected in the cell surface of freshly isolated PBLs. The functional activity of IL-7 receptor is demonstrated by the utility of recombinant IL-7 to stimulate the growth of lamina propria lymphocytes, and conversely inhibit CD3-dependent proliferation of these cells. In contrast, IL-7 caused no significant increase in DNA synthesis and cell numbers when added to PBLs. These findings suggest that human intestinal epithelial cells and epithelial goblet cells produce IL-7, and locally produced IL-7 may serve as a potent regulatory factor for intestinal mucosal lymphocytes.

Adult↗
PubMed1995

Helicobacter pylori urease inhibition by rabeprazole, a proton pump inhibitor.

We investigated the inhibitory effects of four gastric proton pump inhibitors (PPIs): rabeprazole, a novel benzimidazole PPI, omeprazole, lansoprazole and AG-2000, on the urease activity of Helicobacter pylori (H. pylori). Their 50% inhibitory concentrations (I50s) were found to be 0.29, 5.4, 9.3 and 0.3 microM respectively. Rabeprazole and omeprazole were also potent inhibitors of Jack bean and Proteus mirabilis cellular ureases. The thioether derivative of rabeprazole, one of its metabolites, had no inhibitory effect on H. pylori urease, despite being reported as a more potent inhibitor of H. pylori growth than rabeprazole. The inhibitory effect of rabeprazole was prevented completely and reversed considerably by the addition of sulfhydryl compounds, such as beta-mercaptoethanol, glutathione and dithiothreitol. Moreover, the addition of beta-mercaptoethanol recovered the urease activity inhibited by rabeprazole. From these results, we expected that rabeprazole inhibited H. pylori urease activity by forming disulfide bonds between it and the active site of the enzyme.

2-Pyridinylmethylsulfinylbenzimidazoles↗