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Biomedical subjects

S Kamiya

Publications and source records attributed to S Kamiya.

At least 73 records · Page 4Linked to original sources

Characteristics of Helicobacter pylori strains isolated from patients with different gastric diseases.

The pathogenesis of chronic gastritis, gastric ulcer, duodenal ulcer, gastric cancer, and remnant gastritis is considered to be related to infection by Helicobacter pylori. The purpose of this study was to investigate the characteristics of H. pylori strains isolated from patients with various gastric diseases. Fifty strains of H. pylori were clinically isolated at Keio University Hospital and were assessed for (i) vacuolating cytotoxin (VT) production, by cytotoxic assay using rabbit kidney (RK) 13 cells; (ii) the expression of cytotoxin associated gene (cagA), by polymerase chain reaction; and (iii) motility on a semi-solid plate. Positivity for VT production by H. pylori strains isolated from patients with chronic gastritis, gastric ulcer, duodenal ulcer, gastric cancer, and remnant gastritis, was 100%, 89%, 92%, 100%, and 60%, respectively. There was no significant difference in the incidence of positivity for cagA among the strains isolated from different gastric diseases. Swarming distances, which indicate motility of H. pylori strains, were 12.7, 13.2, 12.8, 16.6 and 9.2mm in strains from chronic gastritis, gastric ulcer, duodenal ulcer, gastric cancer, and remnant gastritis, respectively, with H. pylori isolated from remnant gastritis showing significantly less motility than H. pylori isolates from other diseases. There were no significant differences in VT production and cagA positivity among the H. pylori strains.

Animals↗

[Analysis of air pollution and prevalence rate of allergic diseases among elementary school children in Kawaguchi and Hatogaya city].

Kawaguchi and Hatogaya City are located on the northern edge of Tokyo. We analysed between air pollution and prevalence rate of allergic diseases among elementary school children in this area. A prevalence rate of allergic diseases in 1996 May and June was as follows; bronchial asthma 13.5%, atopic dermatitis 24.5%, allergic rhinitis and/or conjunctivitis 22.8%, urticaria 12.4%, food allergy 7.8% and drug allergy 2.2%, respectively. Air pollution of this area was analysed to check the levels of nitrogen dioxide (NO2), sulfur dioxide (SO2) and suspended particulate matters (SPM). NO2 pollution was relatively high in urban area, and SPM pollution was especially high around the highways. SO2 pollution was lower than the environmental standard. No relationship was found between the prevalence rate of bronchial asthma, atopic dermatitis, allergic rhinitis and/or conjunctivitis and air pollution, but it was found that these diseases are slightly related to population density (p < 0.1, p < 0.01, p < 0.1, respectively).

Air Pollutants↗

Differential involvement of synaptic vesicle and presynaptic plasma membrane proteins in Alzheimer's disease.

Alzheimer's disease (AD) is characterized by progressive cognitive decline. Recent studies have shown that synaptic loss in the cortex is the major correlate of cognitive decline in AD. In the present study we assessed synaptic proteins such as synaptobrevin, synaptophysin, synaptotagmin, synaptosomal-associated protein 25 (SNAP-25), and syntaxin1/HPC-1 in control and AD brains to determine whether synaptic proteins are equally or differentially affected in AD. Western analysis showed that in AD levels of synaptobrevin and synaptophysin were decreased by some 30% from amounts in controls, while those of synaptotagmin, SNAP-25, and syntaxin 1/HPC-1 were decreased by only about 10%. As synaptobrevin and synaptophysin are localized mainly in transmitter-containing synaptic vesicles while SNAP-25 and syntaxin 1/HPC-1 are found in presynaptic plasma membranes, these results suggest differential involvement of synaptic components in AD.

Alzheimer Disease↗

Flow cytometric analysis for adhesion of Vibrio cholerae to human intestinal epithelial cell.

The adhesion of Vibrio cholerae O1 strains to human intestinal epithelial cell, Intestine 407, was analyzed by flow cytometer. According to positive percentages of Intestine 407 cells adhered by V. cholerae, two groups of V. cholerae strains were classified as follows: more adhesive (more than 50%), less adhesive (less than 50%) strains. In addition, the fluorescence intensity after attachment of V. cholerae was directly correlated to the number of the microorganisms. It was concluded that flow cytometry is a useful and objective method for analyzing adhesion of V. cholerae to cultured cells.

Bacterial Adhesion↗

Cytotoxicity and motility of Helicobacter pylori.

To clarify the relationship between interleukin-8 (IL-8) production and virulent factors, we examined the motility and cytotoxicity of H. pylori, suggested to be a major cause of chronic gastritis and peptic ulcers. Our results demonstrated that among cytotoxic strains of H. pylori, high-motility strains induced more IL-8 than low-motility strains. There was no correlation between cytotoxicity and motility of H. pylori. Four restriction fragment length polymorphism (RFLP) patterns were observed in the flaA PCR products. There was no correlation between flaA RFLP and motility. In conclusion, our findings suggest that both cytotoxicity and motility are virulent factors in the pathogenesis of gastric mucosal injury.

Animals↗

Effect of sofalcone on adherence, production of vacuolating toxin, and induction of interleukin-8 secretion by Helicobacter pylori.

We examined the effect of sofalcone, a mucosal protective agent that has been reported to inhibit growth of Helicobacter pylori, on adherence, production of vacuolating toxin (VT), and induction of interleukin-8 (IL-8) secretion by H. pylori. Mixing VT with various concentrations (1.5-100 micrograms/ml) of sofalcone resulted in a 50% decrease in the VT titer. When toxigenic H. pylori strains were incubated in the presence of sofalcone (20 and 40 micrograms/ml), although bacterial growth was not inhibited significantly there was significant inhibition of VT production. Notable inhibition of IL-8 secretion by human gastric cancer cells (MKN 45) was detected in the presence of sofalcone (20-100 micrograms/ml) after co-incubation with H. pylori strains. In flow cytometric analysis, adherence of H. pylori strains to MKN 45 cells was significantly inhibited by treatment with sofalcone (20-60 micrograms/ml), indicating at least one reason for the inhibition of IL-8 secretion. These results show that sofalcone is an effective mucosal protective agent that inhibits both production of VT and induction of IL-8 secretion.

Anti-Ulcer Agents↗

Production and characterisation of monoclonal antibodies to heat-shock protein 60 of Helicobacter pylori.

Two monoclonal antibodies (MAbs), designated as H9 (IgG2a) and H20 (IgM), directed against heat-shock protein 60 (HSP60) of Helicobacter pylori strain TK1029 were established. Affinity-purified antigens cross-reacted in immunoblots with MAb H9 and MAb H20 respectively. These antigens also reacted with the 3C8 MAb previously established in this laboratory, which recognised Yersinia enterocolitica HSP60. By amino-acid sequence analysis, the N-terminal amino-acid sequence of the protein recognised by both H9 and H20 MAbs was confirmed as the amino-acid sequence of H. pylori HSP60 reported previously. Both MAbs reacted with nine strains of H. pylori in enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. In addition, MAb H9 reacted with extracts of other bacteria including H. mustelae, Pseudomonas aeruginosa, Vibrio cholerae, Serratia marcescens, Proteus mirabilis, Escherichia coli and Shigella sonnei. In contrast, MAb H20 reacted only with strains H. pylori. These results suggest that both the species-specific epitope recognised by MAb H20 and the common epitope recognised by MAb H9 exist on HSP60 of the bacterial cell. Both MAbs also reacted with the 60-kDa protein in the lysate of human gastric carcinoma (MKN45) cells. It was shown by immunohistochemical staining that gastric epithelial cells of four out of six biopsy specimens examined stained positively with MAb H20. These results suggest that there is a common epitope in H. pylori HSP60 and human gastric epithelial cells.

Amino Acid Sequence↗

Heat-shock protein 60 homologue of Helicobacter pylori is associated with adhesion of H. pylori to human gastric epithelial cells.

A previous study reported a relationship between the expression of heat-shock protein 60 (HSP60) by Helicobacter pylori and its adhesion to human gastric carcinoma (MKN45) cells. To examine whether the HSP60 homologue of H. pylori is associated with the adhesion of H. pylori to human gastric epithelial cells, an inhibition assay of adhesion of H. pylori to MKN45 cells was performed by flow cytometric analysis with monoclonal antibody (MAb) designated as H20 recognising HSP60 of H. pylori. The rate of adhesion of H. pylori pretreated with MAbH20 to MKN45 cells was lower than that of untreated H. pylori. Primary human gastric epithelial cells from a patient with gastric cancer were also prepared for comparison in the inhibition assay with MAbH20. H. pylori adhered to the primary human gastric epithelial cells, and this adhesion was significantly inhibited by MAbH20. These results suggest that the H. pylori HSP60 homologue recognised by MAbH20 might be associated with the adhesion of H. pylori to primary human gastric epithelial cells as well as to cultured gastric cancer cells.

Adhesins, Bacterial↗

Studies on the relationship between adhesive activity and haemagglutination by Helicobacter pylori.

The adhesion of Helicobacter pylori to gastric carcinoma cells (MKN45, KatoIII and MKN28) and Intestine-407 cells was tested by flow cytometric analysis. The mean adhesion rates of H. pylori strains to MKN45, KatoIII and Intestine-407 cells were 90.5, 42.7 and 15.1%, respectively. There was no statistical correlation between the adhesion rates to MKN45 cells and haemagglutination (HA) activity of H. pylori strains, although H. pylori strains with high HA activity with human type O erythrocytes tended to adhere effectively to MKN45 cells. No correlation between adhesion and production of vacuolating toxin was observed.

Animals↗

Growth inhibition of Helicobacter pylori by monoclonal antibody to heat-shock protein 60.

The H20mAb recognizing the 60-kilodalton protein, which existed in the outer membrane and was induced by heat shock at 42 C, was established. The molecule recognized with the mAb was a heat-shock protein 60 (HSP60) of Helicobacter pylori. To understand the role of HSP60 on the cell surface of H. pylori, whether or not H20mAb affects the growth of H. pylori was investigated. When bacteria were cultured with H20mAb, growth was markedly inhibited after 24 hr, although an initial 5 hr-incubation with the mAb induced no significant inhibition of H. pylori growth. The 24- and 48 hr growth of the bacteria after washing to remove the mAb at 5 hr was also inhibited though the inhibitory effect was not strong. In electron microscopical analysis, the spots with high electron density in the cytoplasm of the bacteria treated with H20mAb were increased, depending on the length of incubation time from 5 to 24 hr. After 24 hr treatment with H20mAb, bacterial destruction was also observed, indicating bactericidal activity by H20mAb. These results suggest that the HSP60 on the cell surface of H. pylori might have an essential role in the growth of the bacteria.

Amino Acid Sequence↗

Analysis of interleukin-8 secretion induced by Helicobacter pylori from the gastric epithelial cell line MKN45: a mechanism independent of the intensity of cytotoxicity.

Interleukin (IL)-8, a potent chemoattractant and activator of neutrophils, has been implicated to have a major role in the pathogenesis of gastric mucosal injury by Helicobacter pylori infection. We examined the relationship between cytotoxicity and IL-8 secretion induced by H. pylori. Furthermore, whether the vacuolating cytotoxin of H. pylori mediates IL-8 secretion from gastric epithelial cell lines was examined. Among the inflammatory cytokines, messages for IL-6, IL-8 and transforming growth factor-beta 1 were produced by gastric cancer (MKN45) cells in response to exposure to the cytotoxic strain of H. pylori. MKN45 incubated with the viable cytotoxic strain of H. pylori secreted IL-8. In contrast, the supernatant of neither the cytotoxic nor the non-cytotoxic strain induced IL-8 secretion. There was no correlation between IL-8 secretion and the intensity of cytotoxicity. In conclusion, these findings suggest that IL-8 secretion from MKN45 induced by H. pylori is mediated by factors other than cytotoxicity.

Bacterial Proteins↗

[Trial for isolation of Helicobacter pylori form fecal specimen by immunomagnetic beads separation].

In order to clarify the possibility of feco-oral transmission of Helicobacter pylori, we have employed an immunomagnetic beads (IMB) separation technique and tried to isolate H. pylori from fecal samples. Microaerophilic cultivation and PCR of H. pylori were positive when H. pylori (10(3) cfu) was suspended in Hanks' balanced salt solution (HBSS) for the IMB separation technique. In contrast, when a lower number of H. pylori (10(2) cfu) was suspended in HBSS, 2nd PCR was positive but cultivation was negative. After IMB separation technique from fecal sample containing H. pylori (2 x 10(4) cfu), 20 cfu of H. pylori was recovered by culture. In this case, 1st PCR was negative but 2nd PCR was positive. From a fecal sample of a patient infected with H. pylori, H. pylori was detected only by combination of IMB separation and PCR, although H. pylori was not recovered by culture. These results indicate that IMB separation technique followed by cultivation or PCR would be useful for estimation of the presence of H. pylori fecal samples.

Enzyme-Linked Immunosorbent Assay↗

The Yersinia enterocolitica GsrA stress protein, involved in intracellular survival, is induced by macrophage phagocytosis.

The Yersinia enterocolitica gsrA gene is a stress protein gene which was originally identified as essential for protecting cells under both extracellular environmental stress and intracellular stress in macrophages due to phagocytosis. The gsrA gene was shown to be a member of the htrA class of genes and to possess a sequence homologous to that of the promoter recognized by a stress-induced sigma factor, sigmaE. In order to study the induction of the potentially sigmaE-controlled gsrA gene in Y. enterocolitica after phagocytosis by macrophages, we identified GsrA by overproducing the protein using a T7 promoter-gsrA fusion. We found that it is translated as an unstable 49,500-Da protein which is processed by removal of an amino acid fragment consisting of 27 residues, resulting in a stable 46,800-Da protein. By radiolabeling proteins specific to bacteria in the J774-1 macrophage-like cell line, we found that the production of GsrA protein is indeed enhanced in bacterial cells growing within macrophage phagosomes. Transcriptional activation of the gsrA gene was determined by using the gsrA promoter-lacZ fusion system. This work provides the first piece of evidence that the sigmaE regulon responds to the stressful environment found in macrophages.

Animals↗

Prevention of Helicobacter pylori infection by lactobacilli in a gnotobiotic murine model.

BACKGROUND: Helicobacter pylori is a bacterium which causes gastric inflammatory diseases. Oral inoculation of H pylori usually results in only a temporary colonisation without a successful infection in the stomach of conventional mice in which lactobacilli are the predominant indigenous bacteria. AIM: To determine whether lactobacilli exert an inhibitory effect on colonisation by H pylori in the stomach. METHODS: The effects of H pylori on attachment to murine and human gastric epithelial cells and the H pylori mediated release of interleukin-8 (IL-8) by these cells were examined in vitro. Lactobacillus salivarius infected gnotobiotic BALB/c mice and control germ free mice were inoculated orally with H pylori to examine whether L salivarius can inhibit colonisation by H pylori. RESULTS: L salivarius inhibited both the attachment and IL-8 release in vitro. H pylori could not colonise the stomach of L salivarius infected gnotobiotic BALB/c mice, but colonised in large numbers and subsequently caused active gastritis in germ free mice. In addition, L salivarius given after H pylori implantation could eliminate colonisation by H pylori. CONCLUSION: These findings suggest the possibility of lactobacilli being used as probiotic agents against H pylori.

Animals↗

Flagellin gene diversity among Helicobacter pylori strains and IL-8 secretion from gastric epithelial cells.

BACKGROUND: To clarify the pathological functions of the virulence factors of Helicobacter pylori, a comparative analysis was carried out on the relationship between motility, flagellar gene polymorphism, vacuolating cytotoxin (VT) production and interleukin-8 (IL-8) induction. METHODS: Twenty-five strains were examined for restriction fragment length polymorphism (RFLP) of the flagellin gene. Motility was measured using semisolid agar plates. Cytotoxicity was assayed using RK-13 cells. IL-8 secretion was assessed by the enzyme-linked immunosorbent assay (ELISA) methods. RESULTS: H. pylori was classified into four groups according to their flagellar RFLP. No differences were noted in motility or VT production among the four groups, but a significant difference was noted in IL-8 induction. In addition, highly motile strains produced more IL-8. CONCLUSION: This flagellar genetic polymorphism may be associated with IL-8 induction.

Animals↗

[The influence of an emotion on the retention of episodic scenes].

This study investigated the relationship between affective properties of stimuli and their retention. Thirty sentences describing various scenes were chosen as stimuli to elicit affects which are experienced in daily life. Ninety undergraduates rated each sentence on the following eight dimensions: anxiety, hostility, boredom, liveliness, calmness, friendliness, concentration, and surprise. One week after participating in the scene rating task, participants were given an incidental free recall test, in which they were instructed to recall a word or phrase describing each scene. Results indicated that memory for affectively pleasant scenes was superior to that for unpleasant scenes. However, a discriminant analysis on the kind of emotion activated indicated that the relaxant-tensive dimension determined the retention of the stimulus sentences more than the dimension of pleasantness.

Adult↗

The effects of intravenously infused catecholamines on hepatic blood flow in conscious dogs with experimental obstructive jaundice.

This study was conducted to examine how the effects of dopamine and dobutamine on hepatic blood flow were influenced by obstructive jaundice in a conscious canine model. Prior to biliary obstruction, portal venous blood flow (PVF) increased in response to the infusion of either dopamine or dobutamine: dopamine infused at 8 micrograms/kg per min produced an increase of 19 +/- 0% in PVF, while dobutamine infused at 16 micrograms/kg per min produced an increase of 30 +/- 2%. Although hepatic arterial blood flow (HAF) decreased dose-dependently in response to the infusion of dopamine, no significant change was observed in HAF in response to any dose of dobutamine. Obstructive jaundice attenuated or completely abolished the PVF-increasing effect of dopamine, whereas it did not significantly alter the effect of dobutamine on hepatic blood flow. In dogs with obstructive jaundice, dopamine at 16 micrograms/kg per min produced a decrease of 17 +/- 3% in PVF. These findings suggest that dobutamine is more effective than dopamine for increasing hepatic blood flow in patients with obstructive jaundice.

Animals↗