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Biomedical subjects

R Kodama

Publications and source records attributed to R Kodama.

At least 19 recordsLinked to original sources

In vitro culture system for iris-pigmented epithelial cells for molecular analysis of transdifferentiation.

Dissociated cells of the iris-pigmented epithelium (IPE) from a 1-day-old chick grew in monolayer culture and stably maintained their differentiated state when cultured with standard culture medium. After replacement of the control medium by EdFPH medium, which is effective in inducing dedifferentiation of retinal pigmented epithelium (RPE) cells, all cells rapidly lost pigment granules, proliferated intensively, and dedifferentiated. By further addition of ascorbic acid, dedifferentiated cells accumulated and formed a large number of lentoids. This system provides a useful opportunity for analyzing cellular and molecular mechanism involved in each step of transdifferentiation. Furthermore, Northern blot data indicates that the up-regulation of pax-6 gene could be an important event during lens regeneration as well as during normal lens development.

Animals

Diversity in protein synthesis and viability of Helicobacter pylori coccoid forms in response to various stimuli.

The viability of the coccoid forms of Helicobacter pylori was evaluated by assessing protein synthesis. Metabolic labeling studies showed the synthesis of proteins and the specific protein profiles of H. pylori coccoids produced under various conditions. Harsh conditions such as aerobiosis and starvation (lack of horse serum) in the culture did not affect the synthesis of proteins in the coccoids. Lowering of the pH to that of gastric secretions induced expression of several proteins in the coccoids. However, the coccoids produced under prolonged microaerobic conditions exhibited a profile of acid stress-induced protein expression different from that induced by aerobiosis or starvation. Our data suggest that coccoid H. pylori exhibits diversity in viability following exposure to different stresses and that the response to acid stress of coccoid H. pylori could be involved in infection of the host stomach.

Acids

[Biocompatibility of polyvinylalcohol gel as a vitreous substitute].

Polyvinylacohol (PVA) gel cross linked by gamma irradiation was assessed as a possible vitreous substitute. In our previous, reports, elevation of intraocular pressure and inflammatory changes in the vitreous cavity after operation were observed in some of the experimental animals. Four types of PVA gel (sol) were produced by a newly refined method. We improved the gel systhesis process to remove the monomer and contamination. Colored rabbits were used for this experiment. Vitreous replacement was performed after vitrectomy. This was followed clinically by ophthalmoscopy, tonometry, fundus photography, electroretinogram, chemotaxis, and laser-flare-cell-metering. Histopathological examination by light microscopy was performed after 3 months. PVA gel has good biocompatibility in the vitreous cavity, and gel with a network similar to that of the vitreous body showed the best biocompatibility. Although it is necessary to investigate the biocompatibility over the long term and to evaluate the tamponade effect, PVA gel will be a bood candidate for a vitreous substitute.

Animals

Toxigenicity of Helicobacter pylori isolates possessing cagA gene and vacuolating cytotoxin.

Inflammatory cells in the gastric epithelium and direct cell damage are important effects of H. pylori infection in causing gastroduodenal diseases. The aim of this study was to determine whether H. Pylori possessing the cagA gene and vacuolating cytotoxin activity (cagA+/tox+) has these toxigenicities. The detection of cagA was performed by the polymerase chain reaction method. Culture supernatants of H. pylori were tested on rabbit gastric epithelial cell culture for intracellular vacuolation. H. pylori isolates were divided into two major types, cagA+/tox+ and cagA-/tox- strains. Ten Japanese monkeys were allocated to two groups, with six animals inoculated with the cagA+/tox+ strain and four animals inoculated with the cagA-/tox- strain. Five other Japanese monkeys served as controls. They were observed endoscopically every week and the severity of gastritis was evaluated in biopsy specimens obtained from the antral mucosa. Histopathological examination of the gastric mucosa revealed a more severe neutrophil infiltration caused by the cagA+/tox+ strain than that caused by the cagA-/tox- strain for 3 months after inoculation. These findings indicate that cagA and vacuolating cytotoxin may be important factors in causing the severe damage of the human gastric mucosa produced by H. pylori.

Animals

Helicobacter pylori infection accelerates human gastric mucosal cell proliferation.

Helicobacter pylori causes chronic atrophic gastritis and intestinal type gastric cancer arises against a background of atrophic gastritis. Increased proliferation of epithelial cells is an important indicator of increased risk for gastric adenocarcinoma. We investigated gastric mucosal cell proliferation in H. pylori-associated gastritis and the effect of eradication therapy on this proliferation in 45 patients endoscopically diagnosed (31 with persistent eradication and 14 in whom H. pylori) recurred. H. pylori status was determined by culture and histology in biopsied specimens from the gastric antrum and corpus. Eradication of the infection was defined as reversal to negative on both tests. In vitro Ki-67 immunostaining of endoscopic biopsy specimens was used to measure mucosal cell proliferation in H. pylori-associated gastritis before and after therapy. The proliferative zone was defined as the distance of Ki-67-positive gastric epithelial cells between the highest and the lowest cells. In patients in whom H. pylori was eradicated, cell proliferation in both the antral and corpus mucosa had decreased 4 weeks after completion of the eradication therapy (P < 0.01, P < 0.001), and 6 months later, it had markedly decreased (P < 0.05, P < 0.05) and returned to normal. In patients in whom H. pylori recurred, only antral epithelial cell proliferation was reduced 4 weeks after eradication therapy, but when H. pylori recurred, determined by culture and histology, cell proliferation level was the same as that before eradication. These results suggest that H. pylori infection accelerates cell proliferation in gastric mucosa and may play a causal role in the chain of events leading to gastric carcinoma.

Amoxicillin

Long-term sequelae of experimental gastritis with Helicobacter pylori: a 5-year follow-up study.

To investigate the long-term effects of Helicobacter pylori gastritis on the gastric mucosa, 13 wild Japanese monkeys (six H. pylori-infected and seven controls) were monitored for 5 years. Colonization with H. pylori, the presence of macroscopic and histological gastritis, pyloric glandular height, and epithelial cell kinetics were investigated, using Ki-67 immunostaining in the gastric mucosa. In the infected group, persistent colonization with H. pylori was demonstrated by culture and histopathologic examination. In this group, the gastritis scores were significantly higher than in controls. Simultaneously, a significant decrease in the height of antral glands and a significant increase in the length of Ki-67-positive cells between the highest and lowest cells were also demonstrated in the infected animals. These experimental results directly demonstrate the effect of H. pylori infection on the gastric mucosa and may explain the potential mechanism for its causal role in the chain of events leading to gastric carcinoma.

Animals

Helicobacter pylori infection delays the healing of acetic acid-induced gastric ulcer in Japanese monkeys.

To clarify the relationship between Helicobacter pylori and the healing of gastric ulcers, we investigated the healing of acetic acid-induced gastric ulcers in the antral mucosa of Japanese monkeys (n = 5) infected with H. pylori and in control monkeys without H. pylori infection (n = 6). Using H. pylori-infected Japanese monkeys as an experimental model, gastric ulcers were induced endoscopically with acetic acid. Healing of ulcers and factors that influenced healing were studied. Continuous colonization with H. pylori was confirmed in the infected group throughout the observation period; no H. pylori were isolated from the gastric mucosa of the control group. White scarring was not observed in any infected monkeys 4 weeks after ulcer formation, but was observed in one (20%) of five monkeys at 6 weeks and in all five monkeys eight weeks after ulcer formation. In the control group, white scarring was observed in one (16.7%) of six monkeys at 4 weeks and in six monkeys at 6 (P < 0.01 vs infected group) and 8 weeks. The ammonia concentration of the gastric secretions and the grade of inflammation were significantly increased in the H. pylori-infected group compared with the control group (P < 0.01 and P < 0.001, respectively). The volume of intracellular PAS-positive substance was decreased (P < 0.025-0.01) at the ulcer margin in the infected group compared with the ulcer margin in the control group. The proliferation of gastric epithelial cells was markedly accelerated at the ulcer margin in the infected group compared with the ulcer margin in control group (P < 0.025-0.01). Our results strongly suggest that H. pylori infection delays the healing of gastric ulcers.

Acetic Acid

Sofalcone for treatment of Helicobacter pylori infection.

It is difficult to regard sofalcone as a single drug for the treatment of Helicobacter pylori infection. However, sofalcone exerts multiple effects against H. pylori: it has antibacterial activity, induces morphological changes, inhibits adhesion to gastric mucin and inhibits lipolytic activity. The safety profile of sofalcone even on long-term administration is well established. Therefore, it may be possible to establish a new triple therapy for H. pylori infection using sofalcone combined with antibacterial drugs and proton pump inhibitors.

Anti-Bacterial Agents

Atopic dermatitis successfully treated by eradication of Helicobacter pylori.

A relationship between allergic diseases and Helicobacter pylori infection has recently been noted. We report a case of atopic dermatitis and H. pylori infection in a 14-year-old girl. She had had widespread diffuse skin erythema with erosions and pigmentation since the age of 3 years. Endoscopically, there was chronic antral gastritis with H. pylori infection and histological eosinophilic infiltration. A high titer of H. pylori-specific IgG was present in serum. She was treated with a proton pump inhibitor (lansoprazole 60 mg), an antibiotic (clarithromycin 800 mg), and plaunotol (a mucosal protective agent, 480 mg) for 2 weeks to eliminate the infection. After 10 days of treatment, erythema and itching were more widespread and vesicle formation was seen on the foot. Generalized skin lesions abated a few days later. After eradication of the bacterium by the treatment, eosinophils decreased from 38.8% to 19.0%, and the clinical signs of atopic dermatitis almost disappeared. Serum gastrin level and the pepsinogen I/II ratio were normalized and histological findings of gastric mucosa showed improvement. H. pylori-specific IgE antibody, analyzed by the Western blot method, gradually decreased with the eradication treatment.

2-Pyridinylmethylsulfinylbenzimidazoles

Isolation of a novel chick homolog of Serrate and its coexpression with C-Notch-1 in chick development.

Intercellular signaling mediated by the transmembrane proteins, Notch as receptor and its ligands, Delta and Serrate, plays essential roles in the developmental fate decision of many cell types in Drosophila. The Notch genes are highly conserved both in invertebrates and in vertebrates, suggesting that Notch pathway regulates cell fate decisions during vertebrates development. Notch, Delta and Serrate homologs in chicken have been cloned (Henrique et al., Nature 375: 787-790, 1995; Myat et al., Dev. Biol. 174: 233-247, 1996). We isolated a novel chick homolog of Drosophila Serrate, named C-Serrate-2, and examined its expression patterns during the early chick development using whole-mount in situ hybridization. C-Serrate-2 transcripts were detected in several tissues including the forebrain, the myotome and the apical ectodermal ridge (AER) of the limb bud of a 4-day-old chick embryo. In most of the regions where C-Serrate-2 was expressed, C-Notch-1 was also expressed. Our observations suggest that Serrate-2-Notch-1 signaling plays a role in a variety of morphogeneses during the chick development.

Amino Acid Sequence

From lens regeneration in the newt to in-vitro transdifferentiation of vertebrate pigmented epithelial cells.

Through studies to clarify the cellular origin of lens regeneration in the newt, the pigmented epithelial cells of the iris and the retina of many vertebrate species have been shown to possess a dormant potency to transdifferentiate into the lens. The method of in-vitro culture of pigmented epithelial cells has been optimized to enable detailed studies of the transdifferentiation process by molecular techniques. Growth factors and extracellular matrix components are found to be important in the control of the transdifferentiation process. New systems for in-vitro culture are introduced, while prospects for renewed in-vivo studies using newts are given.

Amphibians

[Preparation of specific antiserum of secretin and its use in time-resolved fluoroimmunoassay].

We have prepared a specific anti-secretin antiserum and developed a sensitive time-relevant fluoroimmunoassay (TR-FIA) for secretin. An anti-secretin antiserum was produced in rabbit by immunization with synthetic secretin C-terminal 8 residue conjugate to chicken serum albumin as antigen. The TR-FIA was based on the competition between free secretin and biotinylated secretin as a tracer for rabbit anti-secretin antibodies on a goat anti-rabbit IgG antibody coated microtiter plate. After separation of bound and free fractions, a solution of europium (III) ion chelate labeled streptavidin (Eu-SA) was added to the biotinylated secretin bound on the microtiter plate. The microtiter plate was washed and then Eu-SA activity was measured by time-resolved fluorometry. In this TR-FIA, the measurable range of secretin was from 2.5 to 100 pg/assay and the specificity was less than 0.01% against other peptide hormones. For the measurement of secretin in the human plasma, a sample was required for the pretreatment in order to separate from interfering substances. The mean recovery of secretin using commercially reverse phase column was 80.2% (n = 11). The plasma secretin concentrations of normal subjects and those with various diseases could be measured by this proposed TR-FIA.

Animals