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Biomedical subjects

M Tsuji

Publications and source records attributed to M Tsuji.

At least 253 records · Page 14Linked to original sources

Assignment of the bacterial agent of urinary calculus in young rats by the comparative sequence analysis of the 16S rRNA genes of corynebacteria.

Comparative 16S rRNA gene sequencing was used to assign four isolates of spontaneous urinary calculus in young laboratory rats. The phylogenetic relationships among the rat isolates and selected species of corynebacteria were also inferred. Based on the homology and evolutionary distance analysis, the 16S rRNA genes of the rat isolates were almost identical with that of Corynebacterium renale ATCC 19412. Also the results of the phylogenetic analysis showed a close relationship among the isolates and C. renale, but they were clearly different from C. pilosum, C. cystitidis, C. kutscheri and Rhodococcus equi. The results of the present study and previously published biochemical data demonstrate that the organism involving urinary infections in young rats is identified to be C. renale.

Animals↗

Morphological, serological and antigenic characteristics, and protein profile of newly isolated Japanese bovine Babesia parasite with particular reference to those of B. ovata.

An intraerythrocytic large protozoan, tentatively designated Babesia sp. 1, was recently isolated from cattle in Hokkaido Prefecture, Japan. This parasite closely resembled B. ovata in shape of piroplasms, but was distinguishable by other morphological, immunological, and biochemical characters. The paired pyriform piroplasm of B. sp. 1 was larger than that of B. ovata. The results from serological and antigenic examination by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis showed that there were cross- but distinguishable-reaction between B. sp. 1 and B. ovata. Protein profiles of both Babesia parasites piroplasms analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) were apparently different from each other. Several major proteins revealed by 2D-PAGE and the immunodominant proteins resolved by Western blot analysis (40 kDa for B. sp. 1 and 29 kDa for B. ovata) were unique to each parasite. The results of the present study indicate the possibility that B. sp. 1 is a species different from B. ovata.

Animals↗

Isolation, characterization and biological activities of novel triprenyl phenols as pancreatic cholesterol esterase inhibitors produced by Stachybotrys sp. F-1839.

Ten triprenyl phenol metabolites were isolated as inhibitors of pancreatic cholesterol esterase from cultures of Stachybotrys sp. F-1839 by solvent extraction and column chromatographies. Combination of spectroscopic analyses revealed that two of these compounds are K-76 (1) and stachybotrydial (2), and that the remaining eight are new congeners (designated F1839-A (3), -B (4), -C (5), -D (6), -E (7), -F (8), -I (9) and -J (10). These compounds inhibited pancreatic cholesterol esterase by 50% at 6 x 10(-5) to 1.1 x 10(-1) M. Inhibition of the enzyme by compound 2, the most potent one among these compounds, was time-dependent and irreversible. When administered to normal rats, 2, at a single oral dose of 100 mg/kg, reduced [14C]cholesterol absorption by 50-60%. In cholesterol-fed mice, dietary supplementation of 2 (0.1%) for 14 days resulted in a 20% reduction in serum total cholesterol level without causing significant change in the high density lipoprotein cholesterol level.

Animals↗

[A case of gastric cancer responding well to MTX/5-FU (methotrexate/5-fluorouracil) and PMUE (CDDP, MMC, UFT, etoposide) therapies upon lymphnode recurrence].

We encountered a case of non-curatively resected gastric cancer (p1, n4) who responded well to sequential MTX/5-FU therapy and PMUE therapy. A 63-year-old man was admitted to our hospital with complaints of nausea and vomiting. Upper gastrointestinal examination and CT scan revealed Borrmann type 3 gastric cancer with pyloric stenosis and multiple paraaortic lymphnodal metastasis. The patient underwent palliative gastrectomy for extensive gastric cancer (H0, P1, N4, T3, Stage IV b). Histological examination of the resected stomach revealed poorly differentiated adenocarcinoma with paraaortic lymphnodes metastasis (n4) and peritoneal dissemination (p1). Chemotherapy with sequential MTX/5-FU was given 13 times. Ten months after the operation, abdominal pain and back pain required analgesic treatment. Abdominal CT scan revealed increased size of paraaortic lymphnodes, suggesting recurrence. Sequential MTX/5-FU therapy was switched by PMUE therapy. Lymphnode size became smaller and habitual analgesics could be discontinued. Since then he was given MTX/5-FU and PMUE therapies alternately on an ambulant basis. The patient resumed his daily activities at 2 years and 8 months after the operation.

Adenocarcinoma↗

In vivo expression of monokine and inducible nitric oxide synthase in experimentally induced pulmonary granulomatous inflammation. Evidence for sequential production of interleukin-1, inducible nitric oxide synthase, and tumor necrosis factor.

The present study examined the temporal pattern and localization of interleukin-1, tumor necrosis factor-alpha, and inducible nitric oxide synthase expression in lung tissue undergoing foreign body granuloma formation. Pulmonary granulomas were induced by the intratracheal injection of dextran beads into genetically high granuloma responder, carrying Bcgs (BALB/c), and low responder, carrying Bcgr (C3H/HeJ and DBA/2), mice. There was a pattern of sequential expression of these molecules in BALB/c mice. Thus, interleukin-1 alpha and inducible nitric oxide synthase were induced mostly in the cells accumulated around the beads and also in some bronchiolar epithelial cells during the early phase (1 to 3 days), whereas tumor necrosis factor-alpha was induced in the cells around the beads at the later resolution phase (3 to 7 days). By contrast, in low responder mice, an increase in the expression of interleukin-1 alpha and inducible nitric oxide synthase was detected in lung macrophages as well as in alveolar cells and bronchiolar epithelial cells on day 1, but that of tumor necrosis factor-alpha was not detected throughout the period. These results together with our previous findings on cytokine activity in granuloma extract suggest that interleukin-1 and nitric oxide produced by recruited macrophages may take part in the early, macrophage-dependent phase of granuloma formation whereas tumor necrosis factor-alpha may be more crucial as a mediator responsible for the difference in innate resistance or susceptibility to granuloma formation.

Animals↗

Taxonomic status of car bacillus based on the small subunit ribosomal RNA sequences.

In an attempt to identify the taxonomic relationship between CAR bacillus and other bacteria, the SSU rRNA gene sequences of two CAR bacillus strains, CBM and CBR isolated from mice and rats respectively were used in the present studies. The SSU rRNA gene sequences, approximately 1.5 kb in size amplified from genomic DNAs from both strains, were determined and 96.8% homologies were found to exist between them. Those sequences were aligned to most eubacteria with a computer search showing high homology with those of Flavobacter/Flexibacter species especially closed to Fx. sancti and Fv. ferrugineum. Phylogenetic analysis indicated that CAR bacillus belongs to a species close to Fx. sancti and Fv. ferrugineum subdivision.

Animals↗

1 alpha-hydroxyvitamin D3 inhibits type II collagen-induced arthritis in rats.

The effects of 1 alpha-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 on Type II collagen-induced arthritis in rats, an experimental model of rheumatoid arthritis, were examined. Oral administration of 1 alpha-hydroxyvitamin D3 significantly suppressed the incidence of arthritis and inhibited hind paw swelling. The level of anti-Type II collagen antibodies was decreased in the 1 alpha-hydroxyvitamin D3 treated-group. In contrast, 24,25-dihydroxyvitamin D3, indomethacin, and gold had no effect on either the incidence of arthritis or the antibody levels. These findings demonstrate a beneficial effect of 1 alpha-hydroxyvitamin D3 on Type II collagen-induced arthritis in rats and indicate that it has an antirheumatic effect.

Animals↗

Gamma delta T cells contribute to immunity against the liver stages of malaria in alpha beta T-cell-deficient mice.

The functional role of gamma delta T cells (expressing the gamma delta heterodimeric T-cell receptor for antigen) in infectious diseases remains largely unknown. We have therefore attempted to define the possible role of these T cells in the immune response against the various developmental stages of malaria parasites. For this purpose, we monitored the immune response and the development of liver and blood stages of Plasmodium yoelii, a rodent malaria parasite, in immunized and nonimmunized alpha beta T-cell-deficient and gamma delta T-cell-deficient mice. Immunization of alpha beta T-cell-deficient mice with irradiated sporozoites induced an immune response that significantly inhibited the development of the parasite's liver stages. This inhibitory immune response was abolished by an antibody-mediated transient in vivo depletion of gamma delta T cells. Two gamma delta T-cell clones were derived from malaria-immunized alpha beta T-cell-deficient mice. The adoptive transfer of one of these gamma delta T-cell clones to normal mice inhibited the development of liver stages, following sporozoite inoculation. These results provide evidence for gamma delta T-cell-mediated protective immunity against parasites, in the absence of alpha beta T cells. As for the blood phase of the infection, both normal mice and gamma delta T-cell-deficient mice cleared the blood stages of the nonlethal strain of P. yoelii, while alpha beta T-cell-deficient mice failed to control the parasitemia.

Animals↗

Trilineage recovery by combination therapy with recombinant human granulocyte colony-stimulating factor and erythropoietin in patients with aplastic anemia and refractory anemia.

Ten patients with aplastic anemia (AA) and seven patients with refractory anemia (RA) were treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and erythropoietin (rhEpo) in combination. rhG-CSF (5-20 micrograms/kg) and rhEpo (120-720 U/kg) were administered by s.c. injection three times a week for at least six months, and the administration was continued as maintenance therapy for as long as possible when hematological responses were observed. Six (60%) of the ten AA patients and four (58%) of the seven RA patients showed multilineage responses. Of these responders, six patients achieved trilineage recovery. While all of the responders were dependent on red blood cell transfusions and eight of them required platelet transfusions before treatment, they now no longer need transfusions of either red blood cells or platelets. A median treatment duration of 9 (range 1 to 28) months was required to achieve multilineage recovery. The responders showed an ability to maintain the multilineage recovery for 9+ to 47+ months and to tolerate long-term treatment. These results indicate that the long-term treatment with rhG-CSF and rhEpo may benefit a substantial percentage of patients with AA and RA and provide an optional therapy for these patients.

Adolescent↗

Inhibitory effect of sialoadenectomy on hepatocellular tumourigenesis in male mice induced by 3'-methyl-4-dimethylaminoazobenzene.

Male C57BL/6xDS-F1 mice that had been treated with 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) neonatally were subjected to sialoadenectomy (removal of the submandibular salivary glands) 60 days after birth. The development of adenomatous nodules and carcinomas in the livers of these mice was compared with the livers of males without sialoadenectomy. The incidence of adenomatous nodules in sialoadenectomized males at 6, 8, 10, and 12 months was significantly lower than that in sham-operated animals. Carcinomas were found in the livers of both sham-operated and sialoadenectomized males that were 10 and 12 months old and were less frequent in sialoadenectomized males although the difference was significant only at the age of 12 months. Sialoadenectomy decreased the serum concentration of epidermal growth factor (EGF) by about 40%. The present results indicate that sialoadenectomy inhibits the development of hepatocellular tumours induced by 3'-Me-DAB in male mice, an effect which may be caused by decrease in the serum level of EGF.

Adenoma↗

Demonstration of heat-shock protein 70 in the sporozoite stage of malaria parasites.

Three monoclonal antibodies generated by immunization of mice with Plasmodium berghei-infected red blood cells were found to react with the 75-kDa heat-shock protein (HSP70) present in liver stages and erythrocytic forms of the parasites. These antibodies were shown to react with a recombinant protein encoding the carboxyl terminal half of PfHSP70 (aa 365-681). Differently from earlier results, we clearly demonstrated that HSP70 was also expressed in the sporozoite stage, using these monoclonal antibodies in an immunofluorescence and Western immunoblot assay. These monoclonal antibodies react not only with sporozoites of P. berghei, the parasites originally used for the immunization, but also with sporozoites of several other rodent and human plasmodial species. Passive transfer of these monoclonal antibodies into naive mice, simultaneously injected with sporozoites, failed to neutralize the infectivity of P. berghei sporozoites and to inhibit the development of liver stages of P. yoelii.

Animals↗

Mapping of Marek's disease virus genome: identification of junction sequences between unique and inverted repeat regions.

Polymerase chain reaction (PCR) to amplify MDV DNA and subsequent sequencing identified the junction of TRL/UL, UL/IRL, IRS/US, and US/TRS. The TRL/UL junction is located 192 bp downstream of the last EcoRI site in the TRL region, while the UL/IRL junction is located 192 bp upstream of the first EcoRI restriction enzyme site in the IRL region. The IRS/US junction is located 950 bp downstream of the second EcoRI site in the IRS region, while the US/TRS junction is located 950 bp upstream of the first EcoRI restriction enzyme site in the TRS region. BamHI restriction enzyme mapping of one of the PCR products identified two novel DNA subfragments, BamHI-U2 and -P4, upstream of the US/TRS junction of the MDV genome. Sequencing of the BamHI-D fragment revealed a novel open reading frame (ORF) encoding a 155 amino acid protein. The TRL/UL junction is located in this ORF. The N-terminal 65 amino acids of this protein is homologous to the N-terminal region of the previously reported pp38, which is located in the UL/IRL region. Computer-assisted analysis indicated that both are transmembrane proteins and that they share an antigenic domain.

Amino Acid Sequence↗

Biochemical properties of collagen from ligaments and periarticular tendons of the human knee.

The biochemical properties of collagens from the periarticular connective tissues of the human knee (ligaments, semitendinosus and gracilis tendons, and the iliotibial band) were investigated in subjects ranging from 20 to 70 years of age. Although the total collagen content of most tissues was more than 90% of the dry weight, the anterior cruciate ligament and the iliotibial band had relatively low collagen contents. There were no significant changes in the tissue collagen content with aging. However, the anterior cruciate ligament and the patellar tendon of the 20-year-old subject had a higher content of soluble collagen than the other ligaments and tendons. The iliotibial band of the 20-year-old subject contained more collagen that was soluble by a combination of salt, sodium citrate extractions and pepsin digestion. Dihydroxylysinonorleucine was the major reducible cross-link of collagen from all the ligaments. The amount of dihydroxylysinonorleucine in the anterior cruciate ligament of the 20-year-old subject was much higher than that in the other ligaments. In contrast, the tendons and the iliotibial band contained a large amount of histidino-hydroxymerodesmosine and hydroxylysinonorleucine, while the patellar tendon was the only tendon with a significant content of dihydroxylysinonorleucine. Hydroxypyridinium non-reducible cross-links were more abundant in collagens from ligaments than in collagens from the other tissues. The cross-link study and the analysis of collagen solubility showed that patellar tendon collagen more closely resembled the collagen from the anterior cruciate ligament than that from periarticular tendons. It was also shown that the anterior cruciate ligament contains relatively immature collagen compared with the other ligaments.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

The role of delta-opioid receptor subtypes in cocaine- and methamphetamine-induced place preferences.

The effects of delta-receptor antagonists on cocaine- and methamphetamine-induced place preferences were examined in rats. Cocaine- and methamphetamine-induced place preferences were significantly attenuated by naltrindole (NTI: a non-selective delta-opioid receptor antagonist). Furthermore, naltriben (NTB: a selective delta 2-opioid receptor antagonist), but not 7-benzylidenenaltrexone (BNTX: a selective delta 1-opioid receptor antagonist), attenuated the cocaine- and methamphetamine-induced place preferences. These results suggest that delta-opioid receptors, particularly delta 2-opioid receptors, may be involved in the reinforcing effects of cocaine and methamphetamine.

Animals↗

Detection of antibodies to Anisakis simplex larvae by enzyme-linked immunosorbent assay and immunoelectrophoresis using crude or purified antigens.

The enzyme-linked immunosorbent assay (ELISA) and immunoelectrophoresis (IEP) were used for the serodiagnosis of larval Anisakis simplex infections in man and immunized rabbits. Sephacryl S-300 gel filtration was used for separating crude antigen. Four fractions were obtained. Sera from patients with other helminth infections sometimes cross reacted with Anisakis larval antigens. With IEP, crude antigen is more sensitive than purified antigens. With ELISA, the third fraction is the most sensitive for detecting antibodies to Anisakis larvae in the sera of humans and immunized rabbits.

Animals↗

Pressure gradients between intraluminal and extraluminal spaces may affect the potassium induced contraction in the rabbit's basilar arteries.

In order to investigate the different contactile effects between intraluminal and extraluminal vasoactive agents, we studied the contractile responses of rabbit basilar artery to selective intraluminal and extraluminal administration of potassium, serotonin and histamine in vitro. We also studied how the physical pressure gradients, such as hydrostatic or osmotic pressure gradients between intraluminal and extraluminal spaces, affect potassium-induced contraction. Intraluminal potassium (30 mM) induced a significantly greater contraction than extraluminal potassium. Serotonin (2 x 10(-7) M) and histamine (10(-5) M) applied intraluminally caused the same magnitude of the contraction as those applied extraluminally and no significant differences were noted between these applications. These differences in potassium-induced contraction were more significant when the intraluminal hydrostatic pressure was elevated by 20 mmHg. Contraction by either intraluminal or extraluminal potassium was significantly decreased when intraluminal pressure was raised by 40 mmHg. As the osmotic pressure gradients between the extraluminal and the intraluminal spaces were increased, these differences in potassium-induced contraction were decreased. Our findings suggest that physical pressure gradients may affect potassium-induced contraction in a different manner from pharmacologically-induced contraction and that free ions can penetrate the vascular wall by physical pressure gradients between the intraluminal and extraluminal spaces of cerebral artery.

Animals↗

Artificial fiber complexes composed of cellulose and guar gum or psyllium may be better sources of soluble fiber for rats than comparable fiber mixtures.

Three fiber complexes composed of cellulose and the same weight of guar gum, cornstarch or psyllium were constructed as three-dimensional networks of cellulose fiber filled with the soluble components. Physiological properties of these fiber complexes were examined in comparison with a mixture of powdered cellulose and the corresponding polysaccharides at a ratio of 1:1 in rats. Another group of rats was fed a fiber-free diet. Body weight gain in the guar gum-cellulose mixture group, but not in guar gum-cellulose complex group, was lower than that of the fiber-free group. The cecal walls in the groups fed the guar gum-cellulose and psyllium-cellulose mixture diets were heavier than those in the corresponding fiber complex-fed groups. Weights of the small intestinal wall and ileal mucosa were also greater in the guar gum-cellulose mixture-fed group than in the guar gum-cellulose complex-fed and the fiber-free diet-fed groups. Fermentable energy in dietary fiber estimated from fecal energy excretion, short-chain fatty acid contents and in vitro short-chain fatty acid production rates in the cecum of the complex-fed groups were similar to those in the fiber mixture-fed groups. However, butyric acid content and production rate in the group fed the guar gum-cellulose complex were markedly higher than in the other groups. The lowering effects of guar gum-cellulose and psyllium-cellulose mixtures on plasma cholesterol concentration tended to be greater than those of guar gum-cellulose and psyllium-cellulose complexes. These findings demonstrate that the artificial fiber complexes supply soluble fibers without increasing mechanical stress to the intestines and decreasing cecal fermentation.

Animals↗