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Biomedical subjects

M Mizuno

Publications and source records attributed to M Mizuno.

At least 181 records · Page 10Linked to original sources

The effects of functional suppression of a membrane-bound complement regulatory protein, CD59, in the synovial tissue in rats.

OBJECTIVE: To investigate the roles of CD59 in the synovial tissue by functional suppression of CD59. METHODS: Rats treated with cobra venom factor to deplete complement or untreated rats were injected intraarticularly with 0.3 mg of the F(ab')2 fraction of a monoclonal antibody, 6D1, that inhibits the function of rat CD59. The circumference of knee joints was measured, and histologic changes in the synovium were studied. RESULTS: Joint swelling, thickening of the synovial tissues, infiltration of inflammatory cells into the synovium, and deposition of membrane attack complex (MAC) on the synovial surface were observed after intraarticular injection of 6D1. The inflammatory reaction reached its peak at 24 hours after injection, and finally subsided to normal within 3 days. It was suggested that functional suppression of CD59 in the synovium induced MAC formation followed by synovitis. Serum complement depletion did not completely suppress this reaction. This indicates that complement existing in the joint space is important for the formation of MAC on the synovial surface and for induction of synovitis. CONCLUSION: The membrane-bound complement regulatory protein, CD59, plays a key role in the protection of joints against MAC-mediated synovial injury and in maintaining the normal integrity of the joint.

Animals↗

Immunohistochemical analysis of intercellular adhesion molecule-1 expression in human gastric adenoma and adenocarcinoma.

In this study, we examined the distribution of intercellular adhesion molecule-1 (ICAM-1) in gastric adenomas and carcinomas immunohistochemically at the light and electron microscopic levels. ICAM-1 was expressed on tumour cells in 12 of 28 gastric carcinomas and in 3 of 11 adenomas but not on most normal gastric epithelial cells. ICAM-1 was localized on luminal sites of neoplastic glands in adenomas and in intestinal-type carcinomas, and rarely on the surface of tumour cells of diffuse carcinomas. Expression of ICAM-1 on the tumour cells was more frequent in intestinal-type than diffuse carcinomas (P < 0.005). At the ultrastructural level, ICAM-1 was present prominently on the apical membrane and weakly on the lateral surface of the tumour cells of the intestinal-type carcinoma and also localized on the perinuclear membrane and the membrane of the endoplasmic reticulum of cancer cells. There was no significant association between ICAM-1 expression and HLA antigen expression or the number of infiltrating lymphocyte subsets. These results may implicate the synthesis of ICAM-1 by gastric cancer cells, but the expression is infrequent and may not be sufficient for host immune surveillance of the tumour cell.

Adenocarcinoma↗

Chemoenzymatic synthesis of a novel glycopeptide using a microbial endoglycosidase.

The chemoenzymatic synthesis of a glycopeptide by chemical synthesis of N-acetylglucosaminyl peptide and enzymatic transfer of an oligosaccharide is described. We synthesized glycosylated Peptide T which blocks infection of human T cells by human immunodeficiency virus. The first step of the chemoenzymatic method is the solid-phase chemical synthesis of N-acetylglucosaminyl Peptide T (Ala-Ser-Thr-Thr-Thr-Asn(GlcNAc)-Tyr-Thr) with an N-acetylglucosamine moiety bound to the asparaginyl residue by a solid-phase method. This product was prepared in high yield by the dimethylphosphinothioic mixed anhydride method without protecting the hydroxyl functions of the sugar moiety using Fmoc-N-acetylglucosaminyl asparagine instead of Fmoc-asparagine. The second step was transglycosylation of complex type oligosaccharide to N-acetylglucosaminyl Peptide T by a microbial endoglycosidase. The endo-beta-N-acetylglucosaminidase of Mucor hiemalis transfer the oligosaccharide of human transferrin glycopeptide to N-acetylglucosaminyl Peptide T. The transglycosylation product was confirmed to be the glycosylated Peptide T with a sialo biantennary complex type oligosaccharide by mass spectrometry. The glycosylated Peptide T was highly stable against proteolysis in comparison to native Peptide T and N-acetylglucosaminyl Peptide T.

Acetylglucosaminidase↗

Application of magnifying chromoscopy for the assessment of severity in patients with mild to moderate ulcerative colitis.

BACKGROUND: A magnifying colonoscope that enables high-power observation of the colorectal mucosa has been recently developed. The aim of this study was to investigate the value of the magnifying instrument in determining the severity of ulcerative colitis. METHODS: Magnifying colonoscopy was performed in 41 patients with ulcerative colitis, and the findings in the rectum were graded according to network pattern and cryptal openings. These findings were correlated with endoscopic, clinical, and histologic severity of the disease. RESULTS: Magnifying colonoscopy did not detect network pattern in 37% and cryptal opening in 24% of the subjects. The clinical, endoscopic, and histologic grades of activity were not different between groups divided by the presence or absence of each finding. However, when the two features were coupled, patients with visible network pattern and cryptal opening had a lower clinical activity index and lower grade of histologic inflammation than those in whom both findings could not be visualized. CONCLUSIONS: Observation under magnifying colonoscopy can be another clue to determining the severity of disease in patients with ulcerative colitis.

Adolescent↗

The effect of arginine supplementation on growth hormone release and intestinal mucosal growth after massive small bowel resection in growing rats.

Four-week-old male Sprague-Dawley rats underwent a 90% small bowel resection. From the fourth day after surgery, they were divided into group 1 and 2, and pair-fed by elemental diets (0.8 kcal/mL, 50 mL/day) with L-arginine (n = 10) or L-glycine (n = 11) as an isonitrogenous and isoenergetic supplement for 3 weeks. They were compared with each other 3 weeks after surgery. A statistical analysis was performed using the unpaired Student's t test and the one-way factorial analysis of variance (ANOVA) using Bonferroni/Dunn multiple comparison test. A Pvalue of < .05 was considered significant. There were no significant differences between the two groups in food intake, body weight, tail length, residual ileal length, and plasma IGF-I level. However, the mean height of ileal villi in group 1 showed higher than that in group 2 (P < .01). Growth hormone-releasing hormone (GHRH) provocative tests (1 microg per rat, intravenously) showed the more significant elevation of growth hormone IGH) secretion in the arginine supplement group than that of glycine supplement group at 5 minutes (P < .05). There were no significant differences between basal levels of plasma rat GH in both groups. It is suggested that arginine has a possible significant role of GH secretion and intestinal mucosal growth after massive small bowel resection.

Animals↗

Effects of oral supplementation of coenzyme Q10 on 31P-NMR detected skeletal muscle energy metabolism in middle-aged post-polio subjects and normal volunteers.

The effects of oral supplementation of 100 mg coenzyme Q10 (CoQ10) for 6 months on muscle energy metabolism during exercise and recovery were evaluated in middle-aged post-polio (n = 3) and healthy subjects (n = 4) by the use of phosphorus-31 nuclear magnetic resonance spectroscopy. The metabolic response to isometric plantar flexion at 60% of maximal voluntary contraction force (MVC) for 1.5 min was determined in gastrocnemius muscles before, after 3- (3MO) and 6-month (6MO) of CoQ10 supplementation. The MVC of plantar flexion was unchanged following CoQ10 supplementation. The resting Pi/PCr ratio in gastrocnemius muscles of all subjects decreased after 3MO- and 6MO-CoQ10 (P < 0.05). The post-polio individuals showed a progressive decrease in this ratio, while less pronounced changes were observed in the control subjects. Similarly, the post-polio individuals showed a lower Pi/PCr ratio at the end of 60% MVC in both 3MO- and 6MO-CoQ10, whereas no change in the ratio was observed in the control subjects. A less pronounced decrease in muscle pH was observed at the end of 60% MVC in both 3MO- and 6MO-CoQ10 in the post-polio individuals, but not in the control subjects. No systematic difference in end-exercise ATP was observed between the three phases in both groups. The half-time of recovery for PCr decreased in all subjects after 6MO-CoQ10 supplementation (P < 0.05). The results suggest that CoQ10 supplementation affects muscle energy metabolism in post-polio individuals to a greater extent than in control subjects. The mechanism for this effect is not clear, but may involve an effect of CoQ10 on peripheral circulation in the calf muscles, its action in mitochondrial oxidative phosphorylation and/or its antioxidant potential.

Administration, Oral↗

Suppression for the proliferation of fibroblasts by external DNA.

Phase-contrast and fluorescence microscopic observation showed that DNA added in the cell-culture medium for fibroblasts localized just on the surface of fibroblasts. The DNA bound to fibroblasts was found to be eluted by treating with collagenase. The suppression for the proliferation of fibroblasts by external DNA was confirmed with microscopic observation for the cells cultured in the presence and absence of DNA. Proliferation of the cells decreased from 412 to 155% by the addition of DNA. These results indicate that DNA has an affinity for collagen, the most major extracellular-matrix produced by fibroblasts, and suppresses the growth of fibroblasts.

Cell Division↗

Differential expression patterns of GABA transporters (GAT1-3) in the rat olfactory bulb.

The localization of GABA transporters 1-3 (GAT1-3) was investigated in the rat olfactory bulb by using in situ hybridization and immunohistochemistry. In the glomerular and the internal granular layers, GAT1 mRNA was expressed in most of periglomerular and granule cells, which are known to be GABAergic. In addition, we compared GAT1 mRNA expression with that of glutamic acid decarboxylase67 (GAD67) mRNA. The expressions were very similar in these two layers, indicating that GAT1 mRNA is mainly expressed in GABAergic neurons. However, in the external plexiform and the olfactory nerve layers, we observed more GAT1 mRNA-positive cells than GAD67 ones, suggesting that GAT1 mRNA is also expressed in non-GABAergic neurons and in glial cells. GAT3 mRNA expression was observed in small glial-like cells which might be involved in GABAergic neurotransmission throughout the olfactory bulb. This was confirmed by double-immunostaining studies which showed the expression of both GAT3 and glial fibrillary acidic protein (GFAP) mRNAs in astrocytes. Moreover, GAT2 mRNA was expressed only in the ependyma and arachnoid. These findings suggest that the expression patterns of GABA transporters differ with the type of cells in the rat olfactory bulb where GAT1 and GAT3 may play an imporatant role in GABA-mediated transmission, such as lateral inhibition.

Animals↗

Localization of molecules involved in cytokine receptor signaling in the rat trigeminal ganglion.

The localization of some cytokine receptors and their downstream intracellular signaling molecules was examined in the trigeminal ganglia of rats. Among cytokine receptor components, we examined signal transduction subchain, gp130, IL-2Rgamma and IL-5Rbeta, which are common to respective groups of cytokine receptors. Most of the sensory ganglion neurons expressed gp130, but not IL-2Rgamma nor IL-5Rbeta. We further examined the localization of Janus kinase (JAK) family members which were reported to be associated with various kind of cytokine receptors and are thought to be implicated in major cytokine receptor-signaling pathways [6,9,11,13]. While JAK1 and Tyk2 were expressed in all the type of neurons, JAK2 was predominantly expressed in the small neurons. In addition, JAK3 immunoreactivity was only found in satellite cells. The present results indicate that most of neurons express gp130, and that the localization of JAK family members differs with the cell type. This also suggests that the cytokine receptor-signaling pathway may be different in neuronal and glial cells.

Animals↗

Attachment of osteoblastic cells to hydroxyapatite crystals by a synthetic peptide (Glu7-Pro-Arg-Gly-Asp-Thr) containing two functional sequences of bone sialoprotein.

We investigated activity of bone sialoprotein (BSP) to mediate attachment of cells to hydroxyapatite using a model peptide, Glu7-Pro-Arg-Gly-Asp-Thr, which contains a putative hydroxyapatite-binding site (poly-Glu) and a cell-attachment site. The peptide has affinity to hydroxyapatite with a dissociation constant of 13.5 microM. The peptide affected in vitro mineralization in a gel system, indicating interaction between this peptide and calcium phosphate. The osteoblastic cell line MC3T3-E1 was incubated with hydroxyapatite powder coated with the peptide or proteins. Attachment of the cells was observed on the powder coated with BSP, but not on the powder coated with serum albumin. The cells were attached to the powder coated with the peptide. The cells were flattened on the powder, and pseudopods developed. The attachment of the cells was inhibited by an excessive amount of Gly-Arg-Gly-Asp-Ser peptide. In conclusion, BSP mediated attachment of osteoblastic cells to hydroxyapatite, and this activity could be accomplished only by the poly-Glu sequence and the Arg-Gly-Asp sequence.

Amino Acid Sequence↗

Osteogenesis by bone marrow stromal cells maintained on type I collagen matrix gels in vivo.

In this study, we demonstrated that bone marrow stromal cells maintained on type I collagen matrix induced bone in vivo. The formed bone contained bone marrow, and the process of bone formation occurred without cartilage formation. Bone marrow stromal cells differentiated into osteoblasts on type I collagen matrix in vitro, but types II, III, and V collagens did not possess this activity. These findings imply that type I collagen matrix offers a suitable environment for the induction of osteoblastic differentiation in vitro and osteogenesis in vivo.

Alkaline Phosphatase↗

Augmentation of anti-Fas antibody-mediated apoptosis on human glioma cells by liposomes associated with the antibody.

We investigated the susceptibility of five human glioma cell lines to anti-Fas antibody. All human glioma cells tested constitutively expressed Fas antigen on their surfaces and the level of the expression varied slightly in each cell line. The cells had a low susceptibility to anti-Fas antibody-mediated apoptosis. There were four moderately resistant cell lines (U251-SP, U251-MG, SK-MG-1, T98) and one highly resistant cell line (U251 nu/nu). For this study we prepared liposomes containing anti-Fas antibody and studied the augmentation of the antibody-mediated apoptosis. The liposomes induced apoptosis significantly more often than did anti-Fas antibody alone. These results indicate that anti-Fas antibody-mediated apoptosis does not require a critical level of cell surface expression of Fas antigen but rather depends on the intensity of Fas signal transduction.

Antibodies, Monoclonal↗

A case of systemic malignant lymphoma with intestinal involvement of lymphomatous polyposis type.

Multiple lymphomatous polyposis is a rare type of intestinal lymphoma characterized by non-Hodgkin's lymphoma of follicular mantle cell origin and extremely poor prognosis. We report a case of systemic lymphoma with the intestinal involvement of multiple lymphomatous polyposis. Although radiographic and endoscopic features of the case were compatible with multiple lymphomatous polyposis, histologic evidence suggested the diagnosis of diffuse large cell lymphoma rather than mantle cell lymphoma. Our case seems to be unique in its histologic findings and also in its prognosis, because the patient has been alive for more than 50 months after diagnosis.

Antineoplastic Combined Chemotherapy Protocols↗

Performance characteristics of chronic schizophrenia on attention tests sensitive to unilateral brain damage.

The aims of this study were to investigate the lateralization hypothesis of schizophrenia, comparing chronic schizophrenics with unilateral brain-damaged subjects and normal controls, using attentional tests sensitive to the unilateral brain damage. Three attentional tests with different modes of stimuli, two vigilance tasks that require the self-paced or nonself-paced responses of subjects and one divided attention task, were administered to 28 chronic schizophrenics, 26 right and 24 left unilateral brain-damaged subjects, and 20 normal controls. The results indicated that schizophrenics performed a possible right-hemisphere damages pattern and also a left pattern in part, with failure of all tasks to show either pattern related to a number of differences between the three tasks. Furthermore, the attention deficits of schizophrenics are less than those of brain-damaged subjects but are clearly abnormal compared with the normal controls.

Attention↗

Interferon-alpha therapy for individuals with normal serum alanine aminotransferase levels before treatment.

While interferon (IFN) therapy is approved for patients with chronic hepatitis C infection, the effectiveness of IFN therapy for anti-hepatitis C virus (HCV)-positive individuals with normal ALT levels remains poorly understood. Twenty-five individuals (M:F = 9:16; age = 51.8 +/- 15.1 years) who showed a normal alanine aminotransferase (ALT) level (< 30 IU/L) at least 6 months before treatment and were positive for both anti-HCV and serum HCV-RNA by reverse transcription polymerase chain reaction (RT-PCR) were studied. Of these, 16 received IFN therapy for 24 weeks (780 million units total IFN-alpha 2b) and nine were followed for 12 months as controls. The serum HCV-RNA level was determined by bDNA assay and HCV genotype was detected by enzyme immunoassay. All were shown to have chronic hepatitis by liver biopsy. Six (37.5%) of 16 individuals lost serum HCV-RNA by RT-PCR 6 months after cessation of therapy (complete response; CR), while the remaining individuals were defined as non-responders (NR). Of the six individuals in the CR group, five had HCV type 2 infection, compared with none of 10 individuals in the NR group (P < 0.01). There was a trend towards a lower pretreatment viraemia level in the CR group than in the NR group. These data suggest that IFN therapy for individuals with HCV type 2 infection and normal ALT levels was effective.

Alanine Transaminase↗

PDGF-BB decreases systolic blood pressure through an increase in macrovascular compliance in rats.

The cardiovascular roles of platelet-derived growth factor (PDGF) were examined in anesthetized rats by monitoring blood pressure and in isolated blood vessels and heart preparations. Intravenous injection of PDGF-BB lowered blood pressure. The decrease in systolic pressure was greater than that in diastolic pressure, so the pulse pressure decreased. PDGF-AA and -AB, other isoforms of PDGF, did not have any effect on blood pressure. Pretreatment of rats with N(omega)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthase, shortened duration of the hypotensive effect of PDGF-BB. The administration of L-arginine with L-NAME partially prevented the effect of L-NAME. PDGF-BB relaxed aortic rings precontracted with phenylephrine with a 50% effective concentration of 3 ng/ml. In contrast, in isolated mesenteric vascular preparations, the vasodilating activity of PDGF-BB was observed only at a high concentration (>12.5 ng/ml). In isolated heart preparations, PDGF-BB had no effect on the beat rate or contractile activity. These results suggest a new role of PDGF-BB that may contribute to the regulation in circulation through the increase in macrovascular compliance mediated by NO.

Animals↗

Delayed signal propagation via CA2 in rat hippocampal slices revealed by optical recording.

Signal propagation from mossy fibers to CA1 neurons was investigated in rat hippocampal slices by a combination of electrical and optical recordings. The slices were prepared by oblique sectioning of the middle part of the hippocampus to preserve fiber connections. The mossy fibers were stimulated to induce population spikes (PSs) and excitatory postsynaptic potentials in the middle part of the CA1 region. Latencies of maximal PSs in CA1 varied widely among slices; they ranged from 7 to 13.5 ms, with two maxima at 9 and 11.5 ms. The fastest PSs probably are evoked by the Schaffer collaterals that connect the CA3 and CA1 regions in the well-known trisynaptic circuit. However, the slower PSs suggest the existence of additional delayed inputs. To determine the source of the delayed input, slices were stained with a voltage-sensitive dye, RH482, and the optical signals relevant to membrane potential changes were detected by a high-resolution optical imaging system. Optical recording of responses to mossy fiber stimulation indicated two distinct types of signal propagation from CA3 to CA1. In preparations evincing the fast type of propagation, signals spread to CA1 within 7.2 ms after the mossy fiber stimulation. During such propagation, activity flowed directly from CA3 to the stratum radiatum of CA1. Other preparations illustrated slow signal propagation, in which optical signals were generated in CA2 before spreading to CA1. During such slow signal transmission, activity persisted in CA2 and its surrounding area for 3 ms before propagating to the strata radiatum and oriens in CA1. In such cases, CA1 activity was detected within 10.8 ms of mossy fiber stimulation. In some slices, a mixture of the fast and slow propagation patterns was observed, indicating that these two transmission modes can coexist. Our data reveal that CA2 neurons can transmit delayed excitatory signals to CA1 neurons. We therefore conclude that consideration of electrical signal propagation through the hippocampus should include flow through the CA2 region in addition to the traditional dentate gyrus-CA3-CA1 trisynaptic circuit.

Animals↗