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Biomedical subjects

S Miyata

Publications and source records attributed to S Miyata.

At least 199 records · Page 11Linked to original sources

Ouabain-induced contraction of vascular smooth muscle in spontaneously hypertensive rats and the effect of hydralazine.

The effect of ouabain (10(-3) M) on contractile responses of SHR (spontaneously hypertensive rat) and WKY (Wistar-Kyoto rat) aortas and mesenteric arteries was studied. Ouabain addition caused a rapid contraction of aortic strips with a steeper rate of rise and a larger maximal force development in strips from SHR than WKY. This difference in contractile response is known to occur in the prehypertensive period of SHR (4-week-old). Phentolamine (10(-6) M) pretreatment had no effect on the ouabain-induced contraction but partially suppressed it in both SHR and WKY aortas when diltiazem (10(-5) M) was also added. The difference in the ouabain-induced contractions of SHR and WKY aortas was more apparent in the residual contraction during suppression by diltiazem. The 45Ca uptake in the presence of ouabain was significantly larger in the early period of incubation in SHR aorta than in WKY aorta. The ouabain-induced contraction of hydralazine-treated SHR aorta from the prehypertensive period was very similar to that of non-treated WKY aorta. These results suggested that the abnormality of the ouabain-induced contraction in SHR arterial smooth muscle could have arisen from an increased Ca2+ movement due to Ca2+ leakage when ouabain inhibited the Na+-pump in the membrane. This abnormality seems to start during the prehypertensive period and continue in the hypertensive stage.

Aging↗

Inhibition of RNA synthesis in embryo of Xenopus laevis by protease inhibitor.

We have studied the role of proteases during the development of Xenopus laevis embryos with the aid of protease inhibitors. The activity of proteases was found to be only minimal in the unfertilized egg and during the initiation of development, but activity began to increase at the morula stage. When the activity of proteases was inhibited by antipain, an inhibitor of endopeptidase activity, RNA synthesis in the embryo was inhibited. To examine the relationship between the inhibitory effect of antipain on protease activity and its effect on RNA synthesis, antipain was reduced with NaBH4 to inactivate its protease inhibitory activity. The reduced antipain did not inhibit RNA synthesis in the embryo. Antipain effectively inhibited synthesis of both rRNA and poly(A)+RNA but not 4S RNA. We therefore suggest that protease activity plays an important role in the initiation and/or continuation of RNA synthesis.

Animals↗

Immunohistochemical examination of Peyer's patches in autoimmune mice.

The distribution of T-cells and B-cells in Peyer's patches was examined in three autoimmune model mice, MRL/Mp-lpr/lpr, BXSB, NZBWF1/J mice and normal BALB/c mice, between one and ten months old. A multiple layering technique was used for immunohistochemical detection of lymphocyte surface antigens of T-cells (Thy1.2, Lyt1, Lyt2) and B-cells (surface IgM) and peanut agglutinin receptor for germinal center cells. The T-cell population of female MRL/Mp-lpr/lpr mice increased markedly with age, and the B-cell population of the male BXSB mouse tended to increase. However, little change was observed with age in the NZBWF1/J mice. The immunohistochemical properties of the Peyer's patches in the three autoimmune model mice were different.

Animals↗

Characterization of proteolytic activities in embryos of Xenopus laevis.

1. Proteolytic activities in early embryos of Xenopus laevis exhibited maximum levels at pH 3.2, 5.6 and 7.2 when 3H-BSA was used as substrate, and the maximum proteolytic activity at pH 3.2 was several thousand-fold higher during the tail bud stage than in the unfertilized egg. 2. The proteolytic activity at pH 3.2 was separated into two fractions by gel chromatography. One fraction corresponded to a mol. wt of about 40,000 and its activity was inhibited by thiol protease inhibitors. The other appeared to be a protease of much higher mol. wt. 3. The maximum activities at pH 5.6 and 7.2 appear to correspond to proteins of mol. wt greater than 1,000,000.

Animals↗

Substitution of porphyropsin for rhodopsin in mouse retina.

Weanling, male albino mice were placed on a vitamin A-free diet for three months to deplete their vitamin A stores. The vitamin A-deficient mice were injected intraperitoneally with all-trans 3-dehydroretinol. 3-dehydroretinol was rapidly incorporated into the liver as a fatty acid ester. The chromophore of visual pigment increased gradually and reached a normal level 13 days after the injection. 3-Dehydroretinal accounted for 95% of the total chromophore in the retina. The high proportion of 3-dehydroretinal was observed also in the long-term experiment which was continued for six weeks with the injection of 3-dehydroretinol once a week. When the animal was injected with a mixture of 3-dehydroretinol and retinol, the ratio of dehydroretinal/retinal in the retina was far lower than the ratio of dehydroretinol/retinol in the liver. These results indicate that 3-dehydroretinol is not converted to retinol in mouse and is used less efficiently than retinol for the chromophore of visual pigment. The synthesis of visual pigment was observed even when the animal was kept in complete darkness after the injection of all-trans 3-dehydroretinol. This fact indicates that light is not required for the production of 11-cis chromophore of visual pigment.

Animals↗

Fatty metamorphosis in hepatocellular carcinoma: radiologic features in 10 cases.

We describe the sonographic, CT and angiographic findings in 10 cases of hepatocellular carcinoma in which extensive fatty metamorphosis occurred within the tumors. Fatty change was diffuse in smaller tumors (less than 3.5 cm) and focal in larger tumors (greater than 3.5 cm). Fatty metamorphosis characteristically caused a low-attenuation area on CT (less than -10 H) and a highly echogenic area on sonography. The sonographic appearance of small hepatocellular carcinomas with fatty metamorphosis was identical to the findings in cavernous hemangioma or focal fatty change of the liver. CT correctly revealed the presence of fat in these hepatocellular carcinomas. In these cases, hepatic arteriography showed no tumor stain; however, CT arteriography (dynamic CT during injection of contrast medium into the hepatic artery) was useful in showing the tumor, its capsule, and its internal septa. In the diagnosis of large hepatocellular carcinoma, the presence of intratumoral fat is not likely to be problematic, but small tumors that are diffusely infiltrated by fat must be distinguished from such benign conditions as focal fatty change, lipoma, and angiomyolipoma.

Aged↗

FR-900520 and FR-900523, novel immunosuppressants isolated from a Streptomyces. II. Fermentation, isolation and physico-chemical and biological characteristics.

FR-900520 and FR-900523, novel neutral macrolide immunosuppressants, were isolated from the cultured broth of Streptomyces hygroscopicus subsp. yakushimaensis No. 7238. Their molecular formulae were determined as C43H69NO12 and C42H67NO12, respectively. The compounds suppressed immune response in vitro. IC50 values of FR-900520 and FR-900523 for mouse mixed lymphocyte reaction were 0.55 nM and 1.6 nM, respectively. FR-900520, the major component, clearly prolonged skin allograft survival in rats.

Animals↗

Regional differences of proteins in isolated cells of early embryos of Xenopus laevis.

Regional differences of proteins were studied by two-dimensional gel electrophoresis in early embryos of Xenopus laevis. Pairs of blastomeres on the dorso-ventral axis were isolated from 16- and 32-cell embryos. Some dorso-ventral differences have been detected at 32-cell embryos. The proteins which were clearly detectable in the vegetal cells of the ventral marginal zone were only faintly detectable or undetectable in those of the dorsal marginal zone, and a regionally specific spot was detected in dorsal blastomeres.

Animals↗

Age-related changes of the function of T cell subsets: predominant defect of the proliferative response in CD8 positive T cell subset in aged persons.

The proportion of CD8 positive cells in the peripheral blood AET-rosette forming T cells from aged persons was significantly reduced than that from young persons. The difference in the proportion between aged and young groups became more significant after proliferative response to a mitogen phytohemagglutinin (PHA) or a specific antigen tuberculin active peptide (TAP). The purified macrophage-deprived T cells (Twp), CD4 (T4) positive cells or CD8 positive cells were prepared from aged or young persons. These cell preparations lost proliferative response to PHA or TAP but showed marked proliferative response to the combined stimulation to 1 microM of ionomycin and 1 nM of phorbol-12-myristate-13-acetate (PMA) at usual culture cell density (2.5 X 10(5)/ml). Proliferative responses of these cell preparations to the combined stimulation were significantly reduced in the aged than those in the young and the magnitude of the difference in the proliferative responses between aged and young groups was more pronounced in CD8 positive cell population than in CD4 positive cell population. Although the cell preparations were relatively independent of exogenous IL-2 for the proliferative response to the combined stimulation of ionomycin and PMA at usual culture cell density, they needed exogenous IL-2 for sustained proliferation at lower culture cell density (5 X 10(3)/ml). These IL-2-dependent proliferative responses to the combined stimulation in the aged were significantly lower than those in the young and again the difference in the proliferative magnitude between aged and young groups was greater in CD8 positive population. The mechanism(s) of age-related change of the proportion and proliferative ability of T subsets were discussed.

Adult↗

Age-associated changes in proliferative and differentiative response of human B cells and production of T cell-derived factors regulating B cell functions.

The abilities of highly purified B cells to repeat replication for clonal expansion and to differentiate into immunoglobulin secreting cells (ISC) were examined in the aged and young groups. B cells from the aged showed twofold less proliferative response to B cell mitogen Cowan 1 (SAC) than those from the young. The original clone size of SAC responding B cells determined by colchicine block and [3H] thymidine [( 3H] TdR) uptake was not significantly reduced in the aged whereas the ability to repeat replication to expand clonal size was significantly reduced. B cells from aged and young persons were induced into ISC by combined stimulation with SAC and partially purified B cell differentiation factor (BCDF) free of IL-2 activity. ISCs for IgG and IgA were rather increased or at least not reduced in number in the aged as compared with those in the young. We also determined the IL-2 and BCDF activity produced by T cells from aged and young persons. Upon PHA stimulation, the aged T cells produced tenfold less IL-2 activity and threefold higher BCDF activity than did young T cells. Approximately threefold increase in spontaneous secretion of BCDF activity by aged T cells was found as compared with young T cells. The inverse correlation between the IL-2 activity and BCDF activity was found when both activities were determined in the same samples.

Adult↗

FK-506, a novel immunosuppressant isolated from a Streptomyces. II. Immunosuppressive effect of FK-506 in vitro.

The immuno-pharmacological profile of a novel immunosuppressive agent, FK-506 produced by a streptomycete, is presented here. We proceeded to test the effect of the agent on various in vitro immune systems. It showed that mixed lymphocyte reaction, cytotoxic T cell generation, the production of T cell-derived soluble mediators such as interleukin 2 (IL-2), interleukin 3 and gamma-interferon and the expression of the IL-2 receptor were suppressed by this agent. The IC50 values of FK-506 and ciclosporin (CS) in all tests were approximately 0.1 nM and 10 nM, respectively. Therefore, the novel agent, FK-506 suppressed in vitro immune systems at about hundred times lower concentration than CS.

Animals↗

Mechanisms of age-related decline in antigen-specific T cell proliferative response: IL-2 receptor expression and recombinant IL-2 induced proliferative response of purified Tac-positive T cells.

Proliferative response of T cells from aged persons was significantly reduced to a specific antigen tuberculin-active peptide (TAP) determined by [3H]TdR uptake and FCM in comparison to that from the young. Cytokinetic analysis for the proliferative response to TAP showed that, in the aged, the clonal size or the number of the first generation responding cells to TAP was not significantly reduced but the ability to repeat replication was more profoundly affected. Neither the delayed entry into the cell replication nor prolongation of the cell cycle time could explain these results. Similar results have been reported on the proliferative response of T cells to mitogen: PHA (Phytohemagglutinin). Expression of Tac-antigen on T cells determined by anti-Tac antibody binding with FACS after stimulation with either TAP or PHA was found to be reduced significantly in the aged. Both the numbers of high and low affinity IL-2 receptors determined by radiolabelled IL-2 binding assay were also reduced in the aged, but the degree of reduction in number of high affinity ones was more pronounced than that in low affinity ones. Tac-positive T cells were isolated with the use of anti-Tac rosette methods and stimulated with recombinant IL-2 (r-IL-2). Their proliferative response was significantly lower in the aged than that in the young at any concentration of r-IL-2 examined. The number of the first generation responding cells to r-IL-2 in purified Tac-positive T cells from the aged was 82% of that from the young whereas the proliferative response by aged T cells was 39% of that by young ones when the cells were allowed to repeat replication for 3 days. The mechanisms of these multifactorial defects in proliferation of T cells from aged persons were discussed.

Adult↗