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Biomedical subjects

H Miyamoto

Publications and source records attributed to H Miyamoto.

At least 217 records · Page 12Linked to original sources

Retinoblastoma gene mutations in primary human prostate cancer.

Structural alterations in the entire coding regions (exons 1 to 27) of the retinoblastoma (RB) gene in primary human prostate cancers were investigated, using polymerase chain reaction and single strand conformational polymorphism analysis of RNA. Of 25 samples obtained from patients, four (16.4%) were found to have RB alterations. DNA sequencing of the PCR products revealed point mutations resulting in single amino-acid substitutions of exons 6 and 19 in two cases, and base deletions of exons 8 and 17 in two cases. Two of four cases with RB mutations were moderately differentiated localized tumors and other two with RB mutations were poorly differentiated tumors with metastases. Our results suggest that RB gene mutation is involved in progression steps of prostate carcinogenesis.

Adenocarcinoma↗

Blood flow velocity in the common carotid artery in humans during graded exercise on a treadmill.

Cerebral blood volume flow and flow velocity have been reported to increase during dynamic exercise, but whether the two increase in parallel and whether both increases occur as functions of exercise intensity remain unsettled. In this study, blood flow velocity in the common carotid artery was measured using the Doppler ultrasound method in eight healthy male students during graded treadmill exercise. The exercise consisted of stepwise progressive increases and decreases in exercise intensity. The peak intensity corresponded to approximately 85% of maximal oxygen consumption. During this exercise, the heart rate (fc), mean blood pressure (BP) in the brachial artery and mean blood flow velocity (vcc) in the common carotid artery increased as functions of exercise intensity. At the peak exercise intensity, fc, BP and vcc increased by 134.5%, 20.5% and 51.8% over the control levels before exercise (P < 0.01), respectively. The resistance index (RI) and pulsatility index (PI) were determined from the velocity profile and were expected to reflect the distal cerebral blood flow resistance. The RI and PI increased during the graded exercise, but tended to decrease at the highest levels of exercise intensity. As vcc increased with increases in exercise intensity it would be expected that cerebral blood flow would also increase at these higher intensities. It is also suggested that blood flow velocity in the cerebral artery does not proportionately reflect the cerebral blood flow during dynamic exercise, since the cerebral blood flow resistance changes.

Blood Flow Velocity↗

Detection of immunoreactive antigen, with a monoclonal antibody to measles virus, in tissue from a patient with Crohn's disease.

Using immunofluorescence (IF), we investigated reactive antigens present in Crohn's disease patients with monoclonal antibodies derived from cells infected with measles virus, but not with the subacute sclerosing panencephalitis virus. During immunoblotting, one monoclonal antibody (mAb 86) reacted with a polypeptide with a molecular weight of 36,000 dalton (M; matrix protein) in measles virus-infected cells. This monoclonal antibody displayed a positive reaction only with tissues from patients with Crohn's disease by the IF test. It did not react with samples from patients with other chronic inflammatory bowel diseases, such as ulcerative colitis. Other monoclonal antibodies to the measles virus protein, and monoclonal antibodies to Herpes simplex virus type 1 did not react with the same tissue samples. The role of measles virus infection and/or a viral antigen (possibly the M protein) as a causative agent in Crohn's disease poses a challenging avenue for further research.

Antibodies, Monoclonal↗

Malignant schwannoma of the liver.

Malignant schwannoma is the most common soft tissue sarcoma in adults, but primary schwannoma of the liver is extremely rare. We report an autopsy case of malignant schwannoma of the liver in a 63-year-old man. The tumor involved almost the entire right lobe of the liver. Microscopically, it was composed of moderately pleomorphic spindle cells with hyperchromatic nuclei with mitotic figures. The spindle cells were stained positively with antibodies to both S-100 protein and vimentin. Necrosis was found in the primary tumor. Only four other cases of malignant schwannoma of the liver were found in a review of the literature. Pertinent features of these tumors are discussed.

Fatal Outcome↗

Serum antibody against Helicobacter pylori assayed by a new capture ELISA.

We developed a highly specific detection technique for serum antibody, using a monoclonal antibody to a specific antigen of Helicobacter pylori. A monoclonal antibody preparation that reacted with the 54-kDa molecule of H. pylori antigens was obtained. Using this preparation, an antigen-capture enzyme-linked immunosorbent assay (ELISA) was established by fixation of the monoclonal antibody, followed by reaction with sonicated whole cell antigens. The serum antibody titers of patients with gastritis and peptic ulcers were significantly higher than control titers, and the antibody titer correlated with the histological severity of gastritis. Patients positive for H. pylori by bacterial culture had higher titers than H. pylori-negative patients. Our new ELISA may be useful for the diagnosis of H. pylori infections and for evaluation of the severity of gastric inflammation.

Antibodies, Bacterial↗

The effect of a naphthalene derivative, TEI-6472, on histamine release and tyrosine phosphorylation in rat basophilic leukemia cells.

It is well known that rat basophilic leukemia cells (RBL-2H3) express high-affinity IgE receptors (Fc epsilon RI) and that the aggregation of these receptors causes the release of chemical mediators. When RBL-2H3 cells are sensitized with IgE antibody and subsequently stimulated by an antigen, significant histamine release and the tyrosine phosphorylation of several proteins are observed. In this study, we examined the effects of a synthetic naphthalene derivative, (7E)-N-(2-carboxyphenyl)-8-(2-naphthyl)-5,6-trans-5,6-methano-7-++ +octenamide (TEI-6472), on the Fc epsilon RI-mediated histamine release from RBL-2H3 cells. Preincubation for 10 min with 100 microM TEI-6472 caused significant inhibition of Fc epsilon RI-mediated histamine release from RBL-2H3 cells. Furthermore, Western blotting analysis using anti-phosphotyrosine antibody showed that Fc epsilon RI-mediated tyrosine phosphorylation of 78 and 92 kDa proteins in RBL-2H3 cells was also significantly inhibited. Tyrosine phosphorylation of these 78 and 92 kDa proteins was not induced by direct activation of protein kinase C (PKC) by phorbol-12-myristate-13-acetate (PMA) and the calcium ionophore A23187. However, the inhibition of histamine release from TEI-6472-treated RBL-2H3 cells was restored by direct activation of PKC. Taken together, these results suggest that tyrosine phosphorylation of the 78 and 92 kDa proteins in RBL-2H3 cells is involved in a signal transduction system for histamine secretion, and that these tyrosine phosphorylations may occur upstream of PKC activation.

Animals↗

Antiphospholipid antibody syndrome in childhood strokes.

The significance of antiphospholipid antibodies was examined in patients with childhood strokes. Eight patients, aged 2-13 years, who presented with acute hemiplegia were studied. On the basis of magnetic resonance imaging, magnetic resonance angiography, single photon emission computed tomography, and cerebral angiographic findings, 3 children were diagnosed as having infarctions due to moyamoya disease, and the others as having idiopathic infarctions. Serologic studies revealed elevated anticardiolipin antibody (ACA) IgG in 3 of the 5 patients with idiopathic infarction; no such elevation was revealed in patients with moyamoya disease. Values for all other tests, including ACA IgM and lupus anticoagulant, were within normal limits or negative in all patients. ACA IgG, therefore, may be a more important causative agent of childhood strokes than was previously considered.

Adolescent↗

Synthesis and biological properties of substituted 1,4-dihydro-5-methyl-4-oxo-3-quinolinecarboxylic acids.

A series of substituted 1-cyclopropyl-6-fluoro-1, 4-dihydro-5-methyl-4-oxo-3-quinoline carboxylic acids was synthesized and tested for their in vitro and in vivo antibacterial activity. The introduction of a methyl group at the 5-position of quinoline nucleus enhanced characteristically the antibacterial activity against Gram-positive bacteria, including Streptococcus pneumoniae, which is a major pathogen in the respiratory tract infection, while retaining Gram-negative activity. Among them, 1-cyclopropyl-6-fluoro-1, 4-dihydro-5-methyl-7-(3-methyl-1-piperazinyl)-4-oxo-3-quinolinecarboxyli c acid hydrochloride (grepafloxacin) exhibited potent in vitro antibacterial activity against Gram-positive bacteria such as Streptococcus pneumoniae and high in vivo efficacy on the experimental systemic infections caused by the Gram-positive and -negative bacteria tested. It also showed a high distribution to the lung and bronchoalveolar lavage fluid in comparison to reference drugs and is now undergoing clinical evaluation.

4-Quinolones↗

Exercise-induced changes in R wave amplitude and heart rate in normal subjects.

An intermittent exercise protocol on a treadmill was used to examine six healthy subjects, and a steady protocol was applied to three of the subjects before and after short-term training. The peak blood velocity in the common carotid artery increased by 73.1% during the intermittent protocol and recovered to resting level within 3 minutes, while the heart rate (HR) remained high even 5 minutes after exercise. R wave amplitude (RWA) increased significantly from 1.40 +/- 0.39 mV at rest to 1.59 +/- 0.33 mV (P < .05) immediately after the start of walking, and decreased gradually to 1.46 +/- 0.36 mV (P < .05) during 3 minutes of walking. Thus, it decreased significantly to 1.31 +/- 0.40 mV (P < .01) during the interphase from exercise to rest, and increased again during recovery or rest periods in the intermittent protocol. The results suggest that an increase in the venous return per heart beat at the start of walking induces the increase in RWA, and that its abrupt decrease at the end of walking induces the decrease in RWA. Subjects with a higher HR response and recovery slopes have smaller abrupt changes in RWA at the interphases between rest and walking. The gradual decrease in RWA during walking may be related to a gradual increase in HR and a gradual decrease in systemic peripheral resistance, and the gradual increase in RWA after walking may be related to a gradual decrease in HR and a gradual increase in systemic peripheral resistance.

Adult↗

Retinoblastoma gene mutations in primary human bladder cancer.

Inactivation of the retinoblastoma (RB) gene is known to be implicated in the pathogenesis of several types of human cancers. Since structural alterations of the RB gene have not been well examined in human bladder cancer, we looked for mutations in the entire coding region of this gene using polymerase chain reaction (PCR) and single-strand conformational polymorphism analysis of RNA. We also examined allelic loss of the RB gene using PCR-based restriction fragment length polymorphism analysis. Of 30 samples obtained from patients with bladder cancer, eight (27%) were found to have RB gene mutations. DNA sequencing of the PCR products revealed five cases with single point mutations and three cases with small deletions. These mutations included one (10%) of ten low-grade (grade 1) tumours, four (50%) of eight intermediate-grade (grade 2) tumours and three (25%) of 12 high-grade (grade 3) tumours. Likewise, mutations were found in four (21%) of 19 superficial (pTa and pT1) tumours and four (36%) of 11 invasive (pT2 or greater) tumours. In 15 informative cases, loss of heterozygosity at the RB locus was shown in five cases (33%), three cases with RB mutations and two without them. These results suggest that RB gene mutations are involved in low-grade and superficial bladder cancers as well as in high-grade and invasive cancers.

Aged↗

The loss of retinoblastoma gene in association with c-myc and transforming growth factor-beta 1 gene expression in human bladder cancer.

PURPOSE: We investigate the roles and possible interactions of the retinoblastoma, transforming growth factor-beta 1 and c-myc genes in bladder cancer. MATERIALS AND METHODS: The expression of these 3 genes was examined in 38 biopsy specimens of human bladder cancer by immunohistochemical analysis or Northern blotting. RESULTS: Loss of the retinoblastoma protein expression was most significantly correlated with high grade cancer. Over expression of c-myc or expression of transforming growth factor-beta 1 was less associated with tumor grade or stage, although c-myc over expression defined stage Ta against other stage tumors, since no stage Ta lesions had increased c-myc expression. Finally, loss of retinoblastoma gene function did not correlate with either c-myc or transforming growth factor-beta 1 expression. CONCLUSIONS: These results further support that retinoblastoma gene inactivation is an important factor in the progression of bladder cancer, and suggest that transforming growth factor-beta 1 and c-myc are not regulators or are not regulated by retinoblastoma gene expression.

Gene Expression Regulation, Neoplastic↗

Low expression of adhesion molecules in a case of cutaneous T-cell lymphoma.

A case of cutaneous T-cell lymphoma (CTCL) with low expression of the adhesion molecules lymphocyte function-associated antigen-1 (LFA-1), intercellular adhesion molecule-1 (ICAM-1), and very late antigen-4 (VLA-4) is described. The patient was a 90-year-old man with red round homogeneous tumors on his scalp, trunk, and extremities. He had no history of definite erythema or plaque stage. A biopsy sample taken from a tumor revealed massive infiltration of atypical lymphocytes in the reticular dermis and subcutis with a definite clear zone. The atypical lymphocytes were medium-sized with slightly convoluted nuclei. Immunohistochemically, the infiltrates showed the phenotype of so-called memory T cells. On the basis of these features, the case was diagnosed as CTCL. Expression of LFA-1, ICAM-1 and VLA-4 on the infiltrates was 9%, 13% and 11%, respectively, which is much lower than that in classic mycosis fungoides. This finding suggests that loss of these adhesion molecules may contribute to loss of epidermotropism in the advanced stage of CTCL.

Aged↗

Temperature effects on Legionella pneumophila killing by and multiplication in phagocytes of guinea pigs.

We examined the effects of temperature on the interaction between Legionella pneumophila and phagocytes of guinea pigs. The body temperatures of guinea pigs infected with a sublethal dose (1.2 x 10(4) CFU) or a lethal dose (1.0 x 10(5) CFU) of L. pneumophila elevated from 38.4 +/- 0.15 C to 40.2 +/- 0.42 C or 40.3 +/- 0.62 C, respectively. The intracellular bacterial killing by and bacterial proliferation in the phagocytes were examined at 33, 37, 40, and 42 C, using in vitro culture systems of peritoneal macrophages or polymorphonuclear leukocytes (PMN) of guinea pigs. In all the macrophages incubated at different temperatures, significant intracellular bacterial killings were observed at 4 hr after in vitro phagocytosis. After 24 hr of incubation, there was about a 100-fold increase of CFU and the number reached a maximum after 48 hr of incubation in the macrophages incubated at 42 C as well as 37 and 40 C, suggesting that macrophages support the intracellular bacterial growth in hyperthermia. In the PMN, L. pneumophila CFU 4 hr or 12 hr after the infection were significantly lower at 42 C than those at 37 C (P < 0.05), indicating that the bactericidal capacity of PMN was enhanced at 42 C compared to 37 C. However, in all the PMN incubated at different temperatures, there were about 10-fold increases of CFU 24 hr after the infection, suggesting that PMN as well as macrophages support intracellular bacterial growth in hyperthermia. The extracellular bacterial growth was examined at 33, 37, 40, and 42 C in buffered yeast extract (BYE) broth or RPMI 1640 medium containing 50% guinea pig serum as a permissive or non-permissive liquid medium for the bacterial growth, respectively. Inhibition of bacterial growth in BYE broth at 42 C, and a decrease of CFU in RPMI 1640 medium containing 50% guinea pig serum at 42 C were observed. In conclusion, hyperthermia may be beneficial by restricting extracellular bacterial survival, but it exerts no beneficial effect on the restriction of intracellular bacterial growth in phagocytes, though PMN showed enhanced initial killing at 42 C. These results suggest that fever, or hyperthermia itself, may not largely contribute as a nonspecific host defense early in the course of legionellosis.

Animals↗

Immunohistochemical quantitation for extracellular matrix proteins in rats with glomerulonephritis induced by monoclonal anti-Thy-1.1 antibody.

Extracellular matrix proteins (type I collagen and fibronectin) in frozen histologic sections of kidney cortex from rats with glomerulonephritis induced by a single intravenous administration of anti-Thy 1.1 antibody were quantified using an immunohistochemical micromethod. Type I collagen and fibronectin contents in renal cortex of rats with experimental glomerulonephritis (4.33 +/- 0.79 and 10.41 +/- 2.01 microgram/mg of total protein, respectively) were 262% and 151%, respectively, higher than in control rats given normal mouse IgG (1.65 +/- 0.16 and 6.88 +/- 0.95 microgram/mg, respectively; p < 0.01 in each case). In the glomerulonephritic rats, the increase in the contents of extracellular matrix proteins, especially type I collagen, correlated with increasing glomeruli with expansion of mesangial areas. The increase in type I collagen content correlated well with increasing urinary protein excretion and blood urea nitrogen and serum total cholesterol levels (r = 0.851, 0.812, and 0.837, respectively; p < 0.05 in each case). The decrease in creatinine clearance correlated with increasing content of type I collagen (r = 0.781; p < 0.05). The immunohistochemical micromethod may make it possible to evaluate the histopathological diagnosis of mesangial proliferative glomerulonephritis quantitatively.

Animals↗

Properties of Ca(2+)-dependent K+ channels of human gingival fibroblasts.

Cells in the oral cavity are normally exposed to different temperatures. Ion transport systems are influenced by temperature in other tissues: In particular, changes in intracellular K+ ion can affect cell growth and synthesis of macromolecules. The purpose of this investigation was to identify K+ channels in human gingival fibroblast cells and analyze the effect of temperature on their K+ conduction properties. Ca(2+)-dependent K+ channels with a large conductance (125 pS in symmetrical K(+)-rich solutions) were identified in human gingival fibroblasts and studied by the patch-clamp technique. The open probability of the channels varied with membrane potential between +40 and -100 mV. When the bath temperature was decreased from 40 to 4 degrees C, channel conductance was reduced, but the mean open time of the channels was increased. The activation energies for the conductance and the reciprocal of the mean open time were estimated to be 9.1 and 22.9 kJ/mol, respectively. These values are lower than those reported for these and other types of channels in cells from other tissues. The open probability of the channels was nearly constant in the temperature range studied. These results suggest that the properties of Ca(2+)-dependent K+ channels of gingival fibroblasts remain relatively unchanged when the cells are exposed to a wide range of temperatures.

Body Temperature↗

Congenital anophthalmia with caudal vertebral anomalies in Japanese Brown cattle.

Anophthalmia and caudal vertebral anomalies such as taillessness or wry tail were morphologically examined in ten Japanese Brown calves obtained in Kumamoto Prefecture. The anomalous calves lacked eyeball bilaterally or unilaterally but had small-sized eyelids and narrow palpebral fissures. A small cystic, solid or spot-like remnant of eyeball (REB) was buried in the mixture of vestigial extraocular muscles, lacrimal gland and adipose tissue of the orbit. The REB was composed of irregularly arranged elements of ocular wall such as sclera, choroid and retina. The retina was often dysplastic and connected to the hypoplastic optic nerve. These morphological changes might represent the defective processes after the formation of the optic vesicle or cup. Therefore, this eye defect may be defined as degenerative anophthalmia. The defects of the vertebral body such as wedge vertebra, hemivertebra, and sagittal cleft vertebra seen in the lumbar, sacral, and coccygial regions and the meandering of the axial line of abnormal vertebrae may suggest the failure of notochord formation in the early fetal period. From the embryological point of view, it seemed possible that the calves were exposed to teratogen at the critical time of optic organogenesis and notochordal formation. The cause of anomalies could not be determined in this study.

Animals↗

17 alpha-Hydroxyprogesterone suppresses neovascularization induced by HPLC-purified ovarian hyaluronic acid-like glycosaminoglycan in mice.

HPLC-purified glycosaminoglycans (hpGAG) prepared from extracts of non-luteal mouse (JcL:ICR strain) ovaries were assayed for neovascularization by implanting Elvax films, containing test samples, on the lateral wall of the sheath of m. rectus abdominis in adult female mice of the same strain. Neovascularization occurred in a dose-dependent manner, and was characterized by capillary outgrowth extending into the tissue surrounding the implant. The single major peak of purified GAG on a column of TSK gel DEAE got out of order after treatment with streptococcal hyaluronidase or nitrous acid. The activity of this fraction was also greatly reduced when treated with streptococcal hyaluronidase or nitrous acid. When hpGAG was embedded in the implant with 17 alpha-hydroxyprogesterone at a dose of 20 micrograms/film, neovascularization induced by means of hpGAGs was suppressed. Progesterone at a dose of 50 micrograms/film did not suppress the neovascularization induced by ovarian hpGAG. These findings suggest that 17 alpha-hydroxyprogesterone suppresses the angiogenic activity of hyaluronic acid-like hpGAG in the ovary.

17-alpha-Hydroxyprogesterone↗