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Biomedical subjects

T Miyata

Publications and source records attributed to T Miyata.

At least 415 records · Page 23Linked to original sources
PubMed1994

Evaluation of the proteolytic activity of factor D accumulated as an active serine protease in patients with chronic renal failure.

Complement factor D, a complement system serine protease, circulating in vivo as its active form, accumulates in patients with chronic renal failure. The pathophysiological role of this active protease in these patients was examined by studies on activities of excess factor D on 10 synthetic peptide substrates for some usual serine proteases. The most sensitive of these substrates to factor D was Boc-Gln-Ala-Arg-MCA, which is used as a substrate for trypsin. The proteolytic activity of factor D (2.17 unit/mg/h) on this substrate was estimated to be 10(-5)-fold that of trypsin (2.18 x 10(5) unit/mg/h). The activities of factor D on other synthetic substrates were lower. Thus the proteolytic activity of factor D is considered to be very specific for its natural substrate, complement factor B bound with C3b, even when it is highly accumulated in vivo. The inhibitory effects of some serine protease inhibitors used clinically (nafamostat mesilate, sepinostat mesilate, camostat mesilate and gabexate mesilate) on the proteolytic activity of factor D on its natural substrate, factor B, were also investigated. Of these synthetic compounds, nafamostat mesilate was the most effective inhibitor (ID50:25 microM) of the activity of factor D on factor B.

Amino Acid Sequence↗
PubMed1994

Fluorogenic assay of activated factor VII. Plasma factor VIIa levels in relation to arterial cardiovascular diseases in Japanese.

Factor VII (FVII) plays an important role in initiation of the tissue factor-induced coagulation pathway. An increase in FVII coagulant activity (FVIIc) has been proposed as an independent risk factor for coronary artery disease. However, it remains uncertain whether high FVIIc levels are due to an increase in the activation of FVII or an increase in the concentration of FVII mass. We developed a new fluorogenic assay for plasma activated FVII (FVIIa) that used soluble tissue factor. The sensitivity of this assay ranged from 0.2 to 1000 ng FVIIa per milliliter of plasma. Plasma FVIIa levels were measured in 110 healthy subjects and 93 patients with hypertension, diabetes mellitus, and/or cardiovascular disease. The mean plasma FVIIa level in healthy Japanese individuals was 2.5 ng/mL, which was lower than that in Western subjects. Gel filtration analysis showed that most of the circulating FVIIa was in a free form, and binding of FVIIa to tissue factor in plasma was not detected. Aging increased both the FVIIa level and FVII mass, whereas menopause increased mainly the FVII mass. Elderly patients with arterial cardiovascular diseases showed increases in plasma FVIIa levels and FVIIa to FVII antigen (FVII:Ag) ratios. Among the elderly, arterial cardiovascular disease was more common in a high-FVIIa than a low-FVIIa group. Plasma FVIIa levels were not correlated with serum levels of total cholesterol or triglycerides. The FVIIa level and the FVIIa-to-FVII:Ag ratio were positively correlated with fibrinogen level and negatively correlated with body mass index and serum albumin level in the elderly. In conclusion, aging, cardiovascular disease, and malnutrition increased plasma FVIIa levels. FVIIa levels were not correlated with lipid levels or hepatic synthesis, suggesting that FVIIa may be an independent risk factor for cardiovascular disease.

Adolescent↗
PubMed1994

Efficient repopulation of denuded rabbit arteries with autologous genetically modified endothelial cells.

BACKGROUND: In an effort to determine whether specific genetic modifications of cells of the vascular system might improve the efficacy of existing clinical procedures such as endarterectomy, atherectomy, and percutaneous angioplasty, we investigated the utility of gene transfer to rapidly and efficiently repopulate injured arteries with genetically modified cells in an animal model. METHODS AND RESULTS: The method involves the harvest of autologous venous-derived endothelial cells, the efficient genetic modification of the cells through the use of recombinant retroviruses, and the subsequent implantation of the genetically modified cells on the surface of balloon-denuded arterial segments. With a rabbit model, freshly isolated endothelial cells were transduced with a recombinant retrovirus encoding the bacterial enzyme beta-galactosidase. The autologous transduced cells were then implanted on the surface of balloon-denuded ileofemoral arterial segments at different cell densities; after 1 to 14 days, the animals were killed, and the vessel segments were examined. Cells expressing the bacterial gene product, as determined by in situ staining for beta-galactosidase, were found to be present on the surface of 28 of the 32 arteries seeded with genetically modified cells. Vessels examined at 4 to 7 days after seeding displayed 40% to 90% coverage with transduced cells, even when seeded at subconfluent density, and an intact endothelial cell monolayer, as evidenced by scanning electron microscopy studies. Vessels examined at 14 days after seeding revealed more variable staining for beta-galactosidase yet, again, in most cases, an intact endothelial cell monolayer. CONCLUSIONS: These studies indicate the feasibility of generating segments of arterial vessels containing genetically modified cells in a rapid and efficient fashion. Further studies are now necessary to determine whether the local expression of specific polypeptides within a region of vessel for a finite period of time will be clinically useful.

Animals↗
PubMed1994

Formation of poly(glucosyloxyethyl methacrylate)-concanavalin A complex and its glucose-sensitivity.

The complex formation between Concanavalin A (Con A) and a polymer having pendant glucose groups was studied in order to design a glucose-sensitive polymer. The polymer having pendant glucose (poly(glucosyloxyethyl methacrylate) or (poly(GEMA)) forms a complex with Con A in tris HCl buffer (pH = 7.5). The solution then becomes turbid due to the multiple associations between poly(GEMA) and Con A. When free glucose or mannose are added to the turbid solution, the solution becomes transparent again. However, the addition of galactose does not cause the solution to be transparent. This indicates that Con A prefers to form a complex with free glucose or mannose (but not galactose) rather than with the pendant glucose in poly(GEMA). Therefore, the complex between poly(GEMA) and Con A is expected to be glucose- and mannose-sensitive. The apparent dissociation constants of the complexes between saccharide (poly(GEMA), glucose, and mannose) and Con A were also determined by affinity electrophoresis.

Biosensing Techniques↗
PubMed1994

Involvement of beta 2-microglobulin modified with advanced glycation end products in the pathogenesis of hemodialysis-associated amyloidosis. Induction of human monocyte chemotaxis and macrophage secretion of tumor necrosis factor-alpha and interleukin-1.

beta 2-Microglobulin (beta 2M) is a major constituent of amyloid fibrils in hemodialysis-associated amyloidosis (HAA), a complication of long-term hemodialysis. However, the pathological role of beta 2M in HAA remains to be determined. Recently, we demonstrated that beta 2M in the amyloid deposits of HAA is modified with advanced glycation end products (AGEs) of the Maillard reaction. Since AGEs have been implicated in tissue damage associated with diabetic complications and aging, we investigated the possible involvement of AGE-modified beta 2M (AGE-beta 2M) in the pathogenesis of HAA. AGE- and normal-beta 2M were purified from urine of long-term hemodialysis patients. AGE-beta 2M enhanced directed migration (chemotaxis) and random cell migration (chemokinesis) of human monocytes in a dose-dependent manner. However, normal-beta 2M did not enhance any migratory activity. AGE-beta 2M, but not normal-beta 2M, increased the secretion of TNF-alpha and IL-1 beta from macrophages. Similar effects were also induced by in vitro prepared AGE-beta 2M (normal-beta 2M incubated with glucose in vitro for 30 d). When TNF-alpha or IL-1 beta was added to cultured human synovial cells in an amount equivalent to that secreted from macrophages in the presence of AGE-beta 2M, a significant increase in the synthesis of collagenase and morphological changes in cell shape were observed. These findings suggested that AGE-beta 2M, a major component in amyloid deposits, participates in the pathogenesis of HAA as foci where monocyte/macrophage accumulate and initiate an inflammatory response that leads to bone/joint destruction.

Adult↗
PubMed1994

[Effects of a novel orally-active antiallergic drug, quinotolast (FK021), on airway clearance].

The effects of a novel antiallergic drug, quinotolast (FK021, sodium 5-(4-oxo-1-phenoxy-4H-quinolizine-3-carboxamido) tetrazolate monohydrate), on airway clearance was studied in comparison with those of tranilast (an orally-active antiallergic drug). FK021 (10 mg/kg, p.o.) did not influence the rabbit airway secretion, whereas tranilast (100 mg/kg, p.o.) caused a slight suppression. Neither FK021 (10(-10)-10(-5) g/ml) nor tranilast (10(-6), 10(-4) g/ml) had any effect on pulmonary surfactant secretion in rat type II pneumocytes. FK021 (10 mg/kg, p.o.) caused a significant increase in the mucociliary transport rate in quails, whereas tranilast (320 mg/kg, p.o.) had no effect. Antitussive effects were examined in normal guinea pigs and guinea pigs made bronchitic by an exposure to SO2. FK021 (10 mg/kg, p.o.) and tranilast (320 mg/kg, p.o.) significantly depressed the cough reflex induced by citric acid in normal animals. FK021 (32 mg/kg, p.o.), but not tranilast (320 mg/kg, p.o.), showed antitussive effects on citric acid-induced cough in bronchitic animals. These results suggest that FK021 may have favorable effects on expectoration and cough reflex observed in the patients with chronic obstructive pulmonary diseases such as asthma.

Administration, Oral↗
PubMed1994

The relationship between the sialic acid concentrations in the serum and whole saliva in rats with naturally occurring gingivitis.

Pocket probing depth was correlated with the amount of salivary sialic acid in pilocarpine-stimulated saliva in ODU plaque-susceptible rats (ODUS/Odu) (r = 0.657, P < 0.01), but not with the content of serum sialic acid. There was no difference in the amount of serum sialic acid content between ODUS/Odu and plaque-resistant rats. These results suggest that the amount of sialic acid in the saliva can be a useful index of the severity of periodontal disease.

Animals↗
PubMed1994

A possible link between molecular evolution and tissue evolution demonstrated by tissue specific genes.

In this paper, we reviewed our recent works on a possible link between molecular evolution and tissue evolution. The evolutionary rates of genes that are expressed tissue specifically were shown to differ widely to one another, depending on tissues: Brain specific genes evolve with significantly slower rate than immune specific genes. The tissue dependence of molecular evolutionary rate strongly suggests the presence of functional constraints against molecular changes from tissue level. A molecular phylogenetic analysis of tissue specific isoforms that are identical to one another in function, but differ only in tissue distribution revealed frequent gene duplications and rapid accumulations of amino acid substitutions during the early evolution of chordates, where rapid evolution at the tissue or organ levels is thought to have occurred. On the basis of functional constraints, a possible explanation for the correlation between evolution at the two levels was presented.

Animals↗
PubMed1994

Mammalian phylogeny inferred from multiple protein data.

On the basis of multiple protein data, the phylogenetic relationships among the major clades of eutherian mammals, Primates, Cetacea, Artiodactyla, Carnivora, Lagomorpha, Myomorpha, and Caviomorpha, were analyzed by the maximum likelihood (ML) method, together with the maximum parsimony (MP) method and the neighbor joining (NJ) method. Using nineteen protein data, we first reexamined the hypothesis of rodent polyphyly proposed by Graur et al. (1991) based on the four taxon system, consisting of human, mouse (or rat), guinea-pig and an outgroup. The ML analysis does not support the rodent polyphyly, but strongly favors the traditional view of rodent monophyly representing the Myomorpha-Caviomorpha association. In the second analysis, the phylogenetic position of Lagomorpha were reexamined using three sets of multiple protein data, 13 mitochondrial DNA-coded proteins from rodents, rabbit, human, seals, bovine, whales and opossum, 25 protein data from human, rabbit and rodents and an outgroup, and 20 protein data from human, rabbit, bovine (or pig) and rodents. Our analysis favors the closer affinity of Lagomorpha to Primates than to Rodentia. The branching sequence of seven mammalian groups revealed by the present analyses is as follows: Myomorpha-Caviomorpha clade (Rodentia), rabbit, Primates, Carnivora, and Artiodactyla-Cetacea clade. Judging from the branch lengths measured by the numbers of synonymous substitutions, a series of divergence of these mammals is likely to be bush-like. The rapid rate of rodent evolution was reexamined in the light of the new phylogeny.

Amino Acid Sequence↗
PubMed1994

[A case of ruptured hepatocellular carcinoma with rapid growth after the interferon therapy in chronic hepatitis].

A 75-year-old man who had had a long history of chronic hepatitis C virus liver disease underwent interferon (IFN) therapy for three months. After three months of the last IFN injection, he presented with a sudden attack of epigastralgia. Abdominal US and CT revealed a tumor about 9 x 10 cm in the right lobe of the liver. Serum AFP level was 8,499 ng/ml. Spontaneous hemoperitoneum due to ruptured hepatocellular carcinoma (HCC) was suspected. Hepatectomy was carried out on 35 days postembolization. Pathological examination revealed HCC of trabecular type. Although the relation between the rapid growth and IFN is not clear, US examination should be performed once a month for the patient.

Aged↗
PubMed1994

Expression cloning of genes for GPI-anchor biosynthesis.

Cloning genes for glycosylphosphatidylinositol (GPI)-anchor biosynthesis is important to further understand its mechanisms and regulation. We have been using expression cloning methods in which a cDNA library was transfected into GPI-anchor-deficient mutant cells. The transfectants which restored surface expression of GPI-anchored proteins were isolated and the plasmids were rescued. In this way we previously cloned cDNAs of genes for complementation classes A and F, and named them PIG-A and PIG-F, respectively. In the present study we have cloned the gene for class B, termed PIG-B. In each case we used different methods. For cloning PIG-A cDNA we used a cDNA library made with an Epstein-Barr-virus-based vector and human class A mutant JY5 which expresses EBNA-1 protein. The EBNA-1 protein allows stable replication of oriP-containing plasmids in the episomal form. For cloning PIG-F cDNA we chose a transient expression method and cotransfected a human T-cell cDNA library made with a vector bearing an origin of replication of polyoma virus with a plasmid bearing polyoma virus large T into the class F murine thymoma mutant. This cotransfection strategy was unsuccessful for cloning PIG-B due to low transfection efficiency of the class B thymoma mutant SIA-b. Thus, we first established large T-expressing SIA-b cells and then transfected them with a cDNA library. PIG-B cDNA restored the surface expression of Thy-1 on SIA-b cells and also synthesis of mature type GPI-anchor precursors in these cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Cloning of the cDNA for a novel receptor tyrosine kinase, Sky, predominantly expressed in brain.

Based on homology to the tyrosine kinase domain of the chick erythroblastosis virus oncogene v-sea, we cloned a cDNA encoding a novel receptor tyrosine kinase from a human hepatoma HepG2 cDNA library. The encoded protein, which we termed 'Sky', contains an intracellular tyrosine kinase domain and a unique extracellular domain with two immunoglobulin (Ig)-like domains and two fibronectin type III (FN III) domains. The overall structure of Sky is homologous to the reported sequence of the oncogenic Axl/Ufo receptor tyrosine kinase. Phylogenetic analysis in the tyrosine kinase domain shows that Sky, Axl/Ufo, Ark, and v-Ryk form a sub-family distinct from other tyrosine kinases. Northern blot analysis revealed that sky mRNA is expressed predominantly in brain and faintly in tissues of other organs. As the combination of Ig-like and FN III domains is often observed in neural cell adhesion molecules and receptor protein tyrosine phosphatases, Sky may be involved in cell adhesion processes, particularly in the central nervous system.

Amino Acid Sequence↗
PubMed1994

Three missense mutations in the protein C heavy chain causing type I and type II protein C deficiency.

We have studied the molecular basis of protein C deficiency in three families with a history of thromboembolic disease. An approximately 50% reduction in both functional and immunologic levels of protein C was detected in the plasma from two unrelated patients, designated protein C Osaka 1 and protein C Osaka 2. An approximately 50% reduction in functional level but normal immunologic level of protein C was detected in plasma from a third patient, designated protein C Osaka 3. DNA sequencing of the amplified DNA revealed one missense mutation in each case. Additional mutations in the coding sequence were excluded by DNA sequencing of all protein C exons. We identified a C-to-T change at nucleotide number 6,218 of the protein C gene in protein C Osaka 1. This results in the amino acid substitution of Arg-169 by Trp at the alpha-thrombin cleavage site. In protein C Osaka 2, a G-to-A change at nucleotide number 8,807 was identified leading to the amino acid substitution of Met-364 by Ile in the protease domain. This substitution may impair the synthesis or stability of protein C Osaka 2. In protein C Osaka 3, a G-to-C change at nucleotide number 8,868 was identified. This results in substitution of Gly-385 by Arg in the protease domain. Based on these, it was concluded that Arg-169-to-Trp mutation and Met-364-to-Ile mutation cause type I protein C deficiency and Gly-385-to-Arg mutation causes type II deficiency.

Adult↗
PubMed1994

Identification of a human cDNA encoding a novel protein kinase with two repeats of the LIM/double zinc finger motif.

By low-stringency screening of a human hepatoma HepG2 cell cDNA library, using the genomic fragment of chick c-sea receptor tyrosine kinase as a probe, we isolated overlapping cDNAs encoding a novel protein kinase, which we termed LIM-kinase (LIMK).* The predicted open reading frame encodes a 647-amino-acid polypeptide containing a putative protein kinase structure in the C-terminal half. In addition, LIMK has two repeats of cysteine-rich LIM/double zinc finger motif at the most N-terminus. To our knowledge, this is the first protein kinase seen to contain the LIM motif(s) in the molecule. Although the protein kinase domain of LIMK has highly conserved sequence elements of protein kinases, phylogenetic analysis revealed that LIMK cannot be classified into any subfamily of known protein kinases. Northern blot analysis revealed that the single species of LIMK mRNA of 3.3 kb was expressed in various human epithelial and hematopoietic cell lines. In rat tissues, LIMK mRNA was expressed in the brain, at the highest level. LIM is suggested to be involved in protein-protein interactions by binding to another LIM motif. As the LIM domain is frequently present in the homeodomain-containing transcriptional regulators and oncogenic nuclear proteins, LIMK may be involved in developmental or oncogenic processes through interactions with these LIM-containing proteins.

Amino Acid Sequence↗
PubMed1993

Human dihydrolipoamide succinyltransferase: cDNA cloning and localization on chromosome 14q24.2-q24.3.

We isolated cDNA for dihydrolipoamide succinyltransferase from a human fibroblast cDNA library in lambda gt11. The cDNA revealed that the human dihydrolipoamide succinyltransferase lacked a sequence motif of an E3 and/or E1 binding site. This suggests that the human dihydrolipoamide succinyltransferase possesses a unique structure consisting of two domains in contrast with the dihydrolipoamide acyltransferases of other alpha-keto acid dehydrogenase complexes. In addition, we found that the human dihydrolipoamide succinyltransferase gene is located on chromosome 14 at q24.2-q24.3 and that a sequence related to the dihydrolipoamide succinyltransferase gene is located on chromosome 1 at p31. Interestingly, the gene for the dihydrolipoamide acyltransferase of the branched chain alpha-keto acid dehydrogenase complex is also located on chromosome 1p31 (Zneimer et al. (1991) Genomics 10, 740-747).

Acyltransferases↗