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Y Urata

Publications and source records attributed to Y Urata.

At least 73 records · Page 4Linked to original sources

Novel separation and amino acid sequences of alpha and beta subunits of pig heart pyruvate dehydrogenase.

Pyruvate dehydrogenase was separated from pig heart pyruvate dehydrogenase complex by gel-permeation chromatography on a TSK G4000SWG column in the presence of 4 M urea, followed by chromatography on a calcium phosphate gel-cellulose column. The pyruvate dehydrogenase was further separated into two nonidentical subunits, alpha and beta, by high-performance liquid chromatography on a Synchropak CM-300 column in the presence of 8 M urea. The complete amino acid sequences of two subunits of pyruvate dehydrogenase were determined. The peptide fragments of S-carboxymethylated subunits were generated by treatment with endoproteinase Lys-C, endoproteinase Asp-N, trypsin, and cyanogen bromide. The subunits alpha and beta contain 361 amino acid residues (Mr 40,294) and 329 residues (Mr 35,787), respectively. The amino acid sequences of subunits alpha and beta in the pig were, respectively, 98 and 96% identical to those in humans. Hydropathy analysis and prediction of the secondary structure of two subunits suggest that the subunit alpha contains the thiamin pyrophosphate-binding domain and that the subunit beta contains segments with a high hydrophobicity.

Amino Acid Sequence↗

[The examination of DNA strand breaks induced by peplomycin-using non-radioactive in situ nick translation method].

In situ nick translation (ISNT) is a method to detect DNA single strand break (nick) at each cellular level. In this study, peplomycin (PEP)-induced DNA strand breaks and its repair were examined using non-radioactive ISNT method. Human fibroblasts were cultured with various concentrations of PEP for different durations, and then DNA strand breaks were evaluated by ISNT method. The signal intensity of DNA strand breaks of the PEP-treated fibroblasts was increased in a dose dependent manner. At the fixed concentration of PEP, it reached to a maximum level after 10 min. of culture and remained unchanged for at least 24 hours. In addition, DNA strand breaks induced by PEP was rapidly repaired in 10 min. after washing. Thus, the non-radioactive ISNT is thought to be a quick, sensitive and specific method not only to detect DNA strand breaks but also to observe its repair.

Bleomycin↗

Amylase-producing multiple myeloma.

We managed a case of amylase-producing multiple myeloma with extensive extramedullary spread. We reviewed five cases of amylase-producing multiple myeloma, including this case. This type of multiple myeloma has shown unique clinicopathologic features. (1) A distinct elevation of the serum amylase activity was demonstrated in all five patients. The amylase isozyme was of the S type without exception. (2) Extensive extramedullary spread with extramedullary tumors and/or myelomatous pleural effusions or ascites was seen in all five patients during the course of illness. (3) In three of four cases in which it was mentioned, extensive destruction of multiple bones was demonstrated roentgenographically. (4) In four patients, excepting one with a solitary bone lesion, the survival from the initial therapy was shorter than 1 year. (5) Myeloma cell lines were established in three cases. A common feature of these three cell lines was a translocation of chromosome 1, which supplied the amylase gene. This finding may be pathogenetically related to this entity.

Aged↗

[Progression from Crow-Fukase syndrome with double gammopathy (IgM-kappa, IgG-lambda) to primary macroglobulinemia].

This report deals with a case of double gammopathy (IgM-kappa, IgG-lambda) with Crow-Fukase syndrome, which developed into primary macroglobulinemia four years after the diagnosis. In May 1980, a 74-year-old woman was admitted to the hospital because of a rapid progression of peripheral neuropathy. The patient was diagnosed as having Crow-Fukase syndrome from the following data: albumin-cytologic dissociation of cerebrospinal fluid, peripheral edema, diffuse hyperpigmentation of the skin, diabetic glucose intolerance, serum double gammopathy (IgM-kappa, IgG-lambda) and hepatomegaly. The administration of prednisolone yielded the improvement of neuropathy. In December 1984, serum IgM level was increased from 104 mg/dl to 3,025 mg/dl. Plasma cells in the bone marrow increased in the percentage from 5.6% to 18.4%, and then Bence Jones protein (kappa type) was excreted in the urine. No antibody activity to myelin antigens was detected in the serum. The patient died of cerebral infarction in 1985. At postmortem examination, lymphomatous involvement was found in the jejunum. At the immunohistological examination of the tumor specimens, the morphology and the distribution of IgM- and IgG-positive cells corresponded to that of kappa- and lambda-positive cells, respectively. A small number of cells containing both kappa and lambda light chains were also demonstrated. It seems likely that IgM (kappa)- and IgG (lambda)-positive cells were derived from the common precursor cells.

Aged↗

Characterization and nucleotide sequence of the gene encoding the human pyruvate dehydrogenase alpha-subunit.

Genomic clones encompassing the entire gene (PDH alpha) encoding the human pyruvate dehydrogenase alpha-subunit (PDH alpha) have been isolated by screening a leukocyte genomic library in the cloning vector, lambda EMBL4. The PDH alpha gene spans 17082 bp and is composed of eleven exons and ten introns. All intron/exon splice junctions follow the GT/AG rule. A total of seven Alu repeats were found in five introns. The entire nucleotide (nt) sequence of the PDH alpha gene has been determined and typical consensus promoter sequences in the 5'-flanking region were found. The results of primer extension analysis imply that the PDH alpha gene transcription start point (tsp) is a thymine residue 124 bp upstream from the ATG start codon in exon 1. The structural organization and the tsp were compared with the recent report [Maragos et al., J. Biol. Chem. 264 (1989) 12294-12298]. Analysis of the PDH alpha gene resolves existing discrepancies among four published sequences of PDH alpha cDNAs. A 93-bp sequence that was missing in our sequence of cultured foreskin fibroblast PDH alpha cDNA [Koike et al., Proc. Natl. Acad. Sci. USA 85 (1988) 41-45] was identified in the gene as exon 6.

Amino Acid Sequence↗

Ph chromosome in a patient with non-leukemic non-Hodgkin B-cell lymphoma.

A standard Philadelphia translocation, t(9;22) (q34;q11), was found in lymph node cells from a patient with non-leukemic non-Hodgkin lymphoma at the time of diagnosis. The rearrangement of the breakpoint cluster region (bcr) was not detected with a bcr-3' probe. The neoplastic clone was of monoclonal B-cell character with E-, CD5-, CD10-, CD13-, CD19+, CD20+, CD21+, CD25-, HLA DR+, and positive surface Ig(kappa). The patient showed no evidence of chronic myelogenous leukemia.

Aged↗

Molecular cloning and characterization of human pyruvate dehydrogenase beta subunit gene.

A genomic clone encompassing the entire gene for the human pyruvate dehydrogenase beta subunit (PDH beta) has been isolated by screening a leukocyte genomic library with a nick-translated human foreskin fibroblast PDH beta cDNA probe. The 18-kilobase clone was characterized by restriction enzyme analysis, extensive DNA sequencing, and primer-extension analysis. The PDH beta structural gene is composed of 10 exons and 9 introns. All intron-exon splice junctions follow the GT/AG rule. The Alu family was found in introns 2 and 8. The 5' flanking region of the PDH beta gene contains a "CAAT" consensus promoter sequence but no "TATA" sequence. Primer-extension analysis indicated that the PDH beta gene transcription start site is an adenine residue located 132 bases upstream from the initiation codon in exon 1.

Amino Acid Sequence↗

[A myeloma (IgG-kappa) terminating in acute myelogenous leukemia].

A 71-year-old man was hospitalized in November, 1983 for a back pain and a diagnosis of multiple myeloma was made, based on the Bence Jones proteinuria, The serum M-component of a IgG-kappa type (3.3 g/dl), and plasmacytosis in the bone marrow (37%). Treatment consisted of melphalan and prednisolone. A blood count in March, 1986 revealed 6000/microliters of WBC with 30% of a blast form and 8% plasma cells, and 20,000/microliters of platelets. A bone marrow aspirate revealed that 14% were myeloblasts and 26% were plasma cells. Distinguishing the myeloblasts from the immature plasma cells in the peripheral blood proved difficult. Studies by electron microscopy and an immunological inspection of phenotypes were helpful in achieving a determination. A karyotypic analysis of the bone marrow cells indicated a hypodiploid cell population, a marker chromosome, and a karyotypic instability. These findings indicate that his multiple myeloma had undergone a leukemic change associated with acute myelogenous leukemia.

Aged↗

[Cellular kinetics and the nuclear ploidy pattern in relation to the gross pathological classification and location of advanced colorectal cancers as analyzed by DNA-RNA cytofluorometry].

Correlated studies on the cellular kinetics in association with the nuclear ploidy pattern and the pathological morphology of advanced colorectal cancers have been carried out using DNA-RNA cytofluorometry with an AO stain. The results showed that advanced colorectal cancers could be separated into the 3 distinct groups that follow: group I, involving a diploid cell population; group II, a polyploid cell population; and, group II', an aneuploid cell population. The group I type was located in the entire colorectal region, where as groups II and II', were in the sigmoid and rectum. Three groups were mostly Borrmann type 3 cancers, and the gross pathological classification was not found to be related to the ploidy pattern.

Adult↗

Acute nonlymphocytic leukemia (M2) with chromosome abnormality trisomy 4 developing eight years after radiation therapy for breast cancer.

We report here the development, 8 years after radiation therapy for breast cancer, of acute nonlymphocytic leukemia (ANLL), type M2 of the FAB classification, in which trisomy 4 was detected as the only chromosomal abnormality. Simultaneous observation of cytologic and cytogenetic features of individual colonies derived from leukemic progenitor (L-CFU) and early progenitor (CFU mix) cultures in this patient revealed that all colonies examined had a normal karyotype, although the clone with trisomy 4 was predominant in the direct bone-marrow culture. These findings suggest that progenitor cells with trisomy 4 were less predominant in colony growth when stimulated by colony-stimulating factors (CSFs) than were stem cells with a normal karyotype.

Breast Neoplasms↗

Disseminated infection of Pneumocystis carinii in a patient with the acquired immunodeficiency syndrome.

This report describes the histopathology of a disseminated Pneumocystis carinii infection in a 24-year-old Japanese male haemophiliac diagnosed as having the acquired immunodeficiency syndrome. He developed respiratory symptoms, and Pneumocystis carinii pneumonia was confirmed by transbronchial lung biopsy. On the 70th day of hospitalization the patient died. Autopsy findings revealed Pneumocystis carinii not only in the lungs but also in the stomach, jejunum, ileum, colon, mesoappendix, abdominal lymph nodes, diaphragm, and thyroid gland.

Acquired Immunodeficiency Syndrome↗

Molecular cloning of cDNAs for alpha and beta subunits of human pyruvate dehydrogenase.

The cDNAs encoding human PDH alpha and PDH beta were isolated from a HeLa cell cDNA library in the lambda gt11 expression vector by immunoscreening, followed by colony hybridization from a human foreskin fibroblast cDNA library. Nucleotide sequence analyses of the positive plasmid clones (pHPDA and pHPDB) revealed an insert of 1.36 kilobases (kb) for PDH alpha and one of 1.69 kb for PDH beta, respectively, allowing us to predict the complete amino acid sequences of the precursor and mature proteins of these two subunits. The amino acid sequences of the amino-terminal regions of the two subunits of human PDH were highly homologous with those of mature porcine PDH. The amino acid sequences of phosphorylation sites determined in PDH alpha of the bovine and porcine enzymes were also conserved in the human PDH alpha. Blot analysis of HeLa cell poly(A)+ RNA and the transcriptional product of the two cDNAs showed a single mRNA of 1.8 kb for PDH alpha and one of 1.7 kb for PDH beta. The precursor proteins of PDH alpha and PDH beta were detected by immunoprecipitation from an 35S-labeled, cell-free translation system. Our sequence of PDH alpha cDNA was compared with those of two other origins. The differences among these three PDH alpha cDNAs have been discussed.

Amino Acid Sequence↗

[CD3-, OKM1+, Leu7-, Leu11+ large granular lymphocyte leukemia with ascites and CNS involvement].

A case of large granular lymphocyte (LGL) leukemia with ascites and CNS involvement was reported. A 39-year-old Japanese female was admitted to our hospital in March, 1987 because of high fever. Her clinical and hematological features were characterized by generalized lymphadenopathy, marked hepatosplenomegaly, high serum LDH level (3,257 mU/ml), marked leukocytosis (71,000/microliters) with 74% LGLs and bone marrow infiltration with 57% LGLs. Despite of chemotherapy, ascites, retroperitoneal mass and CNS involvement developed and she died of sepsis after three months. LGLs from the patient's blood, marrow and ascites, stained positively for acid phosphatase. These LGLs were E rossete+ and Fc (IgG) receptor+ and were positive for CD2, OKM1, HLA-DR and Leu11, but were negative for CD1, CD3, CD4, CD8 and Leu7 as well as for terminal deoxynucleotidyl transferase activity. The natural killer activity against K562 target cells was high and was significantly augmented after stimulation by recombinant human interleukin 2. These LGLs also demonstrated normal antibody-dependent cytotoxicity activity. Cytogenetic study on bone marrow cells and ascitic cells revealed clonal chromosomal abnormalities. These clinical, hematological, immunological and cytogenetic findings suggest that this patient had a neoplastic proliferation of natural killer cells.

Adult↗

[Spontaneous remission of four months' duration in hypoplastic leukemia with tetraploid chromosome after blood transfusions and infection].

Spontaneous complete remission of four months' duration was observed in a 51-year-old male with hypoplastic leukemia. Cytogenetical analysis revealed that leukemia cells of this patient were tetraploid. The diameter of leukemia cells involving myeloid cells ranged from 30 to 50 mu. The remission was apparently associated with repeated blood transfusions and severe infection. Complete remission was confirmed by normal morphology and karyotype of the bone marrow cells, although in vitro marrow stem cell growth did not return to normal. Thus, normal hematopoiesis may not have recovered when the diagnosis of spontaneous remission was made.

Aneuploidy↗

Cloning and sequencing of cDNAs encoding alpha and beta subunits of human pyruvate dehydrogenase.

The cDNAs encoding fragments of the alpha and beta subunits (PDH alpha and PDH beta) of human pyruvate dehydrogenase (PDH, EC 1.2.4.1) were isolated from a HeLa cell cDNA library in the lambda gt11 expression vector by immunoscreening. Phage cDNA fragments were subsequently used to screen a human foreskin fibroblast cDNA library by colony hybridization. Nucleotide sequence analyses of the positive plasmid clones (pHPDA and pHPDB) revealed an insert of 1.36 kilobases (kb) for PDH alpha and one of 1.69 kb for PDH beta, respectively, allowing us to predict the complete amino acid sequences of the precursor and mature proteins of these two subunits. A putative leader sequence of 29 amino acid residues was identified in pHPDA, resulting in a precursor protein of 392 amino acid residues (Mr 43,414) and a mature protein of 363 residues (Mr 40,334). A similar leader sequence of 30 amino acid residues in pHPDB was also identified, resulting in a precursor protein of 359 amino acid residues (Mr 39,046) and a mature protein of 329 residues (Mr 35,911). The amino acid sequences of NH2-terminal regions of the two subunits of human PDH were highly homologous with those of mature porcine PDH. The amino acid sequences of phosphorylation sites determined in PDH alpha of bovine and porcine enzymes were also conserved in the human PDH alpha. Blot analysis of HeLa cell poly(A)+ RNA showed a single mRNA of 1.8 kb for PDH alpha and 1.7 kb for PDH beta, respectively. The precursor proteins of PDH alpha and PDH beta were detected by immunoprecipitation from an 35S-labeled cell-free translation system.

Amino Acid Sequence↗