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Biomedical subjects

T Yamane

Publications and source records attributed to T Yamane.

At least 289 records · Page 16Linked to original sources
PubMed1995

[An operative case of the dumbbell type retromediastinal schwannoma].

Neurogenic tumors of the mediastinum with an intraspinal component connected by a narrowed segment in the intervertebral one are generally described as dumbbell or hour-glass tumors, which need cautious and precise diagnoses and remedies, compared with other neurological tumors. A 62-year-old male was admitted to our hospital for abnormal tumor shadow in the chest X-ray film. We diagnosed this case as dumbbell type neurogenic tumor by MRI and CT. An operation was performed by modified Grillo's method: One-stage operation. With patient in prone position, L-shaped skin incision was made. Through total laminectomy of T-5 and T-6 and resection of the 6th rib, paravertebral portion of the tumor was removed, and thoracotomy in the same position under the same view enabled us to remove the residual tumor. Histopathological diagnosis was schwannoma. After the operation, no neurological complications were detected.

Humans↗
PubMed1995

[Detected Epstein-Barr virus genome using polymerase chain reaction in peripheral mononuclear cells--subacute necrotizing lymphadenitis].

A 23-year old woman was admitted to our hospital because of fever, leukocytopenia, thrombocytopenia and cervical lymphadenopathy. From pathological findings of the cervical lymphnode she was diagnosed as subacute necrotizing lymphadenitis (SNL). Bone marrow aspiration was hypocellular with proliferation of mature histiocytes with hemophagocytosis. Lactate dehydrogenase was elevated to 1863 IU/l and mild liver dysfunction was observed. Hepatosplenomegaly, lymphnode swelling in abdominal cavity, pleural effusion and pericardial effusion were detected. After glucocorticoid therapy all such disorders were normalized. In this course, Epstein-Barr (EB) virus genome was detected by a polymerase chain reaction method from peripheral mononuclear cells. After normalization, EB virus genome became not to be detected. From present case we concluded that EB virus was expanded in clinical course and was associated with pathogenesis of SNL.

Adult↗
PubMed1995

Anomaly of the ulnar artery arising from the brachial artery.

A high origin of the ulnar artery was encountered in the body of an 87-year-old Japanese man in the course of normal anatomic dissection at Nara Medical University. The left ulnar artery originated from the brachial artery at the height of the left inferior angle of the left scapula and passed medially to the brachial artery and the median nerve. In the forearm, the ulnar artery passed deep to the flexor carpi ulnaris muscle. Thereafter, the two branches from the ulnar artery contributed to form the superficial and deep palmar arterial arches. The left brachial artery was divided into two terminal branches of the radial and common interosseous arteries.

Aged↗
PubMed1995

Acute non-lymphoblastic leukaemia with t(16;21): case report with a review of the literature.

A rare case of acute non-lymphoblastic leukaemia with chromosomal t(16;21)(p11;q22) translocation was studied at the molecular level. Most of the previous reports about the translocation have been described at a karyotypic level. Blast cells of this patient expressed the TLS-ERG chimeric mRNA. The clinical, morphological, karyotypic, and immunohistochemical aspects of this leukaemia are also presented with a review of the literature.

Adult↗
PubMed1994

Purification and properties of phosphatidylinositol-specific phospholipase C from Streptomyces antibioticus.

A novel type of phosphatidylinositol-specific phospholipase C (PI-PLC) was purified from culture supernatant of a strain of Actinomycetales, Streptomyces antibioticus. The purified enzyme showed a single band on native polyacrylamide gel electrophoresis (native PAGE) with a molecular weight of 32 kDa, but showed two polypeptides, named alpha- and beta-peptides, on sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) with molecular weights of 23 kDa and 15 kDa, respectively. From the results of both electrophoretic analysis and N-terminal amino acid sequencing, it was estimated that the enzyme was composed of alpha- and beta-peptides. The enzyme could hydrolyze phosphatidylinositol, but not any other glycerophospholipids. The enzyme had pH and temperature optima at around 7.0 and 30 degrees C, respectively, and was stable up to 50 degrees C when incubated at pH 8.0 for 30 min. The PI-PLC was strongly activated by SDS, sodium deoxycholate (SDC) and diethyl ether, but not by Triton X-100, and inhibited by cetylpyridinium chloride (CPC). The enzyme was activated a little by Ca2+ and was inhibited completely by a chelating agent such as ethylenediaminetetraacetic acid (EDTA) and glycoletherdiaminetetraacetic acid (EGTA). Their inhibitions were restored by the addition of Ca2+, suggesting that a certain amount of Ca2+ is essential for the enzymatic activity.

Amino Acid Sequence↗
PubMed1994

A differential cloning procedure of complex genomic DNA fragments.

We have developed a differential cloning procedure designed for cloning of anonymous restriction DNA fragments whose molecular sizes differ between two genomic DNA preparations from higher organisms. The procedure, which was extensively revised from the original one, consists of several steps as summarized below. (i) Digestion of two DNA preparations (target and reference DNA) with the same restriction enzyme (4 base cutter). (ii) Biotinylation of target DNA and conversion of reference DNA to nonamplifiable form by terminal dephosphorylation. (iii) Electrophoresis of the two DNA preparations through a synthetic gel with a large excess of reference DNA as a competitor. (iv) In-gel alkaline dissociation of DNA, followed by reassociation (in-gel competitive reassociation). (v) Elution of DNA from the gel and PCR after adapter ligation and adsorption of DNA onto streptavidin-coated matrix. By repeating these steps, we attained substantial enrichment (approximately 10,000-fold) of DNA fragments which were originally present at one copy or less per complex mammalian genome. The details of the procedure and its unique characteristics in cloning of altered genomic DNA fragments, particularly from mammalian genome, are discussed.

Adenine Phosphoribosyltransferase↗
PubMed1994

High-performance liquid chromatographic determination of 3'-hydroxy-5'-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin (KRM-1648) and its deacetyl metabolite in plasma, whole blood, urine and tissue samples in rats.

A reversed-phase high-performance liquid chromatographic method was developed for the determination of 3'-hydroxy-5'-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin (KRM-1648, I), a new rifamycin derivative, and its 25-deacetyl metabolite (KRM-1671, II) in plasma, whole blood, tissues and urine from rats. I and II were coextracted with an internal standard from each sample matrix by solid-phase extraction (Bond Elut). Plasma and urine were directly loaded onto Bond Elut, while whole blood and tissues were homogenized and extracted with methanol or dichloromethane-chloroform prior to Bond Elut extraction. The extracts were chromatographed on Shim-pack CLC-ODS(M) using acetonitrile-0.02 M citrate buffer containing 0.1 M sodium perchlorate (2:1, v/v), and peaks were detected at 643 nm. The validation data showed that the assays for I and II in plasma, whole blood, tissues and urine were selective, accurate and reproducible.

Animals↗
PubMed1994

Transplantation of allogeneic peripheral blood stem cells after myeloablative treatment of a patient in blastic crisis of chronic myelocytic leukemia.

A 48-year-old man in blastic crisis of chronic myelocytic leukemia received a transplant of allogeneic peripheral blood stem cells. The donor was his HLA-identical sister, who refused to donate bone marrow cells, but agreed to donate peripheral blood stem cells. The patient received standard transplant conditioning with cyclophosphamide (120 mg/kg) and busulfan (16 mg/kg). Peripheral blood stem cells were mobilized with granulocyte colony stimulating factor and collected by apheresis. After transplantation, the white blood cell count and the result of microscopic analysis of the bone marrow became normal, and the leukocyte karyotype became 46XX. DNA fingerprinting showed complete chimerism. Graft-versus-host disease was suppressed with cyclosporine and methyl-prednisolone. The patient died of recurrence of leukemia on day 102+.

Blast Crisis↗
PubMed1994

Risk analysis of carcinogenesis in the remnant stomach with measurement of ornithine decarboxylase activity.

Gastric ornithine decarboxylase (ODC) activity was measured as a biomarker of tumor-promoting activity in the remnant stomach of rats and humans. Gastrectomy of Wistar rats utilizing the Billroth I method caused a significantly high induction of ODC, and use of the Billroth II method caused a significantly higher induction of ODC than the Billroth I method. In humans, ODC activity of remnant gastric cancer tissue, normal-appearing mucosa of remnant gastric cancer patient, and remnant gastric mucosa without cancer after the Billroth II method were significantly higher than that of normal gastric mucosa without gastrectomy. ODC activity of remnant gastric mucosa without cancer after the Billroth II method was significantly higher than that after the Billroth I method. Risk of carcinogenesis was high in the remnant stomach, especially after the Billroth II method.

Animals↗
PubMed1994

Role of cysteine residues in esterase from Bacillus stearothermophilus and increasing its thermostability by the replacement of cysteines.

Bacillus stearothermophilus esterase contains two free cysteine residues at positions of 45 and 115, which react with sulfhydryl reagents resulting in a significant decrease in the enzymatic activity. To understand the role of the cysteine residues in catalytic regions of the esterase, the residues were replaced with serine or alanine by site-directed mutagenesis to construct four single-mutated enzymes (C45A, C45S, C115A, C115S) and two double-mutated ones (C45/115A and C45/115S). Wild-type and mutant enzymes were produced in Escherichia coli cells and purified to homogeneity to examine their chemical and kinetic properties. These mutant enzymes had esterase activity, which suggested that none of the cysteines were required for its activity. Moreover, replacement of both two-cysteine residues made the enzyme insensitive to p-chloromercuribenzoic acid and extensively stabilized it at high temperatures of around 70 degrees C. These results demonstrate that replacement of free cysteine residues by site-directed mutagenesis can improve the thermostability of thermophilic enzymes.

Base Sequence↗
PubMed1994

Phospholipase D from Streptomyces antibioticus: cloning, sequencing, expression, and relationship to other phospholipases.

The extracellular phospholipase D (PLD) gene from Streptomyces antibioticus was cloned, sequenced, and expressed in Escherichia coli. Analysis of DNA sequence data revealed a putative ribosome-binding site and an open reading frame encoding a 556-amino-acid protein that included amino acid sequences obtained from the purified enzyme. The protein was expressed in an insoluble form in E. coli, but reacted with antibody against PLD. After solubilization of the protein with guanidine-HCl and 2-mercaptoethanol, subsequent dialysis restored the PLD activity. Comparison of the nucleotide sequence data with the N-terminal protein sequence indicates that this secreted protein is synthesized as a larger precursor with a 47-amino-acid N-terminal extension to the mature enzyme of 509 amino acids. The amino acid sequence of the S. antibioticus PLD was extensively compared with other PLDs and phospholipase C (PLC). The deduced amino acid sequence of the cloned PLD was highly homologous to PLDs from S. acidomyceticus and Streptomyces sp., and contained a conserved region with S. chromofuscus PLD. From comparisons of the structural similarity and properties of the various PLDs, a classification of PLDs into two subgroups has been proposed and the highly conserved region designated tentatively region XPLD, which may be important in the catalytic function, has been identified. The homology comparison between our PLD and phosphatidylinositol-specific phospholipase C (PI-PLC) is also discussed.

Amino Acid Sequence↗
PubMed1994

Prolonged cell-free protein synthesis in a batch system using wheat germ extract.

Reaction conditions of cell-free protein synthesis using wheat germ extract were examined to prolong the period of protein synthesis in a batch reaction. By optimizing conditions for ATP regeneration system involved in the cell-free system, protein synthesis continued about 4 hours, so that about 17 micrograms dihydrofolate reductase protein was obtained in 1 ml of a reaction mixture. It suggests that maintaining ATP concentration is the primary requirement for long-life cell-free protein synthesis.

Adenosine Triphosphate↗
PubMed1994

High speed polymerase chain reaction in constant flow.

A new simple reactor of the tubing type was developed for polymerase chain reaction (PCR). A thin Teflon capillary tube was used as a tubing reactor in which the reaction mixture of PCR was driven by a pump at a constant flow rate. The sample was treated with three successive thermal stages for denaturation, annealing, and elongation of DNA and primers as a function of the position in the tube. The amplification yield was about a half of that obtained by a commercial thermocycler. Moreover, the total reaction time from 12 to 18 min, which was one-tenth of the time generally required by conventional thermocyclers using metal blocks, assured substaintial amplification of a DNA fragment. In addition, this reactor could be also used for rapid cycle-sequences. This new device will be easily incorporated into automated and rapid DNA analysis systems for DNA sequencing.

Base Sequence↗
PubMed1994

An increased rate of cell-free protein synthesis by condensing wheat-germ extract with ultrafiltration membranes.

Wheat-germ extract for cell-free protein synthesis was condensed with ultrafiltration membranes of which the molecular cut-off values were 10 kDa, 100 kDa, and 300 kDa. Reaction conditions of the cell-free system were optimized for the condensed extracts, which needed a higher concentration of creatine phosphate than the uncondensed one, probably due to the increased activity of degradation of ATP and GTP. By using the condensed extract and optimized reaction conditions, the rate of protein synthesis was increased 2- to 3-fold compared with using an uncondensed extract, and about 10-fold compared with conventional conditions. Condensation of the extract with the 300-kDa membrane showed the highest productivity, which was about 30 micrograms dihydrofolate reductase protein ml-1 h-1. The final amount of synthesized protein was one third of that of a continuous-flow cell-free (CFCF) system reported by Endo et al. [J. Biotechnol., 25, 221-230 (1992)] but the productivity was 5-fold higher than that obtained by the CFCF system.

Adenosine Triphosphate↗
PubMed1994

Treatment with cytosine arabinoside and granulocyte colony-stimulating factor in patients with myelodysplastic syndrome and its leukemic phase.

Twenty-one patients with myelodysplastic syndrome (MDS) or overt leukemia resulting from MDS were treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and cytosine arabinoside (Ara-C). Ara-C was administered in a dose of 20 mg/m2 every 12 h for 5 days and after 2 days 125 micrograms of rhG-CSF was administered for 10 days. After recovery of the leukocyte count the therapy was repeated, doubling the dose of Ara-C serially when possible. Of 13 patients with MDS, four achieved complete remission (CR), two good response (GR), two minor response (MR), and five no response (NR). Of eight patients with overt leukemia from MDS, only one with hyperplastic bone marrow achieved a partial response (PR) and the remaining seven achieved NR. The efficacy of the combination of rhG-CSF and Ara-C in the treatment of MDS and its leukemic phase is discussed, including at which time rhG-CSF should be administered: before, after or concomitantly with Ara-C. Multicenter randomized studies are needed in the evaluation of this combination therapy.

Adult↗