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Biomedical subjects

T Ohshima

Publications and source records attributed to T Ohshima.

At least 199 records · Page 11Linked to original sources
PubMed1995

Postmortem degradation of administered ethanol-d6 and production of endogenous ethanol: experimental studies using rats and rabbits.

Deuterium-labeled ethanol-d6 was employed to study the metabolism and postmortem change of ethanol in putrefied organ tissues. First, 4 ml/kg body weight of 25% (w/v) solution of ethanol-d6 was administered orally to each of 15 rats. The heart blood and organs were collected 15-90 min after the administration and the ethanol-d6 was analyzed by head space gas chromatography/mass spectrometry. The ethanol-d6 concentration in the organ tissues reached its maximum at 15 min after the administration and then gradually declined, showing the same pattern as human ethanol metabolism. Ethanol-d6 (3 ml of the same solution/kg body weight) was injected into the vein of a rabbit's ear (total of 12 rabbits). The rabbit was killed with carbon monoxide 30 min after the administration and the carcass was allowed to stand for 1-4 days at 30 degrees C in a moist chamber. The concentration of ethanol-d6 decreased moderately. Postmortem ethanol and 1-propanol concentrations, in contrast, showed marked increases 2.5 days and more after sacrifice in line with the degree of putrefaction of each organ tissue including skeletal muscle. This suggests the postmortem activation of micro-organism activity. These results indicate that ethanol concentrations in cadaver tissues must be carefully assessed with due consideration of postmortem degradation and production.

1-Propanol↗
PubMed1995

Structural organization and expression of the mouse gene encoding alpha-galactosidase A.

alpha-Galactosidase A (alpha-D-galactoside galactohydrolase, EC 3.2.1.22; alpha GalA) is a lysosomal enzyme that hydrolyses the alpha-D-galactosyl residues from glycosphingolipids. Fabry disease, an inhibited X-linked recessive human metabolic disorder, results from a mutation in the alpha GalA gene at Xq22. As a prerequisite for generating a mouse model of Fabry disease by gene targeting, we have isolated and characterized the mouse alpha GalA gene and cDNA. A cloned mouse alpha GalA cDNA encoded a putative precursor protein of 419 amino acids (aa), including a 31-aa signal peptide (SP). The deduced aa sequence showed high homology (79%) with the human alpha GalA protein. Nucleotide sequence analysis of genomic clones revealed that the overall structure and organization of the gene was very similar to that of human alpha GalA. All exon-intron splice junctions conformed to the GT/AG consensus sequence. Comparison of genomic and cDNA sequences revealed the occurrence of two putative polyadenylation signals whose alternative use results in the two mouse alpha GalA transcripts of 1.4 and 3.6 kb. The 5'-flanking region of mouse alpha GalA had no typical TATA box. Several putative promoter-associated elements including Sp1, AP1 and a potential cAMP-responsive element (CRE) were identified. Northern blot analysis revealed the widespread tissue distribution of mouse alpha GalA transcripts. Lower expression levels, however, were observed in some tissues, implying tissue-specific differences in alpha GalA promoter function.

Amino Acid Sequence↗
PubMed1995

Sex determination by polymerase chain reaction on mummies discovered at Taklamakan desert in 1912.

Sex determination was performed by the polymerase chain reaction (PCR) on eight adult mummies and one child mummy which were discovered at Taklamakan desert in 1912 and now belong to the Lüshun Museum in China. Archaeologically, these mummies were humans living in the seventh century, that is, more than 1300 years ago. Putative sex determination was performed based on external morphology for six of the eight adults, but it was impossible for the other two adults and one child mummy due to marked destruction on the external morphology. Hair, muscle and skin samples were then collected from each adult mummy, and skin and rib samples from the child mummy. Forty PCR cycles were performed as follows: denaturation at 94 degrees C for 40 s, annealing at 55 degrees C for 30 s and extension at 72 degrees C for 1 min. The primer and PCR reaction mixture were prepared according to the report by Witt and Erickson (M. Witt and R. P. Erikson, A rapid method for detection of Y-chromosomal DNA from dried blood specimens by the polymerase chain reaction. Hum. Genet., 82 (1989) 271-274)). Two different pairs of primer were used. One was X1, X2 (X1: 5'-AATCATCAAATGGAGATTTG-3'; X2: 5'-GTTCAGCTCTGTGAGTGAAA-3') to flanking the 170 bp fragment of the alphoid repeats on the human X chromosome, and the other was Y11, Y22 (Y11: 5'-ATGATAGAAACGGAAATATG-3'; Y22: 5'-AGTAGAATGCAAAGGGCTC-3') to flanking the 130 bp fragment of the alphoid repeats on the human Y chromosome. Extracted DNA solutions from mummy samples was purified using a spin column (T. Yoshii, K. Tamura, T. Taniguchi, K. Akiyama and I. Ishiyama, Water-soluble eumelanin as a PCR-inhibitor and a simple method for its removal. Jpn. J. Legal Med., 47 (1993) 323-329 (in Japanese with English abstract) for removing PCR-inhibitors, and bovine serum albumin (BSA) was employed to inhibit the remaining impurities even after the purification with the column. In six adult cases where the putative sex was determined from external morphology, the sex in five cases was consistent with that by PCR using hair, muscle, and skin samples, but the other one was inconsistent. In two adult cases where sex estimation was externally impossible, the sex was determined to be male because both X-specific and Y-specific bands were clearly detected. The child mummy was definitely male. This study shows that the sex determination was possible by the PCR method even with very ancient human samples > 1300 years old, that spin column was useful for removing impurities in the DNA solution from ancient human samples and that the BSA of optimum concentration suppressed the action of the PCR-inhibitory factors.

Adult↗
PubMed1995

Molecular cloning and chromosomal mapping of the mouse cyclin-dependent kinase 5 gene.

Cyclin-dependent kinase 5 (Cdk5) is predominantly expressed in neurons. In vitro, Cdk5 purified from the nervous tissue phosphorylates both high-molecular-weight neurofilament and microtubule-associated tau. The mouse gene encoding Cdk5 (Cdk5) was found to be 5 kb in length and divided into 12 exons. All of the exon-intron junctions matched the expected consensus sequence with the exception of the splice junction for intron 9, which has AT and AC dinucleotides instead of the usual GT and AG bordering sequence. In the 5'-flanking region of mouse Cdk5, several putative promoter elements were present, including AP1, Sp1, PuF, and TATA motifs. A metal regulatory element was also identified at position -207 to -201. Nucleotide sequence analysis of mouse Cdk5 showed high identity to the homologues of other vertebrate species, indicating that this kinase is highly conserved during evolution. Mouse Cdk5 was mapped to the centromeric region of mouse chromosome 5.

Amino Acid Sequence↗
PubMed1995

A new surgical method from the viewpoint of personal identification: a case of identification based on the presence of a metallic Z-stent.

To the best of our knowledge, the first case of successful personal identification based on the characteristic finding of a metallic Z-stent, unexpectedly discovered in the common bile duct, is reported. An unknown human male cadaver was found at the seaside. The age was estimated to be approximately 50-70 years and the postmortem interval was considered to be approximately 2 weeks. There was a small surgical scar of 1.5 cm in diameter in the epigastrium. The cadaver was group A in the ABO blood group system. Internally, the following severe injuries were observed: the dislocation of the occipito-atlantal joint, fractures of the cervical and thoracic portions of the spinal column, multiple fractures of the bilateral ribs, and ruptures of the spleen, left kidney and liver. Furthermore, a 4.5 cm long metallic stent was found in the common bile duct, and the gallbladder was not thoroughly identified because of the severely tight adhesion of the surrounding tissues. In every cut surface of the liver, multiple bile plaques, signifying the presence of long-standing cholestasis, were conspicuous. A microscopic examination revealed the invasion of cancer cells into the liver, which was diagnosed as well or moderately differentiated adenocarcinoma, although the primary origin of the adenocarcinoma could not be determined. The cause of his death was diagnosed as traumatic shock. According to the police investigation after the autopsy, it was revealed that a 60-year-old male, suffering from gallbladder carcinoma, had disappeared.(ABSTRACT TRUNCATED AT 250 WORDS)

Common Bile Duct↗
PubMed1995

Gene cloning, purification, and characterization of thermostable and halophilic leucine dehydrogenase from a halophilic thermophile, Bacillus licheniformis TSN9.

A halophilic and thermophilic isolate from the sand of Tottori Dune was found to produce a thermostable and halophilic leucine dehydrogenase (EC 1.4.1.9). It was identified to be a new strain of Bacillus licheniformis. The enzyme gene was cloned into Escherichia coli JM109 with a vector plasmid pUC18. The enzyme was purified to homogeneity from the clone cell extract by ion-exchange column chromatography with a yield of 31%. The enzyme was found to be composed of eight subunits identical in relative molecular mass (43,000). The amino acid sequence of the enzyme, deduced from the nucleotide sequence of the gene, showed an identity of 84.6% with that of the B. stearothermophilus enzyme [Nagata S, Tanizawa K, Esaki N, Sakamoto Y, Oshima T, Tanaka H, Soda K (1988) Biochemistry 27:9056-9062], although both enzymes were similar to each other in various enzymological properties such as thermostability, substrate and coenzyme specificities, and stereospecificity for hydrogen transfer from the C-4 of NADH. However, they were markedly distinct from each other in halophilicity; the B. licheniformis enzyme was much more stable than the other in the presence of high concentrations of salts.

Amino Acid Oxidoreductases↗
PubMed1995

Toxicological analysis of the psychotropic drugs chlorpromazine and diazepam using chemically fixed organ tissues.

Toxicological analysis for chlorpromazine and diazepam was performed using chemically fixed organ tissue specimens. After chlorpromazine and diazepam had been injected into rabbits, organ tissues (brain, lung, liver, kidney and skeletal muscle) were collected and fixed in 3 fixative solutions: buffered 10% formalin solution (pH 7.4, 10% BF), non-buffered 10% formalin solution (pH 5.1, 10% non-BF), buffered 4% paraformaldehyde solution (pH 7.4, 4% BPA). Chlorpromazine and diazepam were determined by GC-MS (gas chromatography-mass spectrometry) after 5 different fixation periods, and were detected even after 28 days of fixation. Recoveries of chlorpromazine and diazepam in 10% BF were within the range 48-86% and 68-171%, respectively after 28-day fixation, those in 10% non-BF were 22-54% and 48-78%, respectively, and those in 4% BPA solution were 13-59% and 14-50%, respectively. Thus, 10% BF was found to be the most suitable fixation medium for analysis of chlorpromazine and diazepam.

Animals↗
PubMed1995

Relationship between responsiveness to colony stimulating factors (CSFs) and surface phenotype of leukemic blasts.

We examined the responsiveness of leukemic cells to colony stimulating factors (CSFs) as determined by 3H-TdR incorporation and surface phenotypes of leukemic blasts. In acute myeloid leukemia (AML), CD13 and/or CD33 positive and HLA-DR negative M1 and M3 cases tended to show high response to G-CSF, GM-CSFs and IL-3, however, all HLA-DR positive M1, M2, M4 and M5 cases were unresponsive to CSFs but showed high autonomous growth. In acute lymphocytic leukemia (ALL), no response was observed to any CSFs but high autonomous growth was found in mixed leukemia cases. Sole T or B lineage cases showed low autonomous growth. These results suggest the varied nature of the proliferative state in leukemia and the existence of a subgroup in M1.

Cell Division↗
PubMed1995

Screening and determination of methamphetamine and amphetamine in the blood, urine and stomach contents in emergency medical care and autopsy cases.

Methamphetamine (MA) and amphetamine (AMP) were screened and their levels were determined using the Toxi-Lab thin-layer chromatography system and gas chromatography-mass spectrometry, respectively, in the blood, urine and stomach contents from 211 emergency medical care and 417 autopsy cases. MA and AMP were detected in 5 emergency medical cases, and the blood MA and AMP concentrations ranged from 0.697-0.041 micromol/100 g and from 0.0944-0.0003 micromol/100 g, respectively. MA and AMP were detected in 19 autopsy cases, in which blood MA and AMP concentration ranged from 14.3-0.123 micromol/100 g and from 0.256-0.0017 micromol/100 g, respectively. The autopsy cases included 5 cases of sudden death with blood MA concentration of less than 3 micromol/100 g. MA and AMP screening and determination in emergency medical care and autopsy cases provide useful information and are indispenable in clarifying the dimensions of MA abuse in Japan.

Journal Article↗
PubMed1995

Screening of volatile substances and determination of toluene (a thinner component) in the blood and urine in emergency medical care and autopsy cases by the pulse heating method.

Screening of volatile substances was performed by pulse heating gas chromatography - mass spectrometry (GC-MS) using a GS-Q column in 211 emergency medical care and 342 autopsy cases. At least 36 standard substances could be separately detected. Six kinds of volatile substances were screened in a total of 553 cases. Toluene and/or hippuric acid were detected in the blood and/or urine in respectively, 4 emergency medical care and 8 autopsy cases. There were 11 abusers (9 males and 2 females) in these 12 positive cases. The ages of the abusers ranged from 13-26 years. There was no particular pattern to the monthly frequency distribution of identification of thinner (toluene) abuse cases, which occurred throughout the year. It is believed that these data at least partly reflect the present status of thinner/glue abuse in Japan. We conclude that pulse heating GC-MS is useful in the screening and quantitative determination of volatile substances including toluene and other thinner/glue components.

Journal Article↗
PubMed1995

The role of an operating microscope in medico-legal practice.

This article describes our experience in the use of a binocular surgical microscope at autopsy for the in situ forensic examination of various types of mechanical skin wounds. The microscopic image was visualised on a closed-circuit television monitor, and colour photographs and video recordings were simultaneously obtained from the monitor. With abrasions, small epidermal tags, which play a key role in the determination of external force direction, were clearly observed. With lacerations, crushing of the epidermis was observed at the wound margin, and continuous bridges of connective tissue and/or blood vessels were also found within the wound cavity. With incised wounds, the epidermis was intact up to the wound margin, and there was no continuous bridge within the wound cavity. Using these differences, it was, therefore, simple to distinguish lacerations from incised wounds. Ligature marks due to hanging were clearly observed, with petechial haemorrhages and minute abrasions readily seen in the lesion. For stab wounds caused by a single-edge knife, the morphological difference between the two ends of the wound caused by the sharp blade and the blunt side could be distinguished. It was also possible to confirm the identification of unknown cadavers by observing old wound scars using the operating microscope. Abnormal lesions of the blood vessels, such as a mechanical tear of the vertebral artery (the origin of a subarachnoid haemorrhage) and the atherosclerotic stenosis of the coronary artery, were detectable. In addition, the microscope was useful as a teaching aid to colleagues and medical students to study the nature of the wounds during or after the autopsy. We believe that this system presents advantages in terms of education and training of forensic pathologists and physicians as well as crime investigation officers.

Journal Article↗
PubMed1995

Antitumour effects and pharmacokinetics of combination of vinblastine with a staurosporine derivative, NA-382, in P388/ADR-bearing mice.

The effects of a staurosporine derivative, N-ethoxycarbonyl-7-oxostaurosporine (NA-382), on the pharmacokinetics of vinblastine were evaluated, compared with those of verapamil, in multidrug-resistant P388/ADR-bearing mice. At first, the in-vitro experiments indicated that NA-382 permeated into the cells better and were more effective in combined cytotoxicity with vinblastine and on accumulation of vinblastine than with verapamil in P388/ADR cells. In combined intraperitoneal injection with vinblastine (200 micrograms kg-1) into P388/ADR-bearing mice, NA-382 in a suspension form (10 mg kg-1) prolonged the life-span of the mice near to that of P388/S-bearing mice treated with vinblastine alone, but verapamil even at the maximum tolerated dosage (30 mg kg-1) barely affected the in-vivo antitumour effect of vinblastine. When simultaneously administered with vinblastine to P388/ADR-bearing mice, NA-382 maintained significantly higher vinblastine levels in the tumour cells for 24 h and gave a larger area under the time-intracellular vinblastine concentration curve (0 to 24 h) than those receiving vinblastine alone, with long retention of the agent in ascitic fluid. Verapamil increased the cellular vinblastine content for only 6 h, accompanying a rapid elimination of the agent from the ascitic fluid. This study indicates that NA-382 is more effective against multidrug-resistance than verapamil, and its suspension is also advantageous for cancer chemotherapy of multidrug-resistant tumours.

Alkaloids↗
PubMed1995

Complete remission in a patient with hypoplastic acute lymphoblastic leukemia induced by granulocyte-colony-stimulating factor.

We report here a case of hypoplastic leukemia with T cell markers in whom complete remission was obtained with granulocyte-colony-stimulating factor (G-CSF) alone. A 23-year-old male was diagnosed with hypoplastic leukemia: Hb 2.6 g/dl, platelet count 29.0 x 10(9)/l after transfusion, WBC 2.9 x 10(9)/l, hypocellular bone marrow with 70.7% blasts. He was given G-CSF 300 micrograms/day by intravenous drip infusion without antileukemic agents for severe pneumonia. After the administration of G-CSF for 15 days, hematological examination and bone marrow findings had improved to normal, and complete remission was obtained. However, the patient relapsed 45 days after discontinuation of G-CSF. The characteristics of the relapsed leukemia cells were similar to those on admission: negative for myeloperoxidase and positive for T cell markers (CD2 and CD7). The possibilities for the differentiation of leukemic cells and the recovery of normal hematopoiesis with G-CSF are discussed.

Adult↗
PubMed1995

Rectus hematoma secondary to vomiting: a complication of conditioning regimen for bone marrow transplantation.

A 41-year-old woman with chronic myelogenic leukemia was scheduled to undergo transplantation of bone marrow. The patient complained of nausea and vomiting following the initiation of chemotherapy. One day prior to the planned termination of chemotherapy, the patient developed left-sided abdominal pain. Physical examination and imaging examination indicated the possibility of acute abdomen associated with bleeding or herniation. For therapeutic and diagnostic purposes, an emergency operation was performed. A 6 x 5 cm hematoma was detected within the left rectus abdominis muscle. It is suggested that the gastrointestinal symptoms should be carefully controlled in patients undergoing bone marrow transplantation.

Adult↗
PubMed1995

[Detection of Mycobacterium tuberculosis by amplification of mycobacterial DNA from pleural fluid and gastric juice: the comparison of IS6110 gene with Amplicor mycobacterium].

The polymerase chain reaction (PCR) using oligonucleotides based on the repetitive sequence (IS6110) of Mycobacterium tuberculosis and Amplicor Mycobacterium, which combines a PCR with the hybridization assay, were evaluated for detection of M. tuberculosis in pleural fluid and gastric juice. The detection limits of these two assay systems for cultured M. tuberculosis were less than 2 cells/ml, as compared with 200 cells/ml by culture. A total of 42 pleural fluid and 94 gastric juice specimens were examined. A total of 5 pleural fluid and 5 gastric juice were culture positive for M. tuberculosis. Only the PCR gave positive results in 2 pleural fluid of which laboratory findings are characteristic of tuberculous pleuritis, and in one gastric juice of the patient who was diagnosed as having pulmonary tuberculosis. Sensitivity, specificity and positive predictive value for IS6110 gene were 75%, 94% and 71.1%, respectively, in pleural fluid. Two of three positive specimens for IS6110 gene from pleural fluid were negative for Amplicor Mycobacterium. These specimens resulted in positive with Amplicor Mycobacterium when solvent extracted DNA was used. Both the PCR systems had the same sensitivity (80%), specificity (98.8%) and positive predictive value (83.3%) in gastric juice. The PCR systems would be useful for rapid detection of M. tuberculosis without long term culture and for detection of non-cultured one from pleural fluid as well as gastric juice. However, when the presence of an inhibitor of PCR is suspected in a specimen, DNA should be purified from the specimen before amplification.

Adult↗
PubMed1995

Congenital deficiency of histidine-rich glycoprotein: failure to identify abnormalities in routine laboratory assays of hemostatic function, immunologic function, and trace elements.

Histidine-rich glycoprotein (HRGP) has many biologic activities, but its physiologic function is still unclear. To show the physiologic function of HRGP, we studied five patients with congenital HRGP deficiency. Hemostatic screening tests, activities of natural anticoagulants and fibrinolytic proteins, markers of thrombin and plasmin generation, plasma levels of platelet-specific proteins, thrombin times with various concentrations of bovine thrombin, prolongation of thrombin time after addition of heparin or demartan sulfate, and contact activation of blood coagulation were normal or nearly normal in these patients. Serum concentrations of immunoglobulin, functional activity of the classical and the alternative pathway of complement, lymphocyte subsets, and serum concentrations of soluble interleukin-2 receptor were approximately normal in all patients, and serum concentrations of copper and zinc were completely normal. These results suggest that the physiologic functions of HRGP are limited when compared with its biologic activities. However, because the patients examined had plasma HRGP levels of 20% to 35% of normal, it is possible that 20% of normal HRGP level is sufficient for its physiologic functions.

Antigens, CD↗