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Biomedical subjects

H Matsuzaki

Publications and source records attributed to H Matsuzaki.

At least 19 recordsLinked to original sources

Inversion of chromosome 16 and bone marrow eosinophilia in a myelomonocytic transformation of chronic myeloid leukemia.

We report a case of chronic myeloid leukemia (CML) in myelomonocytic transformation associated with bone marrow (BM) eosinophilia. At diagnosis, all BM cells showed a Ph chromosome. At the time of blastic phase, more than 50% of Ph+ cells had a pericentric inversion of chromosome 16, inv(16)(p13q22). This case confirms that blastic transformation of CML can involve any committed progenitor, and myelomonocytic leukemia with BM eosinophilia is specifically associated with rearrangement of chromosome 16 at band p13 and q22.

Blast Crisis

Site-specific recombinase, R, encoded by yeast plasmid pSR1.

The R gene product (R protein) of Zygosaccharomyces rouxii plasmid pSR1 catalyzes site-specific recombination within a 58 base-pair (bp) sequence present in the 959 bp inverted repeats of this plasmid. The R protein was produced in Escherichia coli and partially purified. The partially purified protein catalyzed site-specific recombination in vitro without the supply of an energy source. Recombination resulted in intramolecular inversion or deletion, depending on whether the orientations of the two recombination sites on the substrate plasmid were the same or opposite. Presumably, R protein is the only protein required for the recombination reaction. A circular DNA molecule appears to be a better substrate than a linear molecule in R-mediated in vitro intramolecular recombination. The R protein binds to a set of six 12 bp elements within the inverted repeats of pSR1. Two of these 12 bp elements are arranged in an inverted configuration with a 7 bp spacer in the 58 bp sequence. The R protein mediates strand cleavage in vitro at the junction between the 12 bp elements and the 7 bp spacer. The cleavage sites on the top and bottom strands are staggered and flanked by polypurine tracts that form part of the 12 bp elements.

Base Sequence

Neutrophil chemotactic factors produced by a cell line from thyroid carcinoma.

A neutrophil chemotactic factor (human interleukin 8, human granulocyte-macrophage colony-stimulating factor)-producing cell line, named KHM-5M, was established from a patient with an undifferentiated thyroid carcinoma, neutrophilia, and malignant pleurisy with many neutrophils and a few malignant cells. The cell line was transplanted into nude rats, and the infiltration of neutrophils was observed in and around the transplanted tumor tissue. Neutrophil chemotactic activity was predicted from the clinical features and pathological findings in this case. The extreme chemotactic activity of the neutrophils was demonstrated in conditioned medium from KHM-5M cells using the modified Boyden chamber technique. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis, at least two neutrophil chemotactic activities in conditioned medium from the cell line were observed. The levels of these activities derived from KHM-5M cells were screened by measuring conditioned medium from the COS cells, which expressed a complementary DNA library from the KHM-5M cells. Chemotactic activities (human interleukin 8, human granulocyte-macrophage colony-stimulating factor) were identified by DNA cloning. These results show that the KHM-5M cells derived from an undifferentiated thyroid carcinoma produce multicytokines and suggest that those cytokines modified some pathological features in this case.

Aged

Crystallization and preliminary X-ray investigation of proteinase A, a non-pepsin-type acid proteinase from Aspergillus niger var. macrosporus.

Proteinase A from Aspergillus niger var. macrosporus is a non-pepsin-type acid proteinase distinctly different in various properties from the family of pepsin-type aspartic proteinases, and so far it remains unknown which residues participate in the catalysis of the enzyme and how the mechanism operates. The acid proteinase A was crystallized from an ammonium sulfate solution by the hanging-drop vapor diffusion method. The space group of the crystals was P2(1)2(1)2(1) with unit cell dimensions of a = 54.7 A, b = 70.4 A and c = 38.0 A. On the assumption that there is one enzyme molecule in the asymmetric unit, the calculated ratio of volume to unit protein mass (Vm) was 1.64 A3 per dalton. Diffraction data were collected up to a resolution higher than 1.5 A, using the Weissenberg camera for macromolecular crystallography with synchrotron radiation. The crystal of proteinase A is, therefore, suitable for the structural analysis with a high resolution.

Aspartic Acid Endopeptidases

Clinical values of intraoperative ultrasonography for spinal tumors.

Intraoperative ultrasonography was conducted in 52 cases of spinal tumor, at 7.5 MHz, mainly by means of linear scanning, to evaluate its clinical usefulness. The procedure was effectively applied in such clinical purposes as: 1) locating the tumor, 2) deciding the resectability of intramedullary tumors, 3) deciding the site for intraspinal biopsy or shunt tube insertion, 4) establishing the topical relationship between the spinal cord and the tumor, and 5) differentiating neurilemoma from meningoma. Of 10 patients with intramedullary tumors, 5 (50%) were removed, because extirpation was possible when the spinal cord and the tumor were well demarcated on the ultrasonogram. Intratumorous cysts were found to exist in 73% of neurilemoma and 14% of meningioma cases, enabling the differential diagnosis between the two tumors. Intraoperative ultrasonography is an uninvasive method to reveal intradural and extradural conditions and thus constitutes a valuable diagnostic means to ensure safe and precise spinal surgery.

Adolescent

Human T-cell leukemia virus-1-positive cell line established from a patient with small cell lung cancer.

A stable cell line, KHM-3S, was established from a patient with small cell lung cancer (SCLC), who had a high serum level of soluble interleukin 2 receptors (sIL2-R) and was seropositive for human T cell leukemia virus (HTLV)-1. KHM-3S cells were positive for IL2-R (Tac) and NKH-1, but negative for other lymphocytic markers such as OKT 11, OKT 4, OKT 8, T cell receptor (WT 31), B 1, and B 4. Moreover, the KHM-3S cells were negative for leukocyte common antigen and strongly positive for neuron-specific enolase (NSE). Secretion of sIL2-R and NSE by the KHM-3S line was detected by an enzyme-linked immunosorbent assay. Rearrangement of the T cell receptor gene and monoclonal HTLV-1 integration were found by Southern blot analysis of KHM-3S DNA. However, Northern blot analysis showed no T cell receptor mRNA. KHM-3S may be useful for studies on the role of HTLV-1 in carcinogenesis and IL2-R expression in SCLC.

Antigens, Surface

Human myeloma cell line (KHM-4) established from a patient with multiple myeloma associated with hyperammonemia.

A cell line of plasma cells with high ammonia (NH3) production (KHM-4) was established from a patient with multiple myeloma complicated by hyperammonemia and abnormal serum concentrations of amino acids. Surface marker studies of KHM-4 cells showed that the cells were positive for cytoplasmic immunoglobulins (IgA kappa), HLA-DR, and T 10. Secretion of ammonia by the KHM-4 cells was detected by the addition of L-glutamine and L-arginine into the culture medium of amino acid-free RPMI 1640. In the presence of L-glutamine, KHM-4 cells secreted a greater amount of ammonia than the T cell line, CEM. However, production of ammonia by L-arginine was not observed in other cell lines. These observations provide evidence for the existence of a peculiar amino acid metabolism in the myeloma cells causing hyperammonemia and serum amino acid disturbance.

Ammonia

Cyclosporin for inflammatory pseudotumour.

Dramatic efficacy of cyclosporin was observed in a patient with inflammatory pseudotumour in the liver. The 30-year-old male patient had suffered several attacks of high fever, polyarthralgia and lymph node swelling since 1981. In February 1989, abdominal CT showed multiple space-occupying lesions in his liver, and histopathology of the biopsied liver showed infiltration of plasma cells and lymphocytes with proliferation of collagen fiber, which was comparable to an inflammatory pseudotumour. In May 1990, a treatment regimen of cyclosporin A along with prednisolone was begun because he had been diabetic since March 1990. Within a few days of treatment, the high fever and polyarthralgia subsided, and leucocytosis of 32,000/mm3 and the increased CRP of 17.7 mg/dl were normalised within one month. A reduction of the size of the space-occupying lesions in the liver was observed on abdominal CT taken one month after cyclosporin treatment.

Adult

[Pharmacokinetic studies of continuous and bolus intraportal 5-fluorouracil infusion].

We studied the pharmacokinetics of continuous and bolus intraportal 5-FU infusion. In continuous intraportal infusion group, 20 mg/kg of 5-FU was infused into ileocecal vein for an hour and the same dose was infused bolusly in bolus intraportal infusion group. Blood samples were collected from portal vein continuously and liver specimens were obtained continuously. The rabbits were sacrificed at 180 minutes from the start of infusion of 5-FU and small intestine was obtained. The results were as follows: A significantly higher concentration of 5-FU in the portal vein was observed in the continuous intraportal infusion group compared with the one-shot infusion group. But the elimination rate of 5-FU was more rapid after cessation of infusion in the continuous infusion group. The rate of FdUMP elimination in the liver tended to be lower in bolus intraportal infusion. The FdUMP concentration in the small intestine was higher in the continuous intraportal infusion group than in the bolus infusion group. In the continuous intraportal infusion group, the FdUMP concentration in the liver was lower than in the small intestine. This depended on the effect of the 5-FU catabolic enzyme, dihydrouracil dehydrogenase, which is contained in the liver in large quantities. The results suggest that in order to increase the FdUMP concentration in the liver with continuous intraportal 5-FU infusion, suppression of the effect of this enzyme is necessary.

Animals

[Optimal branching of hepatic artery in patients with liver cirrhosis].

Optimality in branching structure of the hepatic arterial vessel in 16 patients with liver cirrhosis was studied. The controls used in this study were 14 patients with gastric or colonic cancer not accompanied by liver dysfunction. Optimality principle was based on the concept that blood vessel size and arrangement provided for blood flow with minimum energy loss. Measurements were made of parent and branch vessel diameters from selective celiac arteriogram. After that we measured the included angles of the branch-points of portal vein using ultrasonography. The branch-points of interest were the third bifurcations of right intrahepatic portal vein and artery. The following results are presented in terms of three parameters (r2(2)/r1(2), r1/r0, and r2/r0). The theoretical equations have been shown by several authors. The curve representing these equations was shown by Zamir. We analyzed the scatter of the data points and compare the theoretical and actual results with them. The extent of the scatter is studied making a comparison between control patients and patients with cirrhosis. The data points in control patients were plotted near the theoretical curve. In patients with cirrhosis, the data points show a definite tendency to be plotted far away from the theoretical curve, when we take account of both vessel diameters and branching angles. This discrepancy between theoretical values and actual data in patients with cirrhosis is considered to be caused by the following factors which may influence the vascular structure; hemodynamic change caused by increase of intrahepatic shunt, and morphological change caused by increased fibrosis or/and parenchymal nodules of the liver.

Celiac Artery

[Münchhausen syndrome with severe iron deficiency anemia].

A patient with Münchhausen syndrome who had severe iron deficiency anemia (IDA) is reported. A 31 year-old female presented with irregular genital bleeding. However, gynecological examination disclosed no evidence of specific disorders causing bleeding. Tests for bleeding tendency and hemolysis were all negative. Although massive bleeding was absent, she had three episodes of a rapid fall in Hb level associated with marked fatigue and weakness, and subsequently rapidly developed serious IDA afterwards. Factitious bleeding was strongly suspected. Blood transfusion performed when the patient was not being watched failed to increase Hb, while transfusions given while the patient was watched were effective. She was not cooperative throughout the hospital course and was discharged on her own request. In outpatient clinic, the diagnosis was confirmed by the evidence that she diluted her own blood samples.

Adult

Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in tobacco cells.

Recombinase encoded by the R gene of pSR1 of Zygosaccharomyces rouxii mediates reciprocal recombination between two specific recombination sites (RSs) to induce excision or inversion of the DNA segment that is flanked by the RSs. We report here that site-specific recombination mediated by this system takes place effeciently in tobacco cells. To monitor the recombination events in tobacco cells, we have constructed two types of cryptic beta-glucuronidase reporter gene in such a way that recombination such as inversion of the construct or excision of the intervening sequence results in their expression. When these cryptic reporter constructs were transiently introduced together with the R gene by electroporation into protoplasts of tobacco cells, beta-glucuronidase activity was detected. The cryptic reporter genes, when stably resident in the chromosome of tobacco cells, were also activated by the R gene. Structural analyses of the genomic DNA isolated from these tobacco cells showed that the R protein did in fact catalyze precise recombination between two copies of RSs in tobacco cells, with resultant activation of the cryptic reporter genes. This observation provides the basis for development of a DNA technology whereby large regions of DNA can be manipulated in plant chromosomes. Potential uses of this recombination system are discussed.

Base Sequence

Purification of megakaryocyte differentiation activity from a human fibrous histiocytoma cell line: N-terminal sequence homology with activin A.

We purified a protein possessing a potent ability to induce the differentiation of a murine megakaryoblastic cell line, L8057, from the supernatant of a human fibrous histiocytoma cell line, KHM-5M. The protein, a homodimer of a molecular weight of 25-kDa, has an N-terminal sequence identical with that of the beta A-chain of inhibin, indicating that it is identical to or highly homologous with activin A. Thus, it is speculated that activin A or its homologue is involved in megakaryocytic differentiation.

Activins

The PHO80/TUP7 locus in Saccharomyces cerevisiae is on the left arm of chromosome XV: mapping by chromosome engineering.

The PHO80/TUP7 locus in Saccharomyces cerevisiae is reported to be located on the right arm of chromosome XV close to its centromere. In the present study, the locus has been reassigned to the left arm of the same chromosome by reciprocal recombination between chromosomes V and XV at URA3 (on chromosome V) and PHO80/TUP7 loci by using the site-specific recombination system of the yeast plasmid pSR1.

Chromosome Mapping

Practical use of optical cards in medical care.

This paper reports excellent results with respect to application of optical cards in medical care, based on improvements in the technology for encoding optical cards and devices used to increase read-write speed, and also reports the issuance of the first Japanese standards for the data format of optical cards.

Information Storage and Retrieval