Studies on anticoagulant-active arabinan sulfates from the green alga, Codium latum.
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Anticoagulants↗
Biomedical subjects
M Takeshita
Publications and source records attributed to M Takeshita.
Enzymatic instability of NADH-cytochrome b5 reductase as a cause of hereditary methemoglobinemia type I (red cell type).
Nucleotide substitutions in the gene for NADH-cytochrome b5 reductase were identified in three independent probands of hereditary methemoglobinemia type I. Patients in Kagoshima and Okinawa in Japan were shown to possess the same base change, from guanine to adenine at codon 57, which results in amino acid substitution from Arg to Gln. This nucleotide change was the same as formerly found in a patient in Toyoake, Japan (Katsube, T., Sakamoto, N., Kobayashi, Y., Seki, R., Hirano, M., Tanishima, K., Tomoda, A., Takazakura, E., Yubisui, T., Takeshita, M., Sakaki, Y., and Fukumaki, Y. (1991) Am. J. Hum. Genet. 48, 799-808). A type I patient in Italy was shown to have a base change from guanine to adenine at codon 105 which causes substitution from Val to Met. To characterize the enzymes of type I patients, Arg-57----Gln and Val-105----Met mutant enzymes were overexpressed in Escherichia coli and purified to homogeneity. kcat/Km values (NADH) of these two enzymes were 25% in Arg-57----Gln and 14.5% in Val-105----Met compared with that of the wild type enzyme, while the value of type II (generalized, severe form of the disease) mutant enzyme was 3% of the normal value (Yubisui, T., Shirabe, K., Takeshita, M., Kobayashi, Y., Fukumaki, Y., Sakaki, Y., and Takano, T. (1991) J. Biol. Chem. 266, 66-70). The type I mutant enzymes were less heat-stable and more susceptible to proteinase treatment than the wild type. From these results we conclude that restriction of enzyme deficiency to red cells in hereditary methemoglobinemia type I may be generally derived from instability and increased proteolytic susceptibility of variant NADH-cytochrome b5 reductases due to a point mutation.
Amino Acid Sequence↗
Stabilization of human neutrophil NADPH oxidase activated in a cell-free system by cytosolic proteins and by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide.
The superoxide-generating respiratory burst oxidase (NADPH oxidase) from human neutrophils can be activated in a cell-free system consisting of plasma membrane and cytosol by anionic amphiphiles such as sodium dodecyl sulfate and arachidonate (McPhail, L. C., Shirley, P. S., Clayton, C. C., and Snyderman, R. (1985) J. Clin. Invest. 75, 1735-1739; Curnutte, J. T. (1985) J. Clin. Invest. 75, 1740-1743; Bromberg, Y., and Pick, E. (1984) Cell. Immunol. 88, 213-221). Herein, the activity thus obtained is shown to be very labile at 37 degrees C. The rate of inactivation varied inversely with cytosol concentration. The stabilizing factor(s) was destroyed by heat and trypsin, indicating that it is protein in nature. Whereas cytosol from normal cells and from a chronic granulomatous disease patient lacking p67phox stabilized the oxidase activity, that from a chronic granulomatous disease patient lacking p47phox did not. Also, dialdehyde NADPH-treated cytosol showed no stabilizing effect, indicating that p47phox and a putative NADPH-binding component both participate in stabilization. The mechanism of inactivation was further explored by examining the stabilizing effect of agents that can act as chemical cross-linkers. Of several tested, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) was the most effective, but others that utilize different chemical mechanisms were also partially effective. EDC extended the half-life at 37 degrees C from 2 to 120 min, protected against the inactivating effects of Triton X-100 and high salt, and did not affect the Km for NADPH. Stabilization required prior activation in the presence of both cytosol and membrane; and EDC treatment of cytosol, membrane, or a mixture of the two prior to the addition of sodium dodecyl sulfate failed to induce stabilization. EDC eliminated the requirement for the continuous presence of cytosol and activator. Dialysis did not cause a loss in activity, whereas control activity was diminished with dialysis and was largely restored with added sodium dodecyl sulfate. In the absence of EDC, the separation of cytosol from the membrane fraction resulted in a significant loss of activity, which was largely restored by the addition of cytosol. However, EDC treatment allowed the isolation of a nearly fully active oxidase in the membrane fraction, the activity of which was not influenced by added cytosol. These results support a model in which the active NADPH oxidase consists of a dissociable complex among membrane and cytosolic components and indicate that the longevity of the activated state requires continuous association of these components.
Cell Membrane↗
Spermine down-regulates superoxide generation induced by fMet-Leu-Phe in electropermeabilized human neutrophils.
Effect of spermine, a naturally occurring polyamine, was investigated on superoxide generation in intact and electropermeabilized human neutrophils. Spermine suppressed N-formyl-methionyl leucyl phenylalanine (fMLP)-induced superoxide generation in permeabilized cells by reducing the rate and shortening the duration time. The inhibition was specific for spermine comparing with its precursor amines, spermidine and putrescine. The inhibition was not observed when cells were preincubated with spermine without permeabilization. Concanavalin A-induced superoxide generation was also down-regulated by spermine in permeabilized cells, but the activation induced by non receptor-mediated agonist (dioctanoylglycerol, phorbol myristate acetate, and arachidonate) was not affected by spermine. On the other hand, GTP-gamma-S-induced activation of superoxide generation was substantially suppressed by spermine. These results indicate that spermine inhibition occurs at a step prior to protein kinase C in signal transduction or in a pathway which is independent of the kinase.
Cell Membrane Permeability↗
Human T-cell leukemia virus type I associated lymphadenitis.
Histopathologic changes in lymph nodes were examined from ten patients with mild lymphadenopathy, a few atypical lymphocytes in their peripheral blood, skin lesions, and proviral DNA of human T-cell leukemia virus type I (HTLV-I) in their nodes. The proviral DNA of HTLV-I was detected by southern blot analysis, in situ hybridization, and/or polymerase chain reaction techniques. The lymph nodes showed preserved nodal architecture with diffuse infiltration of small to intermediate-sized lymphocytes in association with scattered transformed lymphocytes and a few immunoblast-like cells in the enlarged paracortex. The infiltrating lymphocytes were positive for CD4, but neither rearrangement nor deletion of T-cell receptors and immunoglobulin heavy chain genes was detected. Eight of ten patients received no therapy, and all patients were alive and healthy more than 5 months after the biopsies. The histologic findings resembled those of a viral infection and could be distinguished from HTLV-I associated lymphomas.
Adult↗
A case of histiocytic necrotizing lymphadenitis with bone marrow and skin involvement.
We report a case of histiocytic necrotizing lymphadenitis (HNL) with bone marrow extension in a 29-year-old male in which many large mononuclear cells infiltrated the bone marrow and mimicked malignant lymphoma. A lymph node biopsy confirmed the diagnosis of HNL. Immunohistologically, the infiltrating cells in the bone marrow were positive for lysozyme, LeuM1, Kp-1 and T-cell markers. The cells did not show haemophagocytosis. A skin biopsy from an accompanying facial skin rash revealed a proliferation of large cells similar to those observed in affected foci of the lymph node in subcutaneous tissue. The infiltrating cells were mainly lysozyme and Kp-1-positive histiocytes, some with phagocytosis of nuclear debris but none characteristic of haemophagocytosis. Transformed T-cells were also infiltrating.
Adult↗
Antitumor effect of RBS (rice bran saccharide) on ENNG-induced carcinogenesis.
We examined whether orally administered RBS (rice bran saccharide), prepared from rice bran by hot water extraction, increases immunocompetence, inhibits gastrointestinal carcinogenesis with N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) or shows an antitumor effect. After the administration of RBS, phytohemagglutinin (PHA)- and pokeweed mitogen (PWM)-stimulated blastogenesis of lymphocytes derived from the mesenteric lymph nodes and peripheral blood was enhanced, and the helper/suppressor T-cell ratio was elevated, and migration activity of peritoneal macrophages was also increased in rats treated continuously with ENNG. ENNG-induced gastrointestinal carcinomas were observed in 43% of those administered RBS (ENNG-RBS) as compared with 88% in the control (ENNG) and 94% in the prednisolone (PRD) group (ENNG-PRD). The 12-month survival rate of rats bearing gastrointestinal cancer was 58% in the ENNG-RBS group as compared with 25% in the ENNG group and 15% in the ENNG-PRD group. RBS prevented the reduction in immunocompetence in the course of carcinogenesis, suppressed carcinogenesis, and prolonged the survival of rats with gastrointestinal cancer. Antitumor activities of RBS are thought to be a kind of host mediated action. The growth inhibition ratio of transplantable ENNG-induced cancer in Wistar rats was 42.1% in the RBS and 51.8% in the 5-FU group. Since little is known about the potent antitumor activity of alpha-glucan, it would be interesting to consider the relationship between the structure and the biological activities of polysaccharides.
Adjuvants, Immunologic↗
Immunohistological study of skin involvement in Kikuchi's disease.
Five patients with histiocytic necrotizing lymphadenitis (Kikuchi's disease) with erythematous or popular skin lesions were studied. All patients healed naturally within a few months like the patients with no skin involvement. Histological findings for the skin lesions mimicked cutaneous malignant lymphoma. The infiltrated mononuclear cells usually demonstrated positive reactions for Ki-M1p (20-63%), lysozymes (13-54%), MT-1 (18-64%), UCHL-1 (22-39%) and LN2 (17-36%). OPD-4 and L26 positive cells were few in number. These results suggest that the infiltrated cells in a Kikuchi's disease skin lesion are composed of the same components as the affected lesion in the lymph node.
Adolescent↗
Aspergillotic aneurysm formation of cerebral artery following neurosurgical operation.
A rare case of intracranial aspergillotic aneurysm following neurosurgical operation for facial spasm is presented. Severe meningitis had persisted several days prior to the subarachnoid hemorrhage which occurred on 70th postoperative day. Angiography demonstrated a fusiform aneurysm at the peduncular segment of the superior cerebellar artery on the other side of operation. As the repeated cultures of CSF were negative, the pathogenetic factor causing aneurysm formation could not be identified before second operation of aneurysmal resection. Pathological study of the specimen revealed the arterial wall being deeply invaded by aspergillotic hyphae.
Adult↗
Risk factors for brain abscess in patients with congenital cyanotic heart disease.
Brain abscess is a serious complication of congenital cyanotic heart disease. We retrospectively evaluated the risk factors for brain abscess in 21 such patients treated between 1975 and 1990 in comparison with a control group. The mean arterial oxygen saturation, arterial partial pressure of O2, arterial blood oxygen content, and base excess in patients with brain abscess were significantly lower than in the control patients. The mean arterial partial pressure of CO2, pH, hematocrit, hemoglobin, and red blood cell content in patients with brain abscess were not significantly different. Patients with congenital cyanotic heart disease may develop minute encephalomalacia due to severe hypoxemia and increased blood viscosity resulting from compensatory polycythemia. The increased blood viscosity and reduced blood flow in the microcirculation may induce cerebral thrombosis or exaggerate minute encephalomalacia during dehydration or cardiac dysfunction, and shunted blood containing infectious organisms at such sites may be followed by focal cerebritis.
Brain Abscess↗
[Immunohistochemical in situ demonstration of cytokines in Hodgkin and non-Hodgkin lymphoma].
Primary biopsies from 149 patients with malignant lymphomas were examined by histological and immunohistochemical techniques. Twenty eight cases were classified as Hodgkin's disease and 121 as non-Hodgkin's lymphomas. The immunohistochemical distribution of cytokine expression (Il-1 alpha, Il-1 beta, Il-6 and TNF-alpha) was demonstrated in neoplastic cells and in tumor-associated macrophages (TAM). Neoplastic cells showed mostly weak positivity for TNF-alpha in 40% of Hodgkin's disease and in 20% of T cell lymphoma cases. Two groups of malignant lymphomas were established which differed in the numbers of cytokine expressing TAM. The first group consisted of malignant lymphomas which contained large quantities of cytokine-possessing TAM. In the second group significantly lower frequencies of cytokine expressing TAM were found. In both groups high and low grade malignant lymphomas were encountered. There was a significantly positive correlation between the number of Il-6 possessing TAM and the proliferation of lymphoma cells in only the non-Hodgkin's lymphomas. Our results suggest a paracrine tumor growth stimulating mechanism which is created by a self perpetuating cytokine production loop. A supposed cytokine produced by neoplastic cells dependent from their proliferative activity may induce Il-6 secretion by TAM. Il-6 in turn may stimulate the proliferation of the lymphoma cells without maturation thus leading to a self-sustaining growth.
Antibodies↗
[The effect of CPT-11 in combination with interferon-alpha against human colon carcinoma heterotransplanted in nude mice: preliminary report].
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Animals↗
[Congenital bicuspid aortic valve: stenotic type and insufficient type].
Two-hundred and eighty-one patients underwent surgical treatment of the aortic valves during a 10-year period of 1981 to 1991, 32 of whom (11%) had bicuspid aortic valve. Bicuspid aortic valve is well known to cause calcified aortic stenosis, however, some of these cases develop pure aortic insufficiency of unknown etiology. In our studies of 32 patients with bicuspid aortic valve, 28 patients had aortic stenosis, 2 were aortic insufficiency and 2 were infective endocarditis. Pathogenesis of aortic insufficiency in patients with bicuspid aortic valves was discussed and compared with that of aortic stenosis.
Adult↗
Analysis of factors in hypertension: change of membrane fluidity in the arteries of spontaneously hypertensive and monocrotaline-injected rats.
1. A change of membrane fluidity of a small piece of artery tissue was monitored by Fourier transform infra-red spectroscopy. 2. The measurement of carotid artery in situ revealed that a significant change in the peak position of the methylene absorbance was detected in the spontaneously hypertensive rat subjected to anoxia, but not in the Wistar-Kyoto rat carotid. The shift of a peak to the higher wavenumber position suggested that the averaged membrane fluidity of the artery was increased by the anoxic treatment. A similar change of membrane fluidity was also observed in a pulmonary hypertension induced by monocrotaline. 3. At 4 weeks after the injection of monocrotaline, the membrane fluidity was increased in the pulmonary artery tissue. 4. The change of membrane fluidity may be caused by the change of activity of phospholipases in the arteries with hypertension.
Animals↗
[Pharmacokinetics of epidurally administered bupivacaine during prostaglandin E1- or trimetaphan-induced hypotension].
The effects of prostaglandin E1 (PGE1) and trimetaphan (TMP) on the plasma concentrations and derived pharmacokinetic parameters of bupivacaine were studied in 14 women after its epidural administration. Patients, whose ages ranging from 35 to 60 years for mastectomy, received 50 mg of bupivacaine without epinephrine injected into the cervical epidural space during PGE1- or TMP-induced hypotension (80-90 mmHg of the systolic arterial pressure) under general anesthesia with nitrous oxide, oxygen and isoflurane 0.3-0.5%. No significant differences in pharmacokinetic parameters for the absorption and distribution of bupivacaine were found between the two groups. However, the mean elimination half-life of bupivacaine was significantly longer in patients with TMP [5.0 +/- 1.7 (SD) hr] compared with those with PGE1 (3.1 +/- 1.4 hr). The total clearance of bupivacaine was greater in patients with PGE1 (345 +/- 150 ml.min-1) compared with those with TMP (248 +/- 66 ml.min-1). The results of this pharmacokinetic study indicate that the plasma bupivacaine concentration decreases more rapidly during PGE1-induced hypotension than during TMP-induced hypotension.
Adult↗
[The management of stage I lymphomas of the B-cell type with special reference to the role of radiation therapy].
Fifty-one patients with clinical stage I B-cell lymphomas were treated between 1980 and 1988. For the entire group, the actuarial 5-year survival rate and 5-year freedom from relapse rate were 78% and 64%, respectively. Primary site, tumor bulk and performance status were the prognostic factors. Twenty-three patients were intensively treated with local radiotherapy alone, because (1) they had histologically low grade (7 patients) or follicular large cell (1 patients) tumors, (2) Waldeyer's ring was the site of the primary (11 patients) or (3) the tumor was less than 4 cm in maximum diameter (4 patients). Among them, only one patient died due to lymphoma (5-year survival rate: 95%), and 19 remained disease free (5-year freedom from relapse rate: 80%). In contrast, of the remaining 23 patients treated with both radiation therapy and combination chemotherapy (VEMP or CHOP), the 5-year survival rate and 5-year freedom from relapse rate were 70% and 54%, respectively. These results suggest that high-risk patients should be treated with more aggressive combination chemotherapy as well as radiotherapy. However, low-risk patients with stage I B-cell lymphomas can be treated by local radiotherapy alone.
Adult↗
Defective provirus form of human T-cell leukemia virus type I in adult T-cell leukemia/lymphoma: clinicopathological features.
Human T-cell leukemia virus type I (HTLV-I) is associated with adult T-cell leukemia/lymphoma (ATLL). To examine the relationship between defective HTLV-I proviruses and clinicopathological features, we examined 95 patients with ATLL showing clonal integration of HTLV-I proviral DNA; 77 patients (81%) showed 1 clonal band, 15 (16%) showed 2 clonal bands, and 3 (3%) showed 3 clonal bands. In addition, the defective proviral form was detected in 28 patients (29%): 23 (30%) of the 77 with 1 clonal band, 4(27%) of the 15 with 2 clonal bands, and 1(33%) of the 3 with 3 clonal bands. The numbers of clonal bands had no association with the presence of defective proviruses. We classified the 95 patients with ATLL into four types according to clinicopathological features (smoldering leukemia, chronic leukemia, acute leukemia, and lymphoma types). The distribution of patients with the defective form was not different among these four types. The HTLV-I genomes must have integrated into the human genome DNA and been deleted partially in the cells. The defective form was kept during the clinical stage. All patients with the defective form showed defect of the gag or/and env region. No patient had a defect of the pX region. These data suggest that the pX region of HTLV-I must have played an important role in ATLL genesis.
Amino Acid Sequence↗
Role of cysteine residues in human NADH-cytochrome b5 reductase studied by site-directed mutagenesis. Cys-273 and Cys-283 are located close to the NADH-binding site but are not catalytically essential.
Human NADH-cytochrome b5 reductase (EC 1.6.2.2) contains 4 cyteine residues (Cys-203, -273, -283, and -297). Cys-283 was previously proposed to be involved in NADH binding by chemical modification (Hackett, C. S., Novoa, W. B., Ozols, J., and Strittmatter, P. (1986) J. Biol. Chem. 261, 9854-9857). In the present study the role of cysteines in the enzyme was probed by replacing these residues by Ser, Ala, or Gly employing site-directed mutagenesis and chemical modification. Four mutants, in which 1 of the 4 Cys residues was replaced by Ser, retained comparable kcat and Km values to those of the wild type. All of these mutants were as sensitive as the wild type to treatment with SH modifiers, while a double mutant, C273S/C283S was resistant. Since inhibition by SH modifiers was protected by NADH, Cys-273 and Cys-283 were implicated to be close to the NADH-binding site. C273A and C273A/C283A mutants showed approximately one-fifth of the enzyme-FAD reduction rate of the wild type as revealed by steady-state kinetics and by stopped-flow analysis. Anaerobic titration has shown that reduction and re-oxidation processes including formation of the red semiquinone of these mutants were not significantly altered from those of the wild type. From these results it was concluded that none of the Cys residues of the enzyme are essential in the catalytic reaction, but Cys-273 conserved among the enzymes homologous to NADH-cytochrome b5 reductase homologous to NADH-cytochrome b5 reductase plays role(s) in facilitating the reaction. A difference spectrum with a peak at 317 nm, which was formerly considered to be derived from the interaction between NAD+ and Cys-283 of the reduced enzyme, appeared upon binding of NAD+ not only to the reduced wild type enzyme but also to the C273A/C283A mutant in which both of the Cys residues close to the NADH-binding site were replaced.
Base Sequence↗