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Biomedical subjects

Y Akiyama

Publications and source records attributed to Y Akiyama.

At least 19 recordsLinked to original sources
PubMed1992

In vitro catalysis of oxidative folding of disulfide-bonded proteins by the Escherichia coli dsbA (ppfA) gene product.

It was shown previously that the Escherichia coli gene ppfA (dsbA) encodes a periplasmic protein, and its inactivation leads to a deficiency in disulfide bond formation of envelope proteins (Kamitani, S., Akiyama, Y., and Ito, K. (1992) EMBO J. 11, 57-62; Bardwell, J. C. A., McGovern, K., and Beckwith, J. (1991) Cell 67, 581-589). The DsbA/PpfA protein was overproduced, purified, and examined for its activities in vitro. Its abundance in a wild-type cell was estimated to be about 850 molecules which probably exist as homodimers as suggested by size exclusion chromatography. Purified DsbA markedly stimulated disulfide bond formation of E. coli alkaline phosphatase, either in vitro synthesized or purified and denatured, as well as of reduced bovine ribonuclease A. The DsbA-catalyzed rapid disulfide bond formation occurred after a lag period which appeared to be determined by the redox state of the reaction mixture and concentration of DsbA. Inclusion of higher concentrations of oxidized glutathione or DsbA shortened the lag period. We propose that DsbA, which proved to directly catalyze disulfide bond formation, may also have a role in maintaining the bacterial periplasm oxidative.

Alkaline Phosphatase↗
PubMed1992

Nuclear magnetic resonance abnormalities of the cerebral white matter in children with acute lymphoblastic leukemia and malignant lymphoma during and after central nervous system prophylactic treatment with intrathecal methotrexate.

BACKGROUND: A prospective study was conducted to test the feasibility of nuclear magnetic resonance (NMR) imaging in the early diagnosis of treatment-induced leukoencephalopathy. METHODS: The study group included 16 patients with acute lymphoblastic leukemia and 4 patients with malignant lymphoma. All were given intravenous and intrathecal (IT) methotrexate (MTX) for central nervous system prophylaxis. Serial NMR studies were performed before and/or during induction-consolidation cycles. RESULTS: NMR imaging disclosed leukoencephalopathy in 8 of the 20 patients (40%) in the early stages of treatment. In six of the eight, the leukoencephalopathy was resolved after temporary or permanent interruption of IT MTX, and chemotherapy was completed successfully. The other two patients are being treated. Transient neurologic abnormalities developed in two of the eight patients. CONCLUSIONS: The possible causal relationship between leukoencephalopathy and the antimetabolic effects of MTX is discussed. This study clearly shows that NMR imaging is valuable in the early diagnosis and management of treatment-induced leukoencephalopathy.

Adolescent↗
PubMed1992

Effects of the chaperonin GroE on the refolding of tryptophanase from Escherichia coli. Refolding is enhanced in the presence of ADP.

The refolding of the tetrameric enzyme tryptophanase was facilitated by the chaperonin GroE. Maximum refolding yield of tryptophanase molecules (about 80%) was attained in the presence of a 15-fold excess of GroE 21-mer over tryptophanase monomer. The GroEL subunit was required for this improvement in refolding yield, whereas the GroES subunit was not. Light scattering experiments of the refolding reaction revealed that GroE bound to tryptophanase folding intermediates and suppressed their aggregation. The presence of ATP was required for the efficient dissociation of tryptophanase from GroEL. However, our experiments indicated that tryptophanase dissociated readily from GroEL in the presence of not only ATP, but also in the presence of non-hydrolyzable ATP analogues such as ATP gamma S (adenosine 5'-O-(3-thiotriphosphate)) and AMP-PNP (adenyl-5'-yl imidodiphosphate) as well. Surprisingly, the release of tryptophanase from GroEL was facilitated in the presence of ADP as well. We concluded that the binding of nucleotides such as ATP and ADP changed the conformation of GroEL and facilitated the dissociation of tryptophanase molecules. The conformation formed in the presence of ADP was distinct from the conformation formed in the presence of ATP, as shown by the selective dissociation of various folding proteins from the two conformations.

Adenine Nucleotides↗
PubMed1992

Rapid determination of sulphonamides in milk using liquid chromatographic separation and fluorescamine derivatization.

A simple and selective method is presented for the multiple residue determination of eight sulphonamides in consumers' milk. The drugs are sulphisomidine (ID), sulphadiazine (DZ), sulphamerazine, sulphadimidine, sulphamonomethoxine, sulphamethoxazole, sulphadimethoxine and sulphaquinoxaline (SQ). The milk sample was deproteinized with the same volume of 2 M hydrochloric acid and filtered. A 1-ml volume of the filtrate was mixed with 1 ml each of 1.25 M sodium acetate solution and a buffer (pH 3.0) for derivatization with 0.6 ml of 0.02% fluorescamine solution in acetone. A high-performance liquid chromatographic analysis was carried out on a C18 column with a mobile phase of acetonitrile-2% acetic acid (3:5) at 55 degrees C using a fluorescence detector at an excitation wavelength of 405 nm and an emission wavelength of 495 nm. Average recoveries at fortification levels of 2, 5 and 10 ng/ml were 114%, 109% and 106%, respectively. Relative standard deviations were 1-4% at 10 ng/ml for ID, 5 ng/ml for DZ and SQ and 2.5 ng/ml for the other five sulphonamides. The method was applied to 25 milk samples and all appeared to be free from the drugs.

Animals↗
PubMed1992

[Single photon emission computed tomography by using fan beam collimator].

A multislice fan beam collimator which has parallel collimation along the cephalic-caudal axis of a patient and converging collimation within planes that are perpendicular to that axis was designed for a SPECT system with a rotating scintillation camera, and it was constructed by the lead casting method which was developed in recent years. A reconstruction algorithm for fan beam SPECT was formed originally by combining the reconstruction algorithm of the parallel beam SPECT with that of the fan beam X-ray CT. The algorithm for fan beam SPECT was confirmed by means of computer simulation and a head phantom filled with diluted radionuclide. Not only 99mTc but also 131I was used as a radionuclide. A SPECT image with the fan beam collimator was compared with that of a parallel hole, low energy, high resolution collimator which was routinely used for clinical and research SPECT studies. Both system resolution and sensitivity of the fan beam collimator were approximately 20% better than those of the parallel hole collimator. Comparing SPECT images obtained from fan beam collimator with those of parallel hole collimator, the SPECT images using fan beam collimator had far better resolution. A fan beam collimator is a useful implement for the SPECT study.

Algorithms↗
PubMed1992

Pre-treatment of a human T-lymphoblastoid cell line with L-asparaginase reduces etoposide-induced DNA strand breakage and cytotoxicity.

The effect of L-asparaginase (L-asp) pre-treatment on etoposide-induced DNA strand breakage and cytotoxicity was investigated. In a T-lymphoblastoid cell line, Molt 4, etoposide-induced DNA strand breaks, DNA-protein cross-links and cytotoxicity were reduced by pre-treatment with L-asp for 15 hr, but it did not cause these changes in a promyelocytic-leukemia cell line, HL-60, which is less sensitive than Molt 4 to L-asp. However, pre-treatment of Molt 4 cells with L-asp did not significantly alter the accumulation of [3H]-etoposide. Cell-cycle analyses showed an increase in G1-phase cells, a significant decrease in both S-phase cells and G2/M-phase cells pre-treated with L-asp in Molt 4 cells, but L-asp exposure did not result in any significant changes in HL-60 cells. On the other hand, L-asp pre-treatment did not affect topoisomerase-I (Topo-I) inhibitor, camptothecin (CPT)-induced DNA strand breaks or toxicity in Molt 4 cells. Our data imply that a decrease in S- and G2/M-phase cells following L-asp treatment may explain the reduction of etoposide-induced DNA lesions and cytotoxicity in Molt 4 cells, since topoisomerase-II (Topo-II) content or activity is a function of cellular proliferation status.

Asparaginase↗
PubMed1992

Regulation of acetylcholine release in vivo from rat hippocampus by monoamines as revealed by novel column-switching HPLC with electrochemical detection.

To clarify monoaminergic regulation of acetylcholine (ACh) release in the rat hippocampus, the effects of administration of monoamines through a dialysis probe on extracellular ACh levels were examined using in vivo brain microdialysis combined with a novel column-switching HPLC system. Infusion of dopamine or 5-hydroxytryptamine, but not noradrenaline, increased ACh levels. The ACh levels also increased following infusion of apomorphine and 5-methoxy-N,N-dimethyltryptamine. These results demonstrate that hippocampal ACh release is regulated by dopamine and 5-hydroxytryptamine.

Acetylcholine↗
PubMed1992

Monoamine-mediated enhancement of acetylcholine release in rat hippocampus by 6R-L-erythro-5,6,7,8-tetrahydrobiopterin.

Recently, we reported that intracerebroventricular administration of 6R-L-erythro-tetrahydrobiopterin (6R-BH4), a common natural cofactor for phenylalanine, tyrosine and tryptophan hydroxylases, enhances in vivo exocytotic release of acetylcholine (ACh) from the rat hippocampus, as monitored by brain microdialysis. In the present study, we developed a method for separation of ACh from 6R-BH4 on small columns of Sephadex G-10, which allowed us to determine by high pressure liquid chromatography (HPLC) the changes of ACh levels recovered in dialysates following addition of 6R-BH4 to the dialyzing fluid. Infusions of 6R-BH4 (final concentration, 0.25, 0.5 and 1 mM in the perfusing fluid) produced concentration-dependent increases in ACh levels in dialysates (1.5-, 3- and 8-fold of the basal levels, respectively), but was without effect on choline levels in dialysates. Infusion of 6S-L-erythro-5,6,7,8-tetrahydrobiopterin (6S-BH4), an unnatural diastereoisomer of 6R-BH4, or L-erythro-biopterin (biopterin), an oxidized form of 6R-BH4, did not affect the ACh levels in dialysates. The 6R-BH4-induced increase in ACh levels persisted after pretreatment with atropine, a muscarinic antagonist, but did not occur in rats pretreated with reserpine to deplete monoamines. These results show that 6R-BH4 stimulates exocytotic ACh release in the hippocampus through action on a local monoaminergic system in the hippocampus.

Acetylcholine↗
PubMed1992

Identification and characterization of an Escherichia coli gene required for the formation of correctly folded alkaline phosphatase, a periplasmic enzyme.

Tn5 insertion mutations of Escherichia coli were isolated that impaired the formation of correctly folded alkaline phosphatase (PhoA) in the periplasm. The PhoA polypeptide synthesized in the mutants was translocated across the cytoplasmic membrane but not released into the periplasmic space. It was susceptible to degradation by proteases in vivo and in vitro. The wild-type counterpart of this gene (named ppfA) has been sequenced and shown to encode a periplasmic protein with a pair of potentially redox-active cysteine residues. PhoA synthesized in the mutants indeed lacked disulfide bridges. These results indicate that the folding of PhoA in vivo is not spontaneous but catalyzed at least at the disulfide bond formation step.

Alkaline Phosphatase↗
PubMed1992

L-dopa administration enhances exocytotic dopamine release in vivo in the rat striatum.

Peripheral administration of L-3,4-dihydroxyphenylalanine (L-DOPA) methylester increased extracellular levels of DOPA and dopamine (DA) in the rat striatum monitored by in vivo brain microdialysis. The increase in DA levels persisted after inhibition of DA reuptake by nomifensine. Administration of blockers of voltage-dependent Na+ (tetrodotoxin) or Ca2+ (NKY-722) channels through the dialysis membrane completely eliminated the increase in DA levels. These results demonstrate that the L-DOPA-induced DA release is exocytotic in nature and hence, derived from neurons in the striatum.

Animals↗
PubMed1992

Hepatoblastoma in a 2-year-old girl with trisomy 18.

A very rare case of mosaic type trisomy 18 associated with hepatoblastoma is described. The patient underwent an extended right hepatic lobectomy at 2 years of age, and the resected tumor was diagnosed as a fetal type dominant hepatoblastoma. The results of chromosome analysis demonstrated that in the peripheral blood and skin, the trisomy 18 cells were 80% and 67%, respectively. On the other hand, although virtually 100% of the cells in the normal liver tissue were 46, XX, about one third of the cells were trisomy 18 in the tumor tissue. At 2 years and 9 months after the operation, the patient was generally healthy and had no evidence of recurrence.

Carcinoma, Hepatocellular↗
PubMed1992

Tumor necrosis factor-alpha stimulates colony formation by a megakaryoblastic leukemia cell line, CMK.

The CMK cell line is an acute megakaryoblastic leukemia cell line established from a patient with Down's syndrome, and is known to possess characteristics of normal megakaryocytes. Several cytokines with the ability to stimulate megakaryopoiesis, such as interleukin-3 (IL-3), interleukin-6 (IL-6) and granulocyte-macrophage colony-stimulating factor (GM-CSF), stimulated colony formation by CMK cells. The present study revealed that tumor necrosis factor-alpha (TNF-alpha) stimulated colony formation by CMK cells; the potency was almost equal to that of IL-3, IL-6 or GM-CSF. Scatchard plot analysis revealed that CMK cells possess two types of specific binding sites for TNF-alpha. The high-affinity binding sites had an affinity constant of 0.18 nM, and numbered 5,000. The low-affinity binding sites had an affinity constant of 1.8 nM and numbered 19,000. These results raise the possibility that TNF-alpha can act as a growth-stimulating agent on megakaryocyte-lineage cell line.

Binding Sites↗
PubMed1992

SecY variants that interfere with Escherichia coli protein export in the presence of normal secY.

As an approach for studying how SecY, an integral membrane protein translocation factor of Escherichia coli, interacts with other protein molecules, we isolated a dominant negative mutation, secY-d1, of the gene carried on a plasmid. The mutant plasmid severely inhibited export of maltose-binding protein and less severely of OmpA, when introduced into sec+ cells. It inhibited growth of secY and secE mutant cells, but not of secA and secD mutant cells or wild-type cells. The mutation deletes three amino acids that should be located at the interface of cytoplasmic domain 5 and transmembrane segment 9. We also found that some SecY-PhoA fusion proteins that lacked carboxy-terminal portions of SecY but retain a region from periplasmic domain 3 to transmembrane segment 7 were inhibitory to protein export. We suggest that these SecY variants are severely defective in catalytic function of SecY, which requires cytoplasmic domain 5 and its carboxy-terminal side, but retain the ability to associate with other molecules of the protein export machinery, which requires the central portion of SecY; they probably exert the 'dominant negative' effects by competing with normal SecY for the formation of active Sec complex. These observations should provide a basis for further genetic analysis of the Sec protein complex in the membrane.

Alkaline Phosphatase↗
PubMed1992

The inhibition of lymphocyte blastogenesis by asparaginase: critical role of glutamine in both T and B lymphocyte transformation.

L-Asparaginase has long been used in the treatment of acute lymphoblastic leukemia or malignant lymphoma in childhood. To determine cell type specific sensitivity to this drug, the L-asparaginase-mediated inhibition of blastogenesis of human peripheral T or B lymphocytes was compared. The rate of incorporation of [3H]-thymidine into the DNA of either T lymphocytes due to phytohemagglutinin (PHA) or B lymphocytes due to Staphylococcus aureus Cowan I (SAC) was measured by the addition of Escherichia coli L-asparaginase in the medium. The blastogenic response of either T or B lymphocytes was also determined in medium depleted of exogenous asparagine and/or glutamine, both of which are hydrolyzed by this enzyme. The in vitro blastogenesis of either human T lymphocytes due to PHA or B lymphocytes due to SAC was inhibited by the inclusion of asparaginase in the medium. The deprivation of exogenous asparagine did not have any inhibitory effect on the blastogenic response of both T and B lymphocytes to each mitogen. On the other hand, the glutamine concentration in the culture medium provided a critical influence on the proliferative response of T and B lymphocytes. The rate of incorporation of [3H]-thymidine into DNA was increased markedly as the concentration of glutamine was increased from 2(-7)-2 mmol/l. It is concluded that the mechanism of inhibition of PHA- or SAC-stimulated lymphocyte blastogenesis by L-asparaginase is not asparagine deprivation but glutamine deprivation. Glutamine, which is the most abundant amino acid, is thought to have an important role in the immune response of lymphocytes.

Asparaginase↗
PubMed1992

Effect of aminophylline on brain tissue oxygenation in patients with chronic obstructive lung disease.

BACKGROUND: It is known that theophylline reduces cerebral blood flow in humans. To quantitatively assess the possible adverse effect of theophylline on brain tissue oxygen tension (PO2) due to decreased cerebral blood flow, two sets of experiments were conducted in mildly hypoxaemic patients with chronic obstructive lung disease. METHODS: Firstly, internal jugular venous PO2 (PjO2) was measured simultaneously with arterial and mixed venous blood PO2 (PaO2 and PvO2) during right heart catheterisation in 10 subjects (mean PaO2 73 mm Hg; conversion factor: 10 mm Hg = 1.33 kPa)) before and after intravenous infusion of aminophylline (6 mg/kg). The PjO2 and PvO2 were considered to reflect the average tissue PO2 for the brain and for the whole body respectively. Secondly, the relation between PaO2 and PjO2 over a wide range, with the PaCO2 similar to that in the first study, was investigated in a different group of 12 subjects by stepwise changes in inspiratory gas composition. RESULTS: The mean PjO2 decreased by as much as 6 mm Hg 15 minutes after an infusion of aminophylline, whereas PaO2 stayed at the same level and PvO2 showed only a small decrease. The low PjO2 value of 29 (SD 6) mm Hg with aminophylline in the first study was similar to the PjO2 value of 30 (2) mm Hg obtained during severe hypoxia (PaO2 45 mm Hg) in the second study. The coefficient of oxygen delivery for the brain decreased by 29% with aminophylline treatment, but did not change significantly during severe hypoxic challenge. CONCLUSIONS: These data suggest that an infusion of aminophylline lowers brain tissue PO2 appreciably when given to mildly hypoxaemic patients with chronic obstructive lung disease.

Aminophylline↗
PubMed1992

Rigid spine syndrome with selective respiratory muscle weakness.

Rigid spine syndrome is a rare myopathy characterized by mild axial and proximal muscle weakness, limitation of neck and trunk flexion, scoliosis and mild joint contractures. We describe a 20-year-old woman with this syndrome who developed severe respiratory muscle failure disproportionate to the well-preserved extremity muscles. Nocturnal home care ventilation has completely restored her arterial oxygen desaturation during a 5-year follow-up.

Adult↗
PubMed1992

Augmented heart rate response to hypoxia in patients with chronic obstructive pulmonary disease.

Effects of acute, progressive isocapnic hypoxia on heart rate (HR) and ventilation were determined in 31 patients with chronic obstructive pulmonary disease (COPD) and in 24 normal control subjects. There was an inverse linear relationship between heart rate and SaO2 in each subject. The slope factor (delta HR/delta SaO2) obtained from the regression line was significantly higher in the patients with COPD than in the normal control subjects (0.888 +/- 0.309 SD beats/min/% fall in SaO2 versus 0.693 +/- 0.287; p less than 0.05), whereas the ventilatory response (delta VE/delta SaO2) was not significantly different between the two groups. To elucidate factors responsible for the augmented heart rate response to hypoxia in the patients with COPD, we examined the relationships of delta HR/delta SaO2 with age, physical characteristics, pulmonary function data, and arterial blood gas data in all the subjects. A weak but significant relationship was found only between delta HR/delta SaO2 and FEV1/VC, %FEV1, RV/TLC, and %RV. Because the HR response to hypoxia correlates only with parameters that reflect the grade of airway obstruction, we believe that the enhanced HR response seen in patients with COPD is a result of the disease process in the airway and tissue, although the precise mechanism was not specified in this study.

Female↗