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Biomedical subjects

H Kanazawa

Publications and source records attributed to H Kanazawa.

At least 145 records · Page 8Linked to original sources
PubMed1995

Hypothalamic hamartoma: anatomic, immunohistochemical and ultrastructural features.

Four patients with hypothalamic hamartoma were examined by CT and/or MR imaging, immunohistochemistry and electron microscopy. The hamartomas arose from the hypothalamus and extended inferiorly. LH-RH neurons were detected in three cases by immunohistochemistry. Electron microscopy revealed large myelinated axons, axon terminals containing dense-core vesicles and axon terminals with clear vesicles forming asymmetrical synapses. The development of hypothalamic hamartoma and its functional manifestations (precocious puberty and laugh attacks) are discussed in reference to the migration of LH-RH neurons from the olfactory placode.

Axons↗
PubMed1995

Mechanism of mutagenesis induced by cytosine analogs bearing N(4)-substitutions.

The mechanism of mutagenesis induced by dihydropyrimido [4,5-c][1,2]oxazin-7-one deoxyriboside, P-nucleoside, was studied. This analog is highly mutagenic toward Escherichia coli and Salmonella typhimurium. In E. coli, it induces GC-to-AT and AT-to-GC transitions specifically. No transversions are inducible. P-nucleoside was highly mutagenic to a wild-type E. coli, but little mutagenic in a strain lacking thymidine kinase. This indicates that P-nucleoside may be phosphorylated by thymidine kinase after its uptake into bacteria. The mutagenesis induced by P-nucleoside was efficiently inhibited by the addition of thymidine. This inhibition further confirmed the involvement of thymidine kinase in the first step of the metabolism of P-nucleoside in the bacterial cells. These findings indicate that P-nucleoside is a mutagen of a nucleoside-analog type, causing mutations by the erroneous incorporation and replication. The experiments to prove its ambiguous nature in DNA synthesis is now under way.

Adenine↗
PubMed1994

Adrenomedullin, a newly discovered hypotensive peptide, is a potent bronchodilator.

The present study was designed to examine the effect of adrenomedullin (AM), a novel vasorelaxant peptide originally isolated from human pheochromocytoma, on histamine- or acetylcholine-induced bronchoconstriction in anesthetized guinea pigs in vivo. AM significantly inhibited acetylcholine-induced bronchoconstriction in a dose-dependent fashion. AM also significantly inhibited histamine-induced bronchoconstriction, but 10(-10) M AM had no significant inhibitory effect on this response. We also found that AM induced a long-lasting bronchodilator response, while isoproterenol induced transient bronchodilation. These preliminary findings suggest that AM may play important roles in airway function.

Acetylcholine↗
PubMed1994

Residues interacting with serine-174 and alanine-295 in the beta-subunit of Escherichia coli H(+)-ATP synthase: possible ternary structure of the center region of the subunit.

The mutation of serine-174 to phenylalanine that causes a defect in the Escherichia coli F1-ATPase beta-subunit is suppressed by further mutations; Gly-149 to Ser, Ala-295 to Thr, Ala-295 to Pro, or Leu-400 to Gln (Miki, J., Fujiwara, K., Tsuda, M., Tsuchiya, T. and Kanazawa, H. (1990) J. Biol. Chem. 265, 21567-21572). We analyzed the effects of these second site mutations and of a newly identified Asn-158 to Tyr mutation on the activities of the ATPase without the original Ser-174 to Phe mutation. The beta-subunit with each amino acid replacement was expressed in the mutant strain JP17, which does not have a beta-subunit. Cells transformed with the plasmid carrying Ala-295 to Pro mutation alone did not grow on minimal medium agar supplemented with succinate as the sole carbon source, and showed 3% of the wild-type ATPase activity, suggesting that this mutation caused structural alterations affecting the catalytic function of the enzyme. Conversely transformants with other mutations grew well and had higher ATPase activities, suggesting that these mutations did not cause extensive structural alterations. From the transformants with the plasmid carrying the Ala-295 to Pro mutation, seven revertants capable of cell growth on succinate plates were isolated and reversion mutations were identified at residues 140, 159, 166, 171, 172 and 184 of the beta-subunits. The results suggested that Ser-174 and Ala-295 do not necessarily interact directly, but that the regions including these suppression mutation sites close to Ser-174, and Ala-295 interact with each other for the proper functioning of the ATPase. The ternary structure of the region surrounded by the residues which were identified as the reversion mutation sites for Ser-174 to Phe and Ala-295 to Pro mutations is important for the catalytic function of this enzyme.

Alanine↗
PubMed1994

High-performance liquid chromatographic determination of catecholamine metabolites and 5-hydroxyindoleacetic acid in human urine using a mixed-mode column and an eight-channel electrode electrochemical detector.

An HPLC system for the simultaneous determination of acidic catecholamine metabolites, related compounds and 5-hydroxyindoleacetic acid (5-HIAA) in human urine was developed. A mixed-mode (C18/anion-exchange) column with isocratic elution using citrate buffer and an eight-channel electrochemical detector were used. Vanilmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC), 4-hydroxy-3-methoxyphenyllactic acid (vanillactic acid, VLA), homovanillic acid (HVA), vanillic acid (VA) and 5-HIAA in urine were determined simultaneously. Detection limits and inter (n = 5) and intra-assay (n = 5) coefficients of variation were satisfactory. The mean of analytical recoveries (n = 3, +/- C.V. (%)) were between 97 +/- 3.2 (VMA) and 105 +/- 4.8 (VA). Correlations between the analytical results for VMA, HVA and 5-HIAA obtained by an established method and the present method were satisfactory. The mean +/- 2 S.D. of the excretion rates of VMA, DOPAC, VLA, HVA, 5-HIAA and VA in urine from healthy adult volunteers were 0.61-4.36, 0.13-1.02, 0-0.35, 0.67-6.55, 0.50-5.14 and 0-0.55 mg/g creatinine, respectively.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1994

Reversion mutations in the beta subunit mutants of Escherichia coli F1-ATPase with defective subunit assembly: implications for structure and function of the amino-terminal region.

We analyzed reversion mutations of the mutants with a substitution of Leu-40 by Pro or Glu-41 by Lys in the beta subunit of Escherichia coli F1-ATPase. These mutants had an altered molecular assembly of the alpha and beta subunit on the membranes and lost the binding activity of the beta subunit to the monoclonal antibody beta 31. For the mutation of Leu-40 to Pro, we found that all reversion mutations Gln, or Ser besides the wild-type Leu. These pseudo reversion mutations restored the cell growth on minimal agar supplemented with succinate as the sole carbon source, the assembly of the alpha and beta subunits, and also the ATPase activities in the membranes. These results suggested that Leu-40 itself is not an essential residue for the function of the beta subunit but that the residue contributes to a conformation around residue 40, which is important for the assembly of the alpha and beta subunits onto the membranes. For the mutation of Glu-41 to Lys, pseudo reversion mutations were found at the original residue 41 as Lys to Gln or Asn and also at Arg-218 to Cys or His in addition to the original Glu-41 to Lys mutation. These suppression mutations recovered the cell growth, indicating the recovery of ATP synthesis. The ATPase activity was high in cells with the Lys-41 to Glu mutation but those were relatively low in those with other reversion mutations. The assembly of the alpha and beta subunits recovered in the revertants to the wild-type level, except for the Lys-41 to Asn mutation, which resulted in a decreased amount of the alpha subunit on the membranes. These results suggested that though Glu-41 is not an essential residue, it plays an important role in the assembly of the alpha and beta subunit on the membranes. The results also suggested that residues Arg-218 and Glu-41 are located close together and interact with each other, either directly or indirectly.(ABSTRACT TRUNCATED AT 400 WORDS)

Base Sequence↗
PubMed1994

Intergenic suppression in a beta subunit mutant with defective assembly in Escherichia coli F1ATPase. Second-site mutation in the alpha subunit.

Substitution of Leu-40 by Pro in the beta subunit (beta L40P) of Escherichia coli F1-ATPase caused a decrease in the amount of the alpha and beta subunits on the membranes. A revertant strain, Re50, carrying no suppression mutations in the uncD gene encoding the beta subunit, was isolated from the beta L40P mutant. The uncA gene from this revertant was amplified by PCR, and cloned into an expression plasmid. The expression plasmid carrying the uncA gene from the revertant was used for genetic suppression assays. The suppression mutation in Re50 was in the alpha subunit, and it recovered the assembly of the alpha and beta subunits into the F1F0 complex and the ATPase activity to 50% that of the wild type. In Re50, Leu-111 was substituted by Gln in the alpha subunit. These results suggest that the regions including Leu-40 in the beta subunit and Leu-111 in the alpha subunit are located close together and interact with each other, either directly or indirectly.

Base Sequence↗
PubMed1994

Amino acid replacements at binding sites of monoclonal antibody in the F1-ATPase beta subunit from Escherichia coli caused altered subunit interactions.

The monoclonal antibodies (mAb), beta 12 and beta 31, against the beta subunit of Escherichia coli F1-ATPase (Miki, J., Matsuda, T., Kariya, H., Ohmori, H., Tsuchiya, T., Futai, M., and Kanazawa, H. (1992) Arch. Biochem. Biophys. 294, 373-381) recognize the amino-terminal portion of the maximal 104 residues that are not exposed to the surface of the F1-ATPase. To identify the epitope residues within these 104 residues, we introduced random mutations in the region, and clones without binding activity to the mAb, beta 12 and beta 31, were screened. beta subunits defective in binding with mAb beta 12 or beta 31 had amino acid replacements at residues 26, 40, 52, 71, and 74 or at residues 40, 41, 43, 44, 46, and 52, respectively. These residues could be part of the epitope and are possibly located close together. We tested the effects of the mutations on oxidative phosphorylation-dependent growth by introducing expression plasmids of the beta subunit gene with the point mutations into the beta-less mutant, JP17. The alpha and beta subunits were missing from the JP17 membranes, and both subunits were assembled functionally after expression of the beta subunit was induced by introducing the expression plasmid. The replacement of Leu-40 by Pro caused the amounts of the alpha and beta subunits on the membranes to be reduced less than 20% of the amounts in wild type. The replacement of Glu-41 by Lys caused a loss of the alpha subunit on the membranes, without any decrease in the beta subunit. These results indicated that the mutation of Leu-40 to Pro affects the essential role of the beta subunit in the assembly of the alpha and beta subunits on the membranes and that the mutation of Glu-41 to Lys partly affects it. The amino-terminal region of the beta subunit, in particular its ternary structure, including residues 40 and 41, plays an important role in the molecular assembly of the alpha and beta subunits on the membranes.

Antibodies, Monoclonal↗
PubMed1994

A critical cytoplasmic domain of the interleukin-5 (IL-5) receptor alpha chain and its function in IL-5-mediated growth signal transduction.

Interleukin-5 (IL-5) regulates the production and function of B cells, eosinophils, and basophils. The IL-5 receptor (IL-5R) consists of two distinct membrane proteins, alpha and beta. The alpha chain (IL-5R alpha) is specific to IL-5. The beta chain is the common beta chain (beta c) of receptors for IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF). The cytoplasmic domains of both alpha and beta chains are essential for signal transduction. In this study, we generated cDNAs of IL-5R alpha having various mutations in their cytoplasmic domains and examined the function of these mutants by expressing them in IL-3-dependent FDC-P1 cells. The membrane-proximal proline-rich sequence of the cytoplasmic domain of IL-5R alpha, which is conserved among the alpha chains of IL-5R, IL-3R, and GM-CSF receptor (GM-CSFR), was found to be essential for the IL-5-induced proliferative response, expression of nuclear proto-oncogenes such as c-jun, c-fos, and c-myc, and tyrosine phosphorylation of cellular proteins including JAK2 protein-tyrosine kinase. In addition, analysis using chimeric receptors which consist of the extracellular domain of IL-5R alpha and the cytoplasmic domain of beta c suggested that dimerization of the cytoplasmic domain of beta c may be an important step in activating the IL-5R complex and transducing intracellular growth signals.

Amino Acid Sequence↗
PubMed1994

Effect of radical scavengers on the inactivation of papain by ascorbic acid in the presence of cupric ions.

Incubation of papain (EC 3.4.22.2) with ascorbic acid (AsA) and Cu2+ in acetate buffer (pH 5.6) results in an irreversible loss of enzyme activity by site-specific generation of free radicals [H. Kanazawa, S. Fujimoto, A. Ohara, Biol. Pharm.Bull., 16, 11 (1993)]. In this study, the effect of some compounds, known free radical scavengers, on the relationship between the inactivation of papain by the Cu(2+)-AsA system and the oxidation of AsA was investigated. Catalase completely protected the enzyme from inactivation by the Cu(2+)-AsA system, although hydrogen peroxide (H2O2) by itself, known to be generated during the autoxidation of AsA, did not inactivate the enzyme. The oxidation of AsA was unaffected by catalase. Both thiourea and sodium thiocyanate completely protected the enzyme from inactivation, while AsA was partially oxidized only in the initial stage. In the presence of potassium iodide, both the inactivation of the enzyme and the oxidation of AsA were characterized by a rapid initial phase followed by a stable phase where no reaction took place and, subsequently, a slower phase. Histidine partially prevented the inactivation of the enzyme and the oxidation of AsA. The present results suggest that H2O2 serves as a source of secondary, highly reactive species, probably hydroxyl radicals, which are responsible for the inactivation, and that the protection from inactivation by some radical scavengers, such as thiourea, sodium thiocyanate, potassium iodide, and histidine, is based on the removal of metal ions (Cu2+ or Cu+) at the specific site of inactivation.

Ascorbic Acid↗
PubMed1994

On the mechanism of inactivation of active papain by ascorbic acid in the presence of cupric ions.

An inactivation mechanism of active papain (EC 3.4.22.2) by the Cu(2+)-ascorbic acid (AsA) system was examined. Incubation of active papain, which contains an active sulfhydryl (SH) group, with the Cu(2+)-AsA system under aerobic conditions resulted in an irreversible loss of enzyme activity. The enzyme was not inactivated at a molar ratio of enzyme to Cu2+ of 1:< 1, whereas at a molar ratio of 1:1-2, the extent of inactivation showed the same dependence on the extent of oxidation of AsA. Saturation kinetics were observed with respect to the concentration of AsA. The degree of inactivation was dependent on the decrease in SH content of the enzyme. Catalase at a low concentration partially protected the enzyme from inactivation, but did not affect the oxidation of AsA. In addition, catalase at a high concentration completely protected both the enzyme from inactivation and AsA from oxidation. The present results suggest that an additional function of H2O2, besides producing hydroxyl radicals (.OH), is to promote the conversion of Cu+ into Cu2+, and that an active SH group of papain is site-specifically modified by the .OH, resulting in inactivation of the enzyme.

Amino Acids↗
PubMed1994

[Simultaneous determination of catecholamines, serotonin, and their precursors and metabolites in body fluid by an HPLC system with multi-electrode electrochemical detector].

We have developed two HPLC systems for the simultaneous determination of CA, 5-HT and their precursors and metabolites in human body fluid. One system consisted of two reversed-phase columns, a column switching device, a pair of electrodes for elimination of interferents and two sets of new electrochemical detectors with four electrodes. Using this system, adequate separation of peaks for 17 kinds of CA, 5-HT and their related metabolites in a standard solution required only 17 min. NE, MHPG, DOPAC, 5-HIAA, HVA and TRP levels in cerebrospinal fluid (CSF) were determined without any pretreatment of the CSF samples. The concentrations of DOPAC, 5-HIAA and HVA in CSF were significantly lower in Alzheimer's disease, vascular dementia and Parkinson's disease than in the controls. The other system was developed for determination of CA acidic metabolites and 5-HIAA in human urine. This system consisted of a mixed-mode column (C18/anion) and 8-channel electrochemical detector with isocratic elution of citrate buffer. Detection limits, precision and analytical recoveries of this method were satisfactory for clinical use.

Aged↗
PubMed1994

[A case of angiocentric immunoproliferative lesions in an ATL-virus carrier].

A 48-year-old woman was admitted to our hospital because of cough and low-grade fever. Chest X-ray films showed multiple ill-defined nodular shadows and laboratory findings showed that she was an ATL-virus carrier. Because the specimens obtained by transbronchial biopsy were not diagnostic, open thoracotomy was performed. Open lung specimens showed proliferation of lymphoid cells composed of polymorphorous lymphocytes, which do not represent cytologic atypia, plasma cells and histiocytes without necrosis. Several vessels were occluded by the infiltration of these cells. A diagnosis of angiocentric immunoproliferative lesions (Grade I), which was originally coined by Jaffe, was made. Complete remission was achieved by treatment with prednisolone and cyclophosphamide. This disease, which corresponds to lymphomatoid granulomatosis, is graded into three categories depending on the severity of cytologic and histologic malignancy. Although it has recently been suggested that its pathogenesis may be related to EB-virus infection, ATL-virus may have been a contributory factor in this case.

Carrier State↗
PubMed1994

[An experience of transjugular intrahepatic portosystemic shunt in the treatment of gastroesophageal varices].

Transjugular intrahepatic portosystemic shunt (TIPS) using an 8-mm Gianturco-Rösch Z stent was performed in 10 patients with liver cirrhosis admitted due to ruptured gastroesophageal varices. The treatment was successful in 9 patients, and the portal pressure decreased form 25 mmHg to 17 mmHg immediately after TIPS and to 15 mmHg 2 weeks after TIPS. The patency of the shunt was maintained adequately in the patients examined 3 months after TIPS. Endoscopy 1 month after TIPS showed improvements in the gastroesophageal varices in all patients. Hemobilia and intraperitoneal bleeding were observed in 1 patient each during procedure. Transient jaundice was observed in 2 and increase in the blood ammonia level were noted in 3 (one of whom had hepatic encephalopathy) postoperatively. All these complications could be treated easily. TIPS which brings about an immediate marked reduction in the portal pressure appears to be promising as a new treatment for portal hypertension.

Adult↗
PubMed1994

[Two case reports of recurrent mediastinitis with chronic mediastinal fistula successfully treated with muscle flap re-transposition].

We experienced two cases of recurrent poststernotomy mediastinitis with chronic mediastinal fistula. Both cases had already received muscle flaps for post operative mediastinitis. However, chronic mediastinal fistula appeared after nine months in the first case, and eleven months in the second case. We removed the infected tissue and the predgets, which were used on the ascending aorta. Then closed the wound by the muscle flap closure. The chronic fistula were closed, and the functional and cosmetic results were excellent.

Female↗
PubMed1994

Total circular annuloplasty with absorbable suture for the repair of left atrioventricular valve regurgitation in atrioventricular septal defect.

To obtain a better control of left atrioventricular valve regurgitation, we applied total circular annuloplasty with the use of absorbable sutures to 14 children with atrioventricular septal defect (six complete forms and eight incomplete forms). In the intraoperative period, a good coaptation of the leaflets was achieved and the regurgitation was minimized or disappeared. Follow-up studies with echocardiography for 13 survivors showed a gradual increase of annular size during the postoperative period. Ten patients of the survivor group (77%) maintained good valvular competency in a long-term period. Total circular annuloplasty is a simple and effective procedure to reduce the regurgitation and prevent the annular dilatation during the immediate postoperative period.

Cardiac Surgical Procedures↗
PubMed1993

Liquid chromatography-mass spectrometry for the determination of medetomidine and other anaesthetics in plasma.

A liquid chromatographic-atmospheric pressure chemical ionization mass spectrometric method is presented for the simultaneous determination of medetomidine and other anaesthetic drugs in solutions and dog plasma. The drugs examined were flumazenil, butorphanol, atropine, ketamine, xylazine, medetomidine, atipamezole and midazolam. The separation was carried out on a reversed-phase column using methanol-0.1 M ammonium acetate (3:2) as eluent.

Anesthetics↗