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Biomedical subjects

H Inoue

Publications and source records attributed to H Inoue.

At least 1,729 records · Page 96Linked to original sources

Specific determination of arylsulfatase A activity.

Arylsulfatase activities in biological materials are too low to be detected by the methods available hitherto. A sensitive and specific assay method for arylsulfatase A (AS-A) has been developed in the present study. Ascorbate-2-sulfate is known to be a specific natural substrate of AS-A; the ascorbic acid liberated by the action of AS-A was quantitatively determined using HPLC equipped with an amperometric detector. The method was used to analyze the activity of AS-A in biological materials.

Animals↗

Complete amino acid sequence of rat L-type pyruvate kinase deduced from the cDNA sequence.

cDNA clones, containing the entire coding region of rat L-type pyruvate kinase, were isolated and their nucleotide sequences were determined by the dideoxy-chain-termination method. The predicted coding region, which spans 543 amino acids, established the complete amino acid sequence of the L-type isozyme of pyruvate kinase for the first time. The deduced amino acid sequence of the L type has one phosphorylation site in its amino terminus and shows about 68% and 48% homologies with M1-type pyruvate kinase of chicken and yeast pyruvate kinase respectively. Domain A exhibits higher homology than domains B and C. The residues in the active site of the L-type enzyme of rats, lying between domains B and A2, are rather different from those of the M1-type enzyme of chickens, but other residues constituting the active site are identical with those of the chicken M1 type except for one amino acid substitution.

Amino Acid Sequence↗

Increased labelling of polyphosphoinositide in chemically transformed cell line C3H10T1/2 CL8.

The effect of malignant transformation of cells on phosphatidylinositol metabolism was investigated using C3H10T1/2 cells and its chemically transformed cell line, MCA CL-16 cells. We found that incorporation of [32P]Pi into polyphosphoinositide was greatly increased in the transformed cells. A similar tendency was observed when myo-[2-3H]inositol was used as a labelling reagent. It is also observed that influx of labelled inorganic phosphate is enhanced 2-fold by the cell transformation. Therefore, promotion of polyphosphoinositide labelling in the transformed cell might be caused not only by the enhanced metabolism of phosphatidylinositol but also by the increased membrane permeability for radioactive labelling reagents.

Animals↗

Epicardial activation patterns of torsade de pointes in canine hearts with quinidine-induced long QT interval but without myocardial infarction.

We studied the epicardial activation sequence during torsade de pointes in canine hearts with quinidine-induced long QT interval. Following a toxic dose of quinidine sulfate (30 mg/kg), polymorphic ventricular tachycardia was induced by extrastimuli. In nine dogs, 22 episodes of nonsustained polymorphic ventricular tachycardia and six episodes of monomorphic ventricular tachycardia were induced. Of 22 episodes of nonsustained polymorphic ventricular tachycardia, 12 episodes showed typical torsion of the spikes of the QRS complex around the isoelectric line (torsade de pointes). Isochronal maps were made from 38 simultaneously recorded bipolar electrograms and showed that each change in QRS morphology was associated with a change in the earliest epicardial activation site and/or a change of epicardial activation sequence. During episodes of torsade de pointes, the earliest epicardial activation site migrated gradually from one site to another site. The transitional QRS complexes had their earliest epicardial activation sites between the new and old site of the earliest epicardial activation. The changes in the earliest epicardial activation site during polymorphic ventricular tachycardia without torsade de pointes pattern was not as great as that noted during torsade de pointes. This study suggests that in a noninfarcted canine model torsade de pointes is produced by alteration in the earliest site of activation rather than by competition from two or more competing foci.

Animals↗

The effect of arcuate nucleus transplantation on the development of the anterior pituitary in monosodium glutamate-treated rats.

Treatment with monosodium glutamate (MSG) during the neonatal period is known to produce a selective lesion of the arcuate nucleus in rat brain, which is the major site of production of growth hormone releasing-hormone (GRH), followed by a secondary reduction in growth hormone (GH) synthesis in the anterior pituitary. Normal arcuate nuclei from hypothalamic areas of newborn rats were transplanted into the third ventricles of 27-day-old rats which were treated with MSG on alternate days for the first 10 days of life. Ninety days after birth, the anterior pituitaries were examined for GH synthesis by immunohistochemical staining with GH antiserum. The results indicated that the impaired GH synthesis in the anterior pituitary treated with MSG was partially restored in some recipients by grafts of arcuate nuclei in which the GRH-containing neurons were clearly detected by immunohistochemical staining with GRH antiserum.

Animals↗

Recombinant human granulocyte colony stimulating factor: molecular and biological characterization.

Human or rodent bone marrow treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) in a CFU-GM assay yield predominantly granulocytic colonies. The specificity for granulocyte progenitors in vitro is also demonstrated in vivo by a five- to six-fold elevation in hamster peripheral blood neutrophils. Other cell types (monocytes, lymphocytes and eosinophils) remain stable. Analysis of mRNA from the bladder carcinoma cell line 5637 (1A6) shows the predominant species of mRNA codes for a mature protein of 174 amino acids. A small fraction of the mRNA can code for an alternative form of hG-CSF containing additional three amino acids between positions 35 and 36.

Animals↗

Double and triple sequential shocks reduce ventricular defibrillation threshold in dogs with and without myocardial infarction.

The role of optimal placement of electrodes and mode of shock delivery from a defibrillator was examined in dogs with and without myocardial infarction. Single, double and triple truncated exponential shocks separated by 1 ms were delivered through various electrode combinations and cardiac vectors after electrical induction of ventricular fibrillation. A single shock through a pathway not incorporating the interventricular septum (catheter electrodes or epicardial patches between anterior and posterior left ventricle) required the highest total energy (22.6 and greater than 26.4 J, respectively) and peak voltage (1,004 and greater than 1,094 V, respectively) to terminate ventricular fibrillation. A single shock through a pathway including the interventricular septum required lower total energy and peak voltage to defibrillate. Combinations of two sequential shocks between an intracardiac catheter electrode and anterior left ventricular epicardial patch, between the catheter electrode and subcutaneous extrathoracic plate and between three ventricular epicardial patches all significantly reduced total energy (7.7, 8.7 and 7.8 J, respectively) and peak voltage (424, 436 and 424 V, respectively) needed to defibrillate. Three sequential shocks exerted no significant additional reduction in total energy of the defibrillation threshold than did two sequential shocks. Infarcted canine heart required less peak voltage but not total energy to terminate ventricular fibrillation than did noninfarcted heart. Therefore, two sequential shocks over different pathways reduce both total energy and peak voltage required to terminate ventricular fibrillation.

Animals↗

Ventricular fibrillation threshold in canine hearts with quinidine induced prolonged QT interval: effects of atrial pacing, isoproterenol, and lignocaine.

Ventricular fibrillation threshold was determined using a train of pulses, 4 ms in duration, delivered at 10 ms intervals, in 20 anaesthetised dogs receiving toxic doses of quinidine sulphate (30 mg X kg-1) injected intravenously over 5 min to prolong the QT interval. The effects of atrial pacing, isoproterenol, and lignocaine on ventricular fibrillation threshold were investigated. Quinidine administration lowered the mean(SD) threshold from 14.7(5.1) to 8.8(3.6) mA (n = 20, p less than 0.001). Atrial pacing shortened the mean(SD) basic cycle length from 652 to 400 ms and the QT interval (378(28) vs 334(19) ms, n = 8, p less than 0.01) but did not alter the mean(SD) threshold (7.0(2.3) vs 7.5(1.5) mA). Isoproterenol infusion to maintain basic cycle length at 421(23) ms shortened the mean(SD) QT interval to 291(14) ms (p less than 0.001) and increased the mean(SD) threshold to 13.6(5.9) mA (n = 8, p less than 0.01). Lignocaine (plasma concentration 1.36 micrograms X ml-1) increased the mean(SD) threshold from 10.9(3.4) to 16.7(6.7) mA (n = 7, p less than 0.02) without affecting basic cycle length and QT interval. Thus atrial pacing shortened the QT interval slightly but did not make the canine heart with a quinidine induced prolonged QT interval less vulnerable to electrically induced ventricular fibrillation, whereas both isoproterenol and lignocaine did, although they affected basic cycle length and QT interval differently.

Animals↗

Effects of epidermal growth factor on the syntheses of DNA and polyamine in isoproterenol-stimulated murine parotid gland.

The effects of epidermal growth factor (EGF) on isoproterenol (IPR)-stimulated DNA synthesis and the activities of the rate limiting enzymes of polyamine synthesis (ornithine and S-adenosylmethionine decarboxylases) in parotid glands were investigated in vitro in cultured rat parotid explants and in vivo in submandibulectomized mice (mice after bilateral removal of the submandibular and sublingual glands). When the explants were cultured on siliconized lens paper floating on chemically defined synthetic medium, IPR caused the increases of both tissue cAMP level and the two decarboxylase activities in the prereplicative period and the stimulation of DNA synthesis with similar time courses to those observed in vivo. Dibutyryl cyclic AMP (DBcAMP) also increased the enzyme activities, but not DNA synthesis. EGF (1-2 ng/ml) had little effect on the IPR- and DBcAMP-dependent increases of amylase secretion and the enzyme activities, but it markedly enhanced IPR-stimulated DNA synthesis. Moreover, increase in DNA synthesis by DBcAMP was clearly observed in the presence of EGF when the explants were treated with this nucleotide analogue only during the early prereplicative period. In in vivo experiments, IPR-dependent increase in DNA synthesis was less in submandibulectomized mice than in intact animals. This decreased response to IPR of DNA synthesis was completely reversed by administration of EGF, though EGF alone did not induce either the enzymes or DNA synthesis. In submandibulectomized mice, although increases in the enzyme activities 8 h after injection of IPR were lower and they were significantly reversed by EGF, the activities at 12 h and the changes in polyamine levels at 8 and 12 h were almost the same as those in intact mice and were not affected by EGF treatment. These results obtained in vitro and in vivo suggest that EGF participates in the maximal response of IPR-dependent DNA synthesis but is not involved in the change of polyamine synthesis induced by IPR in murine parotid glands.

Adenosylmethionine Decarboxylase↗

Developmental variation and amino acid sequences of cytochromes c of the fruit fly Drosophila melanogaster and the flesh fly Boettcherisca peregrina.

The amino acid sequences of cytochromes c purified from the fruit fly Drosophila melanogaster and the flesh fly Boettcherisca peregrina were determined. In contrast with the case of the housefly, isocytochromes c were not detected in these flies at any developmental stage. The sequence of fruit fly cytochrome c differed from that reported previously but was identical with that predicted from the nucleotide sequence of the fruit fly cytochrome c gene (DC4) (Limbach, K.J. & Wu, R. (1985) Nucl. Acids Res. 13, 631-644). Isocytochrome c of the fruit fly, reported to be encoded by the DC3 gene, was not detected as a functional cytochrome c molecule.

Amino Acid Sequence↗

Giant-cell reparative granuloma of the thoracic vertebra.

Giant-cell reparative granuloma occurring in the thoracic vertebra of a 16-year-old girl is presented, and the literature concerning the subject is reviewed. This report is of the second documented case with this essentially rare lesion arising in the vertebra. Microscopically, the lesion was characterized by a reactive granulation tissue with formation of reactive osteoid, collagenization and vascularization, and focal accumulations of giant cells, particularly around hemorrhagic portions. The patient is doing well without further recurrence 38 months after the second therapeutic curettage. The authors describe the clinicopathologic features of this particular case because of its rarity in location, and briefly discuss the difference from other giant-cell lesions of bone mainly in view of differential diagnosis.

Adolescent↗

Ventilatory response to phasic contraction and passive movement in graded anesthesia.

The ventilatory response to electrically induced contraction (EIC) and passive movement (PM) of hindlimb muscles at different levels of anesthesia was studied in 11 chloralose-urethan anesthetized dogs with and without rhizotomy. The level of anesthesia was assessed by corneal reflexes and measurements of the ventilatory response to hypercapnia. Muscle contraction was induced by electrically stimulating the peripheral cut ends of the sciatic and femoral nerves for 4-5 min, and PM was induced manually at the same frequency and amplitude as during EIC. In spinal intact dogs (n = 7), initial rapid increases in minute ventilation (VE) during EIC and PM were found in both light and deep anesthesia. Further increases in VE above the initial rise were seen during EIC but not PM. The initial rapid increases in VE did not differ between the two anesthetic levels. The steady-state ventilatory response during EIC decreased as anesthesia deepened but did not do so during PM. Rhizotomy (n = 4) abolished the initial rapid increase in VE during EIC and PM and the steady-state VE response to PM at both anesthetic levels. These results suggest that the transitional ventilatory response is neurally mediated from the muscles and not affected by the level of general anesthesia. Additionally, the anesthesia-induced reduction of ventilatory response may be due to depression of responsiveness to CO2 rather than to the inspiratory motoneuron pathway.

Anesthesia, General↗

Direct-writing recorder of the frequency dependence of dynamic compliance analyzed from one cycle of breathing and pulmonary resistance: effect of fenoterol on asthmatic subjects.

We developed a new method of direct-writing recording of the frequency dependence of dynamic compliance analysed from one cycle of breathing and pulmonary resistance. Both pulmonary resistance (RL) and frequency dependence of dynamic compliance (Cdyn.f) are calculated by Fourier-series analysis of flow and transpulmonary pressure in a single cycle of breathing. RL was obtained from fundamental harmonics. Cdyn.f was calculated from 1st and 2nd harmonics and estimated by the ratio of Cdyn at 0.5 Hz to Cdyn at zero frequency, C0.5/C0. We used fenoterol aerosol which contained 0.2 mg of fenoterol with one puff of aerosol. Two puffs of fenoterol aerosol were used in each subject and followed changes of RL, C0 and C0.5 as long as 30 min. After fenoterol inhalation RL decreased considerably and C0.5/C0 cont [C0.5/(control C0)] increased. With time, while RL was kept stable, C0.5/C0 cont increased further. We suggest that fenoterol has a potent effect on the small airways which is enhanced with time.

Administration, Inhalation↗

Changes in bronchial reactivity to acetylcholine with type C influenza virus infection in dogs.

To investigate the mechanisms involved in increasing bronchial reactivity, we made a model of airway reactivity increase in dogs after Type C influenza virus infection. Five beagle dogs were inoculated intranasally with the virus, and the time courses of changes in the hemagglutination inhibition (HI) antibody titer and the bronchial reactivity were determined. To assess bronchial reactivity the dogs were anesthetized, and dose-response curves of pulmonary resistance were obtained against increasing concentrations of acetylcholine aerosol. The dogs infected with the virus showed mild symptoms of rhinorrhea and cough. The HI antibody titer was significantly increased in all dogs, with peak values at 1 to 3 wk after infection. The bronchial reactivity to acetylcholine began to increase towards Day 3, reached a peak at 1 to 2 wk, and returned to a normal level at 4 wk. The airway reactivity to acetylcholine at 2 wk after infection was increased by 2.3 to 6.5 times the normal value in terms of the acetylcholine provocative concentration. The mean increase was significant at 1 wk (p less than 0.05), 2 wk (p less than 0.01), and 3 wk (p less than 0.05). Although both the HI antibody titer and the airway responsiveness increased together towards 1 to 2 wk, no close relationship between these factors was observed thereafter. The present dog model of airway hyperreactivity may be useful for further investigation of the mechanism governing increase in bronchial reactivity with respiratory viral infection in normal subjects as well as in patients with asthma.

Acetylcholine↗