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Biomedical subjects

H Sakurai

Publications and source records attributed to H Sakurai.

At least 541 records · Page 30Linked to original sources
PubMed1986

Purification of vanadium binding substance from the blood cells of the tunicate, Ascidia sydneiensis samea.

Vanadium binding substance has been partially purified through chromatographies on Sephadex G-25 and SE-Cellulose at pH 2.3. The binding substance was colorless, relatively stable and maintained vanadium ion. The vanadium ion in the substance existed in vanadyl form (VO(IV)). Furthermore, the substance had an apparent affinity for exogenous vanadium ion(V) and contained a reducing sugar.

Animals↗
PubMed1986

Levels of topoisomerase II and DNA polymerase alpha are regulated independently in developing neuronal nuclei.

A possible correlation between activity levels of topoisomerase II and DNA polymerase alpha was studied in neuronal nuclei from developing rat brain. A high level of canonical topoisomerase II activity was detected in neuronal nuclei throughout the development even at a late stage (28 days after birth) when the activity of DNA polymerase alpha decreased to less than 2% of the fetal level. Thus, in contrast to other systems, topoisomerase II does not change in parallel with DNA polymerase alpha during neuronal development. Our results suggest that topoisomerase II is required to maintain some fundamental processes in differentiated cells, including transcription.

Age Factors↗
PubMed1986

[Clinical assessment of posterior pituitary function by direct measurement of plasma vasopressin levels during hypertonic saline infusion].

Clinical usefulness of a radioimmunoassay of plasma arginine vasopressin concentration (AVP) during hypertonic saline infusion for the assessment of posterior pituitary function was studied in comparison with the conventional water deprivation test. Infusion of 5% saline at a rate of 0.05 ml/kg/min for 120 min in 15 normal subjects induced an elevation of plasma osmolality (Posm) from 290.3 +/- 0.7 to 307.5 +/- 2.1 mOsm/kg with a resultant increase in AVP from 2.4 +/- 0.4 to 9.9 +/- 2.2 pg/ml. During the infusion, a highly significant correlation between AVP and Posm was observed with a regression line expressed as AVP = 0.40 (Posm - 283.0). In 22 polyuric patients, on the other hand, the infusion induced a marked elevation of Posm from 302.6 +/- 2.5 to 321.3 +/- 2.9 mOsm/kg, but caused a slight (less than 5.8 pg/ml) or no increase in AVP from the basal levels (0.5 +/- 0.1 pg/ml). A conventional water deprivation test was carried out in ten patients with neurogenic diabetes insipidus, including one who had coincidental nephrogenic diabetes insipidus. As would be expected, urine osmolality (Uosm) did not rise beyond Posm in seven of them. However, two of three other patients, who had a complete lack of AVP response to the hypertonic saline, were able to concentrate their urine with a maximal Uosm/Posm of 1.3 and 1.1 respectively. The concurrent decrease in creatinine clearance to 49 and 57% of the initial values, respectively, indicated that a marked reduction in glomerular filtration rate due to severe dehydration was responsible for the unexpected concentration of urine in the patients with totally impaired AVP secretion. Based on these results, we conclude that the direct measurement of AVP during hypertonic saline infusion is an essential procedure for the accurate evaluation of posterior pituitary function.

Adolescent↗
PubMed1986

DNA strand scission by hemin-thiolate complexes as models of cytochrome P-450.

Hemin-thiolate complexes, as chemical models for cytochrome P-450 monooxygenases, have been shown to cause strand scission of DNA. Circular super-coiled DNA was degraded to open-circular and linear forms by these complexes in 30 min at pH 7.8 under aerobic conditions, the degradation depending on the structure of the thiol ligand and the ratio of thiol ligand to hemin concentration. The relationship between the structure of the thiol ligand and DNA strand cleaving activity was examined. Complete cleavage of DNA was observed by complexes containing TGE and ME at 400-600 moles excess of thiol ligand to hemin, those containing Cys, CysMe, and CysEt at 50-200 moles excess, and those containing MPG, GSH, penta- and nona-peptides at 5-20 moles excess. Complexes containing NACys and MEA caused no cleavage of DNA. Inhibition experiments suggested the involvement of active oxygen species in the cleavage.

Cysteine↗
PubMed1986

A possible model of hemoprotein-peroxide complexes formed in an iron-tetraphenylporphyrin system.

A ferric low-spin species with an anomalously small g3-g1 separation generated by the reaction of Fe(III)tetraphenylporphyrin with t-butylhydroperoxide in the presence of tatramethylammonium hydroxide was characterized by ESR spectroscopy. The reaction kinetics, investigated by monitoring ESR intensity, indicated that this low-spin complex is highly reactive and easily changed to non-heme type iron complexes. The rhombic parameters of this complex are very similar to those of heme-peroxide adducts such as [Fe(II)-hemoglobin-O2]- and [Fe(II)-horseradish peroxidase-O2]-.

Electron Spin Resonance Spectroscopy↗
PubMed1986

Isolation, characterization and thiol exchange reaction of penicillamine selenotrisulfides.

Penicillamine selenotrisulfides formed by the reaction of selenite with D-, L-, and DL-penicillamines in the aqueous solution have been isolated and submitted to the spectroscopic and chromatographic analyses. FAB-MS spectra indicated the molecular ion peak (M+1)+ at m/z 377, which confirmed the molecular formula (C10H20N2O4S2Se) for all these selenotrisulfides. The [77Se]NMR and HPLC results showed that symmetric selenotrisulfides (DD and LL) were formed from D- and L-penicillamines, respectively, while asymmetric selenotrisulfide (DL) in addition to symmetric ones was formed from DL-penicillamine. The occurrence of thiol exchange reaction of selenotrisulfides was verified.

Chromatography, High Pressure Liquid↗
PubMed1986

Effect of serotonin and beta-endorphin on the release of luteinizing hormone in the hen (Gallus domesticus).

Serotonin (5-hydroxytryptamine) and beta-endorphin administered into the third ventricle of the hen blocked normal and progesterone-induced ovulation, and suppressed the release of LH in normal and progesterone-injected hens. p-Chlorophenylalanine, an inhibitor of serotonin synthesis, caused the release of LH and diminished the effect of beta-endorphin. Naloxone, an antagonist of opiate peptides, diminished the effect of beta-endorphin but not the effect of serotonin. The results suggest that both serotonin and beta-endorphin are involved in the control of LH release in the hen as an inhibitory agent, and serotonin is predominant while beta-endorphin is subsidiary to the inhibition of the LH release.

Animals↗
PubMed1986

Plasma renin activity, active and inactive renin concentrations, and their responses to beta 1-adrenoceptor blockade with metoprolol in hyperthyroidism.

Plasma renin activity (PRA), plasma renin concentration (PRC), inactive renin concentration (IRC) and total renin concentration (TRC) were measured in 31 normal controls and in 8 patients with hyperthyroidism. TRC was determined as angiotensin I generated with sheep renin substrate after an acid activation of plasma. The angiotensin I of non-acidified plasma was expressed as PRC. IRC was calculated as TRC minus PRC. The mean values for PRA, PRC, IRC and TRC were significantly (P less than 0.05 to P less than 0.01) higher in the hyperthyroid patients than in the normal or euthyroid controls. The administration of a beta 1-adrenergic blocker, metoprolol (120 mg/day for 14 days), produced a significant (P less than 0.05 to P less than 0.01) fall in levels of T4, PRA and TRC, and reduced the active renin ratio calculated from PRC/TRC significantly (P less than 0.025), as compared to the pretreatment values. Our observations support the idea that the higher PRA in hyperthyroidism is due to an increased secretion of renin. Furthermore, the results may indicate that the conversion of inactive to active renin is accelerated in hyperthyroidism, possibly by an increased sympathetic activity.

Adult↗
PubMed1986

ADP binding to TF1 and its subunits induces ultraviolet spectral changes.

Adenine nucleotide binding sites on the coupling factor ATPase of thermophilic bacterium PS3 (TF1) were investigated by UV spectroscopy and by equilibrium dialysis. When ADP was mixed with TF1 in the presence and in the absence of Mg2+, an UV absorbance change was induced (t1/2 approximately 1 min) with a peak at about 278 nm and a trough at about 250 nm. Similar spectral changes were induced by ADP with the isolated beta subunits in the presence and in the absence of Mg2+, and with the isolated alpha subunits in the presence of Mg2+ although the magnitudes of the changes were different. From equilibrium dialysis measurement we identified two classes of nucleotide binding sites in TF1 in the presence of Mg2+, three high-affinity sites (Kd = 61 nM) and three low-affinity sites (Kd = 87 microM). In the absence of Mg2+, TF1 has one high-affinity site (Kd less than 10 nM) and five low-affinity sites (Kd = 100 microM). Moreover, we found a single Mg2+-dependent ADP binding site on the isolated alpha subunit and a single Mg2+-independent ADP binding site on the isolated beta subunit. From the above observations, we concluded that the three Mg2+-dependent high-affinity sites for ADP are located on the alpha subunit in TF1 and that the single high-affinity site is located on one of the beta subunits in TF1 in the absence of Mg2+.

Adenosine Diphosphate↗
PubMed1986

Cleavage of DNA by model complexes of cytochrome P-450.

Thiolate-hemin complexes as chemical models for cytochrome P-450 have been shown to cause cleavage of DNA. The cleavage of DNA to open-circular and linear forms depended on the structure of thiol ligand and the thiol ligand:hemin ratio at pH 7.8. Complete cleavage of DNA was observed by complexes containing thioglycolate ethylester and mercaptoethanol at 400-600 moles excess of thiol ligand to hemin, those containing cysteine, cysteine methylester and cysteine ethylester at 50-200 moles excess, and those containing mercaptopropionylglycine, glutathione, glutathione dimethylester, penta- and nonapeptides at 5-100 moles excess. Inhibition experiments suggested the involvement of active oxygen species in the cleavage of DNA.

Cytochrome P-450 Enzyme System↗
PubMed1986

[Clinical effects of Chinese herb medicine (hochu-ekki-to) on infertile men].

Hochu-Ekki-To, a Chinese herb medicine, was administrated orally to 45 patients with primary male infertility due to oligozoospermia (sperm density was below 40 million/ml) for over 12 weeks. After the treatment, significant increases in sperm concentrations, rate of motility and total counts of normal spermatozoa, which was calculated as seminal volume by sperm density and by rate of normal form of spermatozoa, were observed, especially, in the group of moderate oligozoospermia (sperm density was ranged from 20 to 40 million/ml). In the clinical effect, the rate of pregnancy was 20.0% (9/45) and the rate of clinical efficiency was 51.1% (23/45). No changes of laboratory examinations were observed. We conclude that this Chinese herb medicine is more effective for the treatment of oligozoospermia, especially moderate oligozoospermia, without any serious side effects.

Drug Evaluation↗
PubMed1985

Atrial natriuretic factor inhibits vasopressin secretion from rat posterior pituitary.

The effects of synthetic atrial natriuretic factor (ANF) were studied in superfused rat posterior pituitary gland. ANF (10(-6)M, 10(-10)M) significantly inhibited basal as well as KC1 (50 mM) or angiotensin II-stimulated immunoreactive arginine vasopressin secretion. The magnitude of inhibition was greater at 10(-6)M than at 10(-10)M. ANF also decreased cAMP secretion and increased cGMP secretion from the posterior pituitary. These results suggest that ANF directly acts on the posterior pituitary to inhibit arginine vasopressin secretion and that this effect is, at least, partly mediated by the changes in cyclic nucleotide production.

Angiotensin II↗
PubMed1985

Kinetic studies on neurotoxin-binding inhibitory antibodies in myasthenia gravis.

A method for measuring antibodies which inhibit the binding of 125I-alpha-bungarotoxin to the acetylcholine receptor was developed. The procedure allows the detection of inhibitory activity in 70% of sera from the patients with myasthenia gravis. Kinetic analysis showed that (1) a linear dose-response exists between logarithmic concentrations of any test serum and inhibitory activity, (2) the slope of the inhibition line is identical among various myasthenic sera and, (3) the slope of line induced by various cholinergic ligands was comparable to that obtained by myasthenic sera. The inhibitory activity correlated with the titer of anti-acetylcholine receptor antibody measured by an immunoprecipitation method (r = 0.76, p less than 0.01).

Adolescent↗
PubMed1985

Partition of divalent and total manganese in organs and subcellular organelles of MnCl2-treated rats studied by ESR and neutron activation analysis.

The possibility that Mn2+ is converted to other valency states in vivo was examined by measuring the ratio of Mn2+, determined by ESR, to total manganese, determined by neutron activation analysis combined with chemical separation, in various organs of control rats and rats treated with MnCl2. In control rats, the total manganese content was high in the thyroid, hypophysis, adrenal, pancreas, liver and kidney, but the Mn2+ contents of these organs were low. In rats treated with Mn2+, the total manganese contents of all organs increased, but the Mn2+ contents still remained low. With regard to subcellular distribution, the total manganese content was high in the nuclear and mitochondrial fractions of the liver and kidney, and in the microsomal and supernatant fractions of the pancreas. The ratio of Mn2+ to total manganese was relatively high in the microsomes of the liver and kidney of control rats, and in the nuclear fraction of the pancreas of Mn2+-treated rats. Thus, the distribution and behavior of manganese in the pancreas were different from those in other organs. Purified liver nuclei and mitochondria were demonstrated to contain manganese, indicating that manganese is tightly bound in each cellular compartment.

Animals↗
PubMed1985

Synthetic analogs of the active site of cytochrome P-450cam characterization of thiolpeptide fragment-hemin complexes by optical and ESR spectrometries.

Thiol-containing penta (Leu-Ala-Cys-Ser-Leu) and nona (Leu-Ala-Cys-Ser-Leu-Ile-Glu-Ser-Leu) peptides corresponding to residues 132-136 and 132-140, respectively, of apo-P450 from Psuedomonas putida were synthesized to examine heme-binding by the enzymes in the oxidized (ferric) form. The peptide-hemin complexes prepared in solution were characterized by their optical and ESR spectra. In these complexes without nitrogenous ligands, no ferric complexes in the low-spin state were observed, suggesting that simultaneous axial coordination of Cys-134 (thiolate) and Ser-139 (hydroxyl) of apo-P450cam to the heme is unlikely to occur. In the presence of nitrogenous ligands, such as py, Im and MeIm, the resulting complexes were good models of apo-P450cam-nitrogenous ligand adducts in the low-spin ferric state retaining a thiolate-Fe(III)-nitrogen axial coordination mode, as judged by their spectral pattern as well as by crystal field analysis of ESR g-values.

Amino Acid Sequence↗
PubMed1985

[A simple and highly sensitive radioimmunoassay for 8-arginine vasopressin in human plasma using a reversed-phase C18 silica column].

A highly sensitive and simple radioimmunoassay for the measurement of 8-arginine vasopressin (AVP) in human plasma has been developed. The dose response curve ranges from 0.025 to 8 pg/tube. This simple extraction technique employing an ODS C18 column recovered 87.1 +/- 10.4 (mean +/- SD)% of AVP in the range of 1-10 pg/ml added to 0.5 ml plasma. Determination of AVP in each fraction of plasma, which was gel-filtrated through a Sephadex G25 (1 X 25 cm), revealed that the fraction of plasma AVP was superimposed on that of authentic AVP, and interference of non-specific substances was completely eliminated by an ODS C18 column. Using the assay, 13 of 16 patients with diabetes insipidus (DI) showed plasma AVP concentrations ranging from 0.03 to 0.21 pg/ml, and the other 3 patients had less than 0.03 pg/ml. The AVP concentrations of DI were clearly distinguished from those obtained in normal subjects (0.30-4.20 pg/ml, n = 65). The within and between assay variability was about 10% each. Plasma AVP concentrations (mean +/- SD) of normal subjects (n = 6) standing, sitting, and supine after an overnight of fluid deprivation were 2.41 +/- 1.15, 1.95 +/- 0.85 and 0.97 +/- 0.48 pg/ml (30 min after) respectively. Plasma AVP concentrations of normal subjects (n = 6) after water load (20 ml/kg wt) were clearly reduced from 1.89 +/- 1.00 (before) to 0.42 +/- 0.21 (standing for 60 min) and also from 0.89 +/- 0.41 to 0.40 +/- 0.22 pg/ml (supine for 60 min).

Adult↗