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Biomedical subjects

T Kokubu

Publications and source records attributed to T Kokubu.

At least 217 records · Page 12Linked to original sources
PubMed1977

A radioimmunoassay of plasma unconjugated and conjugated estetrol.

A radioimmunoassay for the measurement of both unconjugated and conjugaged estetrol in plasma has been developed. The antiserum obtained after 6 months of immunization with 6-oxoestetrol-6-(O-carboxy-methyl)oxim-BSA was used at a final dilution of 1:90,000 and showed almost no cross reaction with other steroids except for estriol at 1.24%. Esterol-glucosiduronate was synthesized by incubating with adrenalectomized rat liver homogenate and uridine diphosphoglucuronic acid. Then, plasma estetrol-glucosiduronate was measured in the same manner for unconjugated estetrol after hydrolysis with beta-glucuronidase. Sephadex LH-20 column chromatography (7X110 mm, benzene:methanol, 85:15) was employed for accurate assessment. The sensitivity was 10 pg and the smallest amount measurable was 40 pg/sample. The method bland was consistently negligible. The intra and inter assay precision was 11.8% and 14.2% for unconjugated estetrol and that for estetrol-glucosiduronate was 13.5% and 17.1%.

Antibody Specificity↗
PubMed1976

Solubilization of angiotensin I-coverting enzyme from rabbit lung using trypsin treatment.

The solubilization of angiotensin I-converting enzyme (peptidyldipeptide hydrolase, EC 3.4.15.1) from rabbit lung was carried out using trypsin treatment. A good recovery of 76% was obtained. The enzyme from solubilized fraction was purified using colums of Sephadex G-200, hydroxyapatite and DEAE-cellulose. The purified enzyme was shown to convert angiotensin I to angiotensin II and also to inactivate bradykinin. The specific activity of the enzyme was 24.3 units/mg protein for hippurylhistidylleucyl hydroxide and 0.182 mumol/min per mg protein for angiotensin I. The enzymic activity obtained after trypsin treatment for 5 h could be divided into two components: (i) an enzyme of molecular weight 300 000 (peak II) and (ii) an enzyme of molecular weight 145 000 (peak III), by Sephadex G-200 gel filtration. The molecular weight of the denatured enzyme was found to be 155 000 by disc gel electrophoresis in the presence of sodium dodecyl sulfate. Km values of peak II and peak III fraction for Hippuryl-His Leu-OH were 2.6 mM.

Animals↗
PubMed1976

The influence of nephrectomy, ureteral ligation, and of estradiol on plasma renin substrate in unilaterally nephrectomized rats.

The effects of three experimental conditions on the concentration of plasma renin substrate were studied with special reference to plasma renin concentration in unilaterally nephrectomized rats. After simultaneous bilateral nephrectomy the maximum increase in plasma renin substrate was 17 times higher than normal within 24 h, while in rats which were unilaterally nephrectomized 10 days previously, followed by the removal of the remaining kidney (two-step bilateral nephrectomy), the maximum increase in plasma renin substrate was markedly suppressed (6-fold of normal). The maximum increases in plasma renin substrate by estradiol treatment in normal and unilaterally nephrectomized rats were about the same, associated with similarly decreased plasma renin concentrations. The similar increase in plasma renin substrate was found after ureteral ligation in unilaterally nephrectomized rats and bilateral ligation of the ureters in normal rats. This was the case where the plasma renin concentrations changed differently after ureteral ligation. After two-step bilateral nephrectomy plus estradiol treatment the maximum increase in plasma renin substrate was found to be higher than that found after two-step bilateral nephrectomy, but was lower than that after simultaneous bilateral nephrectomy. It is suggested that under the pathological conditions that stimulate renin substrate production, the plasma rein substrate concentration is less affected by circulating renin.

Angiotensin II↗
PubMed1976

Affinity chromatography of angiotensin I-converting enzyme from rabbit lung using hippurylhistidylleucyl-OH.

Approximately 50-fold purification of angiotensin I-converting enzyme (Peptidyldipeptide hydrolase, EC 3.4.15.1) from rabbit lung was achieved by affinity chromatography using the synthetic substrate Hippuryl-His-Leu-OH. The specific activity of the enzyme was increased from 0.044 units/mg protein to 1.911 units/mg protein for Hippuryl-His-Leu-OH and from 0.33 nmol/min per mg protein to 13.8 nmol/min per mg protein for angiotensin I.

Angiotensin II↗
PubMed1976

Possible existence of prorenin in rabbit renal cortex.

The possible existence of prorenin (inactive form of renin) in rabbit renal cortex was studied. Renal cortex was homogenized with 0.25 M sucrose. Following centrifugation at 800 g for 10 min, the supernatant (Fraction H) was centrifuged at 20,000 g for 20 min. The pellet (Fraction G) was separated from soluble fraction (Fraction S). Intravenous injection of non-treated Fraction G caused a little blood pressure elevation in rats. However, the pressor activity of the same amount of Fraction G which was frozen and thawed 5 times previously was more than 10 times as that of non-treated one. The renin activity in preincubated fraction G with Fraction S was 2-fold the summation of the activity of renin in each fraction, when measured using indirect method. Fraction S lost not only the renin activity but also the renin activating activity when heated for 5 min at 100 degrees C. Exposure to pH 3.5 made no influence on renin activity in either Fraction G and S.

Angiotensin II↗
PubMed1976

Biological activities of tyrosine-containing somatostatin analogs on inhibition of secretion of thyrotropin and growth hormone.

The following five tyrosine-containing analogs of somatostatin (GIF) were synthesized by the solid-phase method: Tyr-GIF: [Tyr6]-GIF; [Tyr7]-GIF; [Tyr8]-GIF; [Tyr11]-GIF. These analogs except [Tyr8]-GIF were demonstrated to possess almost the same potency to inhibit thyrotropin release stimulated by thyrotropin-releasing hormone as that of synthesized GIF in vivo. [Tyr8]-GIF had potencies less than 0.5% of GIF. They also had the activity to inhibit Nembutal-induced growth hormone rise. The structure-activity relationship and availability of these analogs for radioimmunoassay were discussed.

Animals↗
PubMed1976

Variety of GH and TSH responses to somatostatin in perfused rat pituitaries in vitro.

One male rat pituitary placed in a chamber was perifused with cyclic somatostatin (GIF) for 30 min. Either 160 nM or 1.6 muM GIF caused a decrease in the release of GH. The release of TSH was also decreased by 160 nM GIF, and paradoxically increased by 1.6 muM GIF. Increasing the dose of GIF to 16 muM resulted in an abrupt rise in the release of both GH and TSH during the perfusion; then the level of GH decreased to the nadir level followed by an elevation above the base line, while that of TSH promptly fell back toward the base line. The release of PRL was not clearly affected by 16 muM GIF. [Tyr8]-GIF did not have such stimulatory activities. These results indicate that GIF not only inhibits the release of GH and TSH, but also stimulates that of GH and TSH in this system, depending on its dose.

Animals↗
PubMed1975

Effects of unilateral nephrectomy on plasma renin substrate and renin concentration in rats.

Effects of unilateral nephrectomy on plasma renin concentration and renin substrate (angiotensinogen) concentration were studied over an experimental period of 10 days in rats. The maximum increase in plasma renin substrate concentration after bilateral nephrectomy was 9 times higher than the preoperative level. Very low but measureable plasma renin concentrations were found in rats nephrectomized bilaterally 24 hrs previously. In unilateral nephrectomized rats the concentration of renin substrate in plasma increased maximumly 3.5-fold within 24 hrs after the operation and on the third day it was decreased to half of the maximum level. Even 10 days after uninephrectomy the concentration of plasma renin substrate was significantly higher than that of normal rats. Twenty four hours after unilateral nephrectomy plasma renin concentration decreased to similar low levels as those found in bilateral nephrectomized rats. The decreased concentration of plasma renin, however, returned to normal 3 days after uninephrectomy. The inverse relationship was seen between the concentrations of plasma renin and plasma renin substrate. However, substrate concentration in plasma rised faster and longer than plasma renin decreased.

Angiotensin II↗
PubMed1975

Extraction and partial purification of prolactin-release stimulating factor in bovine hypothalami.

Partial purification of prolactin-release stimulating factor (PRF) was performed by Sephadex G-25 gel filtration of bovine hypothalamic extracts. PRF activity was evaluated on the basis of the measurement of immunoreactive prolactin released from the isolated rat hemipituitary in vitro. PRF activity was found in the fractions with Kav=0-0.49 and prolactin-release inhibiting activity was also detected in the fractions with Kav=0.69-0.89. The dose-response relationship was established between the partially purified PRF and its activity. The elution position of the partially purified PRF preceded that of TRH on Sephadex G-25. TRH at the dose of 100 nM stimulated the release of TSH in vitro, but not the release of prolactin. These results may indicate that there exists PRF with a relatively high molecular weight in the bovine hypothalamus.

Animals↗