In this study, human uterine endometrial estrogen receptor (ER) and progesterone receptor (PR) in normal menstrual cycle were estimated, and biochemical characterization of ER and PR in normal endometrium and endometrial carcinoma were also investigated.
Following results were obtained in this study.
1) In normal menstrual cycle, ER and PR levels in endometrial cytosol gradually rose to peaks in the late proliferative phase, but PR in nuclear fraction rose to a peak in the early secretory phase.
2) Scatchard analysis of ER in normal endometrium and myometrium contains two estradiol (E₂) binding sites with dissociation constants (Kd) of 10^-9M and 10-_-10, but endometrial carcinoma contains a single population of E₂ binding site with Kd's 10-_-10.
3) Total binding sites for ER and PR of normal endometrium have 2 3 times much more than those of endometrial carcinoma.
4) In normal endometrium, specific binding of 40nM ³ HE₂ on isoelectric focusing (IEF) indicated three binding activities with elution pH's (EPH) of 4, 6 and 8. But specific binding of 2 nM 3 HE₂ indicated only one binding activity with EPH of 6 in endometrial carcinoma. 5) Specific binding of EPH 6 indicated high affinity ER (type 1 ER) and specific binding of EPH 8 indicated low affinity ER (type 2 ER) in the result of IEF and Scatchard analysis. Loss of type 2 receptor is important result in endometrial carcinoma.
The above results suggest that increase in blood E₂ level increases endometrial ER and PR, and increase in blood progesterone level after ovulation decreases endometrial ER and PR for anti-ER and PR effect of progesterone. If type 2 ER could transport hormone receptor complex to the nucleus, loss of type 2 ER would be the important cause of ER and PR decrease and get resistance of hormone therapy to endometrial carcinoma.

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A quantitative analysis of steroids in serum, dried blood samples on filter paper and amniotic fluid with high-performance liquid chromatography has been developed and applied to the diagnosis of risk infants on neonatal screening for congenital adrenal hyperplasia. The present method is simple, rapid and accurate. The detection limits of cortisol and 17a-hydroxyprogesterone are 0.2 ng and 0.3 ng, respectively. The twenty-two steroids can be analysed within 60 minutes using the isogradient mobile phase. The method is highly correlated with radioimmunoassay and enzymeimmunoassay.
In 4 patients with the salt-losing form, 17a-hydroxyprogesterone was higher and cortisol lower than in 4 pateints with the simple-virilizing form.
This method seems suitable for the routine clinical elucidation of congenital adrenal hyperplasia.

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An enzyme-linked immunosorbent assay (ELISA) for 17α-hydroxyprogesterone (17- OHP) in dried blood collected on filter paper has been developed. The ELISA method is very easy and rapid and has sensitivity, accuracy and precision. The detection limit of the ELISA method is 0.5 ng/ml blood. Intra-and interassay coefficients of variation are 7.0 “- 8.3% and 6.7”-10.5%, respectively. Correlation between the ELISA method and previously reported enzyme immunoassay is good (r = 0.961).
17-OHP values of neonates with congenital adrenal hyperplasia (CAH) are extremely high compared with normal neonates (11.0 ± 5.9 ng/ml blood).
The present study demonstrates that the ELISA method for 17-OHP is applicable to neonatal screening for CAH.

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Hyperresponsiveness of plasma aldosterone to metoclopramide (MCP) was found in the patients with secondary aldosteronism. To understand the mechanisms of aldosterone reactivity to MCP, we examined the changes of its responsiveness to MCP and ACTH in various states of endogenous renin-angiotensin system which were induced by administrating two antihypertensive agents to patients with essential hypertension.
Aldosterone responsiveness to both stimulants were accentuated during diuretic anti-hypertensive therapy in comparison with those before therapy. After adding converting enzyme blocker to diuretic agent, aldosterone reactivity to MCP decreased.
These results indicates that aldosterone responsiveness to MCP which was thought to be controlled by changes of endogenous dopamine may be also influenced by the state of endogenous renin-angiotensin system.

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Effect of ergot alkaloids on (³H) -nucleosides uptake by dispersed cells from fresh female bovine anterior pituitary glands was examined, because we had interested in the mechanism of pituitary tumor regression following bromocriptine treatment and considered nucleic acid synthesis as an important process in the metabolism of these cells.
The anterior pituitary cells were implanted on culture tubes using D-valine minimal essential medium with serum to suppress the overgrowth of fibroblasts and then maintained in L-valine Dulbecco's modified Eagle medium.
(³H) -Uridine uptake by these cells was suppressed by bromocriptine, @-ergocriptine or ergotamine at a concentration varing from 10^-6M to 10^-5M, but not 10^-5M of lergotrile. Among these alkaloids, bromocriptine had most strong inhibitory effect and suppressed the uptake to below 25% of control value at the concentration of 10^-5M. Bromocriptine also inhibited the uptake of (³ H) -thymidine, (³ H) -cytidine, (³ H) -adenosine or (³ H) -guanosine in the same manner as (³H) -uridine. But neither (³H) -uracil, the base of uridine, nor (³H) -galactose uptake by cells was affected by bromocriptine. The incorporation of (³ H) -uridine or (³ H) -thymidine into TCA-insoluble fraction of the cells was also inhibited by bromocriptine as that in the total cells.
It was suspected that bromocriptine acted on distinct transport site of both ribonucleoside and deoxyribonucleoside, because a high concentration (3.3 X 10^-5M) of radio-inactive thymidine did not modify (³H) -uridine uptake by these cells as well as inhibitory effect of bromocriptine on it.
These effect of bromocriptine were not blocked by haloperidol, known as dopamine antagonist, at the same concentration as bromocriptine, and dopamine had no effect on (³ H) -uridine uptake by the cells. In addition, by adding 5 X 10^-4M of dibutyryl cyclic AMP into the medium, the effect of bromocriptine was also not affected. These data suggest that the effect of bromocriptine on nucleoside transport in anterior pituitary cells may be dependent on the other binding site than D-2 dopamine receptor in the anterior pituitary cell membrane.

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A possible involvement of oxytocin (OT) has been indicated in regulation of water and electrolyte metabolism, based on findings that the secretion of OT is increased by either water deprivation or sodium loading. However, to date, no informations have yet been obtained about the role of OT in hypertension. The present study was therefore undertaken to elucidate the role of OT for abnormalities of fluid and electrolyte metabolism in essential hypertension (EH) in comparison with normotensive subjects (NT).
The major results were as follows.
1) Plasma level of OT was 3.7 ± 2.1 pg/ml (mean ± SD) in EH, not significantly higher than that in NT (3.2 ± 1.7 pg/ml). Plasma OT in low-renin EH (4.8 ± 2.5 pg/ml) was significantly different from that in high-renin EH (2.9 ± 1.4 pg/ml, p<0.05) and NT (p<0.05), but not in normal-renin EH (3.8 ± 2.0 pg/ml).
2) Plasma OT was inversely correlated with plasma renin activity (PRA) in EH (r =- 0.384, p<0.01), but not in NT (r = 0.102).
3) No significant correlation was found between plasma OT and plasma aldosterone concentration (PAC), plasma concentration of antidiuretic hormone (ADH), serum sodium and potassium, blood pressure and renal function in either EH or NT.
4) I.m. injection of OT (0.04 IU/kg) increased significantly urinary excretions of sodium and potassium in EH and NT. However, the increment in sodium excretion was greater in low-renin EH than that in normal-renin EH (0.05<p<0.10), high-renin EH (p<0.05) and NT (p<0.05). PRA, PAC and ADH were significantly decreased after OT injection, but blood pressure, serum sodium and potassium were not altered in both EH and NT. 5) I.v. administration of OT (0.1-0.2 IU/min) suppressed angiotensin II-induced in crease of PAC and elevation of blood pressure in both EH and NT. The decrease in PAC by the OT administration was the greatest in low-renin EH. The reduction of blood pressure was significantly greater in EH than in NT (p<0.05).
6) I.v. administration of hypertonic saline (5%) resulted in a significant increase of plasma OT in EH and NT, and the increment in OT was the greatest in low-renin EH. Serum sodium concentration was increased by the infusion, positively correlated with plasma OT in both EH (r = 0.458, p<0.05) and NT (r = 0.830, p<0.05).
7) High sodium intake (340 mEq/day for 5 days) increased plasma OT in EH and NT. The increment of plasma OT was significantly correlated with that of serum sodium concentration in EH (r = 0.497, p<0.02) and NT (r = 0.831, p<0.05). By contrast, low sodium intake (50 mEq/day for 5 days) caused reduction of plasma OT in EH and NT with high plasma OT level more than 5.5 pg/ml.
8) Water deprivation (15 h) resulted in a significant increase of plasma OT in EH and NT. The magnitude of increase in plasma OT, which was significantly greater in EH than in NT (p<0.05), was correlated with increment of serum sodium concentration (r = 0.529, p<0.05 for EH; r = 0.745, 0.05<p<0.10 for NT) and also plasma osmolarity (r = 0.479, p<0.05 for EH; r = 0.742, 0.05<p<0.10 for NT).
In conclusion, the secretion of OT is stimulated by a high circulating sodium level to display natriuretic effect on the kidney. Also, OT seems to be related, in part at least, to water and electrolyte metabolism in low-renin EH presumably with predisposition of fluid retention, through its natriuretic action and suppression to the renin-angiotensin-aldosterone activity.

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Hypertonic saline test is indispensable for the evaluation of posterior pituitary function. However the test is not simple, including water loading, urine sampling and at least 45 min of hypertonic saline infusion, mostly because the test relies on urinary osmolality as an index of ADH secretion. The object of this study is try to simplify the test by directly measuring plasma ADH concentration before and after 10 min of hypertonic saline infusion. Intravenous infusion of hypertonic saline (5% NaCl, 0.24 ml/kg/min, for 10 min) was performed on normal subjects, patients with diabetes insipidus and patients with renal failure under chronic hemodialysis. Venous blood samples were obtained seriously including just before and after 10 min of the infusion. ADH was extracted from plasma using Sep-Pak C18 column and assayed by specific RIA. Minimum sensitivity of the assay was 0.25 pg/ml.
The hypertonic saline infusion resulted in an increase of plasma osmolality by about 8 mOsm/kg H2 0 and plasma sodium concentration by 4 mEq/l. Plasma ADH increased from 0.77 ± 0.09 to 3.42 ± 0.73 pg/ml (m ± SE, n = 8, p<0.01) in normal subjects of ad lib. water drinking and from 0.55 ± 0.33 to 2.34 ± 0.33 (m ± SE, n = 4, p<0.05) in water loaded normal subjects (20 ml/kg of water, 60 min before hypertonic saline infusion). In patients with diabetes insipidus, plasma ADH level was low (0.63 ± 0.24 pg/ml, n = 5) relative to plasma osmolality (295.4 ± 4.6 mOsm/kg H2 0) and showed no significant increase after hypertonic saline infusion. In patients with renal failure, plasma ADH increased from 2.47 ± 0.28 to 4.14 ± 1.09 pg/ml. Thus basal ADH level was significantly higher than normal subjects but response to hypertonic saline was reduced.
These results suggested that this simplified test with 10 min infusion of hypertonic saline, no water loading and no urine sampling may be useful for the evaluation of ADH secretion and differential diagnosis of polyuria.

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