To elucidate the role of endogenous prostaglandin F_2αin spontaneous and induced labor, plasma concentrations of13, 14-dihydro-15-keto-prostaglandin F_2α (PGFM) were determined before the onset of labor, at onset of labor, during active labor, at the crowning of the fetal head, and 1 and 2 hours after delivery. Patients in spontaneous labor and labor induced by amniotomy, oxytocin, and prostaglandin E₂were studied. The levels of plasma PGFM in patients who entered spontaneous labor fell2to3weeks before delivery, whereas those in the induced labor group did not change until the time of induction. Although the levels of PGFM rose gradually with the progress of labor in all cases, the levels in the spontaneous labor were significantly lower in each stage than in the corresponding stage of induced labor. These results suggest that endogenous prostaglandin F_2α (PGF_2α) production decreases2-3 weeks prior to the spontaneous onset of labor and is increased again as labor progresses, that the patterns of PGF_2αproduction are similar to each other during spontaneous labor and labor induced by various methods. Therefore, it is felt that endogenous PGF_2α may participate in the progress of all kinds of labor.

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Analysis of urinary metabolites of [1, 2-³H]-aldosterone was performed in the male guinea-pig. Separation of urinary metabolites was carried out by countercurrent distribution followed by DEAE-Sephadex A-25column chromatography. A major component was obtained which was both hydrolyzable with sulphatase from Helix pomatia and solvolyzable. Paper chromatography of freed steroids revealed the presence of at least two components and the major aglycone cochromatographed with 3β, 5α-tetrahydroaldosterone.
In order to get more information about the structure of urinary metabolites, a total of68mg of aldosterone was injected into three male guinea-pigs and separation of urinary metabolites was performed in a similar manner. A major component obtained showed the color reaction positive for sulphate (modified rhodizonic acid test) and negative for glucosiduronate (naphthoresorcinol test). Gas chromatographic-mass spectrometric analysis of aglycones released from this conjugate revealed the presence of 3β, 5α-tetrahydroaldosterone and an another aglycone, tentatively identified as21-deoxy-tetrahydroaldosterone.
Taken together, it was concluded that 3β, 5α-tetrahydroaldosterone-monosulphate and21-deoxy-tetrahydroaldosterone-monosulphate comprised most of urinary conjugated metabolites of aldosterone in the male guinea-pig.

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Combination of two thyroglobulin monoclonal antibodies (monoAbs) recognizing epitopes which are rarely recognized by an antibody enabled us to develop a rapid one-step enzyme immunoassay of serum Tg. Of 87 monoclonal antibodies, 20 were selected for the purpose. The method is a sandwich technique employing a monoAb covering microplate and horse-radish peroxidase monoAb conjugate. A combination of monoAb 7A7A solid phase and 31A2E for the conjugate gave the best results. The assay takes60min and the minimal detectable amount is 2ng/ml. Intraassay variation is from 4 to 7%. Interassay variatoin is 5 to 12%. The recovery rate for Tg added to normal sera is between 89 and 111%.
The correlation coefficient with the polyclonal antibody method in Tg hemagglutination negative sera is0.98.
The presence of autoantibody in sera up to 10×2⁴hemagglutination titer does not affect the recovery rate to a statistically significant extent.

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A rapid method for simultaneous preparation of cytosol and nuclear estrogen (E) and progestin (P) receptors and their in vitro determination is described. The method was applied to several uterine or ovarian surgical specimens to evaluate their steroid hormone “dependence”. The results suggest that low cytoplasmic E receptor levels (ERc) are associated with higher nuclear E receptor (ERn) levels but no apparent correlation was observed between PRc and ERn levels. The method appeared to be suitable for screening steroid hormone receptor content in tumor tissues and may provide better estimation of steroid dependence since boh cytoplasmic and nuclear compartments can be studied simultaneously.

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The relationship between glucose-induced insulin secretion and metabolism of inositol phospholipid was investigated by means of an islet perifusion method and direct measuring of inositol phosphates after sonicating the islets. The results showed that the time course of inositol phospholipid breakdown is coincident with the first phase of glucose-induced insulin secretion. Analysis of the effluent perifusate as well as the water soluble inositol-containing substance after sonication of stimulated islets revealed that most of the metabolite of inositol phospholipid is inositol-triphosphate, the hydrolysis product of phosphatidylinositol-4, 5-bisphosphate. On the other hand, perifusion of islets with exogenous inositol-triphosphate showed a monophasic and dose-dependent response of insulin secretion. Thus, the initial process of glucose stimulation is accompanied with the formation of inositol-triphosphate, which is a possible candidate for the triggering of first phase insulin secretion.

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We have investigated the ability of lymphocytes from normal subjects and patients with autoimmune thyroid diseases to respond to a thyroidal antigen (human thyroglobulin, hTG) and a non-thyroidal antigen (Keyhole limpet hemocyanin, KLH) in vitro, using a hapten (trinitrophenol, TNP)-carrier system. This system is based on the concept that the T helper cells which respond to hTG or KLH should stimulate anti-TNP antibody producing B cells in the presence of TNP conjugated hTG (TNP-hTG) or KLH (TNP-KLH). After 5 or 6 days of culture of peripheral blood mononuclear cells with pokeweed mitogen (PWM), PWM and TNP-hTG, or PWM and TNP-KLH, IgM anti-TNP and IgM anti-sheep red blood cell (SRBC) plaque forming cells (PFC) were enumerated. The results showed that (1) in normal controls, hTG caused only suppression in both TNP and SRBC response, and KLH caused doserelated enhancement and suppression in TNP response without a change in SRBC response, and (2) in patients, both hTG and KLH resulted in doserelated enhancement in TNP response without a change in SRBC response. These data suggest that patients with autoimmune thyroid diseases have regulatory cell abnormalities confined to a thyroid antigen.

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Old (28 months) male Wistar rats were treated chronically for two weeks with ginsenoside Rgi or with vehicle delivered via sc implanted Alzet minipumps (rate of ginsenoside release 2.4 μg/0.5 μl/h). The number of Type 1 corticosterone-preferring receptor sites (CR) and Type 2 glucocorticoid receptors (GR) was measured in the cytosol of hippocampus tissue of rat brain with an in vitro binding assay. In old rats the B_max of Type 1 CR and Type 2 GR was reduced by 51.5% and 28.3% respectively. Following the two week treatment with Rgi the B_max of Type 1 CR increased by 60% and a receptor concentration was reached which was 21% lower than that observed in the young control animals. Minor differences in affinity of steroid binding to both receptor systems were observed in the groups of rats. The possible binding of ginsenosides to brain corticosteroid receptors in vitro was investigated as well. The inclusion of a 500 fold molar excess of Rg₁ in hippocampus cytosol did not displace ³H-corticosterone from its soluble receptor sites. The affinity of Rgi with these sites in vitro is therefore negligible. In conclusion, the binding capacity of Type 1 CR and Type 2 GR is reduced in the hippocampal brain region of aged rats. Upon chronic infusion of ginsenoside Rg₁, only Type 1 CR capacity is restored towards the level observed in young control animals. This finding suggests that in old rats the ginsenoside enhances the CORT signal via Type 1 CR on the function of the hippocampus, which is a limbic brain structure involved in cognition, mood and affect.

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Progesterone is said to play an important role in the secretory transformation of the endometrium, maintenance of gestation and relaxation of the myometrium. Progesterone is also said to be involved in ovulation and fertilization, and is contained in large amounts in the follicular fluid and peritoneal fluid. As with other steroid hormones, progesterone exists in the blood in proteinbound and free forms, and it is the free form which exhibits biological activity. In order to evaluate the biological activity of progesterone, we determined the percent free progesterone, employing the method used by Vlahos et al. for the determination of testosterone. No significant difference was noted between the percent free progesterone in the follicular phase (4.44±0.30%) and luteal phase (4.77±0.26%) of the menstrual cycle. However, a tendency for the percent free progesterone to rise was recognized in the first, and the value significantly increased to 5.54±0.51% in the third trimester (p<0.005).
The percent free progesterone during the follicular phase in the peritoneal fluid was 4.95±0.30%, and in the follicular fluid, 5.39±0.22%, both values being larger than the value in the serum during the same phase.

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Histochemical activity of malate dehydrogenase (MDH: E. C. 1. 1. 1. 37) in rat sperm mitochondria decreased during the epididymal transit of sperm. In an electrophoretic study two major MDH isozymes (MDH-A and MDH-B) were demonstrated in the sperm. The epididymal sperm showed two minor isozymes associated with MDH-A, while the testicular sperm did not.

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A 55 year-old female had suffered from 3 consecutive diseases for a year. The diseases were ulcerative colitis, primary hypothyroidism and idiopathic thrombocytopenic purpura, and had been treated with L-thyroxine (50μg daily) and betamethasone (0.5 to 1.5mg daily). On examination, the thyroid gland was not palpable at all, thyroid 99 mTc pertechnetate uptake was 0%, and an echogram revealed the existence of an atrophic gland. Thyrotropin binding inhibitor immunoglobulin (TBII) in the serum was elevated to 58.0% and her IgG almost completely inhibited the in vitro cAMP increase due to bTSH. After 5 months TBII turned out to be negative and the inhibitory IgG activity was reduced significantly. The thyroid gland also became visible scintigraphically. Thyroid medication was then stopped. Four months after the cessation of thyroxine, she felt quite well and her thyroid functions remained within the normal ranges. Antibody to Yersinia enterocolitica was positive at a low titer (×20) in the early stages, but elevated reciprocally with the fall in TBII and finally reached ×320.
In conclusion, evidence of the disappearance of blocking type TBII from the serum was demonstrated for the first time. Steroid might have caused some favorable effects, and this clinical report indicates the possibility that remission of hypothyroidism due to blocking type TBII can be expected.

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The Effect of thyroid hormone on peroxisomal enzyme activity was studied in thyroidectomized-and T₄-administered-thyroidectomized rats. In liver, the activities of isozyme A of L-a-hydroxyacid oxidase, D-amino acid oxidase, urate oxidase and catalase were decreased by thyroidectomy, and the diminished enzyme activities were restored by T₄ administration to rats. These modifications induced by thyroidectomy or by T₄ administration, however, were prominent only in immature animals (20-day-old rats). Although the changes in-a-hydroxyacid oxidase and D-amino acid oxidase activities, induced by thyroidectomy or by T₄ administratration, were also observed in 40-day-old rats, those in urate oxidase and catalase activities were not significant in 40-dayold rats. Acyl CoA oxidase activity was not affected by thyroidectomy or by T₄ administration in either 20-or40-day-old rats. In the kidney, isozyme B of L-α-hydroxiacid oxydase activity was reduced by thyroidectomy and thediminished enzyme activity was restored by T₄ administration in both20-and 40-day-old rats. D-Amino acid oxidase and catalase activities in kidney, however, were not significantly modified by throidectomy or by T₄ administration in either20-or40-day-old rats. The results suggest that thyroid hormone can modify the peroxisomal enzyme activity, which is prominent in immature animals.

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Plasma concentrations of somatostatin-like immunoreactivity (SLI) were determined in uremic patients on maintenance hemodialysis. Plasma SLI levels were significantly (p<0.001) elevated in26diabetic uremic patients (67.1±6.8 pg/ml, mean±SE) and in 24 non-diabetic uremic patients (43.5±7.2pg/ml), when compared with60healthy subjects (5.0±0.7pg/ml).
Paired pooled plasma from uremic patients before and after hemodialysis was subjected to a reverse-phase octadecasilyl-silica (C-18) cartridge and then the extract was gel filtered on a Sephadex G-25column (1.6×90cm). Both elution profiles showed two peaks of SLI which coeluted with synthetic somatostatin (SS)-28and SS-14 markers, respectively. The SS-28-like immunoreactivity (LI) peak, which was estimated by using SS-14as a reference standard, was 3-fold larger than that for SS-14 LI. On the basis of immunoequivalency of the two components in the present assay, SS-28 LI constitutes approximately 75% of circulating somatostatin.
In conclusion, plasma SLI is substantially high in uremic patients of both diabetic and non-diabetic etiology and the SS-28 is a predominant form of circulating SLI in these patients, probably, in part, for a lower clearance of this molecule.

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In frozen sections of thyroid glands with Hashimoto's thyroiditis (HT) and Graves'disease (GD), infiltrating lymphocytes were tested for their expression of HLA-DR antigens as a marker of in situ activation. In a combination of indirect immunoperoxidase and direct immunofluorescence staining, most of the immunoglobulin D positive mature B cells were found to be DR positive (DR+) in both diseases. In HT, sizable portions of both helper/inducer T (Leu3+) and suppressor/cytotoxic T (Leu2+) cells were DR+in interfollicular regions as well as in lymphocyte clusters and lymphoid follicles. In GD, the proportion of DR+cells in the interfollicular Leu2+population was significantly lower than that of HT.DR+thyrocytes were seen in all14cases of HT and in14out of16cases of GD, especially in the vicinity of lymphocyte aggregates. The extent of their DR expression was not correlated with the percentage of DR+cells in either T subset. These results indicate that a significant portion of infiltrating T cells are activated in autoimmune thyroid diseases, and there may be bidirectional interaction between DR+thyrocytes and DR+T cells. The difference in frequency of Leu2+DR+cells may account for the difference between the immunopathological features of HT and GD.

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The significance of PTH-radioimmunoassay was assessed in63surgically proven cases of primary hyperparathyroidism. To detect hyperparathyroidism, an increase in the free serum Ca index, {1-(8×A1b+2×G1b+3)/100}×Ca, has the highest predictive value (100%) for a positive test but a low predictive value (11.8%) for a negative test. It was necessary to improve the criteria to rule out the diagnosis of hyperparathyroidism. We therefore carried out a PTH radioimmunoassay employing kits made by CIS. The criteria for ruling out the diagnosis of hyperparathyroidism were that free serum Ca index was lower than3and PTH was lower than4mU/ml. The predictive value of these criteria for ruling out the diagnosis was 83.3%, significantly greater than the previously described criteria.
Furthermore, the PTH concentration was significantly correlated to the weight of pathological parathyroid tissue surgically excised and to the free serum Ca index, although to a lesser extent than the latter. Thus, PTH-RIA is of value in improving diagnostic accuracy and in determining the severity of the disease.

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Differences in the secretion of estrogens by follicular polycystic ovaries of androgen-sterilized rats and by normal follicular ovaries of early proestrous rats were compared. Some rats were injected i.v. with LH30min before bleeding. This injection of LH did not influence the secretion of estrogensby normal ovaries, but greatly increased that by polycystic ovaries, suggesting that there was abnormal steroidogenesis in cystic ovaries. In the ovaries of such androgen-sterilized rats, two types of enlarged abnormal follicles were seen. One of these was truly cystic with few or no granulosa cells (1st type). The other had a hyperplastic and infolded layer of granulosa cells with a papillary appearance (2nd type).
Because it is known that the preovulatory LH surge is not found in androgen-sterilized rats, a classical approach was taken to circumventhe probable deficit in cyclic release of LH by giving an i.v. injection of LH every4days for16days, and ovarian venous blood was collected4days after the last injection. In consequence the 2nd type of abnormal follicle disappeared as did the abnormalities of estrogen production. These results suggest that the abnormalities of estrogen production by the polycystic ovaries of androgensterilized rats may be due to the 2nd type of abnormal follicle.

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Content of androgen receptor, retention of injected testosterone and karyotype of SC115, androgen-dependent tumor, were compared with those of CS2, an androgen-independent subline derived from SC115. Although Bmax was less than that of SC115, androgen receptor was present in the cytosol and the nuclear extract from CS2. To examine the ability for androgen retention, a large amount of testosterone was injected into tumor-bearing mice, and the amount of androgen in the crude nuclear and postnuclear fractions of tumors was compared. In both fractions, retention of injected androgen was higher in the SC115 than in the CS2. Since most of the injected testosterone was not metabolized in the tissues and the injection of testosterone 5α-reductase inhibitor showed no significant influence on the growth rate of the SC115, intracellular active androgen was assumed to be testosterone in these tumor cells. As the CS2was tetraploid, the androgen independency of the CS2 seems to be related to chromosomal changes.

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Twenty-one patients with pituitary dwarfism were treated with methioninefree hGH (r-hGH) for6months with a dosage of 0.5IU/kg/week. The height of newly treated patients (N=14), increased from 2.4 to 5.0cm during treatment, which corresponded to from 4.8 to 10.0cm with a mean of 8.1±0.5cm/year. In switched patients (N=7), height increased from 2.2 to 3.8cm during the treatment, which corresponded to 4.4-7.6cm with a mean of 6.1±0.5cm/year, which was similar to that observed in previous treatment with pituitary extracted hGH (p-hGH). Anti-hGH antibody was observed in two patients (9.5%) at the end of6months of treatment with a titer of 10. These data indicate that r-hGH has a growth promoting effect and low antigenicity.

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A 64-year-old woman was admitted for evaluation of hyponatremia. She was maintained on hypertonic saline administration. Without this therapy, the serum Na concentration decreased progressively to 127mEq/L and the plasma osmolality to 254 mOsm/Kg H₂O, on Day 3. At that time, the concentration of antidiuretic hormone (ADH) was as high as 3.5pg/ml. A skull radiogram revealed an enlarged sella turcica. Computed tomography (CT) revealed a low density in the sella, and magnetic resonance imaging revealed equal intensity of the sella turcica and the cerebrospinal fluid. A diagnosis of empty sella syndrome was made by metrizamide cisternography in conjunction with CT scanning. A diagnosis of panhypopituitarism was made by endocrine function tests. ¹²³I-thyroidal uptake was 6% when her serum TSH was10.9μU/ml, suggesting that she might also have primary hypothyroidism. When this patient was given glucocorticoid before levothyroxine replacement, her serum Na concentration rose up to about 140mEq/L and a normal relationship between her plasma ADH level (2.4pg/ml) and plasma osmolality (281 mOsm/kg H₂O) was restored. Therefore, it was suggested that ADH hypersecretion induced by the glucocorticoid deficiency might in part contribute to the development of hyponatremia. This is the case of primary empty syndrome associated with panhypopituitarism, in whom initial symptom was caused by hyponatremia.

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The study was conducted to examine the effect of somatostatin on activated renin-angiotensin-aldosterone system in a case of Bartter's syndrome. After 60minutes of 500μg of somatostatin infusion, the plasma aldosterone concentration was reduced from the basal level of 250pg/ml to 140pg/ml, whereas plasma renin activity remained at the basal level. This result suggests that somatostatin may specifically inhibit aldosterone secretion in Bartter's syndrome and the agent can be applied to a treatment of this syndrome.

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To investigate the role of intracellular Ca²⁺ in the mechanism of cellular proliferation of vascular smooth muscle cells (VSMC), the effects of Ca²⁺-antagonists and calmodulin (CaM) inhibitors on DNA synthesis stimulated by serum-derived growth factors were studied in cultured VSMCs derived from rat aorta.
DNA synthesis assessed by incorporation of [³H] thymidine into the cells was significantly stimulated by epidermal growth factor (EGF), platelet-derived growth factor (PDGF) or fetal bovine serum (FBS), of which the effects were dose-dependently inhibited by a variety of Ca²⁺-antagonists, such as verapamil, diltiazem and nicardipine. Trifluoperazine and W-7, both specific CaM inhibitors, similarly inhibited DNA synthesis stimulated by EGF, PDGF or FBS in a dose-dependent manner, whereas W-5, a less specific CaM inhibitor, was minimally effective.
These data suggest that the Ca²⁺-CaM system plays an important role in the mechanism of growth factor-induced DNA synthesis in VSMCs

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Morphological investigation of the endogenous digitals-like substance (EDLS) characterized as a natriuretic humoral agent has not been carried out to date; but EDLS can be detected indirectly by immunohistochemistry using crossreactivity with digoxin-antibody. In this report, the brain and other organs of the rat and macaque were immunostained using commercially supplied digoxin-specific antibodies. The immunoreactive substances were mainly observed in the neurons of hypothalamic paraventricular and supraoptic nuclei, and varicose fibers in some areas of the hypothalamus and infundibulum. No immunopositive substances were detected in the pituitary anterior lobe, adrenal gland, or kidney of the rat.

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