This study was designed to examine whether 8S protein as progesterone receptor exists in the human endometrium which has been primed with estrogen. The kinetic study showed that 8S-progesterone binding was specific with Kd of 2.0×10^-9M. 5Sprogesterone binding was inhibited competitively by cortisol. The study of ligand specificity also showed that progesterone and its related steroids had much stronger affinityfor 8S component than for 5S component. Therefore, 5S protein may be CBG. Progesterone-8S protein binding was easily dissociated during the 5-20% sucrose gradient centrifugation, but such a protein from which progesterone had been dissociated could be sedimented at 8S region. Glycerol could stabilize progesterone-8S protein binding. These results indicate the existence of 8S protein as a progesterone receptor under the low salt medium in the estrogen primed human endometrium.

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We had an opportunity to treat a patient who had been suffering from prolonged. malnutrition. The patient was a 22-year-old male. He was noted to have hypopituitarism clinically. In order to determine the role of malnutrition in the hypopituitarism, several substances and hormones in serum were measured before and after the surgery of intestinal blind loop.
At the time of admission he was noted to have reduced pigmentation, growth retardation, weight loss and delayed adolescence. X-ray examination of his bones showed osteomalacia. Fecal excretion of ¹³¹I-triolein for three days was 26.2% of total triolein, indicating that the absorption of fat was impaired.
Significantly low values were found for a basal metabolic rate, 24-hr-¹³¹I uptake of the thyroid, serum T₄, serum T₃-RU, urinary excretion of 17-OHCS and 17-KS.
The thyroidal radioiodine uptake was increased by consecutive injections of TSH.
Plasma TSH was undetectable without TRH, which induced an increase in the plasma level of TSH. Therefore, hypothalamic hypothyroidism was most likely for a diagnosis.
The impaired response of plasma GH to a hypoglycemia induced by insulin was observed. Also, the impaired response of urinary 17-OHCS to metyrapone was observed, although plasma ACTH was not determined.
After surgery for the intestinal blind loop, he was apparently improved in the signs and the symptoms of malnutrition. He also developed a cracked voice, comedos on the face, pubic and axillary hair, and a beard. In addition, he gained 16kg in weight and 4cm in height for 9 months without any other treatment than the surgery. In the determination after surgery, serum lipids and protein were significantly increased, and then a marked increase in plasma levels of hormones was observed.
Consequently it was considered that malnutrition played an important role in this hypopituitarism maybe through a decrease or cessation of the releasing factors, namely, hypothalamic hypofunction.

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The hypothalamic content and concentration of thyrotropin-releasing hormone (TRH) were determined by radioimmunoassay in normal, thyroidectomized, hypophysectomized and cold-exposed rats treated with or without thyroxine.
In normal animals, the single administration of thyroxine (1, 5 and 20μg/100g B. W.) altered neither the content nor the concentration of TRH in the hypothalamus. However, seven days' administration of this hormone resulted in the dose-dependent increase in the hypothalamic TRH levels. In thyroidectomized rats, the hypothalamic TRH levels were slightly reduced in spite of the marked increase of plasma TSH levels and decrease of pituitary TSH levels. In the animals given thyroxine (10μg/100g B. W.) for 7 days in addition to thyroidectomy, however, the TRH levels exceeded that in the animals which underwent thyroidectomy alone. The hypothalamic TRH levels were markedly reduced in hypophysectomized rats. Conversely, in hypophysectomized rats given 7 days' thyroxine (1 and 5μg/100g B. W.), the levels were increased dose-dependently. In cold-exposed rats, the plasma TSH levels roughly doubled, but the TRH levels remained unchanged.
These findings strongly suggest that the feedback site of thyroxine extends not only to the pituitary gland but also to the hypothalamus, and that thyroxine has an increasing effect of the hypothalamic TRH level, though the mechanism (s) remain to be clarified.

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Partial separation of human urinary substances which have properties to suppress the ovulation induced by PMS and HCG in mice was attempted by gel filtartion on Sephadex G-100 and ultrafiltration using the Amicon Diafio membranes UM-2 and UM-10. In addition to a thermostable inhibitor which has a molecular weight more than 10, 000, the presence of a heat-labile inhibitor with a molecular weight less than 1, 000 was newly demonstrated.

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Anterior pituitaries of male rats were electron-microscopically observed 12hr, 1, 3, 5, 7 and 14 days after thyro-parathyroidectomy in order to investigate the origin and destiny of Siperstein's corticotrophs (S-cells). The formation of a reticular framework by the folliculo-stellate cells may result from the general disaggregation of the solid mass of follicular cells with the pseudolumen into which microvilli project, by losing the junctional complex and pseudolumen. The S-cells, characterized by a row of secretory granules along the cytoplasmic membrane, have properties identical with those of the free agranular folliculo-stellate cells. The star-like shape, numerous ribosomes and polysomes, and elongated narrow rough endoplasmic reticulum, are common characteristics of both cells. Within 12hr to 5 days, the free or reticular folliculo-stellate cells appear in large numbers, and minute secretory granules about 50nm in diameter develop sporadically at their margins. In the course of the 5 days, secretory granules tend to increase in number and grow in size, measuring 100-200nm in diameter. However, the degree of granulation varied considerably from cell to cell during the same interval without complete equilibration with the time course after thyroidectomy. Detection of various transitional cells between folliculo-stellate cells and S-cells suggests that the former may be a stem cell of the latter. Concurrently with hypergranulation, some S-cells hypertrophy and develop a prominent Golgi apparatus, sometimes engulfing a part of the adjacent cell. The other S-cells, however, udergo degranulation associated with excessive vesiculation despite the formation of a circular arrangement of the closed Golgi lamellae. Within 7 days, there are some large vesiculated cells of stellate shape resembling the TX-cells, in which a small number of secretory granules whose diameter was reduced to 50-100nm, remain distributed along the cytoplasmic membrane. Within 14 days, the big TX-cells often appear, being characterized by the deposit of granules in the small cisternae and by the closure of Golgi lamellae which are in the circular arrangement. They are still stellate in shape, being extraordinarily enlarged to an extent 5-6 times that of an acidophil. In addition to the thyroidectomy cells (TX-cells) which are believed to result from the so-called thyrotrophs, the other developmental process of TX-cells in association with the folliculostellate cells was observed in this study. The two kinds of TX-cells are intrinsically different in their fine structures. It is postulated in this study that the folliculostellate cell is a progenitor of the S-cell which is destined to be transformed into the TX-cell, and that the S-cell, for this reason, represents a kind of basophil.

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To examine the properties of androphilic proteins in human benign prostatic hypertrophy, the binding capacity and affinity of the proteins were determined after acetone-treatment, ammonium sulfate precipitation and chromatographies of DEAE and Sephadex G-200. Androphilic proteins in the extract of acetone-dried cytosol from the hypertrophic human prostate was precipitated at 30-50% saturation of ammonium sulfate. The binding of this fraction to dihydrotestosterone and testosterone was high affinity, but the binding to estradiol-17β was the one of non-specific. Androphilic proteins in the 30-50% fraction were eluted from DEAE cellulose column by buffer containing 0.05M KCl.
On Sephadex G-200 chromatography of 30-50% fraction, the androphilic proteins were observed in three peaks; one was eluted in the void volume and other two were eluted at the sites of IgG and albumin. The amount and ratio of proteins eluted in the void volume and the site of IgG from Sephadex G-20Q column were variable in individual tissue samples. The chromatographic behavior of the 30-50% fraction in Sephadex G-200 was not changed significantly by introducing 0.4M KCl in the system. Polyacrylamide gel electrophoresis was applied for further separation of the proteins.

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Insulin-releasing activity of porcine gastrointestinal glucagon-like immunoreactive materials purified by affinity chromatography was examined in the perfused rat pancreas. When glucose concentration of the perfusate was raised from 60 to 100mg/dl, augmented insulin release was observed. The mean incremental area of immunoreactive insulin (ΣΔIRI) during the first 10min thus observed was 19.07±3.76ng/10min. Pancreatic glucagon and the extract from the gastric fundus showed the enhancement of insulin release in this system when they were added to the perfusate at the rate of 100ng/min for 5min;ΣΔIRI were 41.92±.47 and 71.70±18.09ng/10min, respectively, which were significantly higher than that of 100mg/dl of glucose alone. However, no significant diffrence in the insulinogenic activity was noticed between the extracts from the small intestine and the control.
These results suggest that the extract from the gastric fundus has insulinogenic activity similar to that of pancreatic glucagon.

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The interaction of parathyroid hormone-(PTH) and prostaglandin E₁ (PGE₁) on cyclic AMP metabolism in the rat renal cortical tissue was examined in vitro, placing an emphasis on the process of release of the nucleotide from the tissue. PTH enhanced the release of cyclic AMP from cells even in the presence of 10μM cyclic AMP added to the incubation medium. This effect of PTH to accelerate cyclic AMP release was suppressed by simultaneous addition of PGE₁. Although PTH and PGE₁ showed an additive effect in stimulating the adenylate cyclase activity of the purified renal plasma membrane, in the experiment in which slices were employed the sum of cyclic AMP in the tissue and medium under simultaneous stimulation tended to be less than that under the stimulation by PTH alone. However, the increase by PTH of specific activity of cyclic AMP produced in slices prelabeled with [¹⁴C] adenine was further enhanced by the simultaneous addition of PGE₁, suggesting that PGE1, by suppressing the release of cyclic AMP, keeps more cyclic AMP intracellularly and offers phosphodiesterase more substrate. It seemed that the release of cyclic AMP from the kidney cortical tissue could proceed against a concentration gradient and was regulated somehow by PTH and PGE₁. It was noted also that the change in the release process influenced the intracellular metabolism of cyclic AMP, indicating the importance of taking such an extracellular release mechanism into consideration in the study on the cellular metabolism of cyclic AMP.

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Sixty-four blood samples, obtained from 25 pituitary dwarfs treated with KABIHGH preparation, were checked for serum HGH concentration by a double antibody radioimmunoassay and for antibodies against HGH by the method of propylethyleneglycol separation. Antibodies to HGH were detected in the samples whose HGH concentration was falsely estimated to be more than 5ng/ml, but not in those whose HGH concentration was 5ng/ml or less. The measurement of HGH concentration by a double antibody technique in the serum obtained from pituitary dwarfs 3-4 days after the last injection can be used to screen the presence of antibodies to HGH preparation used.

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Implants of paraffin micropellets containing about 5μg 5α-dihydrotestosterone (DHT) into the hypothalamus of 5-day-old female rats were without effect on the sexual differentiation of the brain. By contrast, approximately the same amount of testosterone propionate (TP) given as subcutaneous or intrahypothalamic micropellets masculinized the female brain. In the light of these results as well as the author's previous findings that an antiestrogen implanted into the hypothalamus of neonatal female rats failed to block masculinization by subcutaneous injection of TP, the possibility cannot be excluded that testosterone is capable of masculinizing the brain of neonatal females without being converted into estrogens.

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Effects of high (K⁺) and dopamine on the release of immunoreactive somatostatin from isolated hypothalamic synaptosomes were studied in rats. High (K⁺)(60 mM) and dopamine (10^-6 M) in the incubation media stimulated the release of immunoreactive somatostatin and the former effect was completely abolished by the removal of Ca⁺⁺ from the media. These suggest that hypothalamic somatostatinergic synaptosomes preserved at least one of the important basic properties of secretory cells. Although it is of interest to note that dopamine stimulated the release of somatostatin. Its physiological significance awaits further studies.

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