This study was undertaken to determine which biological site or immunological site in the HCG molecule is essential for uptake of HCG by the ovaries. HCG was obtained from the urine of patients with trophoblastic dieseases by This study was undertaken to determine which biological site or immunological site in the HCG molecule is essential for uptake of HCG by the ovaries. HCG was obtained from the urine of patients with trophoblastic dieseases by adsorption on kaolin. This preparation gave two distinct immunological fractions on Sephadex G-100 column chromatography. One fraction (antigen-1;Ag-1) had both gonadotropic and immunological activities present in HCG, while the other (antigen 2; Ag-2) had only immunological activity of HCG. ¹³¹I Ag-1 and ¹²⁵I Ag-2 were injected into rats intravenously and the localizations of ¹³¹I and ¹²⁵I in each organ and their subcellular fractions were measured by the paired label technique. After injection of HCG, Ag-1 and Ag-2, the activities of Δ⁵-3β-hydroxysteroid dehydrogenase (3β-HSD) in the adrenals and ovaries were measured as an index of steroid biosynthesis. The results obtained were as follows:
1. ¹³¹I Ag-1 was taken specifically in the ovaries and the ratio of the ¹³¹I Ag-1 to ¹²⁵I Ag-2 in the microsomal fraction was 14:1, 3hours after injection of the mixture containing cold HCG. The heated ¹³¹I Ag-1, which possessed only the immunological activity of HCG was not taken specifically in ovaries. These results suggest that the biological site in HCG molecule might be essential for picking up HCG by ovaries.
2. The percentage of ¹²⁵I Ag-2 localization in kidney gradually increased, but no significant change was found in adrenals, thyroid, uterus, liver and blood.
3. 3β-HSD activity in the ovary was stimulated by the administration of HCG and Ag-1, but slightly inhibited by Ag-2. 3β-HSD activity in the adrenals was clearly inhibited by Ag-2. These results suggest that Ag-2 possibly has some other biological activity.

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In the cyclic rats, progesterone secretion increased twice a cycle, on the afternoon of proestrus and on the first day of diestrus. 20α-Hydroxypregn-4-en-3-one (20α-OH-P) secretion elevated after LH surge and remained high by the first day of diestrus. The level of plasma LH increased once a cycle on the afternoon of proestrus and remained low through the estrous and diestrous stages. Plasma prolactin level also increased on the afternoon of proestrus, and it occurred earlier than the onset of LH release. A single injection with 5mg per rat of progesterone at 11.00hr on the day of proestrus (designated Day 1) caused the following effects; a) about 3hr advancement in the release of both LH and prolactin, either amount of which, however, was almost comparable with the controls, b) a marked increase and an advancement in preovulatory progesterone secretion, c) an increase in diestrous progesterone secretion on Day 3, and d) a lower preovulatory progesterone secretion on Day 5 than the respective controls, the occurrence of which delayed about 3hr. In the following estrous cycle, however, both increments of progesterone secretion at diestrus (Day 7) and proestrus (Day 9) and gonadotropin release occurred similarly as in the controls. 20α-OH-P secretion was high during the period from proestrus to early diestrus and low at late diestrus in both control and treated rats. The highest value in 20α-OH-P secretion was seen at 15.00 to 16.00hr on Day 1 in the treated rats. Numbers of ova and newly formed corpora lutea counted in the mornings of Day 2 and Day 6 were similar to those in the controls. An additional treatment with progesterone at 11.00hr or with exogenous LH at 15.00hr on Day 5 increased ovarian secretion of progesterone at 16.00hr, although the responses were lower.
These data suggest that the exogenous progesterone given on the morning of proestrus not only advances the release of both LH and prolactin but also sensitizes the ovarian steroidogenic response to LH, which results in “supersecretion” of progesterone in the proestrous afternoon and larger output of progesterone in the diestrus stage. Endogenous progestins hypersecreted in the previous cycle slightly suppress the ovarian steroidogenic response, but neither the ovulatory response to gonadotropins nor the release of pituitary LH and prolactin is altered.

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The experiment was designed to study which part of the brain, the extrahypothalamic or hypothalamic area, are most involved in the release of LH and FSH in prepuberal rats. Electrical stimulation (100Hz, 0.1msec, 100μA S. Q.) was applied for 30min (30 sec on and off) to the several regions of the brain through implanted electrodes, and pituitary or serum contents of gonadotropin were measured by radioimmunoassay. 1) Rats were stimulated on 27 days of age and killed immediately after the stimulation. Stimulation of the arcuate nucleus (ARC), suprachiasmatic nucleus (SC), medial preoptic area (MPO), medial amygdala (AMYG) and the hippocampus (HPC) elevated the pituitary LH and FSH. Marked increases of pituitary LH and FSH were observed in the rats stimulated at the ARC, SC and HPC, but there was less of an increase in the rats stimulated at the AMYG. Serum FSH increased only by stimulation of the HPC. 2) Rats were stimulated beginning at the same age for 3 days successively and killed immediately after the 3rd stimulation. The 3-day stimulation of the ARC, MPO, and AMYG elevated the pituitary LH, while that of the HPC induced neither increase nor decrease. Both pituitary and serum FSH were elevated by the 3-day stimulation of the AMYG and MPO, while these were depressed by the 3-day stimulation of the ARC. 3) Rats of the same age were injected with estrogen (estradiol bonzoate, 0.5μg s. c.) about 48 hr before the stimulation and killed immediately after the 1-day stimulation. The marked elevations of pituitary LH and FSH were observed 48hr after the estrogen priming. Either stimulation of the MPO or of the AMYG increased pituitary LH, pituitary FSH and serum FSH, while stimulation of the HPC did not increase these in the animals. Therefore, at the first stage of puberty, the HPC may start to stimulare the release of FSH which may, in turn, be responsible for the growth of ovaries and estrogen secretion. After the increase of circulatory estrogen occurs, the AMYG and/or a more non-specific widely spread area may accelerate the further release of FSH to bring the vaginal opening and the first ovulation; however, the mechanism of the latter is unknown.

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Androgen-induced constant-estrous rats (50μg of testosterone propinate was subcutaneously injected at 5th day of age) were stimulated through implanted electrodes. Electrical stimulation (100Hz, 0.1msec, 100μA) was applied for 30min at 30sec period alternating with 30sec rest period. Immediately after the stimulation animals were anesthetized by ether and blood samples and anterior pituitary glands were taken for radioimmunoassay of gonadotropin. Results were as follows: 1) In the androgenized rats pituitary LH was slightly lower, but prolactin was higher than those of normal cycling rats. Pituitary FSH in the androgenized rats was almost equivalent to that in the 4-day cyclic rats. Serum FSH was lower in the androgenized rats, while serum LH was on the same level. 2) Electrical stimulation of the medial preoptic area increased serum LH and FSH, but decreased serum prolactin. The same tendency as induced by medial preoptic stimulation has been observed in the normal cycling rats. 3) Electrical stimulation of the medial part of the amygdala decreased pituitary LH, FSH and serum FSH, and did not increase serum LH. These changes were just the contrast of those found in normal cycling rats in which amygdaloid stimulation increased serum LH and FSH. 4) Hippocampal stimulation increased serum FSH, as was observed in 4-day cyclic rats. However, hippocampal stimulation slightly increased serum LH level in contrast to the change in normal cycling rats, in which the hippocampal stimulation never increases serum LH, but rather inhibits its increase with following to the medial preoptic stimulation. Furthermore, the weakening of inhibitory influence of the hippocampus on ovulation, which was induced by medial preoptic stimulation, was observed in androgenized rats.
In conclusion, when the function of the brain of androgenized rats was compared to that of normal cycling rats, quite a difference was observed in the amygdala and hippocampus rather than in the medial preoptic area.

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The localization of peroxidase in pig thyroid gland was investigated by staining histochemically thyroid slices previously fixed with glutaraldehyde-formaldehyde or unfixed. The reaction product was localized mainly at the membranes of endoplasmic reticulum. Golgi apparatus and microvillous apical cell border did not seem to show a significant amount of the reaction product. Nuclear envelopes and nucleoplasm never displayed the dark material. These results are in good agreement with our biochemical results obtained previously by fractionating the thyroid homogenate. Significance of these findings relating to the site of peroxidase is discussed in connection with the iodination of thyroglobulin in the thyroid follicular cells.

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A female patient with a solitary functioning nodule of the thyroid was operated upon. She had clinical symptoms of thyrotoxicosis and the thyroid scintigram showed a “hot” nodule corresponding to the palpable nodule. A cold area was noted in the “hot” nodule. At operation, a 1.6 by 2cm papillary carcinoma was found to be embedded within a functioning follicular adenoma, and metastatic foci were detected histologically in the ipsilateral paratracheal lymph nodes.

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Physiological effect of two major ovarian hormones, estradiol and progesterone, was examined on the uptake of ³H-leucine in the brains of ovariectomized adult rats. The animals ovariectomized previously were divided into 4 groups as follows according to the subsequent hormonal treatments, group 1; control animals, group 2; animals treated with estradiol benzoate plus oil, group 3; animals treated twice with estradiol benzoate, and group 4; estradiol benzoate-primed animals treated with progesterone. Animals in each group were given a single se injection of ³H-leucine 2 hr after the last hormonal treatment. Animals were sacrificed 2hr after the injection. Intensity of the uptake of ³Hleucine was measured by counting the number of reduced silver grains over cells in various brain regions, using an autoradiographic technique.
Group 1 showed a relatively high uptake in SO and PV, as compared with that in the remaining hypothalamic regions. Group 2 slightly induced an enhancement of the uptake in SO and PV, but not in the remaining hypothalamic regions, as compared with that in group 1. Group 3 showed a signifiant enhancement of the uptake in all the hypothalamic regions except PVA, as compared with that in groups 1 and 2. Group 4 enhanced significantly the uptake in all the hypothalamic nuclear regions except PV and MM, as compared with that in groups 1 and 2. The uptake in cells of the cerebral cortex and ependyme remanined unchanged after hormonal treatments in groups 2, 3 and 4.
The present results suggest that two major hormones stimulate protein synthesis in most of the hypothalamic nuclear regions, but not in the cerebral cortex and the ependyme. It is also suggested that, in connection with protein synthesis in the hypothalamus, response of the hypothalamus to progesterone is not affected by estrogen-priming. A possible role of both hormones in their positive feed-back control of the hypothalamus was discussed, with special reference to gonadotrophin releasing factors produced in the hypothalamus.

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Melanotropic (MSH) activities of synthetic octadeca and tetracosa-peptides were compared with those of α-MSH, β-human-MSH, and porcine ACTH. The MSH activity was determined by in vivo approach using a newly devised reflectance meter to measure the skin colour of Xenopus frog. From the shapes of their time-response curves, α-MSH, Gly¹-ACTH (1-18) amide (I), and β-Ala¹-ACTH (1-18) amide (II) werefound to be of short action, whereas two long chain peptides, porcine ACTH and ACTH (1-24)-OH were of comparatively long action.β-human-MSH, Orn¹⁵-ACTH (1-18) amide (III), and βA-Ala¹, D-Phe7, Orn¹⁵-ACTH (1-18) amide (IV) possessed extremely prolonged activity. The MSH potencies of these peptides were calculated from the dose required to give a minimal melanotropic response and were ranked as IV>β-human-MSH=II=III>ACTH=ACTH (1-24) OH>I.Compound IV was found to be about 400 times as potent as native ACTH. This ranking differs slightly from that found in the in vitro MSH assay proposed by Inouye et al.(1970 b) and is markedly different from the ranking in steroidogenic and lipolytic activities proposed by Tanaka (1971). The structure-activity relation and possible mechanism for these properties are discussed.

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Newborn male rats were castrated on the day of birth (=day 1) and injected subcutaneously with either 100μg of 5α-dihydrotestosterone (DHT), 500μg of DHT, 50μg of testosterone propionate (TP) or oil vehicle on day 5. At 45days of age, they received subcutaneous ovarian transplantation. At autopsy on day 90, ovarian grafts from the TP group contained numerous large vesicular follicles but no corpora lutea (CL). However, ovarian grafts from DHT-treated and control rats were characterized by the presence of numerous CL and a number of developing follicles. Similar results were obtained in the experiments on female rats. Treatment with DHT on day 5 could not alter the postpubertal ovarian function. All DHT-treated rats showed normal vaginal cycles. They became pregnant when placed with normal males. In contrast, treatment with TP resulted in anovulatory sterility. All rats showed persistent vaginal estrus untill autopsy on day 90. Futhermore, DHT did not prevent this sterilizing action of TP when given in combination with TP. These results suggest that DHT is not normally involved in the differentiation of the masculine type of the hypothalamus in the rat.

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In vivo MSH activities of three decapeptides, Gly¹-ACTH (1-10) OH, Ibu¹-ACTH (1-10) OH, β-Ala¹-ACTH (1-10) OH, and a tetradecapeptide Gly¹-ACTH (1-14) OH were compared with those of three octadecapeptides having N-terminal amino acid residues common to the respective shorter peptides; Gly¹-ACTH (1-18) NH₂, ACTH (1-18) NH2, and β-Ala¹-ACTH-(1-18) NH₂. The three decapeptides had almost the same minimum effective dose in assays in African frog, Xenopus laevis D. The potencies of three decapeptides were about 1/1, 000 of the potency of α-MSH on a weight basis and the tetradecapeptide was 1/640. The octadecapeptides, however, showed differing MSH potencies: Glyi-ACTH (1-18) NI12 was 1/48 as potent as α-MSH;β-Ala¹-ACTH (1-18) NH₂ and Ibu¹-ACTH (1-18) NH₂ exhibited 1/1.5-1/2.5 of the potency of α-MSH respectively. A possible mechanism explaining why the effect of substitution of the Nterminal residue on MSH activity is different in decapeptides and octadecapeptides is considered.

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An observation about height, weight and right mid-upper arm circumference (UAC) of 470 male subjects without apparent diseases was made. The subjects were 150 males at 40-49 years of age, 204 at 50-59 of age, and 116 at 60-69 of age. The weightcorrected UAC was calculated as the following formual.
Weight-corrected UAC (cm) =Measured UAC-0.15 W
where W=±percent weight deviation from the standard.
The weight-corrected UAC was 26.7±0.1 (mean±standard error) cm in subjects aged 40-49, 26.5±0.1 in subjects aged 50-59, and 25.9±0.1 in subjects aged 60-69. The difference between those at 50-59 of age and at 60-69 of age was statistically significant. This decrease is due to the decrease of muscle mass with advancing age. On the basis of these findings it is concluded that the weight-corrected UAC is a good index for body muscle mass and declines with advancing age.

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A weight-corrected UAC of 109 males and 292 females with untreated thyrotoxicosis was compared with that of the same number of control subjects. Control subjects were chosen from the subjects without apparent diseases; one control for one patient with about the same age, height and weight. The mean value of the weight-corrected UAC in thyrotoxic patients was 25.4cm both in males and females. The mean value of the controls was 26.7cm in males and 26.0cm in females. These findings suggest that UAC of thyrotoxic patients is smaller than that of controls even in the same height and weight, and that there is a significant decrease of body muscle mass in thyrotoxic patients.
Furthermore, it was suggested that the body muscle mass was decreased in 4 cases which had a long term steroid therapy, because the weight-corrected UAC was smaller than that of the controls.

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