Turnover experiments of¹⁵N-labeled amino acids were carried out to clarify the mechanism of anabolic steroid action upon protein metabolism. Ten patients with gastric ulcer, duodenal ulcer, tinnitus, panhypopituitalism, hypogonadism, chronic nephritis, hypothyroidism, Addison's disease and chronic hepatitis were studied.
Two grams of 30 atom% excess ¹⁵N-glycine or aspartic acid were given orally to each subject, and total nitrogen and ¹⁵N-atom excess in urine, stool and serum were determined within 120 hours. The same loadings of ¹⁵N-amino acids and determinations again were carried out 7 days after the beginning of anabolic steroid, thyroid hormones or glucocorticoid treatment, and metabolic processes before and after the treatment were compared. The size of the metabolic pool was calculated according to the method of Rittenberg and others.
I have analyzed by a new method which was independent of the steady state of protein metabolism. For the purpose of clarifying the shift of ¹⁵N in the body after ¹⁵N-amino acids administration, a series of exponential functions was determined after analyzing the curve of retained ¹⁵N in the body.
An index Kp showing the ¹⁵N utilization rate in the protein synthesis was worked out.
1) In all cases of 5 patients with normal liver and kidney function, decreased urinary nitrogen excretion and increased metabolic pool and Kp value were observed after the administration of anabolic steroids.
2) In patients with liver dysfunction, urinary nitrogen excretion was decreased, while no significant change was seen in metabolic pool and Kp value.
3) In the case of chronic nephritis there was a slight increase in metabolic pool but no significant change in Kp value after anabolic steroid treatment.
4) After administration of 1-Triiodothyronine for hypothyroidism or of dexamethasone for Addison's disease, acceleration and increase of protein synthesis as well as degradation were seen.
From the results of this experiment and former experiment regarding the effect of anabolic steroids upon protein metabolism using ¹³¹I-human serum albumin, it may be concluded that anaboic steroids do not indirectly influence protein metabolism by inhibiting its catabolism but directly accelerate protein synthesis, and that the integrity of liver and kidney function plays an important role in the acceleration of protein synthesis in the body.

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Deuterium turnover studies were carried out on 8 normal subjects, 1 esophagus cancer with ascites and 1 diabetes insipidus.
Following the oral administration of 0.1 ml per kilogram of body weight of 99.8% pure deuterium oxide, accurately timed urine samples for deuterium determinations were collected for 10 days. And then, anabolic steroid or other treatments were begun. One week after the beginning of treatment, each subject was restudied by the same loading test and determination. Deuterium concentration in urine was measured with a massspectrometer, and total body water and biological half-life of deuterium oxide were calculated for both periods. Total body water was calculated using the deuterium concentration in urine and serum four hours after the administration of deuterium oxide.
1) No significant differences of total body water per kilogram of body weight were observed among children, young adults and elderly subjects. Bilolgical half-life was prolonged with age. Thus, in the elderly subjects water metabolism appeared to be slowed.
2) In esophagus cancer with ascites the retardation of deuterium turnover and increase in the body water were observed. On the contrary, accelaration of deuterium oxide turnover and decrease in body water were seen in diabetes insipidus.
3) During anabolic steroid trsatment in 8 normal subjects, a body weight gain of 1 to 3 kilograms was noted in each subject, but edema did not appear in any case. Deuterium turnover did not markedly change before and during anabolic steroid treatment. However, in 1 of the young adults being administered anabolic steroid for a long term and in 2 of the elderly subjects a slight increase in total body water occurred during anabolic steroid treatment.
4) A return to the normal condition of deuterium turnover and of the total body water was seen in the diabetes indipidus and esophagus cancer with ascites after trichlormethiazide treatment.

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The effects of large doses of iodine on thyroid weight and the hormone synthesis was studied in chick and rat with the aid of ¹³¹I and chromatographic analysis.
Male and female Whiet-Rock chicks weighing 900 to 1500 g. were fed on a standord diet throughout the experiments. In order to determine the effect of a single dose of excessive iodide, chicks were given an intraperitoneal injection of 100-200 μc ¹³¹I with carrier 1.0 mg. iodide. Thyroids were removed and analysed at an appropriate time after the injection. Control chicks were injected 10-20 μc ¹³¹I alone and sacrificed at the corresponding time. Long-term treatment with iodide was carried out as follows : Chicks were given about 1.0 mg. iodide every day for the last 4 weeks. and each one was injected 1.0 mg. iodide together with 200 μc ¹³¹I. The thyoids were removed at 4 and 24 hours 1 after ¹³¹I.
For the comparison among different species, male Wistar rats weighing 100-150 g. were also fed on a low iodine diet throughout the experiments. The method was the same as used in chicks, but the quantity of iodide was 500 sg. and the duration of iodide treatment was 3 weeks.
Other experiments were studied with chicks at 1, 3 and 9 weeks after withdrawal of a long-term treatment of iodide for the study of residual effect of the treatment. In these experiments, chicks were sacrificed at 4 hours after the injection of carrier free ³¹¹I.
Radiochromatographic analysis of the thyroid tissue after enzymatic hydrolysis was performed either by paperchromatography or columu chromatography using Pileggi's modification.
Long-term treatment of iodide for 4 weeks induced a significant increment of the thyroid weight in chick; about 1.8 times greater than the controls. Histology of the goiter showed flat epithels and marked colloid storage within large follicles. While the 3-week treatment of iodide failed to produce goiter in rats.
In both animals, it was shown on chromatogram of the thyroid that a single of carrier iodide resulted in a significant but transient reduction of organic ¹³¹I and thyronine ¹³¹I proportion as well as an elevation of ¹³¹I MIT/DIT ratio. After a long-term treatment of rat with iodide, an “escape” occured from this inhibitory effect of iodide; formation of thyronines was resumed with recovery of organification of iodide and MIT/DIT ratio, as previously reported by Pitt-Rivers et al. On the contrary, in the goitrous chicks continuo usly treated with iodide, synthesis of thyronine ¹³¹I was significantly depressed notwithstanding the presence of normal proportion of organic ¹³¹I and rather lower MIT/DIT ratio than controls.
After a 3-week withdrawal of iodide feeding, MIT/DIT ratio recovered first. The depressed thyronine ¹³¹I synthesis as well as increased thyroid weight had not returned to control levels until 9 weeks after stopping the iodide treatment.
It appears that the depressed rate of thyronine synthesis would be a cause of the goiter through the negative feed-back mechanism in iodide-fed chicks.

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This report attempts to study the effect of adrenocortical steroids on the thyroid gland. 1) The effect of adrenocortical steroids (dexamethasone : 0.125 mg./100 gm. body wt., hydrocortisone : 5 mg./100 gm. body wt.) and ACTH (ACTH-Z : 3 I.U./100 gm. body wt.) on the thyroid gland in a rat, was studied.
2) Wister male rats weighing 110 to 150 gm. were maintained on a standard diet, and iodine solution (9.6, μg. /day) was injected throughout the experiment.
A tracer dose of 10 μc. of radioiodine was administered subcutaneously and 24 hrs. later, thyroidal radioactivity was measured once a day using a collimated scintillation counter over the thyroid gland. Radioactivity was corrected for physiological decay and the rate of the I¹³¹ release was calculated by the method of least squares.
Following the determination of the rate of the I¹³¹ release in a control period, adrenocortical steroids (subcutaneously twice a day) or ACTH (intra muscularlly once a day) was administered and the rate of the release was compared with that in a control period.
3) The mean rate (-λ) calculated of the thyroidal I¹³¹ release in a control group was 0.262/ day and a daily injection of dexamethasone or ACTH induced a decrease of the rate while that of hydrocortisone an increase of the rate in two out of three rats.
4) Then, saline solution or hydrocortisone to a control group was injected for 16 days and dexamethasone or ACTH injected for 18 days in subsequent experiments.
The thyroid was weighed and a chromatogram was developed in BAW and BDA solvent after termination of the enzymic hydrolysis. Autoradiography was performed and radioactivity of each fraction was measured by a well-type scintillation counter.
5) The mean thyroid weight was 8.2 mg/100 gm. body wt, in a control group and the weight increased in a dexamethasone and ACTH group. The mean values of serum total iodine revealed 6.8 μg./dl. and those of serum PBI 4.5 μg./dl. in a control group, and these values were not affected by the treatment with dexamethasone, hydrocortisone or ACTH. The weight ratio of iodine, 3-monoiodotyrosine (MIT), 3, 5-diiodotyrosine (DIT), 3, 5, 3',-triiodothyronine (T₃) and thyroxine (T₃) to all iodinated materials in the thyroid gland were 1.8%, 41.8%, 48.3%, 1.8% and 6.2%, respectively, in a control group.
Dexamethasone decreased the ratio of T₃ + T₄/MIT+ DIT and hydrocortisone increased the ratio of MIT/DIT and decreased T₃ + T₄/MIT + DIT.
6) On histological examination, the mean acinar cell hight was 8.3μ, e2 and the mean diameter of follicles was 35.4μ in a control group. The mean acinar cell height increased in the dexamethasone or hydrocortisone group and decreased in an ACTH group. The mean diameter of follicles decreased in the hydrocortisone group.
7) These findings suggest that ACTH suppresses the thyroid functions but the effect of dexamethasone or hydrocortisone on the thyroid functions is a little different from that of ACTH. Moreover, it is likely that although dexamethasone and hydrocortisone may suppress either TSH secretion or action, they have also some effects on the level of the thyroid gland.

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The author intended to investigate how the adrenal cortex of healthy infants and children respond to the successive administration of adrenocorticotropin (ACTH), especially according to their ages. The subjects consisted of 22 healthy infants and children, aged from 2 months to less than 15 years. Daily doses of 20 units of ACTH-Z were given intramuscularly for 5 successive days. The urinary total 17-hydroxycorticosteroids (17-OHCS) excretion was determined by the modified Porter-Silber method and the results were evaluated by three age goups, divided into infants (from 2 months to 1 year of age), young children (from 2 years to 6 years of age) and older children (from 7 years to 14 years of age).
1) During the successive administration of ACTH, the urinary total 17-OHCS excretion increased significantly in all subjects, the maximum values during the observation (mean values, confidence limits) were 12.74±6.07 mg./day in infants, 13.05±4.45 mg./ day in young children, and 14.61±2.97 mg/day in older children, respectively. No significant differences due to ages were found among these absolute values.
2) However, when these absolute values were calculated by the body surface area, the maximum values showed 44.42-±16.46 mg./m²/day in infants, 21.42±6.93 mg./m²/ day in young children, and 16.20±4.22 mg./m²/day in older children, respectively. The maximum value obtained in infants was significantly higher than those obtained in the other two groups of children. Furthermore, the values of increased excretion during ACTH administration as compared with those of basal excretion were also higher in infants.
3) The values in young and older children reached the maximum often on the 3rd day of the administration without a remarkable increase afterwards. However, they in-creased in most cases of infants continuously during the whole period of the observation and reached the maximum just on the 5th day of the administration.
4) These results are considered to be of significance in order to recognize adrenocortical reserve, especially its maximal response, in infancy and childhood. It is concluded, that the adrenocortical reserve in infants is not inferior, as compared with those of young and older children, and it seems, furthermore, to be possible even to interpret that this reserve power may be comparatively superior in infants.

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Following the previous report, the author intended, in this study, to investigate how the adrenal cortex of children with various diseases respond to the administration of adrenocorticotropin (ACTH). The subjects consisted of 32 cases with endocrine or metabolic diseases, 11 cases with central nervous disease and 22 cases who were previously treated with corticosteroids. Daily doses of 20 units ACTH-Z were given intramuscularly for 2 to 5 days, and the urinary total 17-hydroxycorticosteroids (17-OHCS) excretion was determined. The results were compared with those obtained from the healthy infants and children which were described in the previous report.
1) Diseases of endocrine and metabolism i) Dwarfism-The response in two of six examined cases with pituitary dwarfism showed the prolonged type (prolonged response). In a patient with progeria no significant elevation of urinary 17-OHCS excretion was observed during the administration of ACTH. However, such abnormal responses were observed in none of six cases with primordial dwarfism.
ii) Obesity-The response in three of eight cases with simple obesity showed the excessive type (excessive response). On the contrary, two of six cases with Laurence-Moon-Biedl syndrome showed the prolonged response.
iii) Miscellaneous Diseases-No abnormal response was obtained in the cases of chondrodystrophia foetalis (two cases), gargoylism (two cases) and leucine sensitive hypoglycemia (one case).
2) Diseases of central nervous system i) Cerebral palsy-Two of six cases examined showed the weak response, which the author described as the “transitory” type. Other types of abnormal response, such as the prolonged response, excessive response were also obtained in this disease.
ii) Brain tumor-One of five cases examined showed the prolonged response and the other one showed the excessive response.
3) Children with a previous administration of corticosteroids. Nine of twenty-two cases examined showed the prolonged response. No significant relation between the appearance of this response and the ages of the subjects was found.
4) The adrenocortical responsiveness of children, subjected for the study, was commented according to the above-mentioned results.

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The previous report from our institute has shown that the long-term treatment of cockerel with iodide indueed a goiter as well as a decrease in thyroxine-¹³¹I and ¹³¹MIT/¹³¹DIT ratio.
In order to elucidate the metabolism of stable iodine in the goitrous cockerels, the following experiments were made.
For this purpose, column chromatography using DOWEX, 1× 2, Cl^-form, 200-400 mesh was employed. This resin is able to absorb large doses of inorganic iodine and this column chromatography is more excellent in recovery of thyronine fraction and is more convenient in chemical analysis of stable iodine than paper chromatography.
The plasma thyronine level was also assayed by using this mathod.
1) After treatment of cockerels with iodine (1 mg/day) for 4 weeks, tracer ¹³¹I was injected and thyroids were removed 24 hours later Thyroid hydrolysate was fractionated by column chromatography and both ¹³¹I and ¹²⁷I in the fraction were determined. Distribution of ¹²⁷I was compatible with that of ¹³¹I, thyronine proportion decreased (0.01<p< 0.05) and MIT/DIT ratio was low. (p<0.001).
2) After treatment with a diet containing both ¹³¹I and ¹²⁷I, decrease in MIT/DIT ratio (0.01<p<0.05) and thyronine proportion (p<0.001) were demonstrated.
Specific activity of ¹³¹I in the gland was identical with that in the diet after equilibrium study for 5 weeks.
3) In goitrous cockerels treated with iodine, total ¹³¹I content increased to 7 times and thyronine ¹²⁷I to 3 times as much as controls. One week after discontinuance of a 4-week-iodine treatment, total ¹³¹I content dereased depending on the decrease of DIT.
4) In iodide-treated cockerels, plasma thyronine-¹²⁷I, half-life of ¹³¹I-labeled thyronine and daily degradation of thyronine were not different from controls.
Decrease rate of thyronine synthesis (especially coupling process) and of hormone release as possible causes for the goiter in the iodide-treated cockerels were discussed.

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