Progesterone is the naturally-occuring progestational steroid which is inactive when administered orally. It has long been believed that orally administered progesterone is inactivated in the liver before it enters the systemic circulation. While such an inactivation actually occurs in the liver, no evidence has been reported on the stability of progesterone in the small intestine. In this experiment, metabolism of progesterone in the small intestine was studied in rabbits.
A dose of 10 or 20 μc of ³H-progesterone (3.1 μc/μg) was injected into the small intestine of rabbits and the regional mesenterial venous blood was collected for 30-120 min. Extraction of steroids from the serum was carried out as indicated in Fig. 1. While, unexpectedly, preliminary experiments indicated the presence of a considerable amount of radioactivities in the conjugated fraction, more critical examination was carried out as in Fig. 2. Following the extraction of free steroids, conjugated steroids were extracted with butanol and the butanol was evaporated under reduced pressure. Partition of the residue between chloroform and water revealed the presence of almost all the radioactivities in the watre. Recovered radioactivities after hot acid hydrolysis were comparable to that of the butanol extract. These data confirm the presence of conjugated progesterone metabolites in the mesenterial blood.
Both the free and the conjugated steroids obtained as shown in Fig. 1 were then chromatographed on alumina respectively. Fig. 3 shows elution patterns of radioactive steroids in these two steroid fractions. A large proportion of radioactivities was eluted in 0.3% methanol in benzene (MB) fraction and very little was found in the initial 0.1% MB fraction, where, when present, progesterone should be eluted. After chromic acid oxidation of the radioactive steroid eluted in 0.3% MB, the Rf value of the steroid on paper-chromatograph was found to be identical with that of progesterone as seen in Fig. 5. The original steroid is most likely 20-hydroxy-4-pregnene-3-one.
To avoid the influence of bacteria in the intestinal lumen, the metabolism of progesterone was also studied by the “Everted Sac Mrthod” as shown in Fig. 6. After 1-hour incubation, the fluid in the sac was extracted with chloroform. Elution pattern of the steroid in the extract from alumina was almost identical with that of the mesenterial venous blood extract.
All these data indicate that when a small amount of progesterone is administered orally, it must be largely metabolized in the small intestine (probably in the mucosa).
Experiments with a practical amount of progesterone will be reported in the near future.

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Though Plasma Inorganic Iodine (PII) levels have been reported by many authors, they differ from each other.
One theoretically reliable method for the estimation of PIT was devised, and experimentally testified for its reliability and compared with Stanley's method which has been believed most reliable. As a result, the author comfirmed that this method is an excellent one. In normal subjects and patients with various thyroid diseases PII was estimated with this method and their iodine metabolism was simultaneously studied.
On the other hand, correlation between PII and other clinical laboratory tests of thyroid function and effects of various drugs on PII were also investigated.
The results were as follows :
This method which estimates PII within 30 minutes only, is not only simple but more accurate. Correlation between this method and Stanley's was statistically significant (P<0.005).
With this method, the mean PII level and the standard deviation for 22 normal subjects were 0.44±0.24 μg/100 ml, and for 21 hyperthyroid patients were 0.21±0.18 μg/100 ml, that is, the latter was apparently lower than the former (P<0.01).
The mean PII level in 11 hypothyroid patients (0.78±0.49 μg/100 ml) was plainly higher and that in 17 patients of simple goiter showed a low level (0.26±0.23μg/100 ml).
PII levels in hyperthyroid patients who had received antithyroid drugs rose significantly to normal range in correlation with the improvement of their conditions.
Following the administration of thyroid hormone preparations, PIT levels in patients with simple goiter had a tendency to become higher but no change was observed in the PIT levels of hypothyroid patients. From these results it was suggested that PIT levels closely depended on the state of the thyroid function.
The PII levels rose following the sdministration of the iodide supplements (300 μg/ day) of a short duration (about a week), but increase was not found in a long-term administration (3 months). The closest negative correlation between PII and PM was found (correlation coefficinet-0.81, P<0.005).
PII levels in the euthyroid patients who had received both adrenocorticosteriod and anabolic steroid preparations were below normal.
Correlation between the PII and the renal clearance of iodide was not found.

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The extracts of human chorionic tissues and hydatidiform moles were prepared by homogenization and high speed centrifugation. The particle free supernatant fraction precipitated by ethanol and purified samples of urinary HCG assaying 5,100 I.U. and 2,406 I.U. per mg. were subjected to zone electrophoresis in starch grain.
Proteins recovered after electrophoresis from portions of the starch grain were tested for gonadotropic activities by the weight of rat ovaries, seminal vesicles and the augmentation test of Steelman & Pohley. There were found at least two biologically active components (G-A and G-B) in all of the extracts and preparations examined. G-A has luteinizing activity and G-B has follicle stimulating activity. Each electrophoretically homogeneous component could be isolated by repeated electrophoresis.
The extracts of human chorionic tissues in the early stage of pregnancy have predominant G-B activity and in the late stage, G-A activity when assayed by the ovarian weight method. The extracts of hydatidiform moles have more predominant G-B activity than that of normal chorionic tissues when assayed by the augmentation test of Steelman & Pohley.
The urea denaturatuion experiments and RDE digestions show that there may be differences between urinary HCG and trophoblastic HCG, between G-A of urinary HCG and G-A of urinary HMG.
The acetone powder of the supernatant fraction of hydatidiform moles has the specific component with gonadotropic activity, assayed by the augmentation test of Steelman & Pohley, in the zone of segment number 35-39, which is not found in the case of normal chorionic tissues.

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The author made some modification in Morris' method for urinary 17-KGS determination. It was found that the use of NaBiO₃ 2.5 gm per 5cc of urine for two huors of oxidation was better than the original Morris' method where NaBiO₃ 1.0 gm per 8cc of urines was used for the determination of 17-hydroxycorticosteroids. By this modification, the fractions of 17-KGS, i.e. 11-deoxy 17-KGS and 11-oxygenated 17-KGS (11-oxy 17-KGS), were successfully determined.
The values of 17-KGS and its fractions in normal adults by this method were as follows : urinary 17-KGS : 8.2±2.7 mg/day, 11-deoxy 17-KGS : 0.9±0.8 mg/day, 11-oxy 17-KGS : 7.3±2.2 mg/day, the ratio of 11-deoxy 17-KGS to 11-oxy 17-KGS (11-deoxy/11-oxy) : 0.12±0.10. There was found no sex difference in normal adults. In elderly subjects, a lower value of each fraction was observed. Increased 11-deoxy 17-KGS and elevation of 11-deoxy/11-oxy due to placental pregnanetriol were observed in pregnant women, and this tendency became definitely apparent in the last trimester of pregnancy.
Among the cases with adrenogenital syndrome, 10 cases with simple virilizing form revealed a marked increase of urinary 17-KGS, which was due to the remarkable increase of 11-deoxy 17-KGS fraction and a mild increase of 11-oxy 17-KGS fraction and high 11-deoxy/11-oxy (2.41±0.61). In 3 other cases with hypertensive form, an equally higher 11-deoxy/11-oxy (6.76±1.71) was observed, but this was due to a relative decrease of 11-oxy 17-KGS fraction. In 9 cases with Cushing's syndrome, an increase of 17-KGS with normal 11-deoxy/11-oxy was observed in all cases. Since various cases with endocrine disorders such as Addison's disease, acromegaly, thyroidal disease or others, all revealed normal 11-deoxy/11-oxy, the elevation of 11-deoxy/11-oxy of urinary 17-KGS is though to be specific for adrenogenital syndrome due to congenital adrenal hyperplasia, besides pregnancy. Thus, determination of urinary 17-KGS fraction is very important for confirming the diagnosis of adrenogenital syndrome.

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ACTH-Z 20 units, was administered intramuscularly for 3 successive days in ACTH test. In normal adults, the values of 17-KGS and its two fractions on the 3rd day of injection were as follows : urinary 17-KGS : 40.2±12.6 mg/day, 11-deoxy 17-KGS : 7.3±3.7 mg/day and 11-oxy 17-KGS : 32.8±11.1 mg/day. These valeus were 6.3±2.0, 9.1±2.6 and 5.6±2.0 times as much as those before the injection, respectively. However, no change was observed in 11-deoxy/11-oxy. In elderly subjects, a slight elevation of 11-deoxy/11-oxy due to relative increase of 11-deoxy 17-KGS was observed, suggesting lowered relative efficiency of cortisol synthesis in adrenal cortex.
Three cases of adrenogenital syndrome due to the incomplete block of 21-hydroxylation showed an excessive reaction in the ACTH-Z test. This is mainly due to the remarkable increase of 11-deoxy 17-KGS. The metabolic disturbance, which was observed in the above-mentioned cases, could not to be corrected by the administration of ACTH. In 7 cases with Cushing's syndrome, there was not observed any difference in the reaction to the ACTH-Z administration between the cases with adrenal adenoma and those with adrenal hyperplasia. However, in case with adrenal hyperplasia which did not show any significant reaction in ACTH-Z test, an excessive reaction was observed to the ACTH intravenously infused. This fact indicates the inadequacy of ACTH-Z test for the differentiation of Cushing's syndrome due to the adrenal hyperplasia from those due to the adrenal adenoma. The decreased or delayed reaction in the ACTH-Z test was observed in cases with acromegaly, Addison's disease or panhypopituitarism, wutithout showing any increase of 11-deoxy 17-KGS in most of them.

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In normal adults, administration of SU-4885 3.0 gm caused an increase of urinary 17-KGS and 11-deoxy 17-KGS to 9.0-22.0 mg/day and 7.0-20.0 mg/day, respectively on the first day after the administration of SU-4885, but did not cause any decrease of 11-oxy 17-KGS. Therefore, 11-deoxy/11-oxy became higher, 1.80-4.00. In the aged, the increase of 11-deoxy 17-KGS was greater than that in adults,. consequently showing a remarkable elevation of 11-deoxy/11-oxy. This finding was similar to the results observed in ACTH-Z test.
One among 5 cases with adrenogenital syndrome showed no reaction at all, revealing a very low reserve of pituitary function, but the other 4 cases showed a remarkable increase of 11-deoxy 17-KGS. This fact indicates that the block of 21-hydroxylation enzyme system is incomplete and many cases with this disorder still retain a sufficient reserve of pituitary function. There was observed a person who had no physical sign of adrenogenital syndrome, although his two children showed typical adrenogenital syndromes, and revealed unusually high 11-deoxy/11-oxy in urine at resting state and showed excessive reaction in SU-4885 test which was similar to that of his children. This is not a phenotype of adrenogenital syndrome, but a suspected genotype. In that event, it is interesting to note that some genotype of adrenogenital syndrome without any of its clinical sign shows similar endocrinological disturbances to the genotype.
Of patients with Cushing's syndrome, those due to adrenal hyperplasia revealed all excessive reaction in the test and those due to adrenal adenoma showed normal or decreased reaction in the test, and thus the differentiation in them was made possible by the test. Decreased reaction was observed in the test in cases with panhypopituitarism, acromegaly or myxedema.
In the cases showing positive reaction in the test, decrease of 11-oxy 17-KGS was frequently observed. Therefore, 11-deoxy 17-KGS was found to be the most reliable and specific index for the positive reaction to SU-4885. In general, 11-deoxy/11-oxy seems to be closely related to the degree of block of 11-hydroxylation by SU-4885. This is supported by the fact that clear reversed correlation was observed between the maximum value of 11-deoxy/11-oxy in SU-4885 test and the maximum value of urinary 17-KGS in ACTH-Z test.

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Reichstein's compounds, C-20 reduced metabolites of cortisol, have been found in human urine after loading with exogenous cortisol or ACTH. No reports fire found at present about urinary daily excretion of these metabolites in normal subjects under basal conditions.
In this paper, by means of thin layer chromatography, the daily excretion of these compounds as well as tetrahydro- (THF + a11oTHF, THE) and hexahydrocompounds (cortols & cortolones), were determined in 5 normal subjects (male) and 6 patients with cirrhosis of the liver, before and after oral administration of 100 mg of cortisol.
Each urine specimen, after hydrolysis with β-glucuronidase, was extracted with ethyl acetate. Crude extract after washing was separated to fraction A and B by the first thin layer ', chromatography (Kieselgel GF₂54, solvent : cyclohexan-isopropanol 6 : 4). Fraction A, containing Reichstein's compounds and hexahydrocompounds, was oxidized with periodic acid and fractionated using the partition thin layer chromatography of Chang's, and finally determined by Zimmermann reaction. Fraction B was rechromatographed on another silicagel plate using solvent stystem methylene dischloride-ethanol to separate each tetrahydro-compounds, and to quantify by blue tetrazolium reaction. Recovery study was carried out using each ¹⁴C-labelled steroid. Regular existence of Reichstein's compounds in normal urine was demonstrated by preparing the autoradiogram of fraction A..
Normal urinary excretions of Reichstein's E & U in the basal state were 0.27±0.08 mg and 0.22±0.09 mg per 24 hours, respectively. The values of tetrahydro-and hexahydro-compounds in normal state were in good agreement with the data previously reported.
In patients, a three-fold increase was observed in the excretion of Reichstein's compounds as compared to normals. Cortols and cortolones also increased significantly, while tetrahydro-compounds decreased. This tendency was greater after loadimng with cortisol.
These results suggest that severe damage of the liver function would remarkably impair the metabolic pathway to tetrahydro-compounds orf cotisol since the reduction of ring-A occurs solely in the liver. And the greater excretion of C-20 reduced metabolites in the cirrhotic patients also suggests the relative predominance of the pathway via Reichstein's compounds, which is recognized as the minor route in normal cortisol metabolism.

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