The author investigated the conjugated form of corticoids in human plasma, hydrolyzed withβ-glucuronidase or with hydrochloric acid and obtained following results.
1. A glucuronide of corticoid, which hardly can be separated from hydrocortisone by means of paper chromatography after the procedure of Zaffaroni, was found to be present in plasma, besides tetrahydrohydrocortisone and corticoids of pregnane-3, 17, 20, 21-tetrol types. By column chromatography after the procedure of Sweat, the above free made corticoid was present in the eluate with 4% ethanol chloroform, in which hydrocortisone as well as tetrahydrohydrocortisone were to be found. The corticoid in question gave a powerful phosphoric acid-induced fluorescence like hydrocortisone, notwithstanding that it was present as glucuronide. And it had no peak at 240mμ, in the ultraviolet absorption. When hydrocortisone was administered, it showed manifest increase in plasma as glucuronide and it seems, therefore that the substance is closely related to the metabolism o f hydrocortisone.
2. The level of the above substance as glucuronide was determined in plasma of normal adults and patients suffering from various diseases by phosphoric acid-induced fluorescence and it was found that especially in liver disease the substance was manifestly decreased: when hydrolyzed, however, with hydrochloric acid, there was no decrease of the substance. In short, the ability of the substance to conjugate with glucuronic acid is inhibited in liver disease but it is conjugated quite in another form.
In liver disease the compound B-like substance of free form generally is increased; a substance, which was found in the same fraction as compound B after hydrolysis with hydrochloric acid gave phosphoric acid-induced fluorescence and was also greatly increased in the same disease.

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The rapid disappearance of injected alloxan from the blood was demonstrated by Leech et al (1945). For the study on the metabolism of alloxan, it is necessary to determine quantitatively the amounts of alloxan or its decomposed and conversed products, which appeared in the blood or were excreted in the urine.
Blood alloxan determination was carried out by its reducing ability in the cold (Ferricyanide method).
Besoder that for alloxan in urine oxomalonate liberated by hydrosis of urine was measured through paperchromatography using p-nitrophenylhydrazine.
The results are as follows:
1) In experiments in vitro where alloxan was hydrolysed, the quantity of conversed urea coincided with the theoretically calculated amount, and that of oxomalonate was about a half of the calculated, though oxomalonate added in the solution can be recoverd completely.
2) Alloxan was not decomposed by incubation with serum, but decomposod, by incubation with liver slice,-after 60 minutes it showed no color reaction of alloxan, and urea increased markedly. Oxomalonate was not observed. It was supposed that it might change to some other substance such as tautronic acid.
3) Small amount of alloxan were observed in blood two hours after alloxan injection, and none eight hours.
4) The amount of alloxan excreted in urine is maximum two hours after injection, and a little after four hours. The total quantity of excreted alloxan was 7.6% of injected alloxan.
5) The amount of excrected alloxan recoverd from that of oxomalonate measured after urine hydrolysis was about 15% of the injected amount of alloxan.
6) Urea and oxmalonate were measured in blood and in urine after alloxan injection to know the decomposition of alloxan in body.
No free oxomalonate was observed in blood and urine, urea increased markedly in blood two hours after injection of alloxan, and also in urine two to four hours after injection. If the increased amount of urea in urine after injection is supposed to be an exogenic urea, a half amount of injected alloxan might have been decomposed in body.

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It is of interest to study the activity of pituitary adrenocortical function in patients with various surgical diseases and surgical manipulations. The urinary excretion of corticosteroids (C-S) was determined to authenticate the function of these systems. Simultaneously, the immediate response upon urinary excretion of C-S to ACTH was compared with those after surgical operations in the same patients.
The results were as follows:
(1) Each 24 hours urine sample was collected, and the amount of urinary excretion of C-S (measured as formaldehydogenic steroids) was determined in 77 surgical patients-(carcinoma 23 cases, surgical liver diseases 10 cases, surgical abdominal diseases 20 cases, and endocrine diseases 24 cases).
Each group of surgical disease excreted higher amount of urinary C-S than the normal subjects, and the highest value was found in patients with carcinoma while those with gastric ulcer were below the control level. This fact may be applicable clinically as a diagnostic aid for the diagnosis of carcinoma and ulcer of stomach.
(2) The amount of daily excretion of C-S after operation showed daily fiuctuations. One or two elevations (peak) of urinary excretion were seen during the 2 weeks after operation. Among 32 cases of operation, 23 cases showed 2 Peaks and in the remaining 9 cases were 1 peak. The first peak was higher than the second peak in many cases.
(3) The immediate responses upon the urinary excretion of C-S to ACTH were measured in 18 surgical patients, and in average a 3 fold increase in C-S excretion were found in these patients with ACTH injection. It has also been found that the responses to ACTH and to operative manipulation upon urinary excretion of C-S were almost the same when the average increased amounts of urinary excretion were compared in both cases.
(4) The effects of ACTH on blood sugar values of 21 surgical patients were tested. Hypoglycemic effects have been seen in 13 cases, while 2 cases were followed by elevation of blood sugar, and 6 cases responded with both hyper and hypoglycemic phase. Therefore, ACTH was suggested to induce the decreased effects of blood sugar to many of surgical patients. However, no correlation between the urinary C-S excretion and the change of blood sugar value was found to exist following ACTH injection.

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We can successfully find a method of hypophysectomy on the mouse by the modification and improvement of Koyama's Method (external auditory canal method).
Our method of hypophysectomy is to be conducted as fellow:
1. The mouse is anesthetized with ether.
2. The needle (1/1 injection needle) is used to fit with the injector being 2cc in size.
3. The injection needle held as its cut surface downwards is inserted in the external auricular opening and after breaking through tympanic small canal run carefly 2mm verticaly to the longitudinal axis of body.
4. Drawing gently the inside cylinder of injector and then hypophysis is intered into the injector.
5. We can probe hypophysis with capillary tubesnet and distinguish its anterior-lobe from posteriorlobe in a drop of glycerine under microscope.
6. The depth of perforation is 2mm plus to the half of distance between both incisura interlobularis.

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The pineal body of the albio rat is right under the confluence of sinuses. Eliminating usual complicated operation, removing some parts of the interparietal bone, ligating and cutting one of the transverse sinus and just holding up it toward the confluence of sinuses we could pick up the pineal body. We applied the peculiarrity of the topographic anatomy for this operation, and paid attention to the follows. 1. The distans between the sutura and transverse sinus. 2. The angle which made by the sinuses. 3. The cranium is easily separated at the suture. 4. The connection of.dura mater to the cranium. 5. On the straight sinus.
From stated before, you will know how to perform the operation.
A dorsal medline incision is made in the skin of the head, Now you are ready to perforate parietal bone with a trepan, but, before perforating it, you may make two incision-line with a knife, running different way from the point of perforation to both right and left parts of sutures, which made by the interparietal and parietalbones. But, you must make carefully two incision-line not reach to the dura mater. And then you perform complete perforation. You can remove easily interparietal flap the round of perforated point to suture along the two incisions, and you may see the surface of the dura mater around the condluence of sinuses.
You ligate one side of the transverse sinus at the two places with needle with ligature, cut the dura mater around the sinuses with a point of injection-needle, and cut down the middle part of the two ligature then you lift up the confluence of sinuses with one of the ligature diagonally to nose side of the rat. The pineal body will be exposed, and you shall be able to pick up easily with a pincett.
We didn't see may obstruction on the rats which we performed operation of cutting dura mater, thrusting cerebellum and cerebrum, and ligating sinuses. We narconized with Tiopental-Sodium 5mg/100gm.

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Estrogen conversion experiments incubating etradiol, estrone and estriol with human placenta homogenate respectively in an atmosphere of oxygen or nitrogen were studied.
The estrogens were determined by Kober reaction and identified by paper chromatography.
The following results were obtained in these experiments.
1) Estradiol incubated with placenta in an atmosphere of oxygen converted to estrone in large amount, but in an atmosphere of nitrogen converted to estrone in small amount. (Presumably incubating medium dissolve few oxygen.)
2) Incubation of estrone or estriol with placenta in an atmosphere of nitrogen resulted in no conversion to other estrogens. The recovery rate of additional hormones incubated in an atmosphere of nitrogen were lower than in case of oxygen.
3) From the above results, it may be summarized that estrogen level declines in urine or blood in toxemia of late pregnancy depend on oxygen deficiency in placenta.

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The present report concerns with the supplementary studies on Silber-Porter's method which is used to estimate adrenocortical function in both clinical and experimental fields as a simplified procedure.
We reconfirmed the importance of purification of reagens, especially that of chloroform and phenylhydrazine, as has been emphasized by previous reporters.
Particularly in purifing the latter, unglazed pottery plate to absorb solvent is considered to be worthy of extensive application.
More clear-cut result were obtained by adding 8μgm. of standard hydrocortisone than those by the original method in which 5μgm. is appended.
According to our modified Silber-Porter's method, the concentration of the cortical steroid was estimated from the peripheral circulating blood of Japanese under normal condition. The value estimated is statistically slightly lower than that of Anglo-Americans, but coincides fairy well with the reports of other investigation of Japanese. It is, therefore, concluded that the concentration of 17-21-dihydroxy-20-ketosteroid contained in blood of the Japanese is generally lower than that of Anglo-Americans.

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When melanocyte stimulating hormone (MSH) is injected into an animal, the blood level of MSH decreases rapidly thereafter (Landgrebe, Reid and Waring, 1943). As only a small portion of injected MSH is excreted in the urine (Lerner, Shizume and Bunding 1954), the hormone is either rapidly inactivated or bound by tissue in the body.
As the inactivation of posterior pituitary hormones by liver and kidney, and gonadotrophic hormones by liver has been reported previously, the role of liver and kidney in the rapid disappearance of MSH from the circulation has been studied.
MSH was administered intravenously to hepatectomized dogs, dogs with injured liver and nephrectomized dogs, and the disappearance rate of injected MSH from the circulation was compared with that in normal dogs. In addition, the level of MSH in the portal vein and in the hepatic vein was compared after intravenous injection of MSH into normal dogs. In the second part of the study, inactivation of MSH was tested after incubation with homogenates of liver, kidney and other tissues.
From these experiments, the following results were obtained. (1) When hog or dog MSH was injected into dogs, the blood level of injected MSH decreased rapidly thereafter. When hog or dog MSH was injected into hepatectomized dogs, the rate of disappearance was definitely retarded as compared with that in normal dogs.
(2) Similar retardation, although in lesser degree, of the disappearance of injected MSH from the circulation, was observed in the dogs whose livers were injured with carbon tetrachloride.
(3) When MSH was injected into normal dogs, the level of MSH in the hepatic vein was always lower than that in the portal vein, and MSH was not found in the bile.
(4) When hog MSH was injected into nephrectomized dogs, the rate of disappearance was retarded as compared with that in normal dogs, although in lesser degree compared with that in hepatectomized dogs.
(5) When MSH was incubated with homogenates of various tissues of the dog at 37°C., homogenate of liver, kidney and muscle inactivated MSH. The degree of inactivation was most marked by the liver, next by the kidney and then by the muscle. When these homogenates were boiled before incubation or incubation was performed at 4°C., no inactivation was observed.
These results indicate that liver and kidney play great roles in the inactivation of MSH and this inactivation is performed by some enzymatic system.

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From the views of hypothalamic-hypohyseal-gonadal system, experimental evidence pertinent to the problem of the functional relationship of the system was performed.
Results obtained were as follows:
(1) Secretion of androgenic hormones increased usually on tne third or forth day after the surgical operations. Urinary excretion of 17-ketosteroids reincreased on this period. Gonadal fractions of 17-ketosteroids showed the marked increase and excretion of hypophyseal gonadotrophin did so in parallel with increase of gonadal fractions.
(2) Neurosecretion of the hypothalamic area resulted in hyperactivity after the surgical stress. It was showed that gomoriphil substance and content of oxytocin and vasopressin in the anterior hypothalamus and neurohypophyse. decreased remarkably immediately after the stress. Administration of oxytocin proved to the increase of gonadotrophin secretion from adenohypophysis and increase of the secretion of gonadal androgens followed to.
Acetylcholin seemed to act as the transmitor in the central nervous system and to stimulate the neurosectetory function of the hypothalamus, resulting in the increase of pituitary-gonadal function in the same manner as the injection of oxytocin.
(3) With the use of potentialized anesthesia, hyperfunction of the neurosecretory system in the anterior hypothalamus was suppressed even after the surgical stress. Excretion of pituitary gonadotrophin, urinary 17-ketosteroids and its gonadal fractions did not increase according to the potentialized anesthesia. Administration of oxytocin or acetylcholin made the gonadotrophin-gonadal system accelerate even under the potentialized anesthesia.
It was concluded that the potentialized anesthesia might depress the release of acetylcholin and suppress the secretion of oxytocin and pituitary-gonadal function.

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