Serum protein, lipoprotein and glycoprotein were analysed by paperelectrophoresis in 12 normal adult rabbits (8 female and 4 male) weighing 2 to 3kg.
Total cholesterol levels in blood were also measured by a modification of Bloor's method using the photoelectric colorimeter ; blood sugar levels by Somogyi's micromethod and total serum protein values by Pulfrich's refractometer. The results were as follows.
1) Average values of serum protein fractions were : albumin 53.5±2.6%, α₁-globulin 9.4±1.3%, α₂-globulin 6.1 ±0.4%, β-globulin 13.2±1.4%, γ-globulin 17.8±1.8%. The relative ratios of migration were : albumin 100, α₁-globulin 73.5±1.2, α₂-globulin 56.0±1.4, β-globulin 41.3±1.4, γ-globulin O. Lipoprotein fractions were : α-lipoprotein 34.5 ± 3.7%, β-lipoprotein 65.5 ± 3.7%, and the lipoprotein index was 1.99±0.84. Glycoprotein fractions were : albumin bound glycoprotein 20.6±3.9%, α₁-globulin bound glycoprotein 17.2 ± 4.9%, α₂-globulin bound glycoprotein 15.3 ± 2.0%, β-globulin bound glycoprotein 24.4±5.9%, γ-globulin bound glycoprotein 22.5±3.5%. Total blood cholesterol levels were 87 ±8.7 mg/dl, blood sugar levels 101±5.3mg/dl and tolal protein values 6.53±0.53g/dl.
2) The reproducibility of protein, lipoprotein and glycoprotein fractions from the same samples was good.
3) In the measurement of total cholesterol, it seems to be important to be careful of the temperature and time after the Liebermann-Burchard reaction is carried out.
Part II. Serum Protein, Lipoprotein, Glycoprotein, Total Blood Cholesterol and Blood Sugar Levels in Offspring of Rabbits (F₁, F₂, F₅, and Spontaneously Occurring Diabetic Rabbits) Descended from Rabbits with Induced Diabetes over Successive Generations, and in Rabbits Made Diabetic with Alloxan.
Serum protein, lipoprotein, glycoprotein, total blood cholesterol levels, blood sugar levels and total serum protein values were studied in the offspring of rabbits (7 cases of F₁, 6 cases of F₂, 10 cases of F₅ which is before the spontaneous occurrence of diabetes) descended from rabbits made diabetic with alloxan or 5- (p-Hydroxyphenylazo) -8-hydroxyquinoline over successive generations. The same studies were also made on 2 rabbits directly after the spontaneous occurrence of diabetes, on 2 young ones born to F₅ rabbits which continued to have spontaneous diabetes for a long time, and on 10 rabbits made diabetic with alloxan. The results are as follows.
1) As compared with normal values, the serum protein fractions in F₁, F₂, F₅ (before the spontaneous occurrence of diabetes) rabbits revealed an increase in α₂-globulin fraction and a decrease in γ-globulin fraction. The lipoprotein index was increased. Glycoprotein fractions showed an increase in α₂-globulin bound glycoprotein and a decrease in γ-globulin bound glycoprotein. These characteristics seemed more and more significant as the generations progressed, that is, from F₁ to F₂ to F₅. Total blood cholesterol and blood sugar levels tended to increase slightly. Total serum protein levels remained normal.
2) There was no significant difference between the values in rabbits one month before and directly after the spontaneous occurrence of diabetes, except for an increase in blood sugar levels and a slight increase in the lipoprotein index, total cholesterol levels and α₂-globulin bound glycoprotein.
3) In rabbits in which spontaeneous diabetes had continued for a long time,

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Alloxan diabetes was induced by means of alloxan administration in the offspring of rabbits (F₁-F₅, 2-3 cases of each generation) descended from rabbits with induced diabetes (persisted for more than one month) over successive generations. Afetr about 100 days, serum protein, lipoprotein, glycoprotein were analysed by paperelectrophoresis. Total blood cholesterol levels, blood sugar levels and total protein values were measured also. Compared with normal values (12 cases), the results were as follows.
1) Serum protein fractions showed a marked decrease in albumin and an increase in α₂-globulin and γ-globulin. The degree of increase in α₂-globulin became more and more significant as the generations progressed. The lipoprotein index increased in each generation. Glycoprotein fractions showed changes analogous to those of the protein fractions.
2) Total cholesterol levels were slightly increased in F₄ and F₅. Blood sugar levels were markedly increased in every generation. Total serum protein values were normal.
3) It is concluded that the serum protein, lipoprotein and glycoprotein patterns in these rabbits showed changes before alloxan administration, and presented moreover changes seen in alloxan diabetic rabbits.
4) These alterations are similar to those in a few cases of human diabetes.

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This work, the insulin sensibility of the pregnant, has been performed as a part of the co-operative investigation in our department concerning the carbohydrate metabolism in the pregnant woman.
In accordance with the modified Radoslav's method the blood was taken every 10 minutes after a single injection of 5 units of insulin intravenously eary in the morning on an empty stomach.
The blood sugar was estimated by the Somogyi's Anthron method. The insulin sensibility coefficient of the pregnant was proved to be lowered, as compared with the non-pregnant, moreover declining much more as the month of pregnancy proceeds. Regarding to the rate of covery it was almost the same in both groups.
Next, the I/G ratio in the modified Himsworth's method, namely the indicator of insulin sensibility, was proved to decline month after month in pregnacy.
The inorganic phosphate in blood was estimated by the Nakamura's method after the intravenous administration of insulin according to the modified Radoslav's method. The decreasing rate of the inorganic phosphate in blood caused by the insulin administration was proved to get lowered in the pregnant comparing with that of the non-pregnant. This may indicate that the vicissitudes of the inorganic phosphate in blood run almost pararell with that of blood sugar.
In conclusion, the results obtained above may suggest that the insulin tolerance must increase in the pregnant, and it could be inferred, as the interpretation of this cause that the fluctuations of the functions of the pituitary-adrenocortical system would have something to do with it.

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In the experimental studies of drug-sensitization, marked differences were often observed in the intensity of skin reaction to the test on the sensitized animals, all of which recieved the same treatment. As for the differences, even among homologous animals, it has been vaguely expressed so far that the differences possibly depend on the variety of individual characters. Recently many reports have been accumulated about the relationship between corticotrophin or adrenal hormones and allergic reactions. The authors too have researched the relation of adrenal cortex to the intensity of allergic skin reactions in guinea pigs sensitized with 2 : 4-dinitrochlorobenzene, and in this paper we are reporting the morphological studies on adrenal cortex of normal and sensitized guinea pigs, some of which were exposed to a stressor, taking out a large amount of lymph from the thoracic duct.
The amount of lipids in adrenal cortex, decreased in general under the stresses such as sensitization and flowing out lymph through the thoracic duct, and this change was most marked at the middle layer of zone fasciculata. Among the lipids the positive substances by Ashbel-Seligman reaction decreased most prominently. As for free cholesterine, fatty acid, congestion and bleeding in adrenal cortex and the ratio of width of zona fasciculata to that of zona glomerulosa, there was seen a series of changes which were ordinarily observed under the stress.
Comparing the histochemical findings of adrenal cortex of various groups with each other, lipids of the adrenal cortex in a weakly sensitized group showed lesser in amount and more minute in size than that in a strongly sensitized group. These differences between both groups became more marked when exposed to another additional stress, e.i. their lymph were drained out, and especially so in the changes of the positive substances to Ashbel-Seligman reaction.
From these histological findings, and by presuming the adrenal function it can be said that weakly sensitized animals show more marked hyperfunction of adrenal cortex than strongly sensitized ones. Consequently the activity of adrenal cortex seems to have intimate relationship with the intensity of allergic reaction.

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Glu agon is known to cause a transient hyperglycemia by glycogenolysis in the liver. Liver glycogen immediately decreases after glucagon administration. On the contrary, some investigators has recently reported that the liver glycogen increased after more than 24 hours following injection of over medium doses of glucagon. On the other hand, a few studies have been published indicating that a negative balance of nitrogen occured by the administration of a large dose of glucagon. That is, the metabolic action of glucagon is similar to that of glucocorticoid. Therefore, it is reasonable to assume that glucagon causes a certain effect on gluconeogenesis. From the above mentioned stand-point, the mechanism of the delayed liver glycogen increase by glucagon was studied as follows.
1. Liver glycogen production due to intraperitoneal injection of 2g/kg. of alanine to 24 hours fa sting rats definitely increased by the pre-treatment of 300γ/kg. of glucagon (intraperitoneal injection). This fact indicates that glucagon accelerates gluconeogenesis from alanine.
2. The general pattern of metabolism in the acceleating condition of gluconeogenesis was investigated. In this case alloxan diabetic and hydrocortisone given animals were adopted as favourable materials for investigating. At first, alanine (200μM) was added as a substrate to the liver slices of these treated rats. After 1 hour's incubation at 37°C in Warburg flask, the formation of glucose, pyruvate or glycogen from alanine in vitro was found to be increased in the liver slices of treated rats fasted for 24 hours. Secondly, the liver transaminase (glutamic pyruvic- and glutamic oxaloacetic transaminase) was confirmed to be increased 3 to 7 fold, especially a pronounced rise of activity was noted in glutamic-pyruvic transaminase.
3. The experiments were performed in the same may with glucagon and the same trend was confirmed. That is, the delayed increase of liver glycogen is concerned with the enhancement of gluconeogenesis, which is due to a rise of activity in liver transaminase.
4. In order to elucidate whether such an accelerating effect of gluconeogenesis by glucagon depends on the adrenal cortex, 20γ/kg. body weight of glucagon was intravenously administered to human adults, and the urinary excretion of 17-OHCS were measured. Of total 8 cases, increased excretion of 17-OHCS were recognized in 6 cases. On the other hand, in adrenalectomized rats, daily injection of glucagon for 3 days did not promote the liver transaminase activity.
Accordingly, it is concluded that glucagon causes the delayed liver glycogen increase mainly by way of the adrenal cortex.

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The effects of several hormones-thyroxine, adrenalin, cortisol, adrenocorticotrophin and thyro-trophin-upon the glucose uptake by the isolated hemidiaphragm were investigated in vitro and the author found that the influences of these hormones in human plasma upon the glucose uptake diminish extremely or may be eliminated practically by the dilution of plasma.
Using this method, the author estimated plasma insulin activities in normal subjects and some endocrine disorders. The following results obtained :
1) The mean of fasting plasma insulin activities in 22 normal subjects was (3.16±0.45) × 10^-4u. per ml.
2) The fasting plasma insulin activities in diabetics were lower than in normal subjects and the activities in the insulin-requiring diabetics were lower than those in obese diabetics. In uncontrolled and insulin resistant diabetics, no insulin activity was found.
3) In hyperthyroids, the activities were reduced, while in hypothyroids, they were within normal limits.
4) In patients with Cushing's syndrome, hyperadrenocorticism, acromegaly, steroid diabetes and Fröhlich's syndrome, the activities were generally reduced.
5) In patients with Addison's disease and anorexia nervosa the activities were somewhat elevated or within normal limits.
6) When 0.1 unit of crystalline insulin per kg. body weight was injected intravenously into a normal fasting subject, the maximum of insulin activities was found in the plasma taken at 20 minutes after the injection.
7) Plasma insulin activities in all cases of normal subjects increased markedly at 1 to 3 hours after the oral administration of 100g. of glucose.
8) The author found that there were two types in diabetics. In the first type, plasma insulin activities increased, while the activities in the second type decreased after the administration of 100g. of glucose. Moreover, in some other endocrine disorders, the authors observed changes in blood sugars and insulin activities after the administration of 100g. of glucose, and the clinical significances of those changes were discussed.

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Many evidences have been achieved since Gomori and Goldner had succeeded to generate alloxan diabetes. In these evidences it was observed that Ca and Mg in serum related closely to carbohydrate metabolism, especially to phosphous to metabolism, and from the fact that Ca in diabetic serum is observed as lower and Mg as higher, it is considerable that the determination of Ca and Mg in serum may be an index to infer the state of abnormal carbohydrate metabolism.
On the ground of these evidences, I have investigated the variation of Ca and Mg in serum, and considered the influences on the alloxan diabetes at dysfunction of thyroid and adrenal gland.
Examined animals were divided into 6 groups i.e., normal, thyroidectomized, thyradin injected, adrenalectomized, cortisone, injected, and cortisone injected and alloxanized group.
Each group was injected with alloxan 200mg per kg of body weight, besides cortisone was injected i.m. 5mg per kg of body weight during 5 days, thyradin was injected i.m. 0.1mg during 5 days.
From each group blood was picked up after allowan injection 2 hours, 5 hours, 8 hours, 1 days, 3 days, and 5 days, i.e., these pickup times are correspond to initial hyperglycemia stage, hypoglycemia stage, and durable hyperglycemia stage, then blood sugar and Ca and Mg in serum are determined about these pickuped blood.
The results are summarized as follows.
1) In normal group, blood sugar curve shows the typical 3 phases, after 5 hours Ca and sugar in blood has mutually negative correlation and Mg has positive correlation with blood sugar.
2) In adreualectomized group, one does not recognize the first phase. Ca and Mg in serum only diminish.
3) In cortisone injected group, the first phase is prominent, and the development of hyperglycemia is easy, Ca and Mg alter like normal group, but the degree of alteration is more than that of normal group.
4) In cortisone injected and alloxanized group, hypoglycemia is slight, decrease of serum Ca appeares in the early stage and the degree of the decrease is larger. Mg in serum increase temporaly at hypoglycemia stage.
5) In thyradin injected group, hypoglycemia is severe and Ca in serum is similiar as that of the cortisone injected and alloxanized group. Mg in serum decreasat first and the 2nd phase then recovered.
6) In throidectmized group, the change of blood sugar is slight and hyperglycemia can not be observed, decrease of serum Ca is slight and after 3 days it recovers. The alteration of Mg goes parallel with blood sugar.
7) Except thyroidectmized group, Ca in serum decreases, and Mg in serum increases.
8) Hormons of thyroid gland and adrenal gland promot the development alloxan diabetes.

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