For the purpose of studying the metabolic variation in the normal human endometrium during the menstrual cycle, Schmidt-Thannhauser method was applied for the determination of endometrial content of phosphorus, namely acid soluble phosphate (inorganic and organic), phospholipid phosphate and nucleic acid phosphate (RNA-P and DNA-P), during different phases of the menstrual cycle.
In order to investigate the RNA metabolism from the viewpoint of disintegration, RNase activity in the normal human endometrium was also investigated various days in the menstrual cycle by the Sam Brody method.
The results were as follows :
1) Acid soluble phosphate (inorganic and organic), phospholipid phosphate and RNA-P reached the peak at about the 13th to 14th day and the 23rd to 24th day in the menstrual cycle, respectively, each curve showing two peaks. On the other hand, DNAP reached the peak at about the 15th to 16th day in the menstrual cycle.
2) RNase activity appeared to increase markedly at about the 19th to 20th day and reached the peak at about the 23rd to 24th day in the menstrual cycle, Thereafter it decreased as menstruation approached.

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Measurement of human growth hormone (HGH) in plasma has much significance in the study of growth and development of children. Since the radioimmunoassay using ¹³¹I-tracer was introduced into the HGH determination in plasma by Hunter and Greenwood, it becomes easy to estimate the very Low level of HGH in plasma accurately. In radioimmunoassay of HGH are four different kinds of methods to separate the bound ¹³¹I-labeled HGH from free ¹³¹I-labeled HGH; that is, chromatoelectrophoresis, double antibody precipitation method, absorption method using ion exchange resin and salt-precipitation method using sodium sulfate. Among them, the double antibody precipitation method is thought to be the best one to determine the plasma HGH levels because of its simplicity, its good reproducibility of the results. It is the purpose of this paper to comment on the double antibody precipitation method and to elucidate the change of the plasma HGH levels in various ages using this method.
The results were as follows :
1. Labeling of HGH
HGH purified by Dr. M.S.Raben was iodinated with Na¹³¹I using the method of Hunter and Greenwood and ¹³¹I-labeled HGH of the high specific activity was obtained (about 150-200 mCi/mg). PH 8.6 was optimal PH of the reaction medium to obtain the best yield of ¹³¹I-HGH.
2. Double antibody precipitation method
Rabbit anti-HGH serum was obtained ten days after the last of three weekly subcutaneous injections of 5 mg human growth hormone (Raben) emulsified in complete Freund's adjuvant. Guinea pig anti-rabbit γ-globulin serum was used as the second antibody. All dilutions except for human plasma and standard solution of HGH were made with 0.1 % bovine serum albumin in a veronal buffer, 0.07M, PH 8.6.
The same HGH diluted with 0.5% bovine serum albumin in the same veronal buffer was used for the preparation of the standard curve according to the method of Schalch and Parker. The concentrations of standard solution were 0.03, 0.06, 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 and 23.0 mμg/ml. 0.1 ml of ¹³¹I-labeled HGH solution (0.2-0.6 mμg of ¹³¹I-HGH) was added to 0.5 ml of standard solution or diluted plasma (1 : 11). 0.25 ml of diluted anti-HGH serum was then added to each tube and the mixture was incubated at 4°C for seven days after the gently shaking the tube. At the end of the incubation period 0.1 ml of guinea pig anti-rabbit γ-globulin serum was added to the mixture and finally 0.1 ml of 1 : 20 normal rabbit serum was added as the carrier protein and incubated again at 4°C for 24 hours. The tubes were then centrifuged at 3.000 r.p.m. for ten minutes and the supernatants were discarded. The precipitates were counted by well type of scintilation counter and the percentage of ¹³¹I-labeled HGH precipitated was calculated.
The recovery of this percedure was 53% to 153% (mean 104%) and the reproducibility of the result was 62% to 166% (mean 104%).
1) The effect of the concentration of ¹³¹I-HGH solution
¹³¹I-HGH solutions of three different concentrations were used to prepare the standard curve. It was possible to determine the smaller amount of HGH in plasma in the lowest concentration of ¹³¹I-HGH.
2) The effect of the dilution of anti-HGH serum
While it was not possible to ditermine the smaller amount of HGH in plasma with the higher concentration of anti-HGH solution, it was not possible to determine the larger amount of HGH with the lower concentration of anti-HGH solution. The appropriate concentration was 40,000 fold to determine between 0.125 mug/ml to 16 mug/ml of HGH.
3) The effect of the incubation time
Five days were required to reach the equilibrium of the first step of antigeantibody reaction, especially in the lower concentration of HGH solution.
4) The effect of pH of the diluent

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It is generally known that vitamin B₆ plays an important role in protein and fat metabolism as a co-enzyme of various kinds of enzymes. On the other hand, a possible role of vitamin B₆ in carbohydrate metabolism has not yet been established, although some evidence has been found suggesting an intimate relationship between vitamin B₆ and diabetes recently.
The present investigations were carried out in order to compare the abnormality of carbohydrate metabolism in vitamin B₆-deficient albino rat with that in alloxan-diabetic rat from a view point of change of enzyme activity in their liver and muscle.
The following results were obtained.
1) Within two weeks of vitamin B₆-dificiency, albino rats showed a decrease of glucose tolerance, which has become more remarkable in the course of vitamin B₆-deficiency. However, glucose tolerance in the alloxan diabetic rats was disturbed more severely than that in the vitamin B₆-deficient rats.
2) The serum insulin-like activity of the vitamin B₆-deficient rats was high before and after glucose loading. On the other hand, their serum immuno-reactive insulin activity was not high but normal. Their insulin-like activity in the pancreas was also the same as in pair-fed controle rats. These facts suggest that the ability of insulin secretion of the pancreas Langerhan's islet remained normal in the vitamin B₆-deficient rats.
3) The vitamin B₆-deficient rats showed increased activities of the liver glucokinase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase and showed decreased activities of the liver phosphorylase and transaminase. There was no significant change in the activities of the liver glucose-6-phosphatase and fructose-1, 6-diphosphatase. The lower liver phosphorylase activity was in good accordance with the higher liver glycogen level in the vitamin B₆-deficient rats. There seems to be a difference between the change of the liver enzyme activity in vitamin B₆-deficiency and that in alloxan diabetes. These results support the hypothesis that there might be increased glycolysis, decreased glycogenolysis, and no increased gluconeogenesis in the liver of vitamin B₆-dificient rats.
4) The vitamin B₆-deficient rats showed decreased activities of the muscle hexokinase, phosphorylase and transaminase, and showed an increased activity of the muscle glucose6-phosphatase. It seems that glycolysis in the muscle of the vitamin B₆-deficient rats was depressed as well as in the alloxan diabetic rats. The lower serum lactic acid level in vitamin B₆-deficiency also supports this hypothesis. The experiment mentioned above clearly indicated that abnormal metabolism of carbohydrate with the decrease of glucose tolerance, in other words, diabetic state, occurred in the vitamin B₆-deficient rats. In vitamin B₆-deficiency, it is very characteristic and moreover, different from alloxan diabetes, that there existed increased glycolysis without increased gluconeogenesis in the liver. It is highly probable that increased glycolysis in the liver of the vitamin B₆-deficient rats occurred in order to prevent a disturbance of energy production as a consequence of the depressed metabolism of tricarboxylic acid cycle. In conclusion, abnormal metabolism of carbohydrate, that is to say, diabetic state in vitamin B₆-deficiency could not be ascribed to deficiency of insulin such as in alloxan diabetes, but could be ascribed to the primary disturbance of energy production in which vitamin B₆ played an important role as a co-enzyme.

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It is well known that in the living body, catecholamines are inactivated by MAO or by COMT to their final product VMA. It is generally considered that the circulating or the exogenous catecholamine is mainly metabolized by COMT, whereas the endogenous catecholamine is chiefly destroyed by MAO.
In the present experiments, dopamine, norepinephrine (including epinephrine), normetanephrine (including metanephrine) and dehydroxy-mandelic acid (DOMA) were estimated in the inflammatory gluteal muscles artificially produced in the normal and the adrenalectomized dogs and the influence of bilateral adrenalectomy on the catecholamine metabolism was studied.
In the normal dogs, the marked increase of dopamine, norepinephrine and normetanephrine were noted but not on DOMA. On the contrary, in the adrenalectomized dogs, DOMA as well as dopamine and norepinephrine showed a remarkable increase, whereas no increase in normetanephrine was noted.
In the adrenalectomized dogs which were given adrenal emulsion intramuscularly, the results were the same as in the normal dogs. Such results were not obtained with cortisone or aldosterone.
Histochemical examination of the MAO activity in the gluteal muscles under various conditions was also conducted. MAO activity increased immediately after adrenalectomy without any relation to the inflammation and the increase of MAO activity produced by adrenalectomy was normalized by the intramuscular injection of adrenal emulsion.

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The variable conditions of dextran-coated charcoal immunoassay of insulin originally presented by Herbert et al were investigated and compared with the two-antibody method of Morgan and Lazarow.
On Dextran-coated Carcoal Method.
1) Dextran-coated charcoal instantly adsorbed free insulin independently of its concentration in reaction mixtures but rejected antibody bound insulin.
2) The PH of 7.4 was most effective for to insulin-antibody reaction.
3) As duration of incubation increased, the amount of bound insulin increased. And the amount of bound insulin was more increased at 4°C for 1-4 days than at 37°C for 1-4 hours. The sensitivity of the standard curve was more increased at 4°C than at 37°C. But the time and the temperature did not influence the serum insulin values obtained with this method.
4) When serum insulin is assayed, the serum protein decreases adsorption of free insulin to dextran-coated charcoal, so correction with control and without AIS of radioactive count are necessary.
5) The reproducibility of this method as well as the recovery of human insulin added to serum was highly satisfactory. The dilution of serum had no effect on the insulin values obtained.
Comparison with the two-antibody method.
1) The standard curve of charcoal method had good agreement with that of the twoantibody method.
2) The approximated 94.7% average of the bound insulin in the supernatant of the charcoal method was precipitated by AGP-GS. And the averaged 95.3% of free insulin in the supernatant of the two-antibody method was adsorped to dextran-coated charcoal.
3) The serum insulin values obtained in the course of GTT showed a good over-all correlation between the charcoal and the two-antibody method. The shapes of the insulin curves in these tests were essentially similar in both methods. The values obtained with the charcoal method were a little lower than those with the two-antibody method, but the difference was not significant.

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