ヒト健常人並びに患者血清におけるLDHの温度換算係数が異なることは, LDHの乳酸生生成反応による測定法によってすでに報告したが, 今回は, 乳酸を基質とした場合の温度補正について検討した。ヒト臓器からのH₄及びM₄酵素標品間における温度換算係数の差異は, 患者血清, 健常人血清で最も大きく, この差異は, H及びMサブユニット含量に基づいていることが考えられた。
Arrheniusの式によるプロットの結果は, 先の報告2)と同じように2相性の直線関係が得られた。しかし屈曲点が37℃となった。また酵素の構造変化と耐熱性は測定pHによってかなり影響を受けることがわかった。
またNAD (450mM乳酸), TNXD (15mM乳酸) を使用してLDH活性を求めた場合が, H及びMサブユニット間におけるLDH活性の開きが最大となった。このNAD/TNXD比を用いた方法は従来の方法である電気泳動法によるLDHサブユニット含量比率とほぼ一致した。
さらにこのNADアナローグを用いたLDHサブユニット含量を温度補正に適用したところ, いずれの疾患血清においても精密なLDH活性値が得られることがわかった。

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1. 脂肪酸活性化酵素 (Acyl CoA Synthetase) 反応により脂肪酸を活性化し, 生成するAMPを紫外部吸収による酵素法により定量し, これより脂肪酸濃度を求める原理を考案した。
2. 本原理は, 血清脂肪酸の新しい測定法へのアプローチを与え, 臨床化学分野での応用に期待がもたれる。

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Studies are reported here of a colorimetric microdetermination of cholinesterase activity in serum. Acetate hydrolyzed from a substrate by cholinesterase, which was coupled with ATP, acetate kinase, phosphoenolpyruvate and pyruvate kinase, was stoichiometrically converted to pyruvate. This pyruvate produced was colored with 2, 4-dinitrophenylhydrazine in the alkaline solution. The activity of cholinesterase was linear until approximately 20 ul of serum. The relationship between this assay and a phenol red method was close (r=0.88). A normal value of the cholinesterase activity was 0.9±0.18 units by the present assay.

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Concentrations of adenine nucleotides in rat brain and liver have been determined spectrofluorometrically by the coupled enzymatic reactions and the value of energy charge ([ATP]+0.5[ADP]/[ATP]+[ADP]+[AMP]) has been calculatated. The value of energy charge of normal rat brain and liver was 0.89 and 0.85, respectively.
When rat was inhaled with 100% nitrogen gas, the value of energy charge of the brain decreased rapidly to 0.48 at 2 min after the inhalation. This was due to a rapid decrease in ATP concentration in the brain and a concomitant increase in AMP concentration. On the other hand, that of the liver was reduced slowly to 0.69 at 2 min under the same conditions.
The results indicate clearly a higher affinity of the brain tissue to oxygen than the liver.

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In the urine of subjects given an oral dose of spironolactone, several metabolites have been found. Metabolites which interfere with the estimation of 17-oxosteroids (17-KS) by the zimmermann reaction and 17-hydroxycorticosteroids (17-OHCS) by the Porter- Silber reaction were analyzed by thin-layer chromatography, and the interference of each metabolite with the estimations of 17-KS and 17-OHCS was investigated. In the estimation of 17-KS, the metabolite of spironolactone had a maximum absorption at 610nm. The values obtained by the formula of Fraser give positive errors, but the values are corrected by using the reference absorption. In the measurement of 17-OHCS, since the interference is caused by the reaction of the metabolites with sulfuric acid, the value is corrected by subtracting the blank value.

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The galactose excretions in urine during oral galactose tolerance test were examined in cases of 17 normal, 31 diabetes and 32 liver diseases.
Galactose excretions in urine for 2 hours in diabetes and various liver diseases were remarkable as compared with normal subjects, especially liver cirrhosis showed the most conspicuous excretion of galactose in urine.
There was no significant difference among normal, diabetes and various liver disease, in excreating amounts of urine for 2 hours. The changes of urinary protein, ketone bodies, urobilinogen and glucose during oral galactose tolerance test were semi-quantitatively examined. Protein test was unchanged and ketone bodies test turned to positive only in each 1 case in diabetes and liver diseases. Urobilinogen test generally turned to negative after galactose loading except 2 cases in liver diseases, it should be considered to depend on dilution of urine concentration after loading. Glucose test in urine after loading turned to positive in 7 cases out of 17 cases in normal subjects. This positive grade was higher percentage in diabetes and liver diseases than in normal. On the urinary excreting amount of glucose and galactose, slightly positive correlation was observed in urine after galactose loading.

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Oral galactose tolerance test was performed on the cases of 17 normal subjects, 28 diabetes and 32 liver diseases, and then blood glucose and galactose levels were examined on the positive and negative group in urinary glucose test after loading. The change of blood glucose level in normal cases during the galactose tolerance test was recognized no significant difference between negative and positive group of urinary glucose reaction. The similar tendencies were observed in the cases of diabetes and liver diseases. It was found that urinary glucose excretion during galactose tolerance test was not concerned to elevation of blood glucose level.
Fositive group on urinary glucose during galactose loading showed more significant increasing of blood galactose level as compared with negative group. Especially in absorption period, 30 and 60 minutes after loading, it was remarkable in all cases. Therefore it was showed that glucose excretion in urine during galactose tolerance test had some relations for the elevation of blood galactose level.
The relationship between the increasing of blood glucose-Fgalactose levels and glucose excretion in urine during galactose tolerance test was significantly recognized in 30 and 60 minutes after loading on the cases of normal and diabetes.
Glucose excretion in urine during galactose tolerance test, was mainly affected by the elevation of blood galactose level, but the elevation of glucose + galactose levels had also some participations.

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In order to avoid the interference of aldoses and ketoses in gas chromatographic analysis of sugar-alcohols in urine, a sample preparation was developed using a Dowex 1×8 (OH-) column.
Urinary excretion rate of sugar-alcohols in man was measured by the method after fasting and taking glucose, glucuronolactone or Sake. Whereas the excretion of xylitol was incresed remarkably after administration of glucuronolactone, a slight increase of sugar-alcohols except xylitol was seen, but not in all subjects. The influence of taking Sake was observed on the excretion of erythritol, arabinitol, xylitol and mannitol.

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