A specific and sensitive method for the simultaneous determination of β-phenylethylamine, tyramine and tryptamine, in their non-conjugated forms in human urine was described.
These amines were extracted with ethyl acetate from 20ml of urine at pH 12.0 and dansylated overnight in the dark place. Dansyl derivatives were separated by the two dimentional thin layer chromatography. Dansyl-β-phenylethylamine, dansyl-tyramine and dansyl-tryptamine were removed from the plate under the ultraviolet light and analyzed by the fluorometric procedure.
The authors determined urinary excretion levels of β-phenylethylamine, tyramine, tryptamine in healthy male and female subjects by this method. Urinary excretion levels of β-phenylethylamine were 11.4 μg/24 hours in male and 13.6 μg/24 hours in female. Tyramine excretion levels were 543 μg/24 hours in male and 621μg/24 hours in female. Tryptamine excretion levels were 69.7 μg/24 hours in male and 121 μg/24 hours in female.
The effect of bitter-sweet chocolate ingestion (85-140g) on the urinary excretion of these amines was also tested, since the chocolate contains a large amount of ft-phenyl-ethylamine (500μg PEA·HCl/100g). The chocolate ingestion caused no remarkable changes in urinary contents of β-phenylethylamine, tyramine and tryptamine in twenty four hours.

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Cholesterol synthesis from acetate and cholesterol uptake from the incubation medium by cells isolated from human primary hepatoma were investigated in vitro, compared with those from the transplanted hepatoma cells of rat or normal human and rat hepatocytes.
The cholesterol synthesis of primary hepatoma cells was as active as normal hepatocytes and was suppresed by the addition of serum or cholesterol-phospholipid dispersion to the incubation mediumsimilarly in the case in normal human and rat liver.
The human hepatoma cells took up cholesterol from serum as well as cholesterolphospholipid dispersion in vitro as do normal hepatocytes. On the contrary the transplanted hepatoma of rats did not take up cholesterol and the cholesterol synthesis was not affected by the addition of serum or cholesterol-phospholipid dispersion.

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Serum cholestanol concentrations in health and various diseases were determined with mass fragmentography.
The range of serum cholestanol in 18 normal subjects was 0.22-0.58mg/dl.
Hypercholestanolemia without hypercholesterolemia were observed in all 5 cases of cerebrotendinous xanthomatosis.
This paper reports also the remarkable elevation of serum cholestanol in the patients with obstructive jaundice or cholestatic hepatitis.
Other diseases with hypercholesterolemia such as nephrotic syndrome showed slight elevation of serum cholestanol, but the ratio of cholestanol to cholesterol was similar to normal control.
Cholesterol oxidases from various origins reacts. also upon cholestanol. However the amount of serum cholestanol was small enough as compared with that of cholesterol, so that the positive error originated from cholestanol in the enzymatic determination of serum cholesterol is usually considered negligible except the cases of cerebrotendinous xanthomatosis and the obstructive liver diseases

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Recently, drug administration to patients has been complicated both qualitatively and quantitatively, and many hazards owing to drug interactions and individual differences in drug metabolism have been reported. Therefore, accurate and full information on drug metabolism should be accumulated for each patient. Effect of concurrent drug administration on aminopyrine metabolism in patients with chronic diseases (pulmonary tuberculosis etc.) was studied. Rate of aminopyrine demethylation was depressed in the patients. The depression was hardly affected by suspending drug administration for 24 hours. This indicates that the diminished capacity may be due to repression rather than to inhibition of the drug-metabolizing enzymes. However, rate of acetylation of aminopyrine metabolites was not depressed in patients. In volunteers of low acetylation, administration of pantethine increased acetylation of aminoantipyrine and concurrent administration of isonicotinic acid hydrazide and aminopyrine decreased the acetylation. Taking alcohol beverages inhibited production of 4-formylaminoantipyrine, one of the metabolites of aminopyrine.

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The authors studied to find optimal conditions for determining Km values of human serum LDH (L-lactate: NAD⁺ oxidoreductase, EC 1.1.1.27) for lactate in order to screen “abnormal” LDH. LDH activity was measured at 25°C in four buffer systems.
1) The activity and its pH dependence differed slightly but clearly depending on the buffers used: glycine, pyrophosphate, Veronal, and Tris systems.
2) The pH-activity curves were essentially different when 1mM and 50 mM lactate were used.
3) LDH showed two different Km values for NAD⁺ in two ranges of its concentration: 79×10^-6M (Veronal) and 44-57×10^-6M (others) in 0.05-0.25mM, and 133×10^-6M (Veronal) and 95-105×10^-6M (others) in 0.25-3.5 mM.
4) Two values of Km were found also for lactate with a critical concentration around 2 mM: 1.2mM (Veronal) and 0.5mM (others) in the lower concentrations, and approximately 2mM (Veronal and Tris) and 1mM (others) in the higher ones.
5) The sera from patients, which had predominantly H or M type isoenzyme showed also two Km values with a critical concentration similar to that observed in sera with normal isoenzyme patterns. Therefore, this phenomenon may reflect a particular intrinsic property of the tetrameric enzyme: for example, subunit-subunit interaction.

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The authors deviced a rapid microdetermination carbon monoxide (CO) in blood using polarographic oxygen (O₂) electrode and light irradiation apparatus. This method consists of following procedures; (1) 15μl of the CO-biood was aded into 1 ml of O₂-depleted water (O₂: 4-5 μg/ml) in the closed glass cell fixed O₂ electrode. By this step, deoxyhemoglobin (HHb) in the CO-blood was saturated with dissolved O₂ in the O₂-depleted water to become oxyhemoglobin (HbO₂). The amount (a) of oxygen corresponding to [HbO₂ (a') formed from HHb and HbO₂ (a') already existed] in the sample was recorded by addition of 10 μl of 10% K₃Fe (CN)₆.(2) The same CO-blood diluted with water was bubbled by O₂ gas under light irradiation. By this procedure, HHb and HbCO in the CO blood were completely changed into HbO₂ The quantity (b) of O₂ combined with total Hb was determined by addition of 10 μl of 10% K₃Fe (CN)₆, successively. The value, [(b-a)/b] ×100, indicates the per cent of HbCO on the total Hb of the CO-blood tested. This method requires only 35 μl of the CO-blood and 6-7 min for each determination. Results showed excellent agreement with those obtained by spectrophotometry (r=0.993, y=0.95× ±2.14). Within-run precision was measured from 30 blood samples containing 50.0% HbCO, and the standard deviation and coefficient of variation were ±1.3% HbCO and 2.6%, respectively.

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A new method for determination of carbon monoxide (CO) in blood by using polarographic oxygen (O₂) electrode and erythrocytes was deviced. This method consists of following procedures;
(1) 15 μl of CO-blood was added into 1 ml of O₂-depleted water (O₂ 4 μg/ml) in the closed glass cell fixed with O₂ electrode. Deoxyhemoglobin (HHb) in CO-blood was saturated with O₂ dissolved in the O₂-depleted water and converted to oxyhemoglobin (HbO₂).
(2) By addition of 10 μl of 10% K₃Fe(CN)₆ solution, HbO₂ and carboxyhemoglobin (HbCO) in the reaction mixture were changed to methemoglobin, and O₂ from HbO₂ and CO from HbCO were released into reaction mixture.
(3) Then, 30 μl of 30% NaCI solution was added to become isotonicity, and 10 μl of suspension of erythrocytes was added. The released CO from HbCO was entered into erythrocytes, and exchanged with O₂ of HbO₂ in erythrocytes. Equimolar O₂ to CO was released to the reaction mixture. From the increase of O₂ in the reaction mixture, the amount of CO in CO-blood was calculated. This method requirs only 15 μl of CO-blood and 4 min for each determination.

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The principle of this simple and rapid method for estimating blood ammonia concentration is the micronization and modification of Conway's method.1) A plastic plate (3×6×0.3cm) which has 5 holes (6mm diameter) is used. The top of each hole is covered with polypropylene film and pH indicator paper (BCG), and the bottom is sealed with metalic foil. 20 μl of a fresh, whole blood specimen is applied into a hole and a buffer tablet (pH 10.3) is added, and the hole is immediately sealed with the foil. Ammonia gas vaporized from blood permiates the film and changes the color of the BCG. After a 15 min. reaction period this color change is compared with the standard color chart and blood ammonia can be determined. Intrassay reproducibility of the method is observed as follows: n=14,x=160 μM, C. V.=8.2%. n=14,x=44 μM, C. V.=13.1%. The correlation between this method and the enzymatic method using GLDH is obtained as follows: n=139, r=0.86, y=0.92×+5.9. Alkaline labile amines have almost no influence on this method. These results show that this method has various clinical uses.

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Interference of Cyproterone Acetate which is an alkyl substituted steroid used as a drug for anti-androgen was observed on urinary 17-ketosteroids assay. This drug used for patients, sometimes for child, with abnormal high androgen activity is metabolized to 17-keto derivatives and interfer on the 17-ketosteroids assay. Color by Zimmermann reaction from patients urine was orange-brown. and it is impossible to correct the value so that it is prefered to stop the administration of the drug before the test.

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The abnormal fraction of lactic dehydrogenase (LDH) isoenzymograms in the sera of patients with acute hepatitis and/or cardiac damage is reported. Investigation showed that, adjacent to a normal LDH₅ (M₄) fraction, the abnormal fraction has apparent LDH activity.
According to its properties, the so-called extra LDH fraction seems to be alcohol dehydrogenase (ADH).

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