one component of molecular weight 30,000 in the supernatant on 65% ammonium sulfate fractionation of the hemolysate was isolated by the method of Sephadex G-75 and CM-cellulose column chromatography, and electrofocusing.
The physicochemical properties of this protein were as follows; molecular weight 30,000±3,000, isoelectric point pH 6.0. Amino acid analysis was performed. lmmunochemical reaction with arlti human carbonic anhydrase B serum was observed. From the above results, this protein was estimated as carbonic anhydrase A.

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A simple, rapid and specific procedure is described for the routine determination of urea nitrogen in serum with an acidified solution of p-dimethylaminocinnamaldehyde in dimethyl sulfoxide. This procedure is the most useful for theemergency testirlg.

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The authors describe a simplified and rapid method for determining L-cystineaminopeptidase ( CAP) in serum with the aid of S-benzyl-L-cysteine-p-nitroanilide (BCN). The liberated p-nitroaniline reacts with p-dimethylaminocinnamaldehyde to give ared dye. The method requires 15min incubation at 37°, followed by 10min colour development.
The proposed method is more sensitive than that with using L-cystine-di-P-nitroaniiinde (CDNl) and using L-cystine-di-naphthylamide (Wada's method). The correlation coefficient between results of this method and those obtained with L-cystine-di-naphthylamide (CDNA) as substrate was 0.97.

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Optimum reaction conditions at 37° for spectrophotometric assay of activity (from lactate to pyruvate) of human LDH isoenzymes were determined with respect to buffer concentration, NAD concentration, substrate concentration and pH.
Human LDH isoenzymes used were separated by column chromatography.
Optimum concentration of Glycine-NaOH buffer were within the range of 0.1M to 0.2M for LDH₁ and LDH₅ isoenzymes. An optimum activity was obtained at a NAD concentration of 4.5×10^-3M. optimum concentration of DL-lactate was 0.25M for total LDH, since it was 0.1M for LDH₁and0.5M for LDH₅.
Activities of LDH₁and LDH₅were enhanced with the increase of pH, when the activities were compared between of 9.0-10.8.
Although Fendley reported that the LDH5 isoenzyme was unable to measure when the assay was performed of pH 10.0, we have found that the LDH₅ isoenzyme activity can measure when a LDH·NAD complex is formed by a preincubation for 5min. and the reaction is started by a substrate buffer.

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The glucose-forming amylase in human urine was assayed simply and accurately without interference of α-amylase, proteinous materials, and reducing substances by the method based on the following principles: A small portion of dialyzed urine was incubated with maltotriitol as the substrate, the enzyme reaction was stopped by adding acid, and the glucose liberated was determined by the Park and Johnson method after neutralizing and deproteinizing the incubation mixture by the centrifugal gel filtration method.
The present paper describes the assay method definitely, presenting a number of experimental data obtained for urine from many healthy Japanese male adults. Also, some discussions are made about the reason of employing maltotriitol as the substrate and Park and Johnson method as the glucose determination method.

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ln vitro cholesterol syntheses from acetate and mevalonate were not observed in the reaction system with a single liver subcellular fraction, mitochondria, microsomesor cytosol, but in the system with microsomes and cytosol. The factor of cytosol is considered to be carrier proteins and an amount of them may be essential for the overall reaction of cholesterol synthesis.
The significance for cholesterolgenesis was studied using groups of old rats and cholesterol fed rats, whose cholesterol synthesis is decreased, and groups of partial hepatectomy, glucose refed rats and growing rats, whose cholesterol synthesis is increased.
The rate of cholesterol synthesis in hepatic microsomes from cholesterol fed rats and old rats increased in combained incubation with hepatic cytosol from normal or growing rats, while those in the microsomes of normal rats and growing rats decreased with the combination of cytosol from cholesterol fed or old rats. These effects were not brought about by the addition of cholesterol or other inhibitor in cytosol. Therefore, they may be caused either by qualitative or quantitative changes of carrier proteins.
An elevated cholesterol synthesis was observed in those groups of glucose refed rats and partial hepatectomized rats, but the rate of cholesterol synthesis in microsomes of each control was not interfered with cytosol of these treated rats, and the rate in microsomes of the treated rats was not interfered with cytosol of each control. These evidence suggest that the factor of cytosol may be have no relation with the increase of cholesterol synthesis in the groups of partial hepatectomized or glucose refed rats and that cytosol from these control groups contains enough amount of carrier proteins.

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acetate membrane as supporting medium and Blue-Starch® as staining material was carried out. This technique could be applied as a routine laboratory procedure because of its simpilcity and reproducibility.
The pattern of migration of isoenzymes in the this method is in good agreement with that of classical method based on agar gel separation. Futhermore, in some particular instances, the separation of isoenzymes with the this method is better than that with agar gel.

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Recently, various types of data processing systems for analytical clinical chemistry, especially for multichannel automatic analyzers, have been developed. However, relatively few of these systems include the function of process control and data correction.
We have devised a computer-controlled and data processing system connected with the SMA12/60 Auto Analyzer. The features of this system are described as having the adoption of: (1) a sample identification conception utilizing a commercial ID reader,(2) a real time correction of on-line data, based on our previous experience on the analyzer, for drift and sensitivity, carryover between specimens, and bilirubin interference for cholesterol value determinded by the Liebermann-Burchard reaction,(3) a feedback control and morlitor system of the multi-channel analyzer when the differences between the values of standards and monitoring samples are outside of the preset tolerance limits.
Here, we are reporting the principal instrumentation and software of this system and over3year experience on the system in our clinical chemistry laboratory.

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Determination of urinary pregnanetriol using 3α-hydroxysteroid-dehydrogenase was described. It is based on the specific enzymatic convertion of 3α-hydroxy group on the steroid molecule into 3-keto group. Quantitation is obtained by measurment of the absorption at340nm from NADH, following a hydrolysis by β-glucuronidase, extraction with methylenechloride and purification by thin-layer chromatography. The main advantage of the method rests in its specificity of 3α-hydroxysteroid-dehydrogenase in comparison with the reaction of sulfuric acid.

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