We found age dependence of amylase activity and amylase isoenzyme pattern in serum of neonate and infancy. Total serum amylase activity was measured by a chromogenic method and serum amylase isoenzyme was determined by electrophoresis on Titan III-Lipo membrane. Amylase activity in cord serum of neonate at birth was on remarkably low level, so that cord serum was concentrated with Liphogel before isoenzy meanalysis. Serum amylase activity was on low level under 4 months of age after delivery, increased with age after that and reached the normal adult level by approximately 10 years of age. A little Amylase-1 of pancreatic isoennzyme was detected in cord serum at birth and age-depenpent increase of Amylase-1 was observed in serum of neonate and infancy after delivery. Movements after delivery of pancreatic and salivary type amylase are different respectively, which show that function of external secretion in pancreas develops and that of salivary gland also develops, and we consider the movements are phenomena of physiological adaptation in growing neonate and infancy.

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The liver microsomal lipid peroxidation consists of a NADPH-dependent, enzymic reaction and an ascorbate-dependent, non-enzymic reaction, but each needs an extremely small quantity of iron. The authors investigated the influence of ferritin, that plays a role in iron storage within the body, on the lipid peroxidation.
Holoferritin promoted areaction approximately 1.5 times for NADPH-dependent reaction and approximately 1.8 times for ascorbate-dependent reaction. Likewise, the promotion effect of the catalyzation was exerted by heat denatured holoferritin. On the other hand, the addition of 1.5 mg of apoferritin to the reaction mixture inhibited the NADPH-dependent reaction by approximately 80% and the ascorbate-dependent reaction by approximately 60%. However, the inhibition by apoferritin disappeared when ferric ions were added to the reaction mixture. Heat denatured apoferritin inhibited the lipid peroxidation as did native apoferritin. It is suggested that these results are due to the fact that ferric ion binds with apoferritin.

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A Statistical distribution model (dislocated lognormal distribution). which is convertible to normal distribution after transformation of the variable by log (x-a), is adopted as a model for normal range calculation. The distribution parameters are calculated from sample statistics (mean, standard deviation and skewness). The distribution conform well to non-normal and non-lognormal distributions. Calculated normal ranges are makedely affected by a few cases of abnormal data, and the method is only applicable to pure normal samples.

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A specific determination method of lipid hydroperoxides by the enzymatic reaction using a peroxygenase from pea seads is proposed.
The standard calibration curve showed high linearity in both calorimetry and fluorometry. A linearity was proven up to 3μ moles of linoleic hydroperoxide in the presence of 6p moles of in dole in the calorimetry. Since the sensitivity of the fluorometry using N-methyl in dole was better than that of the calorimetry, the former might be favorable for the clinical application.
The minimum detecting sensitivity was 50 n moles/ml as linoleic hydroperoxide in the fluorometry.
The variaton coefficient ranged within±3.02% and the average percentage recovery was as 101.9% in the fluororometric determination.

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L-Leucylglycylglycine hydrolyzing aminopeptidase was electrophoretically shown to be present in human serum using the leucine dehydrogenase coupled staining technique. Before investigating the clinical significances of this enzyme, it was partially purified from normal human plasma by ammonium sulfate fractionation, column chromatography on DEAEcellulose, gel filtration on a Sephadex G-100, and preparative isoelectric focusing.
The molecular weight of the enzyme was estimated to be 56,000 by gel filtration on a Sephadex G-75, and the isoelectric point of the enzyme was pH 5.01 by isoelectric focusing. This enzyme specifically hydrolyzes tripeptides, such as L-leucylglycine and L-valylglycine, at am inoterminals of peptides. Moreover, the enzyme does not act on dipeptides and amino acid amides. The enzyme is slightly inhibited by EDTA, EGTA and HEDTA, and strongly by o-phenanthroline. Bestatin, di-L-leucine and tri-L-leucine competitively inhibit the enzyme, and the inhibition constants were estimated to be 0.18×10^-6, for bestatin, 70×10^-6, for di-Lleucine, and 6.0×10^-6 mole/liter for tri-L-leucine. These physical and kinetic properties closely resemble those of aminotripeptidase (Tripeptide aminopetidase, EC 3.4.11.4) obtained from monkey brain and swine kidney.

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Hereditary xanthinuria is a rare disorder of purine metabolism that results from a marked decreased of xanthine oxidase (EC 1. 2. 3. 2) activity. In Japan, only 2 cases with xanthinuria have been reported. For last tow years (1980-1981) we experienced four Japanese patients with hypouricemia, hypouricosuria and xanthinuria. We report here the biochemical features of four cases with hereditary xanthinuria.
Case 1 was a 50-year-old woman who was initially diagnosed Graves's diseas in 1968, but otherwise well. Case 2 was a 67-year-old man who had a renal stone and died of renal tumor in the middle of this study. Case 3 was a 39-year-old man who was combined chronic renal failure. And his younger brother was case 4 who has been healthy.
A revers phase high-performance liquid chromatography (HPLC) was used to the determination of hypoxanthine, xanthine, uric acid and allopurinol (4-hydroxypyrazolo-(3, 4-d) pyrimidine) in human urine Urinary hypoxanthine concentration was 1.3-5.4 mg/100ml, urinary xanthine concentration was 11.3-26.1 mg/100ml and the ratio (by moler) of xanthine to oxypurine (hypoxanthine+xanthine) was 77-89% in four cases with xanthinuria.
In three cases (1, 3, 4) xanthine oxidase activity in extracts from jejunal mucosa was assayed by a ultraviolet spectrophotometric method, which depends on the enzymatic conversion of hypoxanthine to uric acid. The enzyme activity was marked decreased in all cases.
The findings of these biochemical features on four cases were suggested the presence of hereditary xanthinuria.

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A new didect radioimmunoassay for the determination of estrone sulfate in unextracted plasma without prior deconjugation has been developed. The procedure involves the use of sodium salicylate and methanol as blocking agents to inhibit the binding of estrone sulfate to plasma proteins and of highly specific anti-estrone sulfate antiserum immobilized on p-arylamine glass beads. The proposed method eliminates extraction of estrone sulfate from plasma, chromatographic separation and prior solvolysis and hence, is much more reliable and favorable than known methods for the routine assay.

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A new method was described for the separation of lipases by electrophoresis utilizing a cellulose acetate membrane.
Triolein or partially purified-endogenous fat from obese mice was used as a substrate of lipases and liberated fatty acids from substrate hydrolysed by enzymes on the membrane were coupled with copper ion in a solution of copper-triethanolamine. Coupled-copper ion was stained by diphenylcarbazide.
Four bands were found in the zymograms of lipases in the aqueous extracts from different tissues in mice, which were designated ae Lpl, Lp2, Lp3 and Lp4, respectively.
Furthermore, since fractionated lipoprotein lipase from postheparin plasma by heparin affinnity column chromatography was also observed by zymography, it was known that a new method wight be useful for investigation of lipoprotein lipase.

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Proteins were separated by high performance liquid chromatography (HPLC) using anion-exchange and gel-permeation type of resin. Human plasma was separated into 4 peaks by gel-permeation HPLC. On the anion-exchange type of HPLC, human plasma was separated into 15-16 peaks in only less than 30 min and elution profile was resemble to the pattern of electrophoresis. Ovalbumin which was eluted in single peak on gel-permeation HPLC, was separated into 5-6 peaks by anion-exchange chromatography. On the ionexchange HPLC of rat urine, esterase activity was detected in 5 peaks and peptidase acti -vity was detected in 4 peaks without enzymatic activity being interfered. These results suggested that this new HPLC was an excellent technique for the clinical chemistry of protein.

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A high sensitive micro assay for Creatine phosphokinase (CPK) isoenzyme activity based on Bondar's method is described.
Three kinds of fraction of CPK isoenzyme, MM, MB and BB, in serum were separated by means of stepwise gradient liquid chromatography through a micro column contained DEAE Sepharose CL-6B (1.5×70mm) utilised three kinds of buffered sodium chloride solution (pH 7.5) that are 30mM for MM, 145mM for MB and 300mM for BB.
The separated fractions were determined its activity by spectrofluorometry (Ex 340nm, Em 460nm) of NADH generated from CPK reaction (Oliver-Rosalki s method).
It is able to measure the activity of CPK about 0.1mU/mleasily, so this method is applicable to a low activity of CPK isoenzyme such as BB and MB in the case of early myocardial infarction and a micro sample of new born baby.

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