In this study, we investigated the superoxide dismutase (SOD) activity in serum of normal human subject and the relation between renal disease and SOD activity, Normal range of the total SOD activity was 1.94±0.90U/ml (mean±2SD) in male (n=105) and 1.63±0.66U/ml in female (n=54), Mn-SOD activity was 0.83±0.42U/ml in male (n=105) and 0.99±0.26U/ml in female (n=54), There were significant difference between male and female.
The correlation between creatinine and Cu, Zu-SOD showed a good correlation (r=0.601), However, Mn-SOD was poorly correlated (r=0.396), A patients with renal insufficiency had markedly high Cu. Zn-SOD activity and creatinine concentration, After renal transplantation, Cu, Zn-SOD activity decreased parallel with the decrease of creatinine concentration.
The urinary protein showed a positive correlation with urnary Cu, Zn-SOD (r=0.569) and negative correlation with urinary Mn-SOD (r=-0.553).

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Guanine deaminase (guanase: EC 3.5.4.3.) was purified using a previously reported method. Anti-human liver guanase rabbit serum formed a single precipitin line with the partially purified enzyme. The antiserum also completely inhibited the guanase activity of the extract.
In the immunohistochemical study employing horse radish peroxidase complexes, the DAB (3.3'-diaminobenzidine tetrahydrochloride) reaction was positive at the same site where the histochemical reaction for guanase was positive; and the control reaction using the nonimmune rabit serum in place of antibody was negative. Moreover, the biochemical data indicated that the guanase activity was located in the proximal tubule. Thus, the DAB reaction appears to accurately show the location of guanase itself. This immunohistochemical procedure for guanase should become a valuable tool in clinical studies on this enzyme.

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A fully automated determination of antiarrhythmic drugs and mono-dealkylated metabolite of disopyramide in human serum has been developed by high-performance liquid chromatography (HPLC) with column-switching. TSKprecolumn BSA-ODS and TSKgel ODS-80TM were used as the precolumn and analytical column, respectively. Fifty μl of serum sample was directly injected onto the precolumn. After washing out the serum proteins from the column by potassium phosphate buffer (30 mmol/l, pH 5.4), the drugs adsorbed on the column were introduced to the analytical column by switching the columnconnection. The drugs were eluted with a step-gradient procedure of acetonitrile/potassium phosphate buffer (30 mmol/l, pH 5.4) both containing sodium 1-heptanesulfonate (500 mg/l), 15/85 by vol (0 to 8 min) to 30/70 by vol (8 to 22 min) at a flow rate of 1.0 ml/min. The effluent was monitored at 210 nm. Analytical recovery (94 to 102%), reproducibility (c. v. less than 3%, withinrun) and linearity indicate that this method is suitable for the therapeutic drug monitoring in clinical laboratories.

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Anti-insulin antibody index (Sebriakova), antibody-bound insulin, and 125I-insulin binding ratio were examined to determine anti-insulin antibody.
(1) Anti-insulin antibody index (sebriakova) gave good correlation with low-affinity insulinbinding capacity (Scatchard analysis)(r=0.89). It also correlated with antibody-bound insulin concentration (r=0.71).
(2) Antibody-bound insulin concentration gave good correlation with high-affinity insulinbinding capacity (Scatchard analysis)(r=0.73).
(3) ¹²⁵I-insulin binding ratio correlated poorly with low-or high-affinity insulin-binding capacity (Scatchard analysis), antibody-bound insulin, or anti-insulin antibody index (Sebriakova). But for ¹²⁵I-insulin binding ratios up to 50%, better correlation was obtained between ¹²⁵I-insulin binding ratio and antibody-bound insulin or anti-insulin antiboy index (Sebriakova).
These results suggest that anti-insulin antibody index (Sebriakova) and antibody-bound insulin are useful to ascertain low-affinity insulin binding capacity, and high-affinity insulinbinding capacity, respectively.

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Plasma protein, α_l-acid glycoprotein from patients with various urogenital diseases, including renal cancer, acute pyelonephritis and urolithiasis, was resolved by highperformance liquid chromatography (HPLC) using an anion-exchange column. Before the HPLC the plasma was pretreated by Blue-Sepharosoer Con A-Sepharose affinity chromatography. The peak of the α₁-globulin fraction containing α_l-acid glycoprotein was high in patients with advanced neoplastic or acute inflammatory disease, but not in patients with non-inflammatorby enign disease. The DNA synthetic activity of normal peripheral blood lymphocytes in vitro was suppressd more by the α_l-globulin fraction prepared by HPLC from patients with neoplastic disease than by that from patients with non-inflammatory benign disease.

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A simple method for glutathione (GSH) and glutathione disulfide (GSSG) was devised, using glutathione reductase (GR) and glutathione S-transferase (GSTR).
The total concentration of glutathione (T-GSH) was determined by an enzymatic assay with the use of GR, and the concentration of GSSG was selectively determined after GSH had been eliminated by treating specimens with 1-chloro-2, 4-dinitrobenzene and GSTR.
Using the present method, the concentrations of GSH in normal human red blood cells ranged between 2.1-3.2μmol/10¹⁰ cells, whereas those of GSSG was lower than 0.1μmol/10¹⁰ cells.

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A rapid and sensitive high-performance liquid chromatographic method has developed for the analysis of guanidinosuccinic acid, guanidinoacetic acid, and creathe in physiological fluids.
These guanidino compounds are separated on a cation-exchange column and detected with a fiuorimeter which monitors fluorescent derivatives produced by the reaction of the eluted constituents with glucose as a reagent. The fiu Orescent derivatives are measured at 420 nm with excitation at 310nm.
This method was successfully applied to serum and urine in clinical tests.

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We studied on the substrate specificities of various fatty acids for acyl-CoA synthetase in the determination of serum free fatty acids, with use of acyl-CoA synthetase, myokinase, pyruvate kinase and lactate dehydrogenase as an assay system.
The formation of acyl-CoA with acyl-CoA Synthetase from free fatty acids in serum and from purified fatty acids of Ci_{16: 0}, C_{18: 0}, C_{18: 1} and C_{18: 2} reached almost the maximum within 10 min incubation at 37°C. On the contrary, it needed to reach the maximum for more than 190 min in C_{12: 0}, 80 min in C_{22: 6}, 65 min in C_{20: 5}, 45 min in C_{18: 3}, 35 min in C_{20: 2} and 22 min in C_{14: 0}. Significantly different apparent Km values were obtained for C_{12: 0} (0.3×10^-2M), C_{20: 5} (0.5×10^-3M) and C_{18: 1} (0.9×10^-4M).
These results suggest that there are consistent substrate specificities for acyl-CoA synthetase among various kinds of fatty acids available in human serum.

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