Exercise-associated protein (EAP) was purified to homogeneity from human serum after 10-km run and their properties were investigated. The purification was achieved by preparative polyacrylamide gel electrophoresis, anti-alb Sepharose chromatography, isoelectric focusing and gel filtration with high-performance liquid chromatography. The isoelectric point of the purified preparation was estimated to be 5.0, and the molecular weights in the absence and presence of 2-mercaptoethanol were respectively determined to be 25Kd on SDS-polyacrylamide gel electrophoresis. Amino acid analysis revealed a high content of glutamic acid and leucine and a low or no content of cysteine and isoleucine. Changes of their concentration in serum under various physical exercises were examined by rocket immunoelectrophoresis.

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We describe, to our knowledge, the third case of deficiency of lactate dehydrogenase (LDH; EC 1.1.1.27) H-subunit. LDH activities in the patient serum and erythrocytes were abnormally low, but the other glycolytic enzyme activities were almost within normal limits in the erythrocytes. The LDH isoenzyme patterns of the patient serum, lymphocytes and saliva showed only one band of LDH₅. On the other hand, the LDH electrophoretogram of the patient erythrocytes and thrombocytes showed two bands consisting of LDH₄ and LDH₅. The patient erythrocytes contained high concentrations of fructose-1, 6-diphosphate, dihydroxyacetone phosphate and glyceraldehyde-3-phosphate. From a family study on two generations, we concluded that the propositus was homozygous of the deficiency, whereas three family members were heterozygous.

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A mathematical treatment of absorbance of translucent membranes was performed for two models of distribution of light path length, namely, semi-Gaussian and exponential distributions. Empirical relationships among absorbances obtained by various densitometers of good quality that the absorbances are proportional to each other were deduced from the treatment. Experimental results of the absorbance measurements of Ponceau 3R-stained cellulose acetate membranes loaded by various amounts of bovine serum albumin suggested that not only the light path length distribution but also a stray light flux may play an important role on the relation between absorbances of the membrane in transparent and translucent states.

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Protein A extracellularly produced by UV2 (a mutant of Staphylococcal aureus Cowan I) in some cultivation conditions was analyzed with two dimensional electrophoresis and immunoblotting. Three major proteins (MW 47K_d, pI 5.2; MW 50K_d, pI 5.1; MW 51K_d, pI 5.0) were indicated on the electropherogram of the fluid of conventional cultivation at 37°C, and these proteins were identified with protein A. The same protein A was observed in the cultivation fluid using cellophane bag at 25°C, but the fluid was characterized by the presence of low molecular weight protein A in the range of MW 18K_d-49K_d and pI 4.2-5.8 and many non-protein A protein. On the other hand, in the fluid of the cellophane bag cultivation at 37°C, there only observed a lot of low molecular weight protein A ranging between MW 35K_d-13K_d and pI 4.5-6.5 and considerable amount of non-protein A proteins that were never found in the conventional cultivation fluid.

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