The Donaggio reaction positive substances from the urine after hard work were identified. These substances obtained from deproteinized urine by heat at pH 5.0 and having the high titer of Donaggio reaction were composed of α-glycoproteins, which were shown to be α1-globulin and small amounts of α2-globulin by means of cellulose-acetate electrophoresis. This α1-globulin nearly composed of serum α1-acid glycoprotein and one of the α2-globulin was composed of serum α2-HS globulin, both of which were shown by immuno double diffusion and by single radial immunodiffusion. The results of im-munodiffusion indicated that the other α1-glycoproteins were not contained, such as tryptophan-poor α1-glycoprotein, α1-antitrypsin, and α1-antichymotrypsin. The levels of α1-acid glycoprotein in urine before and after running, which were measured by single radial immunodiffusion, paralleled the Donaggio-titer. Serum mucoprotein purified by perchloric acid from human serum contained Donaggio positive substances. From these results, we can conclude that Donaggio positive substances were composed of at least two sorts of mucoprotein, α1-acid glycoprotein as a major part and α2-HS globulin as a minor.
In order to study the factors which have influenced to β2-microglobulin levels in urine, the effect of hard loading on the concentration of urinary β2-microglobulin was studied. Increase in both actual and corrected β2-microglobulin concentration and slight increase in albumin was observed with volunteers in the urine after marathon running. Maximum excretion of urinary β2-microglobulin was observed 30min after cession of exercise. And increase in actual and corrected β2-microglobulin concentration, and the ratio of β2-microglobulin to creatinine in urine of volunteers loaded 210 steps/min for 1 hour were recognized 30 min after cession of exercise by training machine. Similar result of urinary excretion to β2-microglobulin was obtained with those of creatinine. Concentration of β2-microglobulin in serum had a tendency to increase, but concentration of creatinine in serum was not increased. Results of Sephadex G-100 gel filtrations indicated that urinary β2-microglobulin after exercise, which measured by radioimmunoassay, was contained in the filtrated fraction having molecular weight of approximately 11, 800, suggesting that similar substance to HL-A did not excreted in urine after hard loading.
Normal human serum was applied to Sephadex G-100 gel chromatography and the eluted fractions were analyzed by radioimmunoassay of gβ2-microglobulin. It was revealed that two fractions eluted by gel chromatography have reactivity to β2-microglobulin antibody, the first fraction A having molecular weight of about 48, 000 and the second fraction B having molecular weight about 11, 600. Amount of β2-microglobulin in fraction B is clearly higher than that of fraction A. The first fraction A was separated several protein bands by polyacrylamide gel electro-phoresis and one of them corresponded to the molecular weight of about 11, 600.
A single dermal application of Fenitrothion (750 mg/kg of body weight) was applied on rats and the effect was observed after 8 hr on organ size and organ levels of calcium and magnesium. Significant decreases in organ size of kidney and heart and increases in liver and spleen were observed. However, there was no change in the size of the brain. Calcium and magnesium registered a decrease in heart, kidney, liver and spleen but showed an increase in plasma suggesting migration of metals from the organs towards the circulatory system.
The present investigation reports on the accumulation and damage caused by copper in the liver and kidney of albino rats. Simultaneous treatment with copper and molybdenum shows that no traces of copper are retained in these tissues. Thus these observations lead to the conclusion that copper is ineffective in the presence of molybdenum exhibiting physiological antagonism. Several hypothesises have been considered for their antagonistic behaviour. Present authors suggest that molybdenum may act on copper as a chelating agent binding with it and forming either cupric molybdate or copper thiomolybdate which is ultimately excreted. Intake of molybdenum with the diet has also been suggested for the occupational victims of copper.
Elevated lysozyme activity was observed in the urine of patients with Itai-itai disease. The urinary lysozyme was isolated from the urine specimens of these patients by gel filtration and ion exchange chromatography. Two lysozyme peaks were found on Sephadex G-100 gel filtration. The first peak (lysozyme-E) had two components on DEAE-cellulose chromatography. One of them showed more acidic chromatographic behavior than the other component which was eluted at the void volume of the column. The second peak (lysozyme-F) on Sephadex G-100 gel filtra-tion had only the component which was similarly eluted at the void volume of the DEAE-cellulose chromatographic column. Slight differences in the chromatographic behavior and in amino acid composition between lysozyme-E and -F indicated that the urinary lysozyme was composed of two components.
3-3' dimethylnaphthidine was dissolved in concentrated sulphuric acid, and the solution was used as the colorimetric reagent for the microdetermination of lead. The analytical procedure required to be performed under strict conditions because the concentration of acid, reacting time, temperature, etc. affected the coloring reaction strongly. The solution, however, showed high sensitivity for determina-tion of lead and the procedure was very simple. Few metals besides lead reacted with 3-3' dimethylnaphthidine under such conditions to disturb the determination. About 0.02 μg/ml of lead in final solution was determined easily.