Intramuscular or oral administration of cytochrome c provided a defense against carbon tetrachloride-induced hepatic impairment characterized by accumulation of triglycerides in the liver and damaged mitochondria. The mechanism may involve activation of the mitochondrial function by cytochrome c with the result that ATP production is increased, hence a normal metabolism is regained.
Examinations were made on the bovine testicular hyaluronidase, partially purified by zone electrophoresis for relationship between enzymic activity and intradermal diffusion of dye. Treatment of this hyaluronidase at 65° for 30 minutes results in disappearance of enzymic activity but the spreadig activity remains and this also occurs when enzymic activity is destroyed by tryptic digestion. The enzymic activity decreases to 25% by acetylation but the spreading activity was found to increase by this treatment. However, iodine treatment of hyaluronidase results in disappearance of both these activities.
A method for efficient extraction of urokinase from human urine was established by using polyacrylonitrile synthetic fiber as an adsorbent. By a combination of this method and known methods for purification of proteins, such as gel filtration and ion-exchange chromatography, urokinase with a specific activity of 224,000 International Units per mg of protein was obtained. This sample showed homogeneity by ultracentrifugation, moving-boundary electrophoresis at pH 4.8 and 9.0 and polyacrylamide gel disc electrophoresis at pH 4.0, but was separated into five active fractions by isoelectric focusing and polyacrylamide gel disc electrophoresis at pH 9.4. This sample showed a single precipitin line in double radial immunodiffusion and immunoelectrophoresis using rabbit anti-urokinase serum. This precipitin line fused with that of the International Standard preparation of urokinase and its immunological identity was established. The molecular weight of this sample was 33,000, agreeing with that of the International Standard preparation. Its optimal pH as a plasminogen activator was approximately 8.8.
Intradermal diffusion of dyes by testicular hyaluronidase is considered to be due to hydrolysis of hyaluronic acid by enzymic action, resulting in the lowering of viscosity and increase in permeability. The hyaluronidase prepared from bovine testicle by ammonium sulfate purification was fractionated by zone electrophoresis with starch and spreading activity of each fraction was examined. It was thereby found that the fractions without any enzymic activity also had spreading activity. Aqueous solution of hyaluronidase, after having lost its enzymic activity by heat denaturation also showed some spreading activity.
Histopathological changes of the liver by carbon tetrachloride included vacuolar necrosis of hepatic cells, cellular infiltration, an increase in fat droplets, a decrease in glycogen particles and a degenetation and a decrease in the number of rough surfaced endoplasmic reticula. These changes were prevented by the administration of cytochrome c. These findings were consistent with the results of previous biochemical studies that cytochrome c prevents an increase in transaminase activity, an enhancement of BSP retention, an increase in hepatic neutral fat and a decrease in hepatic glycogen induced by carbon tetrachloride. Meanwhile, histopathological characteristics of the liver of aged rats included an increase in number of vacuoles in the cells, disintegration of and decrease in hepatocyte component, a decrease in the number of glycogen particles and an increase in fat droplets and consumptive substance. The histopathological changes of aging were either alleviated or inhibited completely with cytochrome c. The conclusion is in conformity with previous reports indicating that cytochrome c improved the liver functions of aged rats and decreased lipids of the liver.
Intramuscular or oral administration of cytochrome c provided a defense against carbon tetrachloride-induced hepatic impairment characterized by accumulation of triglycerides in the liver and damaged mitochondria. The mechanism may involve activation of the mitochondrial function by cytochrome c with the result that ATP production is increased, hence a normal metabolism is regained.
Uracil was previously shown by us to be the active principle of Trionyx carapax. In this study, we show that pretreatment with uracil protects rats against hepatic injury induced by allyl alcohol and by D-galactosamine.
No report has been made on the protease contained in testicular tissue but its presence was detected in the crude hyaluronidase prepared by ammonium sulfate fractionation. This protease had optimal pH in the weakly acid range and was most stable at around pH 6. When left at pH 9 and 37°, in the presence of sodium chloride, the majority of protease activity could selectively be inactivated practically without any harm on the hyaluronidase activity.
The metabolic fate of 131I-urokinase in rats and dogs was investigated. The radioactivity in the blood of both animals after intravenous administration rapidly decreased in early stage, followed by a slow decrease of the radioactivity. When 131I-urokinase was given into rats by intravenous infusion, the peak of radioactivity in the blood was found at the end of the dosing period. The radioactivity in the liver and kidneys of the rats 15 min after intravenous administration of 131I-urokinase was found to be 43 and 32% of the administered radioactivity, respectively. In the rats approximately 80% of the administered radioactivity was recovered, mainly in the urine in 3 days. The radioactivity derived from high-molecular compounds in the serum of rats receiving 131I-urokinase intravenously was approximately 95% of the radioactivity detected 1 min after dosing, 70% at 20 min and 40% at 60 min. An artificial thrombus was prepared by the method of Chandler and radioactivity of the thrombus was determined by incubating with radioiodinated urokinase, plasminogen, plasmin or reference compounds. After a 6 hr incubation the ratio of the radioactivity concentration of the thrombus (cpm/mg) to that of the blood (cpm/mg) was 2 for 131I-urokinase, 3 for 131I-plasminogen, 3 for 131I-plasmin, 0.7 for 131I-human serum albumin and 0.8 for Na131I. When the blood added with 131I-plasminogen was employed for the preparation of an artificial thrombus, approximately 15% of the added radioactivity was detected in the thrombus.
