Cholestatic liver injury was experimentally induced in rats by administration of α-naphthylisothiocyanate (ANIT) and the peak activity of mitochondrial L-aspartate: 2-oxoglutarate aminotransferase (m-GOT) released in the serum was found to precede the peak of total GOT activity. To investigate the permeability of the mitochondrial inner membrane to m-GOT, liver mitochondria obtained from rats given ANIT were fractionated into two subfractions : one containing the matrix and the inner and outer membranes, and the other containing the intermembrane space, and the m-GOT in these fractions was determined. As a result, 12 hours after ANIT administration, the relative activity of GOT in the subfraction containing the matrix and the membranes was significantly lower than the control value. In the same period, the ratio of GOT activity to the activity of glutamate dehydrogenase, which is a marker enzyme for the matrix, and the ratio of GOT activity to the activity of cytochrome c oxidase, which is a marker enzyme for the inner membrane, were both decreased by half. In contrast, the relative GOT activity for the subfraction containing the intermembrane space was significantly increased 12 hours after administration. Also, the ratio of GOT activity to the activity of adenylate kinase, a marker enzyme for the intermembrane space, was doubled. These results suggest that m-GOT, which is originally located in the mitochondrial matrix, transmigrated to the intermembrane space via the inner membrane under the effect of ANIT administration.
^<125>I-Labeled batroxobin was prepared and following its intravenous and subcutaneous administrations to rats and dogs, the blood radioactivity was determined. In the both species following the intravenous injections, the decrease in radioactivity was biexponential. Following subcutaneous administration, radioactivity became maximal at 6h and decreased in a manner similar to that of the P-phase of the intravenous injection. The blood concentration of fibrinogen in dogs was also determined. After the intravenous injection, fibrinogen became undetectable 1h later, and appeared again in the blood at 24h. After the subcutaneous injection, the decrease was not so rapid. Fibrinogen resumed its original levels at 7 day after the administration in both the routes. Radioactivity after the both injections was excreted generally in the urine in about the same amounts. The total urinary and fecal excretions in rats and dogs were 80 and 95%, respectively. The distribution of radioactivity in the tissues was examined by counting technique and whole-body autoradiography. Radioactivity predominantly accumulated in the thyroid and stomach and could also be found in the kidneys and liver in fair amounts. The distribution patterns of radioactivity for both the routes of administrations and also for male and pregnant rats were basically the same. In fetus rats, a slight distribution was noted. From the results of gel filtration chromatography and trichloroacetic acid fractionation, [125I] batroxobin was metabolized soon after the administration to afford low molecular substances such as 125I-ion in the plasma and urine.
Environmental substances were examined for their effect on interferon induction and delayed type hypersensitivity (DTH) responses in mice. Amaranth, safrole, phenacetin and nicotine suppressed the DTH response, and suppressed the serum interferon titers induced by virus infection. However, they did not affect the interferon titers which were induced by tilorone, a chemical inducer. The peak of interferon titer was 12 hours after infection with herpes simplex virus type 2 (HSV-2). Therefore, amaranth, safrole, phenacetin and nicotine were given to mice intraperitoneally 24 hours before, and 2 and 18 hours after infection with HSV-2. It was found that safrole and nicotine shortened the mean survival time of HSV-2 infected mice when they were given to mice 2 hours after virus inoculation. Amaranth and phenacetin showed similar effects. However it was not definite statistically. In biochemical and hematological tests, these four substances did not affect the functions of liver, kidney and carbohydrate metabolism in normal mice. These results suggest that substances which may often be taken into the body have the potential to affect the onset of virus infectious diseases as a result of the suppression of the host defense reaction.
Aminoguanidine (AG) sulfate was injected into the thin albumen of fertile eggs on the different days of incubation. The lethality of AG sulfate on the developing chick embryos became lower when the treatment was done at later developmental stage. When AG was injected on the 5th day of incubation, the frequency of retardation of development of the body and liver, aplasia of gallbladder and enlarged spleen was highest without high mortality compared with the other experiments. Embryos with liver damage could not hatch. 14C-AG hydrochloride injected on the 5th day of incubation easily distributed into the embryo through the yolk from the thin albumen of the injection site, but when injected on the 9th day, the agent transferred very little into the yolk and embryo. The change of transport in the eggs with development may elucidate the difference of lethality of AG. Additionally, it was confirmed using thin layer chromatography that a large part of the radioactivity in the embryo might be an unknown metabolite of AG and the substance was accumulated with extremely high concentration in the liver at 24-48 hr after the injection of AG on the 5th day of incubation. This is probably the main cause of the peculiar abnormalities in the liver of chick embryos.
t has been reported that aminoguanidine (AG) sulfate injected into hen's eggs at the earlier incubation stages induced retardation of development of body and liver, aplasia of gallbladder and enlarged spleen in high frequency without high mortality. The present study deals with the influence of AG against rodent animals and their fetuses. AG sulfate showed low lethality at the intraperitoneal administration against young mice and rats. When the maximum tolerated dose of AG sulfate (750 mg/kg in mice and 500 mg/kg in rats) was intraperitoneally injected into pregnant mice and rats on day 0 to 6 or day 7 to 13 of gestation, the incidence of resorption was comparatively high in the day 7 to 13 group of both species, but severe abnormalities as observed in the chick embryos and other external abnormalities were not induced in rodent fetuses. 14C-AG hydrochloride intraperitoneally injected to pregnant mice on day 12 was widely distributed in the maternal body and fetuses within 1 hr and rapidly excreted. Furthermore, AG might be hardly metabolized in mice, while a metabolite clearly appeared in chick embryos. Therefore, the lower toxicity of AG on rodent animals as compared with chick embryos may be due to the fact that the agent is not metabolized and rapidly excreted from maternal body and fetuses.
Acute toxicity, eye irritation, primary skin irritation, skin sensitization, phototoxicity, photosensitization and mutagenicity of sophorolipid derivatives were studied in rats, mice, rabbits, guinea pigs and Salmonella typhimurium strains. The acute oral toxicity of sophorolipid (SL) which Torulopsis bombicola produces, and its derivatives (PSL, Ethyl-SL and Oleyl-SL) were shown to be very low. The LD50 values of PSL ranged from 10 g/kg to 16 g/kg on oral administration in rats and mice, and from 5.8 g/kg to 6.6 g/kg on subcutaneous administration in mice. The oral LD50 values of Ethyl-SL and Oleyl-SL were estimated to be greater than 15 g/kg and that of SL was 12.5 g/kg. In eye irritation study, PSL failed to produce any reactions at 50% concentration even when the rabbit eye was not subsequently washed. SL, Ethyl-SL, Oleyl-SL and Tween 20 were "no irritant" or "slight irritant" to the rabbit eye at 20% concentration. PSL showed no irritancy to both the intact and abraded guinea pig skin at 50% concentration. And in other examinations, it was also indicated that PSL had no potentials of skiri sensitization, phototoxicity and photosensitization in guinea pigs and had no potentials of mutagenicity in Salmonella typhimuritum TA98 and TA100.
Five groups of 12 male and 12 female rats each were fed diets containing 0, 0.06, 0.25, 1.00 and 4.00% PSL for a period of one month. Food consumption of PSL-fed groups did not differ from that of control. Urinalysis and autopsy findings were within normal in every group of rats treated. With 4.00% in the diet, body weight gain was significantly retarded and water consumption was increased, and soft stool occurred. In the hematological examination, decrease of red blood cells and increase of white blood cells were observed at the levels of 1.00 and 4.00% PSL. Changes of white blood cell differentials were also seen at the same levels. Serum Na+ concentration was slightly decreased at the 0.25, 1.00, 4.00% levels and serum glucose was also decreased at the 1.00, 4.00% levels, but the values were within the normal limits. Significant increase of relative liver weight, without histopathological changes, was observed at the 4.00% level. Histopathological examination revealed slight errosion, necrosis or intestinitis in small intestine, at the levels of 0.25, 1.00, 4.00% PSL. It was considered that these findings were attributed to the irritation potential of PSL or its metabolite. These results indicated that the non-effect level was 0.06% (53 mg/kg/day) and the level causing no toxicological effect was 0.25% (208 mg/kg/day), but no deleterious effects was observed in the levels greater than 0.25%.
Protective effect of malotilate on the liver injuries induced by several hepatotoxins was studied in mice and rats. Malotilate suppressed the elevation of plasma glutamate pyruvate transaminase (p-GPT) activity induced by chloroform (CHCl3) in rats When the animals were treated with 25 mg/kg or more dose of malotilate at 6 hours prior to the treatment with CHCl3. The effect was observed even in the rats treated with malotilate 24 or 48 hours prior to the treatment with CHCl3. Malotilate, when orally administered 6 hours prior to treatment with hepatotoxins such as CHCl3, allyl alcohol, bromobenzene, dimethylnitrosamine or thioacetamide, suppressed the elevation of p-GPT activity, liver triglyceride content and/or the decrease of bromosulphalein cleardnce induced by these hepatotoxins in mice. Anethole trithione, which was used as a possible protective agent against chemical-induced hepatotoxicity, tended to normalize changes in the parameters induced by the most of these hepatotoxins, but enhanced the elevation of p-GPT activity induced by CHCl3. In a case of CHCl3-induced liver injury, the protective effect of malotilate was histopathologically confirmed. Malotilate and anethole trithione reduced p-nitroanisole 0-demethylation activity in rat liver 6 hours after the administration but increased or tended to increase the activity 48 hours after the administration. Malotilate showed a protective effect on the liver injury induced by CHCl3 even when the activity of drug metabolizing enzymes in the liver was increased, although anethole trithione enhanced the CHCl3-induced liver injury regardless of the activity of drug metabolizing enzymes.