The effect of a-tocopherol in endotoxicosis was studied. The a-tocopherol level significantly decreased in mouse liver 18 hr after endotoxin administration, thereafter tending to increase to approach the normal range. In endotoxin-tolerant mouse liver, the lipid peroxide level was reduced to less than half of that in nontolerant animals following endotoxin challenge. The liver lipid peroxide level and serum lactate dehydrogenase or acid phosphatase leakage were studied in mice fed a vitamin E-deficient (ED) diet and a vitamin E-supplemented (ES) diet for 40 days. ED mouse liver exhibited a higher formation of lipid peroxide after endotoxin was given while there was a markedly lower level in ES mouse liver. There was significantly more serum lactate dehydrogenase or acid phosphatase leakage in ED mice than in ES mice after endotoxin administration. There was about a 25% decrease in liver superoxide dismutase (SOD) activity in endotoxin-poisoned mice fed both the normal and the ED diets, while the activity was at a higher level in ES-fed mice. These results suggest that atocopherol may be helpful in preventing membrane instability in endotoxin poisoning.In the previous paper (42), we reported that administration of Salmonella endotoxin resulted in marked formation of lipid peroxide in mouse liver compared to that in the fasting controls. On the other hand, the activities of superoxide dismutase and glutathione peroxidase, which are the scavengers of free radicals, decreased in mouse liver 18 hr postintoxication. The lipid peroxide level in endotoxinpoisoned mice given a-tocopherol, the most important form of vitamin E, was lower than that in the controls. Furthermore, it was found that a-tocopherol prevents completely the membrane protein damage which arises from endotoxin challenge.Most, if not all, of the biologically functional a-tocopherol is associated with the membranous portions of cells. Many investigators have shown that a-tocopherol plays an important antioxidant role by blocking the peroxidation of polyunsaturated fatty acid (PUFA) in biomembranes (31,43,50). Lucy proposed that a-tocopherol is intimately associated with the hydrocarbon portion of PUFAs of membranes, and may play a physicochemical role in the stabilization of biological membranes (12,26). Cook et al (9) observed that PUFA-deficient rats were significantly more resistant to bacterial endotoxic shock than normal rats, 787

A possible role of intracellular Ca2+ and participation of calmodulin in cellular metabolism in endotoxin‐poisoned mice were investigated. The levels of calcium in liver cytosol and liver mitochondria fractions in poisoned mice were markedly higher 18–48 hr after endotoxin injection than in the control mice. On the other hand, the levels of serum calcium in the poisoned mice were about 20% lower at 18 hr than in the controls. The serum calcium levels in mice injected with 50 and 100 μg of endotoxin showed no dose‐response effect, but a dose‐response effect was observed at a dose of 200–400 μg. The serum Ca2+ levels in endotoxin‐tolerant mice were similar to those in the control mice. The levels in mice injected with glucocorticoid‐antagonizing factor mice were about 14% lower at 3 hr than in the controls. The mice fed a vitamin D3‐ and calcium‐free diet showed a higher mortality rate in the early stage (12–18 hr) of endotoxication than that of the mice fed a normal diet.The lipid peroxide levels and Ca2+‐ATPase activity in the liver mitochondria fraction in endotoxin‐poisoned mice showed a higher level than those of the control mice. There was little or no difference in the levels of serum glucose between the mice injected with calmodulin antagonist (trifluoperazine, TFP) plus endotoxin and those given endotoxin alone. However, the liver glycogen levels in TFP plus endotoxin‐treated mice were markedly higher than that in mice given endotoxin alone. Furthermore, calcium antagonist (verapamil) plus endotoxin‐treated mice had about a 40% higher survival rate after 72 hr than those given endotoxin alone.The findings suggest that there is a possibility of participation of the Ca2+‐calmodulin system in carbohydrate metabolic disorders during endotoxemia and that the changes in intracellular Ca2+ may result in various metabolic disorders.

The administration of endotoxin to mice rendered hypersensitive by lead acetate resulted in profound lipid peroxide formation in the liver 6 hr postintoxication. Endotoxin plus lead acetate administration depressed glutathione peroxidase and superoxide dismutase activities in mouse liver, whereas superoxide anion generation significantly increased in the livers of endotoxin plus lead acetatetreated mice compared with that in mice treated with endotoxin alone. Serum acid phosphatase and lactate dehydrogenase isozyme exhibited much more leakage in endotoxin plus lead acetate-injected mice than in sera of mice given endotoxin alone. Nonprotein SH level in the liver was reduced markedly in endotoxin-lead treated mice compared with those receiving endotoxin alone. The plasma vitamin E level was found to decline by 6 hr postintoxication in both endotoxin-lead and endotoxin alone-treated mice, and the transient elevation of the plasma level at 18 hr may be considered to indicate mobilization from other tissues into the blood.

About 1200 urine samples were collected, mostly in winter seasons in 1982-1984, from adult women in 7 nonpolluted areas in widely separated parts of Japan, and analyses for cadmium (Cd-U) were conducted in a single laboratory. The geometric mean (GM) by decades of age groups of Cd-U, after adjustment for a specific gravity of urine of 1.016, increased from 0.88 microgram/l in the twenties to reach a maximum of 1.78 micrograms/l in the fifties followed by gradual decrease to 1.31 micrograms/l in the eighties. The effect of smoking (about 8 cigarettes/d as a mean) was absent. Analyses of additional 125 urine samples from men revealed that Cd-U in men was not higher than that in women. When classified geographically, Cd-U was higher in the area on the coast of the Sea of Japan, as suspected in preceding studies on blood cadmium levels and dietary cadmium intakes. The Cd-U levels observed in the present study are similar to the values in previous publications on the Japanese and are apparently higher than the counterpart values from Europe, the United States, and New Zealand.

In a previous paper (7) we reported that administration of endotoxin plus lead acetate to mice resulted in marked formation of lipid peroxide, and the activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in the liver were lower than those in mice treated with endotoxin alone. It has been noted that a blockade of the reticuloendothelial system (RES) increases the sensitivity of animals to the lethal effect of endotoxin (12). However, because of its numerous biological effects on cell metabolism, lead acetate, an RES depressor, can be considered to be a specific blocker different from the other agents. The present study was carried out to observe the effect of endotoxin administration under RES depression by trypan blue.Male ddY mice (Shizuoka Experimental Animal Farm Co., Japan), 6 to 7 weeks old and weighing 22 to 25 g, were purchased and maintained in this laboratory for at least one week. Salmonella typhimurium endotoxin (Westphal preparation obtained from Difco, Detroit, MI, U.S.A.) was used throughout this study. This toxin was suspended in normal saline prior to use. A dose of 50 mg/kg or 200 mgjkg of 1.25% trypan blue (Wako Pure Chemical Industries Ltd., Osaka) was injected intravenously into mice 24 hr before endotoxin administration unless otherwise noted. Control mice were injected intraperitoneally with 0.2 ml of a 0.9% saline solution. Mice were fasted after endotoxin administration.Lipid peroxide in liver tissue homogenates (in 8.1 % sodium dodecyl sulfate[SDS] containing 20% acetate buffer, pH 3.5) was determined fluorometrically with thiobarbituric acid (TBA) by a modification (4) of Vagi's method (14). Isozymes of lactate dehydrogenase were estimated electrophoretically according to the method by which NADH is formed in the presence ofphenazine methosulfate and NBT is reduced to a strongly colored formazan using the "LDH Isozyme-Test Kit" (Wako Pure Chemical Industries Ltd., Osaka). Serum lipoprotein was separated by disk-gel electrophoresis in 3.75% polyacrylamide gel performed with a previously stained lipoprotein according to the method of Narayan et al (3). Table 1 shows the percentage of survival after endotoxin injection in trypan blue (TB)-pretreated mice. TB was injected intravenously into mice at doses of 50 mg/kg, 100 mg/kg, and 200 mg/kg. Twenty-four hours after TB injection, 18 mg/kg of endotoxin was administered intraperitoneally to the mice. TB (200 973