Mice were injected intraperitoneally with Salmonella typhimurium endotoxin to examine changes in the activity of δ-aminolevulinate (ALA) synthetase or ALA dehydrase, which palys an important role in initial steps of heme synthesis. A maximal decrease in the serum-iron level was observed 16hr after injection. The level returned to a normal range 48hr after injection. The activities of ALA synthetase and ALA dehydrase decreased significantly in the liver 16hr after injection, but increase to a normal range about 32hr after injection. They also showed a transitory decrease in the spleen 16-24hr after injection, but increased thereafter. They were at higher levels during two weeks than those in the control mice. On the other hand, the blood ALA dehydrase level decreased markedly 1-2 days after injection. The administration of pentobarbital in the endotoxin-poisoned mice resulted in an appreciable prolongation of time of sleep which had been induced by the drug in these mice. It was, therefore, assumed that the hepatic durg-metabolizing enzyme activity might have been affected with a decrease in the activities of hepatic ALA synthetase and ALA dehydrase in these mice.
An experiment was carried out to determine whether the cellular amino acids and proteins of Bifidobacterium bifidum harbored in the intestinal tract could be utilized by the host. In it germfree and conventional mice were administered orally with live cells of B. bifidum labelled with [3H] leucine, as well as heat-killed cells and cytoplasmic extracts derived therefrom. Radioactivity in the serum was followed in these animals over a 7-day experimental period. Four different types of samples were prepared from radiolabelled B. bifidum for administration by the aid of a gastric tube. They were (i) suspension of live cells, (ii) suspension of heat-killed cells, (iii) cytoplasmic extract, and (iv) heated cytoplasmic extract. In conventional mice, radioactivity was recovered from the serum at appreciable rates within 1 hour after administration, reaching a maximal level at the 4th to 8th hour with a subsequent gradual decrease. The recovery rate in these animals was dependent on the type of samples. It was the highest in mice administered with sample (ii) and rather low in mice administered with sample (i). Radioactivity was the lowest in mice administered with samples (iii) and (iv). On the other hand, the recovery rate was higher in germfree mice administered with sample (i) than in those administered with sample (ii). These results led to the following conclusions. When mice were administered orally with live cells, heat-killed cells, or cytoplasmic extracts of B. bifidum, amino acids and proteins contained in this organism were absorbed from their intestinal tract and utilized efficiently. The efficiency of their intestinal absorption depended significantly on the type of cells or extract administered and on the state of the intestinal flora.
The reaction between polysaccharide extract from Actinomyces israelii (APS) and CRP-positive serum was studied. The results obtained are summarized as follows. 1) CRP was precipitated with APS in the presence of Ca ions and 68-93% of it in serum. absorbed with APS. 2) APS contained galactose, glucose, glucosamine, phosphate and choline in the molecular proportion of 4:1:2:1:1. 3) CRP-APS precipitation was inhibited by various phosphate monoesters of low molecular weight, of which phosphoryl choline was the most active inhibitor. 4) It was presum that choline-phosphate might be present at the end of APS-polymer, providing a site of reaction for APS. 5) CRP was precipitated with polysaccharide extracts from A. israelii, A. odontolyticus and A. bovis, but not with that from A. naeslundii or A. viscosus.
Essentially the same simplified methods were devised for the purification of staphylococcal enterotoxins D and E. Each of them consisted of two steps, chromatography on SP-Sephadex C-25 and gel filtration on Sephacryl S-200. Each purified toxin formed a single band in disk electrophoresis at pH 4.3. It formed one fused precipitin line on agar against homologous anticrude serum, reference anti-toxin serum from Dr. Bergdoll and anti-purified toxin serum prepared from rabbits immunized with the purified toxin. Each anti-purified toxin serum formed one fused precipitin line against crude toxin, reference toxin from Dr. Bergdoll and purified toxin. Anti-toxin D serum formed a precipitin line against toxin D, but not against toxin A, B, C2 or E. Anti-toxin E serum formed two precipitin lines against toxin E and A, respectively. Each line formed a spur. By SP-Sephadex C-25 chromatography, each toxin was rasolved into two or three fractions. By isoelectric focusing in 5% polyacrylamide gel with 2.5% Pharmalyte, pH 3-10, each was focused into five to seven components with different pI values.