The lipids of Bacteroides melaninogenicus were readily extractable with chloroform-methanol. Three per cent of the fatty acids were not extractable. The neutral lipids contained 4% of the extractable fatty acids, the stench characteristic of these organisms, and 0.5 ,urmole of vitamin K2 isoprenologues K2-35, K2-40, and K245 per g (dry weight). This is one-fifth to one-tenth of the vitamin K2 level found in other bacteria. Ninety-six per cent of the extractable fatty acids were associated with the phospholipids (60 umoles of lipid phosphate/g, dry weight), which consisted of the diacyl lipids phosphatidic acid, phosphatidyl serine, and phosphatidyl ethanolamine (with phosphatidyl glycerol and cardiolipin in one strain). The unusual phosphosphingolipids ceramide phosphorylethanolamine, ceramide phosphorylglycerol, and ceramide phosphorylglycerol phosphate accounted for 50 to 70% of the lipid phosphate. In protoheme-requiring strains, the protoheme concentration in the growth medium regulated the growth rate and the amount of enzymatically reducible cytochrome c. There were no gross changes in the lipid composition in cells containing different levels of enzymatically reducible cytochrome c. the bacteria by a modified Bligh and Dyer procedure (2). A 30-ml suspension of bacteria in 50 mm phosphate buffer, pH 7.6, containing about 14 mg (dry weight) of cells was mixed with 75 ml of methanol 84 on July 16, 2020 by guest

Female B6C3F1 mice were exposed to graded doses of nickel sulfate to determine a threshold response for myelotoxicity and immunotoxicity, and to identify which of the populations of lymphoreticular cells were most sensitive to the toxic effects of nickel. Animals were given free access to the chemical in the drinking water at 0, 1, 5, or 10 g/l for 180 d. Water consumption, blood and tissue nickel concentrations, body and organ weights, histopathology, immune responses, bone marrow cellularity and proliferation, and cellular enzyme activities were evaluated. There was no mortality. Mice in the 5-g/l and 10-g/l dose groups drank less water than controls; the responses measured in the 10-g/l group may have been due to a combination of dehydration and chemical toxicity. Decreases in body and organ weights were confined to mice in the 10-g/l dose group, except for the dose-related reductions in thymus weights. Blood nickel was measured at 4, 8, 16, and 23 wk of exposure. The mean blood nickel values showed increases between 4 and 8 wk that were proportional to time and dose; thereafter there was no substantial increase in blood nickel in any of the dose groups, except for an increase in the mean blood concentration in the 10-g/l group at 23 wk. The kidney was the major organ of nickel accumulation. The primary toxic effects of nickel sulfate were expressed in the myeloid system. There were dose-related decreases in bone marrow cellularity, and in granulocyte-macrophage and pluripotent stem-cell proliferative responses. In unfractionated bone marrow cells glucose-6-phosphate dehydrogenase enzyme activity from the hexose monophosphate shunt was more sensitive to nickel sulfate than were representative glycolytic or Krebs cycle enzymes, with 25-35% maximum inhibition at 5 g/l and 10 g/l. Aliquots of bone marrow cells were separated into enriched bands of lymphocytes, granulocyte-macrophages, and erythrocytes; enzyme inhibition that occurred in unfractionated bone marrow cell aliquots was only expressed after cell separation in the enriched granulocyte-macrophage cell population, suggesting that these committed stem cells were a primary target of nickel sulfate toxicity. There was one example of systemic immunotoxicity, reduction in the lymphoproliferative response to lipopolysaccharide, and it was regarded as secondary to the primary effect of nickel sulfate on the myeloid system, since this was the only significant change among a panel of seven immune parameters that were evaluated.

Logarithmically growing Haemophilus parainfluenzae lost 15 to 20% of the phospholipids, demethyl vitamin K2, cytochrome b, and cytochrome c, and 50% of the lipopolysaccharide when incubated in ethylenediaminetetraacetic acid (EDTA)-tris-(hydroxymethyl)aminomethane (Tris) for 10 min. This loss of membrane components occurred without loss in viability, and the lost components were recovered as membrane fragments in the surrounding buffer. The phospholipids recovered in the membrane fragments had a slightly lower specific activity than the phospholipids in the residue. Lysis of a portion of the cells could not account for the release of membrane components, as the cells lost neither glucose-6-phosphate dehydrogenase activity nor deoxyribonucleic acid. The treated cells were osmotically stable and contained the same proportions of the individual phospholipids as pretreatment cells. Prolongation of the EDTA-Tris treatment did not induce further loss of phospholipid or demethyl vitamin K2, but caused a decrease in viability. If the cells were returned to the growth medium after 10 min, the cells immediately resumed growth at the pretreatment rate. During growth in the recovery period, the phospholipids increased logarithmically in the pretreatment proportions, although there was a marked decrease in the turnover and a shift from the use of extracellular lipid precursors to the use of intracellular pools of precursors.

Heterogeneity in the metabolism of cardiolipin (CL) has been detected in Haemophilus parainfluenzae . Pulse-chase experiments showed that a portion of the total CL incorporated and then lost 32 P much more rapidly than the rest of the CL in the cells. The metabolism of each phosphate of the CL differed. The phosphate of the phosphatidyl glycerol (PG) portion of the CL had a more active metabolism than the phosphate of the phosphatidic acid portion of the molecule. Only a portion of the PG pool contributed to the formation of CL. Ethylenediaminetetraacetic acid inhibited the CL-specific phospholipase D in vitro and, when added to growing cells, resulted in more rapid PG metabolism, suggesting that CL hydrolysis contributed to the PG pool.