Calcium-binding to non-ionic lipids: crystal structure of a calcium chloride complex of geraniol.

X-ray diffraction data were used to determine the crystal structure of a calcium chloride complex of geraniol. The geraniol molecules assume a bilayer arrangement, with channels of calcium and chloride ions separating the bilayers. Each calcium ion is coordinated to the hydroxyl groups of two symmetry-related geraniol molecules and to four chloride ions. Our results demonstrate that hydrophobic interactions within a lipid bilayer can lead to an arrangement of hydroxyl groups suitable for binding calcium ions. Similar interactions may be involved in the calcium-binding sites on membrane surfaces.     

Blockade of histamine-induced contractions of guinea pig ielum by beta-haloalkylamines.

In the longitudinal muscle strip of guinea pig ileum phenoxybenzamine (POB) produces a maximum parallel shift of 0.7 log units in the dose-response curve to histamine. In the presence of sodium thiosulfate in the wash fluid the parallel shift whith retention of maximum response increases to about 2 log units, and a similar value is obtained for Nethyl-N-(2-bromoethyl)-1-naphthylamine. The The agent N-ethyl-N- (2-chloroethyl)benzylamine produces a significantly smaller shift of dose-response curve of 1.53 log units before the maximum response becomes depressed. The receptor-specific depression of maximum response produced by higher doses of POB is reversed by sodium thiosulfate and by bovine serum albumin, while the parallel shift in dose-response curve is unaffected by both treatments. These findings may be explained by a hypothesis involving interaction of 2-haloalkylamines at two sites.     

The elimination of urease activity in Streptococcus faecium as evidence for plasmid-coded urease.

A strain of Streptococcus faecium from the sheep rumen showed spontaneous loss of urease activity when subcultured at the normal rumen temperature of 38 degrees C, although in mixed cultures in vivo or in vitro loss of urease was not apparent. The rate of loss of urease in pure cultures was increased at incubation temperatures above 38 degrees C, but loss was never complete. However, at temperatures below 38 degrees C loss was greater, and at 22 or 18 degrees C the urease was completely eliminated. Incubation with sodium dodecyl sulphate (0-002%) or ethidium bromide (2-5 X 10(-5)M) caused complete loss of urease activity. The urease activity was also eliminated when the streptococcus was grown aerobically, and this loss of activity was irreversible. It is suggested that the urease activity is controlled by a plasmid gene and that aeration, low growth temperature and chemical agents 'cure' the streptococcus of the plasmid. Attempts to demonstrate the presence of covalently closed circular extrachromosomal DNA by caesium chloride-ethidium bromide equilibrium density-gradient centrifugation were unsuccessful.     

Endotoxic lipopolysaccharides stimulate steroidogenesis and adenylate cyclase in adrenal tumor cells.

Lipopolysaccharides (endotoxins) from Escherichia coli, Serratia marcesens and Salmonella typhosa stimulated steroid production in Y-1 adrenal tumor cells in culture with a latent period of 3-4 h. Lipid A, derived from Escherichia coli lipopolysaccharide, also stimulated steroidogenesis. Lipopolysaccharides and lipid A also stimulate adenylate cyclase activity and cause rounding of the cells. In contrast, lipopolysaccharides do not stimulate steroidogenesis in receptor-deficient adrenal tumor cells (OS-3) or Leydig tumor cells (I-10). This tends to rule out contamination by enterotoxin to which these lines respond. Although both hormone and lipopolysaccharide responses are lost in these lines, there was no interaction between these sites as judged by the failure of lipopolysaccharides to block, during their latency, the response to corticotropin in Y-1 cells. The possibility that the lipopolysaccharide effect is one on membrane conformation is discussed.     

Interaction of two mechanisms regulating alkali cations in HeLa cells.

The alkali cation content of HeLa cells is independent of culture density and of whether the cells are grown in suspension or attached to the culture vessel. With a cell doubling time of 28 hours, the cell K content turns over approximately once per hour. Following partial blockade of the alkali-cation transport system with ouabain, two distinct but interrelated mechanisms operate in the cellular response: (a) an increase in intracellular Na stimulates the pump so that the short-term alteration in electrolyte compostition is less than would be expected from the fraction of pump sites inhibited, and (b) there is a cycloheximide-sensitive recovery in transport capacity reflecting a restoration of functional transport sites to their normal density on the cell surface. Experimental manipulations that mimic the effect of ouabain lead to a stimulation of transport, but they do not result in an increase in the number of ouabain-binding sites on the surface. The data are consistent with a four-to-six hour turn-over of transport sites at the surface, but there is no evidence for a speicific induction of the transport system within this short-term recovery period.     

Mode of formation of cholesta-5,7-dien-3β-ol from cholest-5-en-3β-ol by larvae of Calliphora erythrocephala

1. The conversion of cholest-5-en-3beta-ol (cholesterol) into cholesta-5,7-dien-3beta-ol by axenic Calliphora erythrocephala larvae was demonstrated. 2. The transformation is probably direct (Delta(5)-->Delta(5,7)) and does not involve a Delta(0) intermediate (Delta(5)-->Delta(0)-->Delta(7)--> Delta(5,7)). 3. Delta(7)-bond formation involves the stereospecific elimination of the 7beta hydrogen atom. 4. The relative amounts of free and esterified sterols were determined in larvae grown on cholesterol as sole sterol source and on 5alpha-cholestan-3beta-ol supplemented with minimal amounts of cholesterol. 5. The significance of the results is assessed in relation to the probable role of cholesta-5,7-dien-3beta-ol as an intermediate in the biosynthesis of ecdysones.     

Field manipulations of tidal flushing,light exposure and natant macrofauna in a Louisiana salt marsh: Effects on the meiofauna

Field manipulative experiments were used to investigate some of the potential regulating factors of the meiofauna of a Louisiana salt marsh. Effects of various combinations of marsh grass clipping, exclusion of natant macrofauna, and tidal flushing on nematode, polychaete, and copepod density, as well as copepod species composition, were determined. Edaphic chlorophyll a was measured simultaneously. Grass clipping consistently affected the meiobenthic copepod assemblage; diversity and evenness dropped by Day 29 when Nitocra lacustris (Schmankevitsch) became dominant. Nematode density relative to controls was lower by Day 29 in clipped plots. N. lacustris abundance increased relative to controls in clipped plots enclosed by a solid Plexiglas box. Nematode density was negatively correlated with chlorophyll a content. No simple explanation of these patterns is possible; concomitant changes in microflora coupled with associated changes in the physical/biological environment of meiofauna must be responsible. Exclusion of only natant macrofauna (fish, shrimp, crabs) had no influence on meiofauna contrary to findings in other marshes. Dense grass cover and short, irregular tidal inundation may normally restrict intertidal grazing in Louisiana marshes.     

Regulation of lipogenesis in isolated hepatocytes by triglyceride-rich lipoproteins.

Very low density lipoproteins, chylomicrons, and remnants caused, within an hour, significant inhibition of fatty acid synthesis but not cholesterol synthesis in hepatocytes isolated from meal-fed rats. In contrast, low density lipoproteins, high density lipoproteins, and the serum fraction of density greater than 1.21 failed to significantly inhibit either fatty acid or cholesterol synthesis within 1 h. The Scatchard plots of specific binding showed that rat and human very low density lipoproteins interact with the high affinity sites on the hepatocytes with the apparent dissociation constants of 64 and 106 nM, respectively. These data also indicated that each hepatocyte was capable of binding 6 X 10(5) molecules of very low density lipoproteins.