Stereochemistry and size of sugar head groups determine structure and phase behavior of glycolipid membranes: densitometric, calorimetric, and X-ray studies.

The role carbohydrate moieties play in determining the structure and energetics of glycolipid model membranes has been investigated by small- and wide-angle X-ray scattering, differential scanning densitometry (DSD), and differential scanning microcalorimetry (DSC). The dependence of a variety of thermodynamic and structural parameters on the stereochemistry of the OH groups in the pyranose ring and on the size of the sugar head group has been studied by using an homologous series of synthetic stereochemically uniform glyceroglycolipids having glucose, galactose, mannose, maltose, or trimaltose head groups and saturated ether-linked alkyl chains with 10, 12, 14, 16, or 18 carbon atoms per chain. The combined structural and thermodynamic data indicate that stereochemical changes of a single OH group in the pyranose ring can cause dramatic alterations in the stability and in the nature of the phase transitions of the membranes. The second equally important determinant of lipid interactions in the membrane is the size of the head group. A comparison of lipids with glucose, maltose, or trimaltose head groups and identical hydrophobic moieties has shown that increasing the size of the neutral carbohydrate head group strongly favors the bilayer-forming tendency of the glycolipids. These experimental results provide a verification of the geometric model advanced by Israelachvili et al. (1980) [Israelachvili, J. N., Marcelja, S., & Horn, R. G. (1980) Q. Rev. Biophys. 13, 121-200] to explain the preferences lipids exhibit for certain structures. Generally galactose head groups confer highest stability on the multilamellar model membranes as judged on the basis of the chain-melting transition. This is an interesting aspect in view of the fact that galactose moieties are frequently observed in membranes of thermophilic organisms. Glucose head groups provide lower stability but increase the number of stable intermediate structures that the corresponding lipids can adopt. Galactolipids do not even assume a stable intermediate L alpha phase for lipids with short chain length but perform only Lc----HII transitions in the first heating. The C2 isomer, mannose, modifies the phase preference in such a manner that only L beta----HII changes can occur. Maltose and trimaltose head groups prevent the adoption of the HII phase and permit only L beta----L alpha phase changes. The DSD studies resulted in a quantitative estimate for the volume change associated with the L alpha----HII transition of 14-Glc. The value of delta v = 0.005 mL/g supports the view that the volume difference between L alpha and HII is minute.(ABSTRACT TRUNCATED AT 400 WORDS)

Changes in the responsiveness of the colonic smooth muscle to carbachol in Hirschsprung's disease.

In vitro experiments were performed on smooth-muscle strips cut out in longitudinal and circular direction from the colon of Hirschsprung's patients who underwent an operation and from patients operated on for tumors of the sigmoid colon and rectum, serving as controls. The changes in the contractile activity of the smooth-muscle strips after carbachol applied cumulatively were examined and dose-response curves were plotted. The EC50 values for the circular strips from the aganglionic part of the colon were 6 x 10(-7) M in Hirschsprung's patients and 3.7 x 10(-8) M in control patients: the pD2 values were 6.23 and 7.43, respectively. This showed that the affinity of cholinoreceptors in the aganglionic part of the colon for carbachol was 16 times lower in Hirschsprung's patients as compared to control patients. The EC50 values for longitudinal strips from the ganglionic part of the colon were 2.6 x 10(-7) M in Hirschsprung's patients and 4 x 10(-8) M in control patients; pD2 values were 6.6 and 7.4, respectively. The affinity of cholinoreceptors in the smooth muscle of the ganglionic part of the colon for carbachol was also decreased (nearly 6.5 times) in Hirschsprung's patients as compared to controls.

Effect of lipid admixtures on the L-dipalmitoylphosphatidylcholine subtransition.

The effect of lipid admixtures on the properties of the L-dipalmitoylphosphatidylcholine (L-DPPC) subtransition is investigated by using high-sensitivity differential scanning calorimetry. The four admixtures used are D-DPPC, L-dipalmitoylphosphatidylethanolamine (L-DPPE), cholesterol, and palmitic acid. In all cases the subtransition decreases in enthalpy until disappearance with increase of the admixture concentrations. About 5-7 mol% of D-DPPC or palmitic acid are sufficient for abolishment (without position shifts) of the subtransition, while, on addition of L-DPPE or cholesterol, it persists up to about 20 mol% of the admixture and its disappearance is accompanied by a slight shift to higher temperatures. These data are tentatively interpreted in terms of lateral mixing of L-DPPC and admixture as indicating compound formation with D-DPPC and palmitic acid, and clustering of L-DPPE and cholesterol.

Maternal diabetes mellitus and changes in neonatal rat lung and alveolar surfactant phospholipids.

We investigated the extent of the influence of maternal diabetes on the phospholipid composition and exchange activity of the neonatal lung alveolar surfactant. The results show that each phospholipid fraction (as well as the total phospholipid content) of the surfactant of neonates with diabetic mothers are decreased to about 30% of the control values. Phosphatidylcholine and phosphatidylglycerol, which are the most important surface active phospholipid fractions, were decreased to 27% and 34% respectively. In lung tissue of the neonates with diabetic mothers, all phospholipid fractions were increased. We found that the phosphatidylcholine-exchange activity in the alveolar surfactant does not exist in neonates with diabetic mothers. This inhibited phospholipid-exchange activity may be the reason for the decrease in the surfactant phospholipids and their increase in the lungs of neonates with diabetic mothers. The cholesterol content in the surfactant of such neonates decreased by almost half in comparison with the controls, while in lung tissue it remained unchanged. Producing an experimental respiratory distress syndrome could permit to study more deeply the causes which provoke it and the accompanying metabolic changes.

Phospholipid composition of subcellular fractions and phospholipid-exchange activity in chicken liver and MC-29 hepatoma.

The phospholipid composition of mitochondria, microsomes and plasma membranes from liver and MC-29 hepatoma from White Leghorn chickens has been investigated. It was established that all mitochondria and microsome phospholipid fractions obtained from MC-29 hepatoma are increased strongly compared to those from liver. The sphingomyelin augmentation was particularly great. In hepatoma plasma membranes only the sphingomyelin quantity was increased. Sphingomyelin- and phosphatidylcholine-exchange activities were observed in avian liver for the first time. These two activities were increased in MC-29 hepatoma cells. Three phospholipid-exchange proteins have been established in chicken liver 105000 X g supernatant. One of them specifically transports phosphatidylcholine, the second one is non-specific for phosphatidylcholine and sphingomyelin, and the third one is specific only for sphingomyelin. In hepatoma cells only a non-specific phosphatidylcholine- and sphingomyelin-exchange protein was found.