Effect of ethanol exposure on Xenopus embryo lipid composition.

Exposure to ethanol is known to alter cell membranes both chemically and biophysically; these effects may be related to the development of tolerance and physical dependence. In the present study, the effects of various concentrations of ethanol (1-250 mM) on the lipid composition after the first 6 days of Xenopus embryo development were investigated, using an in vitro fertilization technique. Lipid analysis revealed: (1) a decrease of the cholesterol/phospholipid molar ratio mainly derived from a higher content of phospholipids; (2) an increase of phospholipid unsaturated fatty acids, especially C20:4 and C20:5, with ethanol concentration of 150-250 mM; (3) a decrease of lipid-bound sialic acid with ethanol concentrations of > or =5 mM. These results underline that sialoglycoconjugates are a more sensitive target of alcohol in comparison with other lipid components. The cultured embryo method certainly represents a useful model for investigation of the direct effects of ethanol on lipid metabolism, excluding maternal interference which can lead to misinterpretation of data.

Rat erythrocyte susceptibility to lipid peroxidation after chronic ethanol intake.

Erythrocytes, from 2- and 7-month-old rats chronically fed with a liquid diet containing ethanol, were analyzed for their susceptibility to lipid peroxidation estimated as thiobarbituric acid reactive substance (TBARS) products. In spite of a lower alcohol consumption, adult rats were the more affected by the ethanol treatment. Erythrocyte membranes prepared from alcoholic animals were more sensitive to lipid peroxidation than those prepared from control rats. In both age groups lipid analysis revealed similar changes: 1) an increased cholesterol/phospholipid molar ratio mainly derived from a higher content of cholesterol that accounts for the lower membrane fluidity and the higher tolerance to the disordering effect exerted by ethanol in vitro; 2) an increase of phospholipid unsaturated fatty acids, especially C20:4; 3) a modification of the phospholipid pattern, characterized in the ethanol group by an increase of PE and decrease of PE and decrease of PC levels; moreover, significant increases of the anionic phospholipids were detected in the adult group.

Age-related effects of chronic ethanol intake on physical properties, lipid composition and galactosyltransferase activity of rat small intestine microsomes.

The effect of a 4-week ethanol administration on: (1) glycoprotein content of brush border membrane (BBM): (2) galactosyltransferase activity; (3) lipid composition and fluidity of intestinal microsomes prepared from young and adult rats was investigated. In spite of a lower alcohol consumption, the more dramatic effects of treatment have been observed in the older rats, where BBM protein-bound hexoses and microsomal galactosyltransferase activity were significantly decreased. On the contrary, these parameters were unaffected in young rats. However, both rat groups were similarly affected in having their microsomal cholesterol contents significantly increased. Microsomal membranes from ethanol-fed adult rats were less fluid compared to control rats: the high fluorescence anisotropy value could be related to the high cholesterol/phospholipid ratio and to the decrease of the unsaturated fatty acids C22:4 and C22:6.

Age related changes in functions and physicochemical properties of rat jejunal brush border membrane after chronic ethanol administration.

1. We investigated the chronic effects of a 4 week treatment with ethanol on functions and physicochemical properties of BBM of young and adult rats (2 and 7 months old respectively). 2. In the ethanol treated groups the cholesterol/phospholipid and the protein/lipid ratios as well as the D-glucose uptake and lactase specific activity and Vmax were increased. In spite of a minor alcohol consumption the adult group was the more affected. 3. Membranes from the ethanol fed rats were less fluid and more tolerant to the in vitro addition of ethanol.

Role of phosphatidylethanol in membranes. Effects on membrane fluidity, tolerance to ethanol, and activity of membrane-bound enzymes.

We investigated the effect of phosphatidylethanol (PEt) on fluidity and membrane tolerance to the fluidization induced by ethanol as well as on the activity of two membrane-bound enzymes, Na+/K+ ATPase and 5'-nucleotidase. PEt was synthesized from 1,2-dimyristoylphosphatidylcholine and phosphatidylcholine from bovine brain and studies were performed to determine the optimal experimental conditions for the insertion of PEt in natural bilayers. The effects of PEt, evaluated by differential scanning calorimetry or fluorescence polarization techniques, were studied in model membranes made of synthetic phospholipids or made of total lipids extracted from rat brain crude mitochondrial fraction (P2 fraction) and from natural membranes (P2 fraction). The presence of PEt increased the fluidity of artificial as well of natural membranes, but tolerance to the addition of ethanol, displayed by dimyristoylphosphatidylcholine vesicles and by natural membranes containing PEt, was lacking in vesicles made of dimyristoylphosphatidylethanolamine and in artificial bilayers reconstituted from total P2 lipid extracts, suggesting an involvement of PC on PEt-induced ethanol resistance. Na+/K+ ATPase activity was enhanced by the addition of small amounts of ethanol (up to 50 mM) and progressively inhibited at higher concentrations, while 5'-nucleotidase was not affected up to 400 mM ethanol. The presence of PEt in the bilayer exerted the opposite effects on the two enzymes, reducing the Na+/K+ ATPase activation induced by ethanol and enhancing 5'-nucleotidase activity. The mechanisms of the PEt-induced modifications are discussed.

L-carnitine and carnitine ester transport in the rat small intestine.

L-carnitine and its esters (acetyl-L-carnitine and propionyl-L-carnitine) at pharmacological doses (1, 5 and 10 mM) are absorbed by the rat jejunum by simple diffusion. Partition coefficients of carnitine esters determined in lipophilic media (diethyl ether/water and olive oil/water) are greater than that of L-carnitine. It would therefore seem that esters diffuse more easily through the lipid component of the intestinal barrier. The transport of acetyl- and propionyl-L-carnitine at pharmacological doses seems to be linearly and positively correlated with K+ transport but not with Na+ transport.

Postnatal maturation of rat intestinal membrane: lipid composition and fluidity.

1. We studied the lipid composition and the fluidity of small intestine brush border membrane (BBM) of rats of different age: 'very young' (5-7 weeks old), 'young' (9 weeks old), 'adult' (30 weeks old) and 'old' (85 weeks old). 2. Fluorescence anisotropy, as assessed by 1,6-diphenyl-1,3,5-hexatriene probe (DPH), was increased from very young to adult rats. 3. In agreement with these results the lipid composition in adult animals showed a lower lipid/protein ratio (derived mainly from a lower content of total polar lipids) and an increase of cholesterol esters and sphingomyelin (SM) saturation index. 4. A marked decrease of the order parameter was observed in the 'old' group, accompanied by a decreased cholesterol/phospholipid ratio. 5. The percentage distribution of membrane phospholipids significantly changed during development, but the modifications were not correlated with the anisotropy of DPH.

Effects of cholesterol and alfa-2-recombinant interferon on cell growth and cell antigen production in the HT 29 cells, an established cell line of human colon adenocarcinoma.

HT 29 cells, an established cell line of human colon adenocarcinoma, were grown in RPMI 1640 medium without or with cholesterol at 25, 50, 100 micrograms/ml concentrations. In some experiments 100 or 200 U/ml alfa-2-A recombinant Interferon were added to the medium. Only in the case of the highest cholesterol concentration there was a reduced number of cells at confluence. Moreover, only the production of CEA increased in the presence of cholesterol. Interferon did not affect cell growth appreciably but stimulated CEA release into the medium during the first three days of culture. Morphological analysis of cells in the presence of cholesterol seems to indicate an attempt of the cells to differentiate.

Effects of semistarvation on transintestinal D-glucose transport and D-glucose uptake in brush border and basolateral membranes of rat enterocytes.

The present work shows that semistarvation (8-10 g of food for 10 days) increases net D-glucose, Na+ and water transport in the everted and perfused rat jejunum. A linear and positive correlation between cell sugar concentration and transport was found in control and semistarved rats, but the phenomenon was more relevant only in semistarved animals. Membrane vesicle experiments showed that semistarvation increases sugar overshoot only in brush border membrane vesicles, while this situation does not occur in basolateral membrane vesicles. The effect of partial food deprivation seems to enhance net sugar transport by increasing sugar entry across the apical membrane of enterocytes.

Intestinal sugar transport during ageing.

Ageing effects on sugar intestinal transport were studied by using the everted sac and the brush-border membrane vesicle techniques. Four age groups of rats were used: very young, young, adult and old animals. Net transintestinal transport of D-glucose and intracellular sugar accumulation were greater in young than in very young, adult and old rats. Net Na+ transport was high in very young and young animals and then it declined with age. In brush-border membrane vesicle experiments D-glucose overshoot was smaller in the groups of animals where net sugar transport was less. In old rats, however, the overshoot did not occur. Short-circuiting of vesicles with valinomycin showed that the driving forces for sugar accumulation, i.e. the chemical potential gradient of Na+ and the electrical potential gradient, played different roles during ageing. In very young animals the chemical potential gradient seems to be responsible for D-glucose overshoot; in young rats both gradients are important while in adult animals the electrical potential gradient represents the main driving force.

Effect of maternal diet on ganglioside distribution in fetal rat brain.

Female rats were fed a diet with low protein content or with low and high amount of essential fatty acids (EFA). Ganglioside content and distribution were analyzed in the brain of animals at two different periods of fetal life (15th or 20th day). In the fetuses from mothers fed the diet with low amount of EFA the content of ganglioside is significantly lower than in the control group. The three diets resulted in the modification of ganglioside pattern, mainly for the animals on the 15th day of gestation.