Altering erythrocyte membrane composition with phospholipid exchange protein

• 1.1. In establishing a protocol for the use of a phospholipid (PL) 2 exchange protein (PLEP) preparation to incorporate fluorescent lipids into membrane fragments, the potential for the PLEP preparation to alter membrane composition was investigated.
• 2.2. Cholesterol was removed from erythrocyte ghosts and PL content was increased after incubation of the membranes with phosphatidylcholine (PC) vesicles and a nonspecific bovine liver PLEP preparation.
• 3.3. The elevated PL/cholesterol ratio of the ghosts decreased fluorescence polarization of diphenylhexatriene (DPH), indicating an increased average membrane fluidity.
• 4.4. The increased PL/cholesterol ratio and reduced DPH fluorescence polarization were prevented by including cholesterol in the PC vesicles during the initial incubation of ghosts with PLEP preparation and lipid.
• 5.5. In addition, the PC content of ghost membranes was elevated after incubation with PLEP preparation and either PC or mixed PC-cholesterol vesicles.

     

Na+-independent sugar uptake by rat intestinal and renal brush border and basolateral membrane vesicles

• 1.1. Uptake of [¹⁴C]-labelled d-glucose, l-arabinose and d-fructose by intestinal and renal brush border and basolateral membrane vesicles was studied in the absence of Na⁺ .
• 2.2. The Na⁺-independent d-glucose transport system in these membrane vesicles was saturable, sensitive to phloretin, stereospecific and accessible only to d-glucose and d-galactose.
• 3.3. Na⁺-independent l-arabinose transport was not saturable even when its concentration was raised to 300 mM and it was insensitive to phloretin.
• 4.4. Na⁺-independent d-fructose transport demonstrated saturation kinetics with only renal brush border membrane vesicles, but it was not inhibited by either phloretin or phlorizin.
• 5.5. These studies indicated that the Na⁺-independent carrier-mediated d-glucose/d-galactose transport system of intestinal and renal brush border and basolateral membranes is clearly not shared by other monosaccharides.

     

Investigation of phycocyanin-membrane interaction: Promotion of membrane interactions

• 1.1. In a continuing investigation of phycocyanin-membrane surface interaction, fluorescence quenching experiments were performed with a mixture of two populations of fluorescence probe-encapsulated phospholipid bilayer vesicles in the presence and absence of phycocyanin.
• 2.2. These membrane vesicles were prepared with 1,2-dimyristoyl phosphatidylcholine (DMPC), cholesterol and a probe molecule.
• 3.3. A fluorophore was encapsulated in one population of membrane vesicles, while a quencher was encapsulated in another population of membrane vesicles.
• 4.4. The result was compared with those of experiments in the presence of other biomolecules, including albumin, cytochrome c, hemoglobin, myoglobin or RNA.
• 5.5. Interestingly, a one-third reduction of the fluorescence intensity was observed in the mixture of these two populations of membrane vesicles in phycocyanin's presence.
• 6.6. In contrast, the other biomolecules caused no significant reduction in the fluorescence intensity.
• 7.7. These findings were evidence of a phycocyanin-induced membrane perturbation.
• 8.8. This was further demonstrated by a phycocyanin-induced change in the thermotropic behavior of DMPC vesicles, as measured by differential scanning microcalorimetry.
• 9.9. Such a unique property of phycocyanin is believed to be associated with its known membrane surface-interacting character.
• 10.10. A possible phycocyanin-modulated membrane-membrane interaction was discussed.

     

The isolation of skeletal muscle plasma membranes from rainbow trout (Salmo gairdneri)

• 1.1. Plasma membranes were isolated from caudal flank skeletal musculature of rainbow trout by discontinuous sucrose gradient centrifugation.
• 2.2. Na⁺−K⁺-ATPase was enriched 8-fold and 5′-nucleotidase activities 4-fold in a fraction isolated at the 8–25% sucrose interface.
• 3.3. A cholesterol: phospholipid ratio of 0.37 in the plasma membrane fraction was 85% greater than that observed in adjacent subcellular fractions.
• 4.4. Electron microscopy provided morphological confirmation of enrichment and integrity of skeletal muscle plasma membranes at the 8–25% sucrose interface.

     

Effect of caffeine,theophylline and nicotine on d-glucose and folate transport in rat jejunal brush border membrane vesicles

• 1.1. Nicotine at 10 mM, but not caffeine or theophylline, reduced by 20% the overshoot of the Na⁺-dependent d-glucose transport in ratjejunal brush border membrane vesicles.
• 2.2. Since nicotine did not affect the transport of Na⁺, its inhibition on Na⁺-dependent d-glucose transport must be due to a direct effect upon the d-glucose transport system.
• 3.3. Folate transport in these membrane vesicles was found to a be a free diffusion process at pH 7.4.
• 4.4. Neither caffeine, theophylline nor nicotine has any effect on folate transport.

     

The influence of calcium and magnesium on sea bass (Dicentrarchus labrax) intestinal brush border membrane purification and activity

• 1.1. The activity of brush border enzymes (alkaline phosphatase, maltase, sucrase, trehalase, leucine amino peptidase) was higher in purified membranes prepared with calcium. The contamination of these membranes with basolateral membranes was also lower (1.27 for Na-K-ATPase activity ratio).
• 2.2. The extraction of brush border lipids was carried out according to Folch adapted method. Two dimensional thin layer chromatography was used to separate the phospholipidic fractions. Fatty acids of phospholipids were analysed using gas chromatography after acid transmethylation (column SP 2330).
• 3.3. Phospholipids are composed of phosphatidylcholine (PC: 33%), phosphatidylethanolamine (PE: 30%), sphingomyeline (SM: 21%), phosphatidylserine (PS: 14%) and phosphatidylinositol (PI: 2%). 4. PC, PE and PS are characterized by high levels of unsaturated fatty acids (monounsaturated MUFA: 21.5% and polyunsaturated PUFA: 34.9%). The most abundant PUFA belong to the (n-3) family [18:3 (n-3), 20:5 (n-3) and 22:6 (n-3)].
• 4.5. Fatty acids from sphingomyelin of purified membranes have low proportions of PUFA (13.5%) but higher proportions of MUFA (39.5%).
• 5.6. No specific differences were found between calcium and magnesium prepared membranes.
• 6.7. The low content in LPC and the absence of LPE confirmed the absence of major structural lipids transformation during the membrane purification with calcium or magnesium.
• 7.8. Glycine transport was measured during 10 sec at different temperatures using the rapid filtration technique. Glycine transport was higher with Na⁺ than with K⁺. In the presence of Na⁺, this transport increases with temperature.
• 8.9. Arrhenius curves were mono phasic without obvious breakpoint and indicated no phase transition in the lipid bilayer.
• 9.10. A significant Na⁺ dependent glycine transport has been characterized at low temperatures (0°C) which suggests a possible role of membrane polyunsaturated fatty acids in the control of glycine transport.

     

Na+/H+ exchange in the kidneys of eels (Anguilla anguilla) adapted to sea water or to freshwater environments: Studies with brush border membrane vesicles

• 1.1. In brush border membrane vesicles isolated from eel kidneys, adapted either to sea water or freshwater environments, a Na⁺/H⁺ antiporter is present.
• 2.2. Using a calibration plot it is possible to evaluate the amount of protons that this antiporter can accumulate inside the vesicular space.
• 3.3. The activity of the antiporter seems to be affected by the salinity of the water; it is higher in animals adapted to seawater.
• 4.4. This adaptation seems to occur by a J_max regulation of the antiporter.

     

Interaction of N-methyl-anthraniloyl-labelled porcine apolipoprotein A-I with porcine lecithin: Cholesterol acyltransferase: An energy transfer study

• 1.1. To investigate whether a direct protein-protein interaction between apoA-I and lecithin: cholesterol acyltransferase (LCAT) is necessary for the activation of the enzyme, apoA-I was labelled with N-methylisatoic anhydride at lysine residues. The intermolecular resonance energy transfer from tryptophan residues of LCAT (donor) to N-methyl-anthraniloyl (NMA)-labelled apoA-I (NMA-apoA-I) (acceptor) was used as a sensitive fluorescence method for studying molecular interactions.
• 2.2. In the absence of lipids no fluorescence energy transfer was measurable.
• 3.3. Fluorescence energy transfer occurred from LCAT to NMA-apoA-I in the presence of liposomes with phospholipid/cholesterol ratios ranging from 5:1 to 18:1 and regardless whether only 1 or up to 5 NMA-apoA-I molecules resided at the liposome surface.
• 4.4. This indicates a preferred binding of the enzyme directly to or in spatial proximity to the activator protein NMA-apoA-I even if enough space at the liposome surface is available to allow LCAT binding at a distance, where no energy transfer is measurable.

     

On the mechanism of decreasing the lithogenicity of bile in Camelus dromedarius

• 1.1. Analysis of camel bile revealed that it is highly concentrated.
• 2.2. The molar fraction of phospholipids in camel bile was low and cholesterol was very high relative to rat bile.
• 3.3. Cholate is the main primary bile acid secreted. Moderate quantities of the secondary bile acid deoxycholate were found but no lithocholate was detectable possibly secondary to rapid recycling of the bile acid pool in the enterohepatic circulation.
• 4.4. Glycoconjugated bile acids predominated over tauroconjugates.
• 5.5. The hepatic bile from the camel may be concentrated as part of the general mechanism of water conservation exhibited by that species. The increased concentration of bile acids helps maintain cholesterol in solution, thus reducing lithogenicity.

     

On the turnover of tissue lipids in the mouse during dietary deprivation

• 1.1. The double isotope ratios of individual lipid fractions in major tissues of mice were determined before and after a period of total dietary deprivation.
• 2.2. Several significant alterations in these ratios were caused by this treatment, with a marked individuality evident in the response of separate tissues.
• 3.3. The increased metabolic flux overall was contributed to by increased rates of lipid degradation in liver, and active mobilization from the peripheral depots.
• 4.4. Significant utilization of phospholipid fractions for energy purposes was also noticeable, however. along with tissue specific variation in the patterns of both double isotope ratios and specific radioactivities.
• 5.5. These data are discussed in relation to the comparative priorities of synthesis and degradation in the different tissue and lipid sources in response to this physiological perturbation.

     

Factors influencing lipid levels and lecithin:cholesterol acyltransfer in plasma of the lizard,Tropidurus torquatos

• 1.1. Lipid concentrations and lecithin:cholesterol acyltransferase (LCAT) activity in the plasma Tropidurus torquatos were remarkably variable.
• 2.2. Both lipid levels and LCAT activity were highest for lizards collected during the early rainy season (March–April) than during other seasons, and were higher for females than for males.
• 3.3. Plasma lipid levels and LCAT activity were significantly and inversely correlated with body weight (age) of male lizards, this being associated with an apparent change to an herbivorous diet in older males.
• 4.4. During prolonged fasting, plasma lipid levels and lecithin:cholesterol acyltransfer (LCAT) and hepatic phospholipids were markedly reduced.
• 5.5. LCAT activity in plasma of fasted and non-fated lizards was significantly correlated with the molar proportion of PC to UC, suggesting that the apparent low LCAT in plasma of fasted lizards is partly due to depletion of PC in the lipoprotein substrates.

     

Biphasic action of growth hormone on intestinal amino acid absorption in striped bass hybrids

• 1.1. Weekly injections of bovine growth hormone (bGH) increased the maximal transport rate of both Na⁺-dependent and Na⁺ -independent l-leucine transport with little effect on the affinity constants in the intestine of striped bass hybrids.
• 2.2. The Na⁺-dependent and the Na⁺-independent transport of the non-metabolizable analog cycloleucine was also stimulated by bGH.
• 3.3. The Na⁺ -dependent active transport was stimulated 2 days after the hormone treatment, while the stimulation of the Na⁺-independent diffusional transport was not observed until after 2 weeks of treatment.
• 4.4. Studies of intestinal morphometry and l-leucine transport using brush border membrane vesicles suggested that bGH affects intestinal amino acid absorption initially by increasing the number of transporters per cell.
• 5.5. This phase is followed by a general increase of the intestinal mass after long-term treatment with the hormone.

     

Phosphatase activity in the hepatopancreas of Helix aspersa

• 1.1. Phosphatase acid (PhA) activity in the digestive gland (hepatopancreas) of the common garden snail Helix aspersa has been investigated using cytochemical methods.
• 2.2. All the cells composing this gland show PhA activity, the distribution pattern differing according to the cell type.
• 3.3. The digestive cells show the most widely distributed reaction product (brush border, phagolysosomes, multivesicular bodies and autophagic vacuoles).
• 4.4. In the excretory cells this activity appears in large sacs, while in the calcium cells the reaction product is abundant in the calcium granules.
• 5.5. Cellular digestion processes performed by each of these cell types is discussed together with their role in the detoxification of heavy elements derived from the environment.

     

Subcellular localization of monoglyceride acyltransferase,xanthine oxidation,NADP: Isocitrate dehydrogenase and alkaline phosphatase in the mucosa of the guinea-pig small intestine

• 1.1. Rate dependent and isopycnic banding in a zonal rotor were used to analyse the subcellular sites of enzymes in homogenates of guinea-pig small intestinal mucosa.
• 2.2. The results demonstrate the following localizations: monoglyceride acyltransferase—microsomal; xanthine oxidase and dehydrogenase—soluble phase, and NADP: isocitrate dehydrogenase—soluble phase and mitochondrial.
• 3.3. Alkaline phosphatase is confined to brush borders and is absent from the basolateral plasma membrane. A variable proportion of the activity, up to 40% is on brush borders which during homogenization break up into particles of reduced density and slow sedimentation rate.

     

Effect of ozone exposure on the compositions of gill and erythrocyte membrane lipids and proteins of Japanese charr (Salvelinus leucomaenis)

• 1.1. Compositions of lipids and proteins of erythrocytes (RBC) and gills from Japanese charr (Salvelinus leucomaenis) which were exposed to 0.4 and 0.7 ppm ozone for 30 min were compared with those of the control.
• 2.2. On exposure to ozone, both RBC and gill membrane phospholipid content, especially phosphatidylethanolamine (PE), dropped.
• 3.3. The decrease of PE was brought about by the decrease of docosahexaenoic acid content which comprised the major component of PE.
• 4.4. RBC membrane protein with 215 and 225 kDa, which is equivalent to cytoskeletal protein, selectively disappeared on exposure to ozone.

     

Effect of dietary coconut oil on lipoprotein composition of young chick (Gallus domesticus)

• 1.1. The composition of HDL, the major lipoprotein fraction from chick serum, drastically changed after 2 weeks of coconut oil feeding. Total cholesterol and triacylglycerols significantly increased following dietary 10 or 20% coconut oil supplementation.
• 2.2. Changes in LDL composition were less profound, cholesterol being the only component that increased by coconut oil supplementation (10 or 20%).
• 3.3. IDL proteins were the only components that increased following the same dietary treatment (20%).
• 4.4. VLDL cholesterol and proteins also increased after 1–2 weeks of 20% coconut oil supplementation to the diet.
• 5.5. Of total lipoproteins, the cholesterol content strongly increased after dietary treatment, while triacylglycerols did not change significantly.

     

Plasma lipoprotein profiles and arylesterase activities in two inbred strains of rabbits with high or low response of plasma cholesterol to dietary cholesterol

• 1.1. Cholesterol feeding for 4 weeks of female and male rabbits of two inbred strains increased plasma cholesterol concentrations by about 11 and 48 mmole/I in the hypo- and hyperresponsive strain, respectively.
• 2.2. On the low-cholesterol pre-experimental diet, the hyporesponsive animals had significantly higher plasma HDL (high density protein) cholesterol levels than hyperresponders.
• 3.3. In both strains, cholesterol feeding caused elevations of cholesterol in all lipoprotein classes, the difference between the hypo- and hyperresponsive strains in essence only being observed in the VLDL (very low density lipoprotein) fraction.
• 4.4. Basal plasma total arylesterase activity was significantly higher in the hypo- than in the hyperresponsive rabbits.
• 5.5. Dietary cholesterol caused an increase in plasma esterase activity in both strains.
• 6.6. We suggest that in rabbits a low plasma arylesterase activity and a low concentration of HDL cholesterol are associated with an increased sensitivity to dietary cholesterol.

     

Lipid changes in blood serum and tissues of the Egyptian Cobra “Naja haje haje” during the hibernation cycle

• 1.Total lipids, free fatty acids, triglycerides, phospholipids and total cholesterol in blood serum, liver, brain, cardiac and skeletal muscles of Naja haje haje were determined during the different phases of the hibernation cycle.
• 2.A sharp decrease in the level of total lipids of blood serum and all tissues occurred during hibernation. Upon arousal, lipogenesis is commonly restored.
• 3.Elevated concentrations of serum free fatty acids predominated in pre-hibernation and hibernation periods, while the tissues recorded highly significant declines during hibernation.
• 4.Occurrence of marked decreases in triglycerides contents of serum and tissues except the cardiac muscles in the hibernation and arousal phases.
• 5.Sharp increases in the phospholipid contents of blood and the selected tissues were recorded during hibernation. The level declined in both liver and cardiac muscles in arousing animals.
• 6.Total cholesterol level was lowered in blood during hibernation. The cardiac muscles showed a highly significant decrease while liver, brain and skeletal muscles showed elevations in the same phase.

     

Water diffusion,ion transport and lipid composition of the skin of Rana cyanophlyctis Schneider under osmotic stress

• 1.1. Analysis of the total lipid content (TL), components of the neutral lipid fraction (NLF), phospholipid fraction (PLF), recordings of electrical potential differences and diffusional permeability were carried out in the skin of the aquatic frog Rana cyanophlyctis subjected to in vivo salt stress (0.9 sodium chloride) for different durations (0, 1, 3 and 7 days).
• 2.2. A general decrease of skin TL and of components of the NEE and PEE was observed.
• 3.3. Stoichiometric ratios for skin PLF components under initial salt stress of different durations reveal an increase of the ratios of sphingomyelin and phosphatidyl choline after osmotic stress.
• 4.4. The diffusional permeability of water increased following exposure to salt stress of I, 3 and 7 days duration.
• 5.5. The transepithelial potential difference measured in vitro after a salt stress of 3 days was considerably higher than the controls.

     

Plasma membrane: Can its structure and function be modulated by dietary fat?

• 1.1. Compositional analysis of plasma membranes from rats fed nutritionally adequate diets different in fatty acid composition establishes that fundamentally different dietary fat intake results in alteration in structural lipid composition of plasma membranes in brain, liver and the intestinal mucosa.
• 2.2. Dietary differences in fatty acid intake altered the fatty acyl tail composition of plasma membrane phospholipids in brain, liver and intestinal mucosa.
• 3.3. Diet altered the phospholipid profile observed in brain synaptosomal and liver plasma membrane.
• 4.4. Feeding high vs low polyunsaturated to saturated fat diets for 7 days altered the fatty acid composition of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin and mono-glucosylceramide isolated from plasma membrane of the intestinal mucosa