Effect of choline,ethanolamine and serine supplementation on the membrane properties ofMicrosporum gypseum

Phospholipid bases, choline and ethanolamine, when supplemented in the growth medium ofMicrosporum gypseum resulted in an increase in the corresponding phospholipid and total phospholipid content. However, when serine was supplemented, marginal changes were observed. The fatty acid profile of phospholipids remained almost unchanged. The uptake of lysine, leucine and aspartic acid in the spheroplasts of choline- and ethanolamine-grown cells was higher as compared to the control. 1-Anilinonaphathalene-8-sulfonate (ANS) binding to the spheroplast membrane, as calculated from Scatchard plots, demonstrated an increase in the number of binding sites in choline- and ethanolamine-grown cells, while a decrease was observed in the serine-supplemented cells. The results are discussed in terms of the effect of phospholipid polar head group composition on the membrane structure and function of this fungus.     

Lipids of dermatophytes. III. Sterol-induced changes in the lipid composition and functional properties ofEpidermophyton floccosum

Sterol supplementation, alone or in the presence of cerulenin, resulted in an increase in the total sterol content ofEpidermophyton floccosum. While the total phospholipid levels ofE. floccosum exhibited only marginal changes with sterol supplementation, the fatty acid profiles of these phospholipids were highly varied. In the presence or absence of cerulenin, the oleic acid content of phospholipids were increased significantly by cholesterol supplementation, whereas linoleic acid levels were enhanced by ergosterol supplementation. These variations resulted in higher unsaturated/saturated phospholipid fatty acid ratios in sterol-supplemented cells. The uptake of labeled amino acids (aspartic acid, lysine, glycine) was influenced by sterol supplementation. Alterations in the number of binding sites for the membrane probe, 1-anilino-naphthalene-8-sulfonate (ANS), were seen based on Scatchard plot calculations. The results indicate a correlation between sterol-induced changes in membrane lipid composition and function.     

The effect of antioxidant deficiency on tissue lipid composition in the rat. II. Liver

The hepatic phospholipids of the antioxidant-deficient rat fed a source of both linoleate and linolenate showed a progressive net decrease in eicosapentaenoate, a progressive net increase in arachidonate, and there was a concomitant accumulation of fluorescent pigment of the lipofuscin or ceroid type in the tissue. An increased incorporation of isotopically labeled acetate into both the tetraenoic and pentaplus hexaenoic acid fractions was also noted, indicating that the disappearance of polyunsaturated fatty acids was partially countered by increased synthesis. Comparable results were obtained on diets containing either suboptimum or adequate levels of biologically available selenium. Vesicular dilation of the endoplasmic reticulum was noted in animals fed the tocopherol-deficient diet. In separate experiments using a necrogenic diet containing torula yeast, these subcellular alterations were found to be prevented by tocopherol but not by selenium, although selenium supplementation did prevent macroscopically observable damage.     

Effects of cyclopropenoid fatty acids on fungal growth and lipid composition

Cyclopropenoid fatty acids (CPE) isolated fromSterculia foetida oil by urea clathration and reverse phase high performance liquid chromatography (HPLC) were introduced into fungal cultures. Stearate levels in phospholipids and triacylglycerols fromUstilago maydis sporidia rose considerably in response to 30 μM CPE. In addition, CPE themselves were incorporated into glycerolipid fractions. Sterol composition was unaffected. Changes in lipid composition were accompanied by inhibition of dry weight accumulation and sporidial number. Treated sporidia showed irregular wall deposition and a branched morphology. Oleate alleviated CPE effects on growth and morphology. Hyphal extension byRhizoctonia solani was inhibited somewhat by 30 μM sterculate, whileFusarium oxysporum showed no appreciable response. Although CPE appeared to inhibit fatty acid desaturation byF. oxysporum, gross increases in the proportion of stearate were limited to the triacylglycerol fraction during 30 μM treatments. The possibility that the CPE synthesized by plants serve as antifungal agents is discussed.     

Lipids of cultured hepatoma cells: I. Effect of serum lipid levels on cell and media lipids

Minimal deviation hepatoma cells were cultured as monolayers to confluency in roller flasks containing modified Swim's medium, supplemented with decreasing amounts of serum, lipid-free serum, and lipid-free serum containing added fatty acids. Good cell growth was observed until serum levels fell below 5% of the medium. Media containing lipid-free serum or lipid-free serum plus linoleic or palmitic acids did not support good growth. Lipids were extracted from cells; media, obtained during the first and last half of the incubation period, resolved into neutral and phospholipid fractions; fatty acid composition of each fraction analyzed by gas liquid chromatography; and lipid class distributions compared by thin layer chromatography. The data showed that the media contained more neutral lipids and phospholipids after incubation than initially, indicating that minimal deviation hepatoma cells excreted lipids into the media. The class composition of the excreted lipids resembled that of the serum. A comparison of media, cells, and serum fatty acid compositions indicated that the lipids secreted into the media were of cellular origin. Although some differences were noted, in general, cells grown on the nine different media had the same ca. neutral lipid and phospholipid class and fatty acid compositions. In contrast, dramatic differences were observed in the class and fatty acid compositions of the serums from that of the cells and media. These results indicate that exogenous serum lipids had little influence on cellular class and fatty acid compositions of the minimal deviation hepatoma cells. This neoplasm did not contain detectable levels of glyceryl ether diesters, indicating that this compound is not characteristic of all tumors. Lipid class profiles and fatty acid compositions of cells grown on various media suggest that the minimal deviation hepatoma cells can synthesize most, if not all, neutral lipid and phosphoglyceride classes found in liver. Presented at the AOCS Meeting, New Orleans, April 1973.     

Effect of medium-chain fatty acid positional distribution in dietary triacylglycerol on lymphatic lipid transport and chylomicron composition in rats

The present study was carried out to examine if the positional distribution of medium-chain fatty acid (MCF) in dietary synthetic fat influences lymphatic transport of dietary fat and the chemical composition of chylomicrons in rats with permanent cannulation of thoracic duct. Four types of synthetic triacylglycerol were prepared: (i) sn-1(3) MCF-sn 2 linoleic acid, (ii) interesterified sn-1(3) MCF-sn 2 linoleic acid, (iii) sn-2 MCF-sn-1(3) linoleic acid, and (iv) interesterified sn-2 MCF-sn-1(3) linoleic acid. A purified diet composed of equal amounts of the synthetic fat and cocoa butter was given to rats with permanent lymph duct cannulation. The positional distribution of MCF in the dietary fat had no significant effect on the lymph flow, triacylglycerol output, phospholipid output, lipid composition of chylomicrons, or the particle size. The positional distribution of MCF in the synthetic triacylglycerol was maintained in the chylomicron triacylglycerol. These results showed that MCF in the dietary triacylglycerol is transported into lymphatics and the positional distribution is well preserved in chylomicron triacylglycerol.     

Effect of fatty acid positional distribution and triacylglycerol composition on lipid by-products formation during heat treatment: II. Trans isomers

This study examined the effect of the fatty acid positional distribution and of the triacylglycerol (TG) composition on heat-induced trans isomerization of linoleic and linolenic acids. For this, we synthesized diacid TG molecules that were acylated only with linoleic acid (L) or with linolenic acid (Ln) along with palmitic acid (P). The fatty acid of interest was positioned either in the central position (PLP and PLnP, respectively) or in one of the two outer positions (PPL and PPLn, respectively). Monoacid TG, i.e., trilinolein and trilinolenin, were also synthesized and mixed with tripalmitin in a 1:2 ratio. This model TG was also compared to another TG model, which consisted of a canola oil and its randomized counterpart whose fatty acid positional distribution and TG composition were determined by means of high-performance liquid chromatography. After heating, the content of trans isomers was determined by gas-liquid chromatography with a polar capillary column. In model TG, polyunsaturated fatty acids in monoacid TG (LLL and LnLnLn) exhibited the highest degree of isomerization, compared to diacid TG, and this effect was greatest at 220°C. At this temperature, an effect of the TG structure was observed only with linolenic acid. In that situation, 18:3n-3 acylated in the central position of the TG molecule (PLnP) displayed the highest sensitivity to trans geometrical isomerization. Although to a lesser extent, the same trends as for the pure TG model were observed with the canola oil model with regard to the influence of the fatty acid positional distribution and TG molecular species.     

Effect of EFA deficiency on lipid transport from liver

Studies are reported of the effect of an essential fatty acid (EFA) deficiency on synthesis of triglycerides (TG) and phospholipids (PL) and secretion of these compounds by livers of male Sprague-Dawley rats. Animals were fed a semipurified diet containing corn oil or hydrogenated coconut oil (HCO) as the sole source of fat or no fat from weaning to 20 weeks of age. Liver function of the animals in each group was compared by an isolated liver perfusion technique with perfusates containing erythrocytes and linoleate, and in vivo experiments via tail vein injection of palmitate-³H. Perfusion experiments showed that an EFA deficiency reduced the ability of the liver to secrete TG and PL. Accumulation of TG in the liver and its diminished secretion into the blood of EFA deficient animals were demonstrated by in vivo experiments with palmitate-³H. The rate of conversion of linoleate to arachidonate and synthesis of PL was greater in livers of EFA deficient rats than in the control, corn oil fed animals. The results suggest a relationship of EFA metabolism to lipid transport. One of five papers to be published from the Symposium “Lipid Transport” presented at the AOCS Meeting, New Orleans, April 1970.     

Lipids of cultured hepatoma cells: II. Effect of media lipids on cellular phospholipids

Minimal deviation hepatoma cells were cultured in a modified Swim's 77 medium supplemented with decreasing amounts of serum, lipid-free serum, and lipid-free serum containing added palmitic or linoleic acids. Cellular phospholipids were extracted and the class distribution determined quantitatively. The fatty acid composition of each phospholipid class was determined, and the percentages from cells grown on each of the various media were compared. Cellular phospholipid class and fatty acid compositions differed from media compositions, indicating that intact serum phospholipids are not incorporated into cellular structures. Phosphatidylcholine percentages decreased as the media serum and lipid levels decreased, while phosphatidylinositol and phosphatidylethanolamine percentages increased. Sphingomyelin of cells grown in medium containing added linoleic acids contained a high level of a 24∶2 acid. All classes, except sphingomyelin, contained elevated levels of 18∶1 acid and decreased levels of polyunsaturated fatty acids, relative to normal rat liver. Cells cultured on lipid-free medium did not contain increased concentrations of 20∶3 acid, suggesting that this hepatoma cell cannot desaturate monoenoic acids. Phosphoglycerides of cells, grown on lipid-free medium, had the highest monoene fatty acid concentration, whereas those cells grown on media containing added linoleic acid had the lowest concentrations, suggesting that linoleate may inhibit or regulate monoenoic acid biosynthesis in this cell. These mass data also demonstrate that monoenoic fatty acid biosynthesis in this cultured hepatoma cell responds to dietary changes.     

Effect of essential fatty acid deficiency on lipid composition of basolateral plasma membrane of pig intestinal mucosal cells

The effect of essential fatty acid (EFA) deficiency on the lipid composition of basolateral plasma membranes (BPM) from intestinal mucosal cells was investigated in weaning pigs fed control or EFA-deficient diets for 12 weeks. The phospholipid and cholesterol contents relative to protein were similar in both groups, showing a cholesterol/phospholipid molar ratio of 0.6. The distribution of phospholipid classes was also unaffected by the diet. In contrast, fatty acid profiles of the two phospholipid main classes, phosphatidylcholine and phosphatidylethanolamine were altered by EFA deficiency. Linoleic acid (18∶2n−6) was largely reduced, whereas arachidonic acid (20∶4n−6) only slightly decreased in EFA-deficient pigs. The unsaturation index was essentially maintained by high levels of oleic acid (18∶1n−9) and by conversion of oleic acid to 5,8,11-eicosatrienoic acid (20∶3n−9). Finally, during the period of EFA deficiency, the lipid composition of BPM of the intestinal mucosal cells was little affected, suggesting a preferential uptake of 20∶4n−6 and (or) precursor mobilized from other tissues. However, an effect of dietary treatment on the function of membrane-associated proteins cannot be ruled out.     

The effect of dietary zinc deficiency on the lipid composition of the rat erythrocyte membrane

The effect of dietary zinc deficiency in the rat on the lipid composition of the erythrocyte membrane was determined. Weanling male Wistar rats were fed an egg whitebased diet containing <1.0 mg Zn/kg dietad libitum. Control rats were either pair-fed orad libitum-fed the basal diet suppelemented with 100 mg Zn/kg diet. A zinc refed group was fed the −Zn diet until day 18 and then pairfed the +Zn diet until day 21. The voluntary feed restriction associated with dietary zinc deficiency resulted in erythrocyte membranes that had depressed phospholipid/protein and elevated cholesterol/phospholipid ratios. Similarly, all feed restricted groups had elevated 22-carbon n−3 polyunsaturated fatty acid (PUFA) and depressed 22-carbon n−6 PUFA concentrations in alkenylacyl and diacyl glycerophosphoethanolamine, phosphatidylserine and phosphatidylcholine; they also had depressed 24∶2n−6 levels in sphingomyelin. The relative concentrations of phospholipids in the membrane was similar between −Zn and +Zn (ad libitum) groups; however, the −Zn group had significantly less phosphatidylserine relative to +Zn (pair-fed) controls.     

Effect of hypothyroidism on the lipid composition of rat plasma and erythrocyte membranes

The effect of hydrothyroidism on plasma and erythrocyte membrane lipid components has been investigated. This pathological state is accompanied by a) a cholesterol increase of about 60% in plasma, and at the same time a 22% reduction in erythroycte membranes; b) 44% and 30% phospholipid level decreases in both plasma and red cell membranes, respectively; and c) almost unaffected phospholipid and fatty acid compositions of both plasma and erythrocyte membranes. All changes were corrected by treatment of the hypothyroid rats with triiodothyronine for two days. These findings suggest that in hypothyroid rats a reduced transfer of cholesterol from plasma to erythrocyte membrane probably takes place. This could explain, at least in part, the increased hematic cholesterol level observed in hypothyroid animals. In red cell membranes, the simultaneous decrease in cholesterol and phospholipid levels does not alter the cholesterol/phospholipid molar ratio, thus avoiding their abnormal function.     

Study on morphology of ternary polymer blends. II. Effect of composition

The composition effect on morphology of polypropylene/ethylene–propylene–diene terpolymer/polyethylene (PP/EPDM/PE) and polypropylene/ethylene–propylene–diene terpolymer/polystyrene (PP/EPDM/PS) ternary blends has been investigated. In all of the blends, polypropylene as the major phase was blended with two minor phases, that is, EPDM and PE or PS. From morphological studies using the SEM technique a core–shell morphology for PP/EPDM/PE and separated dispersed morphology for PP/EPDM/PS were observed. These results were found to be in agreement with the theoretical predictions. The composition of components affected only the size of dispersed phases and had no appreciable effect on the type of morphology. The size of each dispersed phase, whether it forms core or shell or disperses separately in matrix, can be related directly to its composition in the blend. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 82: 1138–1146, 2001     

Effect of the environment and fasting on the lipid and fatty acid composition ofDiplodom patagonicus

Some effects of food, habitat and temperature on the lipid composition of a freshwater mollusk,Diplodom patagonicus, were studied. Animals kept and fasted up to 60 days in an aquarium at 9 C and 20 C showed a decrease of the total lipid content that corresponded to a decrease of triacylglycerols and diacylglycerol ethers. This decrease evoked an increase of polar-to-nonpolar lipid ratio. However, no significant change in the total fatty acid composition was shown. Moreover, a decrease of temperature from 20 C to 9 C decreased the incorporation of labeled linoleic and α-linolenic acid into the lipids, but did not modify the unsaturated: saturated acid ratio of the mollusk lipids during this period. A change of habitat from lake to estuary changed very significantly the fatty acid composition of the animal. The ω6 acids, linoleic and arachidonic, typical ofD. patagonicus living in the lake, were partially replaced by ω3 acids. That this change was due to a change of food composition was indicated by the fatty acids of corresponding sediments. Therefore, the fatty acid composition ofD. patagonicus is highly sensitive to food composition and varies little with temperature and seasonal changes. Member of Carrera del Investigador Científico de la Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Member of Carrera del Investigador Científico del Consejo Nacional de Investigaciones Ciéntíficas y Técnicas, Argentina.     

Effect of dietary fats on some membrane-bound enzyme activities,membrane lipid composition and fatty acid profiles of rat heart sarcolemma

The effect of various dietary fats on membrane lipid composition, fatty acid profiles and membrane-bound enzyme activities of rat cardiac sarcolemma was assessed. Four groups of male weanling Charles Foster Young rats were fed diets containing 20% of groundnut, coconut, safflower or mustard oil for 16 weeks. Cardiac sarcolemma was prepared from each group and the activities of Na⁺,K⁺-ATPase, 5′-nucleotidase, Ca²⁺-ATPase and acetylcholinesterase were examined. ATPase activities were similar in all groups except the one fed coconut oil, which had the highest activities. Acetylcholinesterase activity was also similar in all the groups, however, it was significantly higher in the group fed mustard oil. No significant changes were observed among the groups in 5′-nucleotidase activity, in the cholesterol-to-phospholipid molar ratio and in sialic acid content. The coconut, safflower and mustard oil diets significantly increased cholesterol and phospholipid contents and the lipid-to-protein ratio of cardiac sarcolemma as compared to feeding the groundnut oil diet. The fatty acid composition of membrane lipids was quite different among the various groups, reflecting the type of dietary fat given. The total unsaturated-to-saturated fatty acid ratio was not different among the various groups; however, the levels of some major fatty acids such as palmitic (16∶0), oleic (18∶1) and linoleic (18∶2) acids were significantly different. Cardiac sarcolemma of the group fed safflower oil had the highest polyunsaturated fatty acid content. The results suggest that dietary fats induce changes not only in the fatty acid composition of the component lipids but also in the activities of sarcolemmal enzymes involved in the regulation of cardiac function.     

Effects of membrane fatty acid composition on sodium-independent phenylalanine transport in Ehrlich cells

We have examined Na⁺-independent phenylalanine transport in Ehrlich cells having different degrees of membrane fatty acid saturation. These differences were produced by growing the cells in mice fed a fat-free chow supplemented with either sunflower seed oil or coconut oil. Plasma membranes isolated from the cells grown on sunflower oil were enriched with polyenoic fatty acid, especially 18∶2, whereas those isolated from the cells grown on coconut oil were enriched in monoenoic fatty acids, primarily 16∶1 and 18∶1. Arrhenius plots of phenylalanine uptake showed two transitions. The temperatures of these transitions were different in the two cell preparations; 17 C and 24 C for the cells enriched in polyenoic fatty acids, 19 C and 28 C for those enriched in monoenoic fatty acids. Therefore, this transport system is sensitive to changes in the fatty acid composition of the lipid phase in which it operates. The activation energies, however, were the same in both cell preparations; 14, 8 and 4 kcal/mol. There also was no significant effect of the lipid modifications on either the K′_m or V′_max of this transport process. The K′_m for phenylalanine uptake from a choline medium remained constant as the temperature was raised from 17 C to 37 C, whereas the V′_max showed about a two-fold increase in both cell types. Phenylalanine exodus from the cells into an amino acid-free suspending medium, analyzed using first-order kinetics, also was not influenced by these membrane fatty acid modifications. The changes in the transition temperatures probably reflect differences in the degree of fatty acid unsaturation of lipids that surround and interact with the phenylalanine carrier. Such differences, however, do not appreciably influence the catalytic activity of this transport system.     

Lipids of cultured hepatoma cells: IV. Effect of serum and lipid upon cellular and media neutral lipids

Minimal deviation hepatoma 7288C cells were cultured in a modified Swim's medium supplemented with decreasing levels of serum, lipid-free serum, lipid-free serum plus fatty acids, and other additives. Cellular and media neutral lipid classes were quantitated, the fatty acids of triglycerides and sterol esters analyzed, and the carbon number distribution of triglycerides determined. Cellular triglyceride biosynthesis virtually was inhibited when the medium was supplemented with bovine serum alone. This inhibition was not observed when the medium was supplemented with fetal calf serum alone or mixtures of fetal calf serum and bovine serum. Cells cultivated on medium supplemented with lipid-free serum plus palmitic or linoleic acids had much lower levels of free and esterified cholesterol. The fatty acid composition of cellular triglycerides and cholesterol esters differed dramatically from the corresponding media lipid classes. Except when linoleic acid was added to the medium, changes in the media serum and lipid levels had only marginal effects upon the fatty acid composition of cellular triglycerides and cholesterol esters. These data, in conjunction with earlier data that showed the media neutral lipid levels did not decrease during cell growth, indicate that these hepatoma cells utilize little or no serum triglycerides and cholesterol esters. Linoleic acid added to the medium dramatically reduced the level of 18∶1 acids in cellular triglycerides and cholesterol esters. Palmitic acid added to the medium did not change the fatty acid compositions significantly. Comparison of experimentally determined and calculated triglyceride carbon number percentages indicated a random distribution of fatty acids in this glyceride. The fatty acid composition of cellular triglycerides was similar to the composition of the cholesterol esters. The lack of characteristic and distinguishable compositions of these two classes that occur in most normal tissues suggests a loss of specificity in the lipid metabolism of this neoplasm at the class level.     

Lipids of maturing grain of corn (Zea mays L.): I. Changes in lipid classes and fatty acid composition

The total lipids of the grain from three strains of corn were compared throughout the growing season. The Illinois High Oil stock and the two inbreds, H51 and K6, represented high, intermediate and low oil-producing lines. In all three strains lipid synthesis was most active between 15 and 45 days after pollination. The lipids were extracted from the grain with a mixture of chloroform, methanol and water and were separated into classes by silicic acid and thin layer chromatography. Triglycerides constituted 10–17% of the total lipids at 10 days after pollination and increased to 75–92% at 75 days. Polar lipids at 10 days represented 70–72% and at 75 days 4–21%. Fatty acid compositions of the triglycerides and polar lipids changed as the grain matured, but the fatty acids of the polar lipids were more saturated than those of the triglycerides throughout the sampling periods. The major polar lipids were digalactosyl diglyceride, monogalactosyl diglyceride, phosphatidyl choline, phosphatidyl inositol and phosphatidyl ethanolamine. Presented in part at the AOCS-AACC Joint Meeting, Washington, D.C., April 1968.     

Effect of chemical composition on gas transport properties of ethylene-based ionomers

Transport properties of oxygen and carbon dioxide in three ethylene–acrylic acid ionomers, characterized by different amounts of acrylic acid groups, percentage of neutralization and counterion type, have been investigated. Gas permeation experiments have been performed in the 25–65°C range. Gas permeabilities, diffusivities and solubilities were evaluated along with activation energies and heats of solution. The transport properties have been analysed in order to correlate the effect of counterion type and its concentration on macromolecular mobility and on polymer–gas interaction, which in turn are the factors determining permeation rate.