Differences in the phospholipid,cholesterol, and fatty acyl composition of 3T3 and SV3T3 plasma membranes

An analysis of the phospholipid, cholesterol, and phospholipid fatty acyl composition of isolated plasma membranes of 3T3 and SV3T3 mouse embryo cells has been performed. The results show that the plasma membrane of SV3T3 cells contain relatively less phosphatidylethanolamine and sphingomyelin and more cholesterol than 3T3 plasma membranes. The fatty acyl composition of individual phospholipid classes as determined by gas liquid chromatography also showed differences between 3T3 and SV3T3 plasma membranes. The plasma membranes of SV40 transformed 3T3 cells contain: (a) a higher percentage of 18∶1 and less 20∶3 and 20∶4 in phosphatidylethanolamine; (b) a higher percentage of 18∶1 in phosphatidylserine; and (c) a higher percentage of 18∶2 and 20∶4 in phosphatidylinositol.     

Effects of ethanol diets on cholesterol content and phospholipid acyl composition of rat hepatocytes

Chronic treatment of adult male rats with ethanol liquid diets resulted in alterations in phospholipid and cholesterol contents as well as the acyl composition of phosphatidylethanolamine (PE), phosphatidylinositol (PI)-phosphatidylserine (PS) mixture, and phosphatidylcholine (PC) of isolated hepatocytes. The influence of ethanol on these lipids was largely dependent on the proportion of dietary fat. Phospholipid and total cholesterol contents were elevated 23 and 27%, respectively, by ethanol when offered in a low-fat diet (5% corn oil). Only the percentage of arachidonic acid from PI-PS was significantly reduced in the low-fat ethanol group. Exposure to a high-fat (34% corn oil) diet in the presence of ethanol for 4–5 weeks resulted in a significant decrease in arachidonate/linoleate ratios of hepatic PE, PS-PI and PC, while total phospholipid content remained constant. In the high-fat, ethanol-treated group, hepatic cholesterol content was increased 2-fold. These results suggest that the level of dietary fat plays an important role in determining the effects of chronic ethanol consumption on hepatic cholesterol content and phospholipid acyl composition.     

Growth hormone and liver mitochondria: Effects on phospholipid composition and fatty acyl distribution

Effects of growth hormone on phospholipid composition and fatty acyl distribution were studied in liver mitochondria of hypophysectomized rats. After hypophysectomy, only cardiolipin showed a 25% decrease. Its fatty acyl distribution, which consisted mainly of linoleic acid (55–60%) and oleic acid (20%), was unchanged. In phosphatidylcholine and phosphatidylethanolamine fractions the contents of docosahexaenoic and arachidonic acids were decreased with a concomitant increase in linoleic acid content. These changes could be accounted for by small but significant decreases in the activities of Δ⁹-desaturase (sucrose-induced), Δ⁵-desaturase and mitochondrial elongation enzymes. The activities of Δ⁶-desaturase NADH cytochrome b₅ ferri-reductase, cytochrome b₅, NADH cytochrome c reductase and microsomal elongation enzymes remained virtually unchanged. Injection of bovine growth hormone daily for seven days restored cardiolipin and fatty acyl distribution and the enzyme activities. From these and other results, we conclude that growth hormone-dependent increase of respiratory activity of liver mitochondria may be partly mediated by the hormonal effects on membrane lipid distribution.     

Dietary ω3 fatty acids and cholesterol modify enterocyte microsomal membrane phospholipids,cholesterol content and phospholipid enzyme activities in diabetic rats

Diabetes-associated changes in intestinal uptake of nutrients are modified by isocaloric variations in the type of dietary lipids, and are associated with alterations in the phospholipid and fatty acyl content of the intestinal brush border membrane. The present study was designed to test the hypothesis that diet- and diabetes-associated changes in enterocyte microsomal membrane phospholipids are due to variations in the activity of two phospholipid metabolizing enzymes, 1,2-diacylglycerol: CDP choline cholinephosphotransferase (CPT) and phosphatidylethanolamine methyltransferase (PEMT). Adult female Wistar rats were fed one of four semisynthetic diets—beef tallow low in cholesterol (BT), beef tallow high in cholesterol (BTC), fish oil low in cholesterol (FO) or fish oil high in cholesterol. In half of the animals, diabetes mellitus was produced by injection of streptozotocin. Jejunal and ileal enterocyte microsomes (EMM) were isolated and analyzed for cholesterol and phospholipids, as well as for CPT and PEMT activities. In control animals, feeding FO reduced EMM total phospholipids including phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol. Feeding FO resulted in a greater than 95% reduction in the activity of CPT. Diabetes was associated with increased jejunal EMM total phospholipids including sphingomyelin (SM) and PE, without associated changes in CPT or PEMT. Dietary cholesterol supplementation did not effect EMM total cholesterol or phospholipid composition in control rats fed BT or FO, but was associated with an increase in EMM cholesterol in diabetic rats fed BT or FO. A decrease in total phospho-lipids due to a decline in SM, PC and PE in diabetic rats fed FO was not associated with changes in the activities of CPT or PEMT in EMM. Thus (i) enterocyte microsomal membrane cholesterol and phospholipid contents are influenced by diabetes, dietary cholesterol and the type of fatty acid in the diet, and (ii) changes in phospholipid composition are not fully explained by alterations in the activities of CPT and PEMT.     

Dietary fat composition and age affect synaptosomal and retinal phospholipid fatty acid composition in C57BL/6 mice

The purpose of this study was (i) to determine whether dietary fat-induced differences in neural and retinal membranes occur when dietary fat treatment is implemented in aged animals and (ii) to characterize the effect of long-term differences in dietary fat on neural and retinal membrane composition. For the first objective, young (six-week-old) and old (95-week-old) mice were randomly assigned to beef tallow (TAL) or soybean oil (SBO) diets for eight weeks. For the second objective, young (four-week-old) mice consumed either TAL or SBO diets for 99 weeks. Young and old mice challenged with a change in dietary fat for an eight-week period showed both diet and age effects on neural and retinal phospholipid fatty acid composition (P<0.05). In addition, significant diet by age interactions were evident. In mice that consumed TAL and SBO diets throughout their life, only retinal phosphatidylethanolamine (PE) 18∶2n−6 and neural phosphatidylserine 22∶5n−6, PE 18∶2n−6 and phosphatidylcholine 18∶2n−6 differed between dietary treatments (P<0.05). Neither the unsaturation index nor the n−6/n−3 ratio was affected by diet. Neural and retinal phospholipid fatty acid composition were responsive to changes in dietary fat even when the treatment was implemented beyond developmental or post-weanling stages. In contrast, when mice consumed TAL or SBO diets throughout their life, fewer differences in phospholipid fatty acid composition were detected, suggesting that the effect of the dietary treatment was mitigated by aging.     

Liver and intestinal fatty acid-binding protein expression increases phospholipid content and alters phospholipid fatty acid composition in L-cell fibroblasts

Although fatty acid-binding proteins (FABP) differentially affect fatty acid uptake, nothing is known regarding their role(s) in determining cellular phospholipid levels and phospholipid fatty acid composition. The effects of liver (L)- and intestinal (I)-FABP expression on these parameters were determined using stably transfected L-cells. Expression of L- and I-FABP increased cellular total phospholipid mass (nmol/mg protein) 1.7- and 1.3-fold relative to controls, respectively. L-FABP expression increased the masses of choline glycerophospholipids (ChoGpl) 1.5-fold, phosphatidylserine (PtdSer) 5.6-fold, ethanolamine glycerophospholipids 1.4-fold, sphingomyelin 1.7-fold, and phosphatidylinositol 2.6-fold. In contrast, I-FABP expression only increased the masses of ChoGpl and PtdSer, 1.2- and 3.1-fold, respectively. Surprisingly, both L- and I-FABP expression increased ethanolamine plasmalogen mass 1.6- and 1.1-fold, respectively, while choline plasmalogen mass was increased 2.3- and 1.7-fold, respectively. The increase in phospholipid levels resulted in dramatic 48 and 33% decreases in the cholesterol-to-phospholipid ratio in L- and I-FABP expressing cells, respectively. L-FABP expression generally increased polyunsaturated fatty acids, primarily by increasing 20∶4n−6 and 22∶6n−3, while decreasing 18∶1n−9 and 16∶1n−7. I-FABP expression generally increased only 20∶4n−6 proportions. Hence, expression of both I- and L-FABP differentially affected phospholipid mass, class composition, and acyl chain composition. Although both proteins enhanced phospholipid synthesis, the effect of L-FABP was much greater, consistent with previous work suggesting that these two FABP differentially affect lipid metabolism.     

Effect of dietary fat on phospholipid class distribution and fatty acid composition in rat fat cell plasma membrane

The effect of dietary fats on phospholipid class distribution and fatty acid composition was studied in rat fat cell plasma membrane. Three groups of male Wistar weanling rats were fed for 8 wk three diets differing in the amount and nature of the fats: 1.5% sunflower oil (low fat control; LFC), 10% sunflower oil (high fat, unsaturated; HFU), 1.5% sunflower oil+8.5% cocoa butter (high fat, saturated; HFS). Plasma membranes were prepared from epididymal adipocytes. The amount and type of dietary fat significantly altered membrane phospholipid distribution. Phospholipid content was lowered with HFU as compared to LFC or HFS diets, but no changes were observed for cholesterol. Phosphatidylinositol (PI) and phosphatidylserine (PS) were less affected by dietary changes than were other phospholipid classes. Major changes were detected for phosphatidylcholine (PC), phosphatidylethanolamine (PE) and sphingomyelin (SM) contents. No large changes in PC and PE fatty acid compositions were observed between the LFC and HFS groups, but the HFU diet induced several changes. Correlations with plasma membrane 5′-nucleotidase activities are discussed.     

Lipoprotein composition and serum cholesterol ester fatty acids in nonwesternized melanesians

In this study, the relationships between dietary fat [as measured by serum cholesterol ester fatty acids (CE-FA)], age, smoking, body mass index, and serum lipids were analyzed in 151 subsistence horticulturalists, aged 20–86 yr, from Kitava, Trobriand Islands, Papua New Guinea. Their diet consists of tubers, fruit, coconut, fish, and vegetables with a negligible influence of western food and alcohol. Total fat intake is low [21% of energy (en%)], while saturated fat intake from coconuts is high (17 en%, mainly lauric and myristic acid). In multivariate analysis, 11–43% of the variation of the serum lipoprotein composition was explained by CE-FA, age, and smoking habits. The proportion of CE20∶5n-3 explained much of the variation of triglycerides (TG, negative relation) and high density lipoprotein-cholesterol (HDL-C, positive) in both sexes and serum apolipoprotein A1 (ApoA1, positive) in the males. CE16∶0 was positively related to TG and negatively related to HDL-C and ApoA1 in both sexes, and in males it related negatively to total cholesterol (TC) and low density lipoprotein-cholesterol (LDI-C). In males, negative relationships were present between CE18∶2n-6 and TC and between CE14∶0 and serum lipoprotein(a). Smoking was independently associated with lower ApoA1 in both sexes and with lower HDL-C and higher TG, TC, LDL-C, and apolipoprotein B in males. In conclusion, marine n-3 fatty acids and linoleic acid showed the same potentially beneficial relationships with lipoproteins and apolipoproteins as in western populations. The relations of palmitic acid to serum lipids may be explained in terms of endogenous fat synthesis at a low-fat intake, rather than reflecting its relative intake.     

The influence of n-6 and n-3 fatty acids on kidney phospholipid composition and on eicosanoid production in aging rats

Changes in eicosanoid production may contribute to some of the complications of the aging process such as atherosclerosis and glomerular sclerosis. Polyunsaturated fatty acids of the n-6 and n-3 series are precursors of eicosanoids. We fed diets containing safflower oil as a source of n-6 fatty acids, fish oil as a source of n-3 fatty acids or beef tallow as a source of saturated fats to three groups of normal rats from 2–18 months of age. We demonstrated incorporation of the n-3 fatty acids, 20:5n-3 and 22:6n-3 into kidney phospholipids. Feeding of the diet containing n-3 fatty acids was associated with a markedly decreased giomerular production of PGE, 6-keto-PGF_1α and TXB₂. It also decreased the aortic production of 6-keto-PGF_1α and platelet production of TXB₂. No significant effect of n-6 fatty acids on dienoic eicosanoid production was observed. There were no adverse effects on kidney function as measured by urinary protein excretion and serum creatinine levels or on renal morphology by any diet. A diet enriched in n-3 fatty acids for 18 months remains effective in decreasing dienoic eicosanoids in the aging rat.     

Dietary fish oil dose- and time-response effects on cardiac phospholipid fatty acid composition

Fish consumption is associated with reduced cardiovascular mortality, and elevated myocardial long-chain n−3 polyunsaturated FA (PUFA) content is implicated in this cardioprotection. This study examined the dose and time responses for incorporation of n−3 PUFA into cellular membranes in rats fed fish oil (FO)-containing diets. For the time course study, rats were fed a 10% FO diet for periods ranging from 0 to 42 d, after which myocardial and erythrocyte membrane fatty acid composition was determined. For the dose response study, rats (n=3) were fed 0, 1.25, 2.5, 5, or 10% FO for 4 wk, with myocardial, erythrocyte, and skeletal muscle membrane FA determined. Myocardial DHA (22∶6n−3) levels doubled in 2 d, stabilizing at levels ≈200% higher than control after 28 d feeding with 10% FO. By comparison, DHA levels doubled after 4 wk of 1.25% FO feeding. In myocardium and skeletal muscle, EPA (20∶5n−3) levels remained low, but in erythrocytes EPA levels reached 50% of DHA levels. The n−3 PUFA were incorporated at the expense of n−6 PUFA in myocardium and skeletal muscle, whereas erythrocytes maintained arachidonic acid levels, and total n−3 PUFA incorporation was lower. This study shows that low doses of FO produce marked changes in myocardial DHA levels; maximal incorporation takes up to 28 d to occur; and while erythrocytes are a good indicator of tissue n−3 incorporation in stable diets, they vary greatly in their time course and pattern of incorporation.     

Lipid and fatty acyl composition of rat brain capillary endothelia isolated by a new technique

A method is described for the isolation of pure capillary endothelia from rat brain and the phospholipid composition of these cells is reported. This method is rapid and requires only a small amount of starting material. It involves: (a) tissue disruption by high speed homogenization, (b) separation of the capillary endothelia from other brain structures using sucrose gradients, and (c) a final purification using a glass bead column. Choline and ethanolamine phosphoglycerides were found to be the predominant lipid classes of these cells amounting to 31.9% and 24.4%, respectively, of total phospholipids. The choline phosphoglycerides consisted almost exclusively of 1,2-diacyl glycerophosphorylcholine, whereas the ethanolamine phosphoglycerides consisted of approximately equal amounts of 1,2-diacyl and 1-alk-l’-enyl-2-acyl glycerophosphorylethanolamine. The composition of the constituent fatty acids of both choline and ethanolamine phosphoglycerides and the alk-1-enyl composition of ethanolamine phosphoglycerides is reported. Saturated fatty acids accounted for 45% of the total fatty acids in choline phosphoglycerides and for 53% in ethanolamine phosphoglycerides. Arachidonic acid accounted for approximately 48% of the total fatty acids in alk-1-enyl ethanolamine phosphoglyceride.     

Relationships between the fatty acid composition of muscle and erythrocyte membrane phospholipid in young children and the effect of type of infant feeding

Muscle membrane fatty acid (FA) composition is linked to insulin action. The aims of this study were to compare the FA composition of muscle and erythrocyte membrane phospholipid in young children; to investigate the effect of diet on these lipid compositions; and to investigate differential incorporation of FA into muscle, erythrocyte and adipose tissue membrane phospholipid, and adipose tissue triglyceride. Skeletal muscle biopsies and fasting blood samples were taken from 61 normally nourished children (15 males and 16 females), less than 2 yr old (means ±SE, 0.80±0.06 yr), undergoing elective surgery. Adipose tissue samples were taken from 15 children. There were significant positive correlations between muscle and erythrocyte docosahexaenoic acid (DHA) (r=0.44, P<0.0001), total n−3 polyunsaturated fatty acids (PUFA) (r=0.39, P=0.002), and the n−6/n−3 PUFA ratio (r=0.39, P=0.002). Adipose tissue triglyceride had lower levels of long-chain PUFA, especially DHA, than muscle and erythrocytes (0.46±0.18% vs. 2.44±0.26% and 3.17±0.27%). Breast-fed infants had higher levels of DHA than an age-matched group of formulafed infants in both muscle (3.91±0.21% vs. 1.94±0.18%) and erythrocytes (3.81±0.10% vs. 2.65±0.23%). The results of this study show that (i) erythrocyte FA composition is a reasonable index of muscle DHA, total n−3 PUFA, and the n−6/n−3 PUFA ratio; (ii) breast feeding has a potent effect on the FA composition of all these tissues; and (iii) there is a wide range in long-chain PUFA levels in muscle, erythrocytes, and adipose tissue.     

Dietary modulation of phospholipid fatty acid composition and lipoxygenase products in mouse lung homogenates

This study investigated the potential of dietary fats to modulate the arachidonic acid content of mouse lung phospholipids and the formation of lipoxygenase products from arachidonic and eicosapentaenoic acids. Prior to breeding, female mice were fed for five months diets with 10 wt% of either olive oil, safflower oil, fish oil, or linseed oil. The same diets were fed to the females during gestation and to the pups from day 18 to day 42 postpartum. On day 42, the phospholipids were extracted from fresh lung tissue and separated into classes [phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylcholine (PC), and phosphatidylinositol (Pl)] by thin-layer chromatography. Methyl esters of phospholipid fatty acids and unesterified fatty acids were analyzed by gas chromatography. At comparable dietary n-3/n-6 ratios, arachidonic acid was reduced 85 and 75% in lungs from mice fed linseed oil and fish oil, respectively, compared to lungs of safflower oil-fed mice. Dietary fats affected the proportion of arachidonic acid in phospholipids in the order: PE>PS>PS>Pl. Following incubation of homogenized lung tissue, the total amount of 12-lipoxygenase products was lowest in lungs from mice fed olive oil, and 12-hydroxyeicosatetraenoic acid was lowest in incubated lungs from mice fed linseed oil. Comparison of the amounts of lipoxygenase substrate fatty acids in the individual phospholipids with the lipoxygenase products suggested that the major substrate pool for the 12-lipoxygenase pathway in mouse lung homogenates was PC.     

Effects of fructose and troglitazone on phospholipid fatty acid composition in rat skeletal muscle

Skeletal muscle phospholipid fatty acid (PLFA) composition is associated with insulin sensitivity in animal models and in man. However, it is not clear whether changes in insulin sensitivity cause a change in PLFA composition or vice versa. The present studies have examined the effects of agents known to increase or decrease insulin sensitivity on PLFA composition of the major phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), in soleus and extensor digitorum longus muscle. Four groups of Sprague-Dawley rats— control, 0.2% troglitazone (Tgz), 60% fructose fed, and fructose + Tgz—were treated for 3 wk. Fructose feeding was associated with a decrease in muscle membrane polyunsaturated fatty acids (PUFA) and n-3 fatty acids in both PC and PE. Administration of Igz alone resulted in an increase in liver (3.75±0.93 to 6.93±1.00 μmol/min/mg tissue, P<0.05) and soleus muscle (0.34±0.03 to 0.67±0.09 μmol/min/mg, P<0.01) elongase activity, which would be expected to increase membrane PUFA. However, Tgz decreased PLFA associated with greater insulin sensitivity (e.g., PUFA and n-3 fatty acids) and increased PLFA associated with decreased insulin sensitivity (16∶0 and n-6 fatty acids) in both PC and PE. Co-administration of fructose and Tgz did not reverse the decrease in PUFA observed with fructose alone. We conclude that the improvement in insulin sensitivity reported with Tgz is associated with an apparently paradoxical effect to decrease PUFA and n-3 PLFA composition in rat skeletal muscle. These studies suggest that Tgz-mediated increases in insulin sensitivity do not result in improved PLFA composition.     

Influence of dietary fatty acid composition on cholesterol synthesis and esterification in hamsters

To investigate the effects of dietary fat quality on synthesis and esterification of cholesterol, Syrian hamsters were fed diets containing corn, olive, coconut or menhaden oils (10% w/w) with added cholesterol (0.1% w/w). After 3 weeks, animals were sacrificed 90 min following IP injection of³H₂O. Synthesis of free cholesterol and movement of free cholesterol into ester pools were measured from³H-uptade rate in liver and duodenum. Plasma total cholesterol and triglycerides levels were highest in coconut oil-fed animals, whereas hepatic total cholesterol and ester levels were elevated in olive oil-fed animals, as compared with all other groups. No diet-related differences were seen in duodenal cholesterol or total fatty acid content. In duodenum, uptake of³H per g tissue into cholesterol was greater compared with liver; however, within each tissue,³H-uptake into cholesterol was similar across groups. Notably,³H-uptake into cholesterol ester in liver was highest in menhaden oil-fed animals. These data suggest that menhaden fish oil consumption results in enhanced movement of newly synthesized cholesterol into ester as compared with other fat types.     

The effect of temperature on the fatty acid and phospholipid composition ofCephalosporium falciforme andCephalosporium kiliense

The effect of temperature on the lipid composition ofCephalosporium falciforme andCephalosporium kiliense, causative agents of maduromycosis, was investigated. The fungi were grown at 28.5 C and 37 C in a chemically defined medium. The lipids were solvent extracted, purified on Sephadex, and separated into their component classes by silicic acid column chromatography. Five lipid classes were found: (a) sterol esters, (b) triacylglycerides, (c) free fatty acids, (d) sterols, and (e) phospholipids. Fatty acids were analyzed by gas liquid chromatography and phospholipids by thin layer chromatography. Temperature induced changes of varying degrees occurred in both the fatty acid phospholipid fractions of each organism.     

Plasma and lipoprotein fatty acid composition in glycogen storage disease type I

Nocturnal intragastric feeding has been shown to be an effective means to improve clinical and biochemical features in glycogen storage disease type I (GSD-I). In this study, we investigated the fatty acid patterns in a whole plasma and in circulating lipoproteins in patients on this therapy. The results demonstrated massive concentration of total fatty acids coupled with higher levels of triglycerides, free cholesterol, cholesterol ester and phospholipids. This hyperlipidemia involved all fatty acids without distinction of carbon or bond numbers. However, the increase was more pronounced for saturated than polyunsaturated fatty acids, as was demonstrated by the ratios of both oleic acid to linoleic acid (1.91±0.40 vs 0.80±0.09 in controls) and of ω3+ω6 to ω9 fatty acid families (0.92±0.11 vs 1.66±0.08 in controls). The fatty acid patterns in very low (VLDL), low (LDL) and high (HDL) density lipoprotein showed substantial differences in composition, reflecting an association between an abnormal lipoprotein pattern and essential fatty acid deficiency. Furthermore, GSD-I patients exhibited a significant increase in VLDL (17±2 vs 47±7 mg/dl) and LDL cholesterol (124±7 vs 206±24 mg/dl), coupled with a decrease in HDL cholesterol (49±4 vs 28±3 mg/dl). These data documenting high LDL cholesterol and low HDL cholesterol associated with an increased concentration and proportion of saturated fatty acids suggest that GSD-I patients on nocturnal intragastric feeding are at high risk for atherosclerosis and its complications.     

Alteration in mouse splenic phospholipid fatty acid composition and lymphoid cell populations by dietary fat

The fatty acid composition of diacyl- and alkylacylglycerophosphocholine (PC), phosphatidylinositol (PI), phosphatidylserine (PS), alkenylacyl-glycerophosphoethanolamine (aPE), and diacyl- and alkylacyl-glycerophosphoethanolamine (dPE) was assessed in isolated splenocytes from C3H/Hen mice fed one of four purified isocaloric diets for six weeks. Diets contained 20% by weight of either a high-linoleate sunflower oil (Hi 18∶2), a high-oleate sunflower oil (Hi 18∶1), a mixture of 17% menhaden fish oil and 3% high-linoleate sunflower oil (Hi n−3), or a mixture of 17% coconut oil and 3% high-linoleate sunflower oil (Hi SFA). Spleen weight and immune cell yield were significantly higher (P<0.05) in mice fed the Hi 18∶1 or the Hi n−3 diets compared with those fed the Hi 18∶2 and Hi SFA diets. Distinctive patterns of fatty acids were observed for each phospholipid in response to dietary fatty acids. Dietary fat significantly affected (P<0.05) total polyunsaturated fatty acids (PUFA) in PC and dPE, total saturated fatty acids (SFA) in PC, total monounsaturated fatty acids (MUFA), and n−3 PUFA in all phospholipid classes examined. In mice fed the Hi n−3 diet, n−3 PUFA were significantly elevated, whereas n−6 PUFA decreased in all of the phospholipids. In these mice, eicosapentaenoic acid (EPA) was the predominant n−3 PUFA in PC and PI, whereas docosahexaenoic acid (DHA) was the major n−3 PUFA in aPE and PS. Interestingly, the ratios of n−3/n−6 PUFA in the phospholipids from these mice were 3.2, 2.4, 1.8, 0.8 and 0.8 for aPE, PS, dPE, PC and PI, respectively. These data suggest a preferential incorporation of n−3 PUFA into aPE, PS and dPE over PC and PI.     

Influence of fasting-refeeding and dietary linoleate on liver glucose-6-phosphate dehydrogenase and phospholipid fatty acid composition in rats adapted to a purified diet

Responses to refeeding after fasting were studied in male rats fed a purified casein-sucrose diet containing 5% safflower oil. After a 48 hr fast, the rats were fed either the same diet or the same diet minus oil (fat-free diet). These experiments were designed to distinguish changes due to fasting and refeeding alone without a change of diet from those changes caused by refeeding a diet of different composition. In the first experiment, rats were refed for 3 or 7 days. In rats refed either diet, liver glucose-6-phosphate dehydrogenase (G6PD) activity was elevated above refasting levels, but after 7 days, activity in rats refed the 5% safflower oil diet was significantly lower than in those refed the fatfree diet. The amount of liver arachidonate in rats refed the safflower oil diet was the same during refeeding as before fasting. In the second experiment, rats were refed the fat-free diet for 1, 2, 3, or 7 days. Liver G6PD and fatty acid synthetase were measured, as well as fatty acids in liver total lipids and phospholipids. G6PD activity increased above prefasting levels after one day refeeding and continued to increase, for 7 days. Fatty acid synthetase activity increased for the first 3 days of refeeding, with no additional increase after 7 days. In all rats refed the fat-free diet, the proportions of arachidonate and linoleate in liver phospholipids diminished with time, and eicosatrienoate appeared. These results show that (a) maintenance of liver phospholipid arachidonate did not prevent increased G6PD activity in early refeeding, but the elevated G6PD activity later declined when phospholipid arachidonate was maintained by feeding a source of linoleate; (b) the metabolic state of fastedrefed rats had not returned to prefasting conditions even after 7 days of refeeding a linoleate-rich diet to which the rats were adapted before fasting.     

The phospholipid and fatty acid composition of platelets in patients with primary defects of platelet function

Platelet phospholipid and fatty acid composition was determined in nine normal subjects and in 11 patients with primary defects in platelet function. Two of the patients had thrombasthenia (Glanzmann) and nine had various types of abnormalities in platelet aggregation and platelet factor 3 availability attributed to impairment of the platelet release reaction. The values observed for platelet lipids in the normal subjects were similar to those reported by others. Four of the patients with a disturbance in the platelet release reaction were in the same family and showed the same abnormal pattern of platelet lipid composition. Phospholipid analysis showed a decrease in the relative amount of phosphatidyl ethanolamine (PE) and an increase in lecithin. Abnormalities in fatty acids consisted of an increase in the relative amounts of 18∶1ω9, 20∶0 and 20∶1ω9 and a decrease in the 22∶4ω6+24∶1 fraction. Similar changes in PE and 18∶1ω9 were also observed in another patient with a similar defect in platelet function. In this patient the relative amount of platelet sphingomyelin was also increased. The platelet lipid composition in the other six patients and in one normal subject given aspirin was essentially normal.