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The linoleic acid and trans fatty acids of margarines.

Fifty brands of margarine were analysed for cis-polyunsaturated acids by lipoxidase, for trans fatty acid by infared spectroscopy, and for fatty acid composition by gas-liquid chromatography. High concentrations of trans fatty acids tended to be associated with low concentrations of linoleic acid. Later analyses on eight of the brands, respresenting various proportions of linoleic to trans fatty acids, indicated that two of them contained still higher levels of trans fatty acids (greater than 60%) and negligible amounts of linoleic acid. It is proposed that margarine could be a vehicle for the distribution of some dietary linoleic acid and that the level of linoleic acid and the summation of the saturated plus trans fatty acids be known to ascertain nutritional characteristics.

Chromatography, Gas↗

Effect of long-chain fatty acids in the culture medium on fatty acid composition of WEHI-3 and J774A.1 cells.

As a first step in determining the mechanism of action of specific fatty acids on immunological function of macrophages, a comparative study of the effect of long-chain polyunsaturated fatty acids (PUFA) in the medium was conducted in two macrophage cell lines, J774A.1 and WEHI-3. The baseline fatty-acid profiles of the two cell lines differed in the % distribution of saturated (SFA) and unsaturated fatty acids (UFA). J774A.1 cells had a higher % of SFA (primarily palmitic acid) than WEHI-3 cells. Conversely, WEHI-3 cells had a higher % of UFA (primarily oleic acid) than J774A.1 cells. Neither cell line had detectable amounts of alpha-linolenic acid (ALA) or eicosapentaenoic acid (EPA). The most abundant polyunsaturated fatty acid in both cells lines was arachidonic acid (AA). The efficiency of transport of fatty acids from the medium to the macrophages by two delivery vehicles (BSA complexes and ethanolic suspensions) was compared. Overall, fatty acids were transported satisfactorily by both delivery systems. Alpha-linolenic acid and doscosahexenoic acid (DHA) were transported more efficiently by the ethanolic suspension system. Linoleic acid (LA) was taken up more completely than ALA, and DHA was taken up more completely than EPA by both cell cultures and delivery systems. A dose-response effect was demonstrated for LA, ALA, EPA and DHA in both J774A.1 and WEHI-3 cells. Addition of polyunsaturated fatty acids (PUFA) to the cell cultures modified the total lipid fatty acid composition of the cells. The presence of ALA in the culture medium resulted in a significant decrease in AA in both cell lines. The omega-3/omega-6 fatty acid ratio (omega-3/omega-6), polyunsaturated/saturated fatty acid ratio (P/S), and unsaturation index (UI) increased directly with the amount of PUFA and omega-3 fatty acid provided in the medium. The results indicate that the macrophage cell lines have similar, but not identical, fatty acid profiles that may be the result of differences in fatty acid metabolism. These distinctions could in turn produce differences in immunological function. The ethanol fatty-acid delivery system, when compared with the fatty acid-BSA complex system, is preferable for measurement of dose-response effects, because the cellular fatty acid content increased in proportion to the amount of fatty acid provided in the medium. Similar dose-response results were observed in a previous in vivo study using flaxseed, rich in ALA, as a source of PUFA.

Animals↗

Preoperative oral supplementation with long-chain Omega-3 fatty acids beneficially alters phospholipid fatty acid patterns in liver, gut mucosa, and tumor tissue.

BACKGROUND: The uptake of omega-3 polyunsaturated fatty acids (PUFAs) into the liver, gut mucosa, and tumor tissue and plasma levels after preoperative administration of supplemented enteral nutrition was investigated in patients with malignancies of the upper gastrointestinal tract. The objective of the study was to evaluate the incorporation of preoperatively administrated PUFAs, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) into cell phospholipids. METHODS: Patients undergoing major gastrointestinal surgery (n = 40) were prospectively randomized to receive a PUFA-supplemented liquid oral diet 5 days preoperatively or an isocaloric control diet. The planned diet intake was 1000 mL/d providing 3.7 g of PUFA. The diet was given in addition to the usual hospital diet. The phospholipid fractions in plasma were analyzed on the day of surgery. Tissue samples of liver, gut mucosa (small intestine), and tumor were taken during surgery and homogenized. EPA and DHA content was analyzed using liquid gas chromatography. RESULTS: Both patient groups (PUFA group: n = 20; control group: n = 20) were similar in age, weight, and surgical procedures. As compared with the control group, the PUFA group had significantly increased levels of EPA in liver tissue (0.4 vs 1.3 weight %), gut mucosa (0.3 vs 1.0 weight %), and tumor tissue (0.3 vs 0.8 weight %). Also, the DHA levels in the PUFA group were significantly higher than the control group: liver tissue (4.1 vs 7.5 weight %), gut mucosa (2.1 vs 3.7 weight %) and tumor tissue (1.9 vs 4.2 weight %). CONCLUSIONS: This study suggests that administration of PUFA-enriched diets leads to increased incorporation of EPA and DHA not only in liver and gut mucosa tissue, but also in tumor tissue in patients with solid gastrointestinal tumors. Thus, preoperative administration of oral PUFA-enriched diets could have an impact on the postoperative inflammatory response after major abdominal surgery.

Administration, Oral↗

Ultrastructure and fatty acid composition of fatty acid-modified Morris 7777 hepatoma cells.

Morris 7777 hepatoma cells, maintained in culture for 5 days in Lewis medium supplemented with 0.1 to 0.35 mM cis-unsaturated fatty acid of the oleic (number of carbon atoms: number of double bonds, 18:1), linoleic (18:2), or arachidonic (20:4) type, were compared to similarly maintained, unsupplemented cells as to ultrastructure and fatty acid composition. The cells of unsupplemented cultures were distinguished by their spherical shape, multilobated nuclei, large nucleoli, and distribution of chromatin. Cellular extensions in the form of pseudopodia and filopodia along with junctional complexes and structures resembling bile canaliculi were evident. The cytoplasmic matrix and cellular organelles appeared normal in morphology. The ultrastructure of fatty acid-supplemented cells differed from unsupplemented hepatoma cells in cell size, location of the nucleus, abundance of endoplasmic reticulum, accumulation of lipid droplets, mitochondrial abnormalities, redistribution of cytoplasmic fibrils, and plasmalemmal extensions. Supplemented cells contained lipid droplets varying in number from a optimum of six to a maximum of greater than 70. The size of these lipid droplets varied from 0.57 +/- 0.34 (S.D.) micrometers for oleic, 1.22 +/- 0.67 micrometers for linoleic, and 0.91 +/- 0.46 micrometers for arachidonic acid-supplemented cells. Variation in the ultrastructure of supplemented cells was also evident. Cytoplasmic vesiculation appeared more frequently and was more prominent in the linoleic acid-supplemented cells. Alterations in the number of surface specializations and nuclear morphology were more pronounced in the arachidonic acid-supplemented cells. Occasional swelling, loss of matrix density, and dilated cristae were evident in mitochondria of oleic acid-supplemented cells. The neutral and phospholipid fractions of arachidonic acid-supplemented cells differed from the unsupplemented cells in the concentrations of 14:0, 15:0, 16:0, 16:1, 17:0, 18:0, 18:1, 18:2, 20:1, 20:3, 20:4, 22:5, and 22:6 fatty acids. The most significant change was detected in the higher level, 26- or 8-fold, of fatty acids 20:3 omega 6 and 22:6 in the phospholipid fraction of arachidonic acid-supplemented cells.

Animals↗

Phospholipid synthesis by chick retinal microsomes: fatty acid preference and effect of fatty acid binding protein.

The acylation of 1-palmitoyl-sn-glycerophosphocholine (1-16:0-GPC) or 1-palmitoyl-sn-glycerophosphoethanolamine (1-16:0-GPE) was measured using the microsomal fraction prepared from retinas of 14-15-day-old chick embryos. Rates of incorporation of exogenously supplied fatty acids into diacyl-GPC were generally 5-7 times greater than into diacyl-GPE. Substrate preferences for incorporation into diacyl-GPC and diacyl-GPE were, respectively, 18:2 greater than 18:3 = 20:5 greater than 20:4 greater than 18:1 greater than 22:6 = 18:0 and 18:2 greater than 22:6 greater than or equal to 18:3 = 18:0 greater than or equal to 20:4 = 18:1 greater than 20:5. The apparent selectivities were not consistent with the reported fatty acid compositions of these lipid classes. The addition of partially purified fatty acid binding protein (FABP) to the reaction had no effect either on overall rates of incorporation or on the substrate preference. When fatty acyl-CoA substrates were used, rates of incorporation of the 18:0 derivative were much higher than with the fatty acid, while rates with other fatty acyl-CoA were similar to those with the respective fatty acid. Substrate preferences for CoA derivatives incorporated into diacyl-GPC were: 18:0 greater than 20:4 greater than 18:2 greater than or equal to 22:6, and into diacyl-GPE: 20:4 = 22:6 greater than 18:0 greater than 18:2. Polyunsaturated fatty acyl CoA (PUFA-CoA) were thus favored for incorporation into diacyl-GPE, and to a lesser extent into diacyl-GPC, a result that is consistent with composition data.(ABSTRACT TRUNCATED AT 250 WORDS)

Acyl Coenzyme A↗

Effect of essential and nonessential fatty acids in complex mixture on fatty acid composition of liver lipids.

Linoleate, linolenate, arachidonate, docosahexenoate and six other fatty acids were major components of 24 ester preparations fed as 5% of the diet for 60 days to groups of male white rats. The experiment was designed so as to provide that all major fatty acid components were independent of each other in the sense that the intake of each was poorly correlated with the intake of any of the others. Fatty acid compositions of liver lipids were determined and were related to the composition of the diet lipids. Linolenate and docosahexaenoate contents of diet and tissue revealed the same relationships reported previously from experiments in which individual pure acid esters were added to a fat-free diet. Linoleate, when fed in lipid mixtures, was more effective in raising the linoleate concentration in liver lipids than when fed alone, but this increase did not change the shape of the dose-response curve or the estimated nutritional requirement. Large amounts of fish oil in the diet tended to depress the arachidonate concentration in tissue lipids.

Animals↗

n-3 and n-6 fatty acid intake and serum phospholipid fatty acid composition in middle-aged women living in rural and urban areas in Okayama Prefecture.

Dietary fatty acids and serum lipids were evaluated in 68 middle-aged women living in the northern, rural area of Okayama Prefecture, and were compared with the values obtained from 65 urban women from the southern part of this prefecture. A higher level in HDL cholesterol and a lower atherogenic index were observed in the rural women. The percent of energy intake as fat was lower (20.4 +/- 0.8% vs. 23.2 +/- 0.7%) and that of carbohydrate was greater in the rural group. Eicosapentaenoic (EPA, 0.41 +/- 0.04 g/day) and docosahexaenoic acid (DHA, 0.70 +/- 0.08 g/day) intakes were significantly higher in the rural subjects than in the urban group. Significantly higher DHA levels and n-3/n-6 fatty acid ratios in serum total phospholipids were found in rural women in their fifties and the sixties compared to urban women. Dietary linoleic acid (LA) amounts were positively correlated with LA (p < 0.05), and negatively with the EPA (p < 0.05) and DHA (p < 0.01) contents of serum total phospholipids. These results suggest that the traditional Japanese diet, containing little fat but enriched in complex carbohydrates and n-3 fatty acids of marine origin, may be related to the low atherogenic index in this rural area.

Adult↗

Effects of exogenous fatty acids and inhibition of de novo fatty acid synthesis on disaturated phosphatidylcholine production by fetal lung cells and adult type II cells.

De novo fatty acid synthesis may be an important source of saturated fatty acids for fetal lung disaturated phosphatidylcholine (DSPC) production. To investigate the roles of de novo fatty acid synthesis and exogenous fatty acids, we incubated dispersed fetal lung cells and freshly isolated adult type II cells with exogenous palmitate and oleate and measured DSPC synthesis. Unlike adult type II cells, fetal lung cells did not increase DSPC synthesis when exogenous palmitate was available; adult type II cells increased DSPC synthesis by 70% in the presence of palmitate. Exogenous oleate decreased DSPC synthesis by 48% in fetal cells but not in adult type II cells. Incubation of fetal lung cells with TOFA [2-furancarboxylate, 5-(tetradecyloxy)-sodium], a metabolic inhibitor of fatty acid synthesis, decreased fatty acid synthesis by 65%. There was a simultaneous 56% inhibition of DSPC production, but no effect on protein, DNA, or glyceride-glycerol production, measured by precursor incorporation. The inhibition of DSPC synthesis associated with TOFA was partially prevented by exogenous palmitate but not oleate. Fetal cells prepared from explants that had been cultured in dexamethasone also had TOFA-associated inhibition of DSPC synthesis that was similar to non-dexamethasone-exposed cells. These studies suggest that under baseline conditions of low fatty acid availability, such as in the fetus, de novo fatty acid synthesis in fetal cells, but not in adult type II cells, provides sufficient saturated fatty acids to support maximal DSPC production. Inhibition of de novo fatty acid synthesis resulting in decreased DSPC production in fetal lung cells in conditions of low fatty acid availability suggests that fatty acid synthesis may be central to maintain DSPC synthesis in the fetus.

Aging↗

Intestinal and liver fatty acid binding proteins differentially affect fatty acid uptake and esterification in L-cells.

Differential effects of intestinal (I-FABP) or liver (L-FABP) fatty acid binding proteins on fatty acid uptake and esterification were examined using transfected mouse L-cell fibroblasts. L-FABP, but not I-FABP, expression increased the initial rate and extent of cis-parinaric acid uptake by 50 and 29%, respectively, compared to control cells. I-FABP and L-FABP expression preferentially increased [3H]-oleic acid incorporation into triacylglycerols by 5.5-fold and 3.8-fold, respectively. While both L-FABP and I-FABP increased esterification of [3H]-oleic acid into ethanolamine glycerophospholipids, these proteins had opposite effect on esterification into choline glycerophospholipids. These data show for the first time that distinct FABP differentially affect both fatty acid uptake and intracellular esterification.

Animals↗

Effect of dietary columbinic acid on the fatty acid composition and physical membrane properties of different tissues of EFA-deficient rats.

The effect of columbinic acid (5 trans, 9 cis, 12 cis, octadeca-trienoic acid) supplemented to a fat-free diet on the fatty acid composition and its correlation to the physical properties of several tissues of rats, was studied. The absence of lipids in the diet produced the typical changes in the fatty acid composition characteristic of essential fatty acid (EFA) deficiency, namely a significant increase in the relative percentage of monoenoic fatty acids with a concomitant decrease in linoleic and arachidonic acids and a rise in eicosa-5,8,11-trienoic acid in liver, kidney, lung and spleen homogenates. Columbinic acid supplemented to a fat-free diet for 24 or 48 hr was incorporated into the different tissues and was partially elongated to 7 trans, 11 cis, 14 cis eicosatrienoic acid, but it was not desaturated. It modified the fatty acid spectrum of the lipids in the different tissues returning it to a similar composition of non-EFA deficient animals, except for a decrease of linoleic acid. The absence of lipids in the diet produced an increase in the 1-6 diphenyl-1,3,5-hexatriene (DPH) steady-state fluorescence anisotropy (rs) in liver microsomes, that was corrected by the administration of columbinic acid for 24 hr. It is concluded that columbinic acid produced a change in the pattern of total fatty acid composition of the different tissues studied which induced a favorable effect on the physical properties of the liver microsomal membranes (rs), leading to an improvement on the fatty acid deficiency in those membranes. Besides, columbinic acid would also exert a favorable effect in the short term, but not in the long-term eicosanoids production.

Animals↗

Effects of L-carnitine administration on short-chain fatty acid (acetic acid) and long-chain fatty acid metabolism during hemodialysis.

The purpose of the study is to investigate the effects of L-carnitine on the exogenous acetate metabolism during hemodialysis together with the triglyceride and free fatty acid metabolism. Sixteen chronic renal failure patients on acetate dialysis were orally administered 1,200 mg of L-carnitine chloride per day for 12 weeks. Plasma triglyceride concentrations at 30 and 60 min following initiation of hemodialysis were significantly lower than before hemodialysis, while the plasma concentrations of free fatty acid at the same points into hemodialysis were significantly higher than before hemodialysis, whether L-carnitine chloride was being given or not. This is attributed to the enhanced degradation of triglyceride and the increased generation of the free fatty acid with heparin administration during hemodialysis. The L-carnitine chloride treatment did not affect the plasma triglyceride concentrations at 30 and 60 min into hemodialysis, whereas the free fatty acid concentrations at the same points into hemodialysis were significantly lower after L-carnitine chloride administration commenced than before it. This is attributed to the fact that fatty acid oxidation was enhanced following the L-carnitine chloride treatment. Compared to before the drug administration, the whole body clearance of exogenous acetate was significantly increased after the drug was given, and the plasma acetate level during hemodialysis fell accordingly. This is attributed to the fact that with L-carnitine chloride administration, the amount of accumulated acyl-CoA in the cytosol decreased and consequently the citrate cycle function increased.

Acetates↗

Effects of dietary alpha- and gamma-linolenic acids on liver fatty acids, lipid metabolism, and survival in sepsis.

The effects of dietary treatment for 3 weeks with soybean oil, linseed- and safflower oil (high alpha-linolenic acid, ALA), or borage oil (high gamma-linolenic acid, GLA) on the liver fatty acid profile and lipid metabolism in fed rats, in normal fasted rats, and septic fasted rats, and on survival from sepsis, were studied. The results were the following: 1) Dietary ALA increased incorporation of alpha-linolenic (18:3w3), eicosapentaenoic (20:5w3), and docosapentaenoic (22:5w3) acids in neutral lipids and phospholipids, and docosahexaenoic (22:6w3), dihomo-gamma-linolenic (20:3w6), arachidonic (20:4w6), stearic (18:0), oleic (18:1w9), and linoleic (18:2w6) acids in phospholipids in the livers of fed, fasted, and septic fasted rats. Dietary GLA increased all w6 fatty acids except 18:2w6, and reduced all w3 fatty acids in neutral lipids and phospholipids. 2) Dietary ALA increased liver phospholipid content in fasted as well as in septic fasted rats and was more potent than GLA in lowering serum cholesterol and liver neutral lipids. 3) Dietary ALA counteracted sepsis-related changes in liver weight, platelet count, body temperature, prekallikrein, serum glucose, beta-hydroxybutyrate, and free fatty acids. 4) Dietary GLA reduced survival from sepsis. The results suggest a role for w3 fatty acids to balance w6 fatty acids in the septic state.

Animals↗

An improved purification and further characterization of the messenger ribonucleic acid for the fatty acid synthetase from rat liver.

High purity fatty acid synthetase mRNA has been prepared from rat liver. The translational purity of the mRNA preparation was at least 27% as judged by the percentage of the radioactivity incorporated into acid-insoluble material that was precipitated by anti-fatty acid synthetase antibody. The specific activity of the mRNA was 220-times greater than that reported previously from this laboratory [1]. The large increase in the specific activity was achieved by the repeated use of high resolution linear-log sucrose density gradient centrifugation and the removal of 28 S rRNA by Sepharose 4B chromatography, as well as by the optimization of the K+ concentration (160 mM) in the reticulocyte lysate translation system. The mRNA preparation showed a single major band on agarose gel electrophoresis under denaturing conditions, and the translational activity of the fatty acid synthetase mRNA on the gel was found to coincide with this band. The molecular weight of the fatty acid synthetase mRNA is 2.5.10(6) Da. The mRNA directed the synthesis of fatty acid synthetase with a molecular weight indistinguishable from that of the authentic enzyme subunit (Mr = 240 000). The copurification of the translation product and authentic enzyme revealed that the fatty acid synthetase polypeptides synthesized in the reticulocyte lysate system are assembled in vitro into dimers, the native form of the enzyme.

Animals↗

Dietary trans-fatty acids alter adipocyte plasma membrane fatty acid composition and insulin sensitivity in rats.

The present study was designed to investigate the effects of dietary trans-fatty acids (TFA) present in Indian vanaspati (partially hydrogenated vegetable oils) in comparison with saturated fatty acids (SFA) on adipocyte plasma membrane fatty acid composition, fluidity, and insulin action. The effects of 3% energy (% en) TFA was studied at 2% and 4% en of linoleic acid (18:2 n-6). WNIN male weanling rats were divided into 4 groups and fed casein-based diet containing 10% groundnut oil control (CON), palmolein (SFA), blend of vanaspati and safflower oil (3% en TFA and 2% en 18:2 n-6, TFA-1), or blend of vanaspati and safflower oil (3% en TFA and 4% en 18:2 n-6, TFA-2) for 12 weeks. Compared with CON, rats fed TFA and SFA diets had high levels of fasting plasma insulin and triglycerides. Both TFA- and SFA-fed groups had low levels of arachidonic acid (20:4 n-6) in adipocyte plasma membrane phospholipids. However, adipocyte plasma membrane fluidity decreased only in TFA-fed rats. Norepinephrine-stimulated lipolysis was high, whereas the antilipolytic effect of insulin and insulin-stimulated glucose transport were low in the adipocytes of SFA- and TFA-fed rats. However, the extent of decrease in the antilipolytic effect of insulin and insulin-stimulated glucose transport was greater in TFA-fed rats. These findings suggest that diet providing approximately 10% en SFA (PUFA/SFA [P/S] ratio 0.2) decreased adipocyte insulin sensitivity in rats. In these diets, replacement of approximately 2% en SFA (16:0) and approximately 1% en monounsaturated fatty acid (18:1 cis) with TFA decreased adipocyte insulin sensitivity to a greater extent. However, increasing dietary 18:2 n-6 did not prevent or reduce the TFA-induced adipocyte insulin resistance.

Adipocytes↗

Questions to the supply of young infants with fat and fatty acids. II. Fat content and fatty acid pattern in milk formulae for healthy infants in the first 6 months of life.

The fat content and the fatty acid pattern were analyzed in 30 commercially prepared milk formulae for healthy infants in the fisrt 6 months of life. We found an average fat content of 3.4 g or 3.6 g/100 ml in "partly adapted" and "adapted" milk formulae, between 1.4 g and 3.3 g/100 ml in "not defined" milk formulae. We regard a fat content lower than 3.0 g/100 ml and more than 4.0 g/100 ml as not advisable. In most milk formulae the ratio of saturated: unsaturated fatty acids is similar to the ratio in human milk fat. This ratio is obtained from a mixture of cow's milk fat with vegetable oils or by a mixture of various vegetable fats. The difference in the fatty acid pattern between the milk formulae and the fatty acid pattern in mature human milk are demonstrated and discussed. The tolerance of milk formulae for young infants is not influenced in an unfavorable manner by butyic acid. A high portion of lauric acid in mild formulae seems undesirable. The importance of the position of palmitic acid in the triglycerides of milk formulae for infants is discussed, and it is referred to the advantage of a mixture of cow's milk fat with vegetable fats. A linoleic acid content of 3 kcal% in the minimum and 7 kcal% in the maximum in milk formulae for infants is regarded as advisable.

Animals↗

Fatty acid transport proteins facilitate fatty acid uptake in skeletal muscle.

In view of the importance of long chain fatty acids (LCFAs) to many cellular processes, it may be desirable to regulate the LCFA disposition in the cell. Such regulation may be present at the level of the plasma membrane, since a number of putative LCFA transport proteins have been cloned. The development of a model system of giant vesicles has proven to be important in identifying the metabolic role of the LCFA transport system. LCFA transport rates and transporters (FABPpm and FAT/CD36) are scaled with oxidative capacity of heart and muscle. FAT/CD36 is a critical LCFA transport protein in muscle. With chronic contraction the increase in this protein also results in an increase in LCFA transport. Most importantly, LCFA transport is also increased acutely by muscle contraction, involving the translocation of FAT/CD36 from intracellular depots to the surface of the muscle cell. The acute (minutes) and chronic (days) regulation of LCFA transporters and transport by muscle may be an important mechanism for LCFA utilization during exercise and adaptable with training and with a metabolic disease such as type 2 diabetes.

Animals↗