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Changes of N-6 and N-3 fatty acids in liver from spontaneously hypertensive (SHR) and normotensive rats after diets supplemented with alpha-linolenic or eicosapentaenoic acids.

In spontaneously hypertensive (SHR) and normotensive rats (WKY), diets supplemented with n-3 fatty acids of different chain length (alpha-linolenic acid, LNA-C 18:3, n-3 with linseed oil and eicosapentaenoic acid, EPA-C 20:5, n-3 with cod liver oil) were fed over a period of 22 weeks. A diet with commercially available pellets served as control. After the LNA-rich diet the augmentation of LNA was most pronounced in liver triglycerides (TG) and free fatty acids (FFA), whereas the increase of EPA was most marked in phosphatidylethanolamine (PE) and phosphatidylcholine (PC) when compared with the controls. Docosahexaenoic acid (DHA) was decreased mainly in neutral lipids. Of the n-6 fatty acids linoleic acid (LA) appeared significantly depressed in TG and FFA, but increased in phospholipids. Arachidonic acid (AA), however, was lower in all lipids. In SHR and WKY fed the EPA-rich diet EPA and DHA were significantly higher as compared to the controls on a pellet diet. On the contrary, LNA was not detectable in all lipid classes. LA and AA were markedly depressed. Docosenoic acids were significantly increased. The p/s-ratio did not reflect the changes in the 20:5/20:4- and n-3/n-6-ratios. The data indicate a differential effect of dietary n-3 fatty acids of different chain length on the supply of other n-3 fatty acids. Moreover, after an LNA-rich diet divergent alterations of LA in neutral lipids and phospholipids occurred. The results are dissimilar to those obtained in adipose tissue. Blood pressure was not influenced by the diets in either SHR or WKY.

Animals↗

Effects on fibrinolytic activity of corn oil and a fish oil preparation enriched with omega-3-polyunsaturated fatty acids in a long-term study.

In addition to their beneficial effects in reducing platelet responsiveness, it has been a matter of controversy whether polyunsaturated fatty acids impair the fibrinolytic system or not. In a double-blind, parallel clinical trial, 40 subjects were randomized to treatment with 6 g/day of corn oil, or to 6 g/day of a fish oil preparation, enriched with omega-3-polyunsaturated fatty acids (2.0 g/day of omega-3-PUFA). Clinical and fibrinolytic variables were measured before and after 5 months of treatment. In the corn oil group, plasminogen activator inhibitor (PAI-1) decreased significantly but in the cod liver oil group, PAI-1 remained unchanged. Activities and mass concentrations of tissue plasminogen activator (t-PA) were unchanged in both groups. It is concluded that, in the doses given here, both these preparations have small or no effects on the fibrinolytic system.

Adult↗

Influence of dietary n-6/n-3 polyunsaturated fatty acid balance on the development of tolerance during chronic ethanol intoxication in rats.

The present study addresses the possible interacting effects of dietary n-6/n-3 polyunsaturated fatty acid (PUFA) balance and chronic ethanol intoxication on the synaptic membrane responses to ethanol and the development of tolerance in rats. Wistar rats were fed either a standard lab chow or various semi-synthetic diets: rich in PUFA (from soya oil: SO), deficient in linolenate (from sunflower oil: SFO) or rich in long-chain (n-3) PUFA (cod liver oil: CLO). Male adult rats from the second specially fed generation were submitted to a 3-week alcoholization by daily intubation. Functional tolerance was quantified by the hypothermic response to a challenge dose of ethanol. Synaptic fluidity and sensitivity to ethanol (variations after acute ethanol addition) were assessed by fluorescence polarization (FP) of DPH, TMA-DPH or PROP-DPH. Membrane fatty acid composition was determined by GLC. The fatty acid composition of the synaptic membranes was influenced by the diet, but rearrangements among the lipids occurred, resulting in an apparent stability in brain membrane fluidity parameters. Nevertheless, clear-cut differences were noted in response to ethanol intoxication according to the diet. In the same period of time, rats fed SFO or CLO diets were unable to develop tolerance to ethanol at the membrane level as well as functionally, contrarily to the rats fed SO or standard diets. The structurally specific roles of PUFA are suggested by the negative membrane effects of the alpha-linolenate deficient diet (SFO) and the positive ones of a diet (SO) rich and well balanced in (n-3 + n-6) PUFA. Furthermore, the n-6/n-3 PUFA balance in the synaptic membrane needs to be kept within very narrow limits to allow normal development of the adaptive response to ethanol.

Alcoholic Intoxication↗

Effects of low-dose marine oils on intimal hyperplasia in autologous vein grafts.

The effects of low-dose cod-liver oil on intimal hyperplasia of vein grafts were examined in 45 adult mongrel dogs undergoing peripheral arterial reconstruction. Fifteen animals served as the control group, 15 animals were fed a fish-oil supplement containing 240 mg of eicosapentaenoic acid daily, and a further 15 animals received 480 mg of eicosapentaenoic acid daily. Segments of undistended external jugular vein were anastomosed to bilaterally divided femoral arteries. The grafts were harvested at 6 weeks and intimal thickness was measured with a computerized interactive image analyzing system. Serum cholesterol level, prothrombin time, partial thromboplastin time, bleeding time, and platelet counts were measured before the operation and at 2, 4, and 6 weeks after the operation. Plasma levels of thromboxane B2 and prostaglandin F1 alpha and serum levels of eicosapentaenoic acid were measured before and 4 weeks after the operation. Serum cholesterol level increased similarly and significantly in all animals. Serum levels of eicosapentaenoic acid rose proportionately with the oral ingestion of fish oil but did not affect coagulation parameters. Plasma thromboxane B2 and prostaglandin F1 alpha were not significantly affected by the ingestion of marine oils. Intimal thickness was 39 +/- 5 microns in the control dogs. Ingestion of 240 mg of eicosapentaenoic acid reduced intimal thickness to 24 +/- 3 microns at 6 weeks (p less than 0.01). Increasing the dose by a factor of 2 did not decrease intimal thickness further, the intima being 23 +/- 2 microns (p less than 0.005). Our data indicate that small doses of fish oil will reduce intimal proliferation in autologous vein grafts and that marine oils may exert their beneficial effects on intimal hyperplasia by a mechanism other than their known effects on prostanoid metabolism.

Animals↗

Polyunsaturated fatty acid deficiencies: effects on hepatic plasma membrane fatty acid composition and enzyme activity.

Research on dietary polyunsaturated fatty acids (PUFA), on the activity of 5'nucleotidase and adenylate cyclase are largely contradictory due, mostly, to the absence of adequate control group. In this study; four different diets have been evaluated on the 5'nucleotidase and adenylate cyclase activities in rat liver plasma membranes. Wistar rats were given a semisynthetic diet in which lipids were supplied by 5% of either peanut oil (n-3 PUFA deficient diet), cod liver oil (n-6 PUFA deficient diet) partially hydrogenated palm oil (total PUFA deficient diet) and a mixture of peanut and rapeseed oil (control group). Liver plasma membranes were separated by using a Percoll gradient in a Beckman JA 20 centrifuge. 5'nucleotidase and adenylate cyclase activities were measured in a liquid scintilation detector by following the degradation of 3HAMP (adenosine monophosphate) and production of 3HcAMP (cyclic adenosine monophosphate) respectively. Animals fed the total PUFA deficient diet exhibited significant lower body weight and lower liver weight than did the control group. Low cholesterol concentrations were observed in animals deficient either in n-3 or total PUFA in relation to the control group. All dietary deficiencies studies provoked reduced phospholipid levels. Phosphatidylcholine and phosphatidylethanolamine were not modified whatever the deficiency studied. Phospholipids fatty acid composition was significantly modified by the diets studied. The specific activity of 5'nucleotidase in hepatic plasma membrane was independent of dietary PUFA. The catalytic unit of adenylate cyclase complex in totally deficient animals was augmented. The unit of the enzyme stimulated by the guanydyl imidodiphosphate (GppNHp) in n-3 PUFA deficient animals was augmented and reduced in animals receiving the n-6 PUFA deficient diet. In conclusion, our results show that each dietary PUFA deficiency modifies differently the proportions of phospholipid classes and their fatty acid composition. The mechanisms responsible for these modification remain to be elucidated. However, the phospholipid fatty acid changes did not influence the 5'nucleotidase activity except in the case of extreme excess which concerns more toxicology than nutritional modifications. Finally, the catalytic unit (Forskoline + GDP beta s) of adenylate cyclase complex and the regulatory unit (GppNHp) may be sensitive to alterations in PUFA composition.

5'-Nucleotidase↗

Application of ethyl esters and d3-methyl esters as internal standards for the gas chromatographic quantification of transesterified fatty acid methyl esters in food.

Ethyl esters (FAEE) and trideuterium-labeled methyl esters (d3-FAME) of fatty acids were prepared and investigated regarding their suitability as internal standards (IS) for the determination of fatty acids as methyl esters (FAME). On CP-Sil 88, ethyl esters of odd-numbered fatty acids eluted approximately 0.5 min after the respective FAME, and only coelutions with minor FAME were observed. Depending on the problem, one or even many FAEE can be added as IS for the quantification of FAME by both GC-FID and GC-MS. By contrast, d3-FAME coeluted with FAME on the polar GC column, and the use of the former as IS requires application of GC-MS. In the SIM mode, m/z 77 and 90 are suggested for d3-methyl esters of saturated fatty acids, whereas m/z 88 and 101 are recommended for ethyl esters of saturated fatty acids. These m/z values give either no or very low response for FAME and can thus be used for the analysis of FAME in food by GC-MS in the SIM mode. Fatty acids in sunflower oil and mozzarella cheese were quantified using five saturated FAEE as IS. Gravimetric studies showed that the transesterification procedure could be carried out without of loss of fatty acids. GC-EI/MS full scan analysis was suitable for the quantitative determination of all unsaturated fatty acids in both food samples, whereas GC-EI/MS in the SIM mode was particularly valuable for quantifying minor fatty acids. The novel GC-EI/MS/SIM method using fatty acid ethyl esters as internal standards can be used to quantify individual fatty acids only, that is, without determination of all fatty acids (the common 100% method), although this is present. This was demonstrated by the exclusive quantification of selected fatty acids including methyl-branched fatty acids, erucic acid (18:1n-9trans), and polyunsaturated fatty acids in cod liver oil and goat's milk fat.

Animals↗

Linoleic acid enhances mitogen response and survival of septic weanling rats.

Immunologic responses to various dietary fats and concentrations remain controversial. The authors examined the effect of dietary omega-3 and omega-6 polyunsaturated fatty acids (PUFA) on the immune system of weanling rats. Male weanling Sprague-Dawley rats were fed special liquid diets based on a standard enteral formula: (I) control: 2.2% calories as safflower oil (linoleic acid), (II) omega-3: 2.2% calories as safflower oil and 30% calories as cod liver oil (linolenic acid), and (III) omega-6: 32.2% calories as safflower oil. Body weight and caloric intake were measured daily. Rats were fed special diets for 18 days, at which time they were subjected to either septic challenge with 2 x 10(3) colony forming units of Streptococcus pneumoniae (eight rats per diet group) or killed and a splenocyte mitogen assay performed (four rats per diet group). Supplementation with omega-6 resulted in prolonged survival after bacterial challenge (control, 30.5 +/- 0.5 hours; omega-3, 31.6 +/- 0.5; not significant; omega-6, 39.8 +/- 5.0; P < .05). A significant difference in mitogenic stimulation was seen with omega-3 and omega-6 in response to PHA (control, 10,856 +/- 3,342; omega-3, 14,605 +/- 3,042; P < .05; omega-6, 35,737 +/- 7,596; P < .05) and with omega-6 only in response to LPS (control, 3,543 +/- 1,083; omega-3, 8,777 +/- 1,269; omega-6, 10,101 +/- 3,008). Previous studies have suggested that diets high in omega-6 PUFA are immunosuppressive because of an increase in the production of the dienoic prostaglandins.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Docosahexaenoic acid is superior to eicosapentaenoic acid as the essential fatty acid for growth of grouper, Epinephelus malabaricus.

Juvenile grouper (Epinephelus malabaricus) were fed seven experimental diets, one control diet and one reference diet for 12 wk to determine the dietary requirement of grouper for docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids. Each of the seven diets contained 1 g/100 g DHA and EPA in various combinations and 9 g/100 g tristearin. The control diet contained 1 g/100 g trilinolenin and trilinolein (3:1, wt/wt), and no supplemental EPA or DHA. The reference diet contained only natural oils from a mixture of cod liver oil, linseed oil and safflower oil at a ratio of 2:1:1 (wt/wt/wt). Significant differences (P < 0.05) in growth were observed among the dietary treatments but not in survival rate or relative liver weight. Only the diet with the highest DHA/EPA ratio (3:1) promoted significantly greater growth than the control diet. Purified EPA and DHA did not perform better in promoting growth than did the impure EPA and DHA oils. Enhanced growth was observed when the dietary DHA/EPA ratio was greater than 1, indicating that DHA was superior to EPA in promoting fish growth. Neutral lipid (NL) was the predominant lipid fraction (>70%) in both liver and muscle. Tissue NL/polar lipid did not differ among groups except the reference diet group that had a higher ratio (P < 0.05). DHA and EPA levels in the grouper tissues, especially muscle, were highly reflective of dietary levels of DHA and EPA, indicating that direct incorporation was likely. In addition, the 20:1(n-9), concentration in NL fractions seems to be an appropriate indicator of dietary essential fatty acid deficiency in grouper.

Animals↗

Fish and healthy pregnancy: more than just a red herring!

In modern Western diets we eat predominantly omega-6 essential fatty acids from vegetable oils, and too little omega-3. The Department of Health recommends doubling the amount of omega-3s we eat. Omega-3 essential fatty acids (EFAs) are derived mainly from fish oils. Omega-3s are believed to help reduce the risks of cardiovascular disease. Among the main materials required for fetal brain and CNS growth in late pregnancy are omega-3s (mainly docosahexaenoic acid [DHA]) and omega-6 EFAs (mainly arachidonic acid [AA]). These come from the mother's diet. Several formulae for preterm infants now contain DHA and AA to aid optimum brain, nerve and retinal development. One manufacturer has begun to include DHA and AA in formulae for term babies. Breast milk contains DHA and AA, derived from the mother's diet. Eating oily fish in pregnancy has been found to have a slight beneficial effect on birthweight and length of gestation. Eating fresh or canned oil-rich fish (e.g. kippers, herring, mackerel, salmon, sardine, pilchards, tuna) twice or three times a week can be encouraged as part of a healthy balanced diet, in pregnancy and for all the family. As well as containing omega-3 polyunsaturates, oily fish is a good source of protein and vitamins A and D. Alternatively, a fish-oil supplement may be taken. Cod liver oil is best avoided during pregnancy, because of concerns over the possible teratogenicity of vitamin A.

Animals↗

[Blood leukotriene B4 response to endotoxin in fish oil fed rats].

Six-week-old male Wistar rats were given a low fat diet containing 15% cod liver oil (FO; n = 30) or 15% safflower oil (SO; n = 30) for 6 weeks. In the polymorphonuclear leukocytes, arachidonic acid (AA) was significantly lower in the FO group than in the SO group (p < 0.001), and eicosapentaenoic acid (EPA) in the FO group was present in large amounts showing the EPA/AA ratio of 1.76. When lipopolysaccharide (LPS; 10mg/kg) was intraperitoneally injected, white blood cell counts in the blood significantly decreased in both groups (p < 0.001) compared to the controls 2 hours later, and at 4 hours, the counts in the SO group (2,033 +/- 151/mm3) were lower than in the FO group (3,189 +/- 624/mm3), (p < 0.01). In both groups, leukotriene B4 (LTB4) in the whole blood increased at 4 hours following LPS-administration compared to the controls (1,219 +/- 167.3pg/ml in the SO group and 600.0 +/- 109.0pg/ml in the FO group, p < 0.001). It should be noted that the level of LTB4 in the SO group was significantly higher than in the FO group (p < 0.001). Changes of the metabolites including decreased LTB4 production in the arachidonic cascade related to the increase of EPA may play a role in the inhibition of the decrease in white blood cell counts.

Administration, Oral↗

On-line clean-up of pressurized liquid extracts for the determination of polychlorinated biphenyls in feedingstuffs and food matrices using gas chromatography-mass spectrometry.

This paper describes a fast and simple pressurized liquid extraction method for the determination of polychlorinated biphenyls (PCBs) in feedingstuffs and food matrices. The method is based on a simultaneous extraction/clean-up step requiring a minimum of sample handling. The final analysis was performed with gas chromatography-mass spectrometry. Seven PCBs (28, 52, 101, 118, 138, 153 and 180) were analyzed, all of which are indicator congeners that, according to European legislation should be included in the analytical monitoring program. The extracted matrices were spiked feed for poultry and two certified reference materials naturally contaminated with PCBs (cod-liver oil and milk powder), which showed excellent conformity with certified data.

Animal Feed↗

Improved resolution of natural diacylglycerols by gas-liquid chromatography on polar siloxanes.

A new cyanoalkylphenylsiloxane (SILAR 5CP) liquid phase is shown to possess sufficient polarity to permit improved GC separations of natural diacylglycerols based on unsaturation and positional placement of fatty acids as well as on molecular weight, which was previously possible only on ethylene glycol succinate polyesters. Unlike the polyesters, the polar siloxane polymer has moderate thermal stability and provides GC columns which can be used for several months without replacing the packing. The GC analyses were made with conventional columns containing 3% SILAR 5CP on Gas Chrom Q at 270 degrees C isothermally. The diacylglycerols were chromatographed as the TMS ethers. Excellent seperations were obtained for the 1,2(2,3)- and 1,3-diacyglycerols derived from corn, linseed, peanut and cod liver oils and for the 1,2-diacyl-sn-glycerols from hepatic glycerophospholipids.

Animals↗

Influence of preeclampsia or maternal intake of omega-3 fatty acids on the vasoactive effect of prostaglandin F-two-alpha in human umbilical arteries.

The vasoactive effect of prostaglandin F(2alpha) (PGF(2alpha)) was studied in in vitro perfused human umbilical arteries following maternal dietary supplementation with omega-3 fatty acids or in pregnancies complicated by a moderate degree of preeclampsia. In most preparations PGF(2alpha) induced a biphasic pressure response with a transient dilatation followed by a constrictory response. The pressure increase was significant in both groups, but no significant differences in the constrictory response or in the proportions of preparations displaying dilatatory responses were observed when compared to appropriate control groups. In conclusion, neither preeclampsia nor dietary supplementation with cod-liver oil had any significant effect on the vasoactive response to PGF(2alpha) in umbilical cord arteries.

Analysis of Variance↗

Spectrophotometric determination of vitamin A in oils and fats.

A spectrophotometric method for determining vitamin A based on interaction with 50% trichloroacetic acid solution in dichloromethane was developed. The blue reaction product had a maximum absorption at 620 nm and obeyed Beer's law over the concentration range of 0.5-5.0 microgram retinol/ml solution. The molar absorptivity of the reaction product was 1.58 X 10(5). As much as 100 microgram vitamin D2, and beta-carotene up to 12 times the vitamin A concentration, did not interfere with the determination. The results obtained from the determination of vitamin A in cod liver oil and butter showed excellent agreement with the Carr-Price method, 43.013(d).

Butter↗

Fish oil feeding lowers thromboxane- and prostacyclin production by rat platelets and aorta and does not result in the formation of prostaglandin I3.

It is demonstrated that feeding cod-liver oil to rats leads to a considerable reduction in the formation of platelet TxA2 and of vascular PGI2. No appreciable formation of TxA3 and PGI3 is observed, although arterial thrombosis is depressed and bleeding time is prolonged. These findings contradict the suggested role of prostaglandins of the 3-series in thromboregulation.

Animals↗

Effect of dietary fish oil on insulin sensitivity and metabolic fate of glucose in the skeletal muscle of normal rats.

The aim of this work was to study the effect of the administration of cod liver oil on the non-oxidative and oxidative fate of glucose metabolism in the skeletal muscle of normal rats. To achieve this goal, the gastrocnemius was examined regarding glucose oxidation, glycogen synthase activity and glycogen storage both at baseline and during euglycemic hyperinsulinemic clamping. The results show that dietary fish oil decreases plasma insulin levels without alteration in glucose homeostasis (at baseline). In addition, the observed enhancement in whole body glucose utilization during clamping suggests an increased peripheral insulin sensitivity. Furthermore, under insulin-stimulated glucose disposal, an enhancement in the glycolytic pathway (increased levels of muscle glucose-6-phosphate and plasma lactate) rather than changes in the oxidation (pyruvate dehydrogenase complex) and storage components of glucose metabolism was observed in the skeletal muscle of rats fed dietary fish oil. These results coupled with the hypolipidemic effects of fish oil may have implications for the prevention and/or management of some pathological states manifested by insulin resistance with or without dyslipidemia.

Animals↗

Identification of fatty acids by electrospray mass spectrometry and tandem mass spectrometry.

Low-energy negative-ion electrospray mass spectrometry (ESI-MS) and ESI-MS/MS were used to characterize saturated and unsaturated fatty acids. The carbon number and degree of unsaturation of fatty acids were determined using ESI-MS, and MS/MS was used to localize some double bond positions of mono-and polyunsaturated fatty acids. For compounds with up to two unsaturated bonds, fragmentation was dominated by loss of H2O from the carboxyl moiety and very low-intensity peaks generated from bonds cleaved at carbons alpha and/or beta to sites of unsaturation. Fragmentation of monounsaturated fatty acids was minimal using this soft method of mass spectrometric analysis, but increased with progressively greater degrees of fatty acid unsaturation. There was extensive hydride migration during ESI-MS/MS of compounds with three or more double bonds. Although this behavior complicated localization of double and triple bonds, the spectra were reproducible. Many peaks could not be definitively assigned to specific product ions, but the spectra of standards and complementary natural products were similar and isobaric compounds could be differentiated. The utility of this technique to examine biological samples was shown by analysis of the fatty acid composition of cod liver oil. Detection limits for negative-ion ESI-MS/MS were at or below 1 pg.

Arachidonic Acids↗