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Influence of dietary fish oil on arterial thrombosis and atherosclerosis in animal models and in man.

In animal models of arterial thrombosis, fish-oil-enriched diets have been shown to have an antithrombotic effect. There is strong evidence, however, that this only holds if the fish-oil consumption is associated with a reduction of the saturated fat intake. In animal models of atherosclerosis the effect of a marine diet is not consistent. Although in some studies with cod-liver oil (which is relatively low in saturated fatty acids) a beneficial effect is observed, studies with other, more saturated fish oils do not confirm this. The influence of fish-oil consumption on some thrombotic complications of human atherosclerosis is not well documented. So far, the few results reported are not very promising, but it should remembered that this mainly concerns short-term studies. Studies with respect to human fibrinolysis are equivocal. Evidence has been obtained that fish consumption may even have an adverse effect on certain aspects of the fibrinolytic process. The studies performed so far do not provide conclusive evidence as to the importance of fish- and fish-oil-enriched food for prevention and therapy of ischaemic cardiovascular disease. Further research is needed, especially with long-term, high-quality animal models. Moreover, long-term, well-controlled, prospective human intervention studies are required to investigate the effect of feasible fish(oil)-enriched diets on morbidity and mortality of ischaemic cardiac and vascular diseases.

Angina Pectoris↗

Intraduodenal fat and amino acids activate gallbladder motility in the rainbow trout, Oncorhynchus mykiss.

The effects on gall bladder motility of intraduodenal injections of fat (cod liver oil) and amino acids were investigated in rainbow trout, Onchorhynchus mykiss, in vivo. Fat (1 ml/kg, 15 min) increased basic tonus of the gallbladder by 79% and frequency of contractions by 53%; both effects were antagonized by pretreatment with atropine (1.2 mg/kg). The amino acid mixture similarly increased tonus and frequency of contractions of the gallbladder smooth muscle by 63 and 109%, respectively. The effects were mimicked by intraarterial infusion of cholecystokinin (100 pmol-1 nmol/kg-hr). It is concluded that the presence of oil and amino acids in the duodenum stimulates rainbow trout gallbladder motility and tension, possibly by a release of cholecystokinin, and that the effect of oil, as for cholecystokinin, involves a cholinergic pathway.

Amino Acids↗

Effects of dietary polyunsaturated fatty acids on rat brain plasma membrane fatty acid composition.

The effects of four different diets on phospholipid fatty acid composition of rat brain plasma membranes were evaluated. Rats were given a semisynthetic diet in which lipids were supplied by 5% peanut oil (n-3 PUFA deficient diet), cod liver oil (n-6 PUFA deficient diet), partially hydrogenated palm oil (total PUFA deficient diet) or a mixture of peanut and rapeseed oil (control group). Animals fed the total PUFA deficient diet had significantly lower body and brain weights than the control group (p < 0.05). Lower brain cholesterol and phospholipids also were observed in animals fed the total PUFA deficient diet, whereas the brain of animals fed the n-6 PUFA deficient diet had higher levels of these lipid components than the control group (p < 0.05). Phosphatidylcholine and phosphatidylethanolamine were mutually replaced in animals fed the n-6 and n-3 PUFA deficient diets, so that the sum of these two membrane constituents was maintained around 77% of total phospholipids. Brain phospholipid fatty acid composition was significantly modified by the diets studied. Thus, despite being a highly protected organ, the fatty acid composition of the brain can be extensively modulated by dietary lipids.

Animals↗

Comparison of effects of fish oil and corn oil supplements on hyperlipidemic diet induced atherogenesis in swine.

The addition of a fish oil supplement rich in n - 3 unsaturated fatty acids to a high cholesterol, high saturated fat (BT) diet for swine has been shown previously to result in modest lowering of plasma cholesterol levels and in marked retardation of atherogenesis. It has been suggested that the effect was due to the change in polyunsaturated (PUFA) to saturated fatty acid ratios (P/S) and that a supplement of PUFA of the n - 6 series might have the same effect as the fish oil. We have tested this hypothesis in swine fed an atherogenic diet by comparing the effect of a fish oil supplement producing a P/S ratio of 0.28 to that of corn oil in the same amount producing a ratio of 0.46. The P/S ratio of the atherogenic diet without supplements was 0.16. Thirteen young male Yorkshire swine were fed either BT alone (n = 4), BT + cod liver oil (n = 4) or BT + corn oil (n = 5) for 6 months and then killed for quantitative studies of atherosclerosis in the aortas and coronary arteries including lesion areas, number of lesion cells, and number of monocytes attached to endothelium. Plasma cholesterol levels were determined periodically and lipoproteins were separated terminally by density gradient ultracentrifugation, Pevikon block electrophoresis and immunoelectrophoresis. The fish oil supplement resulted in a 30% reduction in time-weighted average plasma cholesterol levels, and a marked shift in terminal lipoprotein patterns from predominantly apo B and E containing ones to predominantly apo B only ones. Atherogenesis was reduced by the fish oil supplement as judged by several morphometric criteria including size of lesions, number of lesion cells, and number of monocytes attached to lesion endothelium. The corn oil supplement produced no significant reductions in any of these variables from those in swine fed the atherogenic BT diet without the supplement. We conclude that the n - 3 fatty acid rich fish oil supplemented diet retarded atherogenesis, but that this effect was not shared by the corn oil supplemented diet which had an even higher P/S ratio.

Animals↗

Determination of organochlorine compounds in fatty matrices. Application of rapid off-line normal-phase liquid chromatographic clean-up.

A clean-up method for organochlorine compounds in fatty samples based on normal-phase liquid chromatography (NPLC) is described. To this end, an existing clean-up procedure which uses column switching for the separation of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) from the fat matrix was simplified to a single silica LC column procedure. The use of an LC column packed with 3 microns silica enables complete fat/OCP separation in a total fraction volume of 12 ml, and results in a fully automated clean-up procedure that takes only 32 min per sample. The method showed average recoveries of 80-110% in the concentration range of 1-510 micrograms/kg, with relative standard deviations of less than 10%. Limits of determination were in the range of 0.5-50 micrograms/kg. The simplified approach has shown its potential for a variety of samples, such as milk, pork fat, animal feed and cod liver oil, showing its general applicability to fatty samples.

Adipose Tissue↗

Prostaglandin I3 is formed in vivo in man after dietary eicosapentaenoic acid.

Greenland Eskimos who live on a traditional marine diet rich in long chain omega 3-polyunsaturated fatty acids have a low incidence of cardiovascular disease and myocardial infarction. In their plasma and platelet lipids, arachidonic acid, the precursor of dienoic prostanoids, is partly replaced by eicosapentaenoic acid (C20:5, omega 3; EPA), the precursor of trienoic prostanoids. Studies with an Eskimo diet or a Western diet supplemented with sea fish or fish oil rich in EPA resulted in an 'Eskimo-like' pattern of plasma and platelet lipids. Moreover, less reactive platelets, a reduced ex vivo formation of proaggregatory thromboxane A2 and a blunted circulatory response to pressor hormones were reported. These favourable functional effects may be induced by a shift of prostanoid formation from the dienoic to the trienoic series. We show here that the major urinary metabolite of endogenous prostaglandin I3 is present in subjects that have ingested either cod liver oil (approximately 4 g EPA per day) or mackerel (approximately 10-15 g EPA per day). Our studies provide the first direct evidence for in vivo formation of prostaglandin I3 in man.

Blood Coagulation↗

Fractionation of fish oil fatty acid methyl esters by means of argentation and reversed-phase high-performance liquid chromatography, and its utility in total fatty acid analysis.

The utility of reversed-phase and argentation high-performance liquid chromatography (HPLC) as pre-fractionation methods in fatty acid analysis is discussed. Both HPLC modes were applied to cod liver oil fatty acid methyl esters. Apart from positional isomers, the fractions obtained by reversed-phase HPLC were analysed by gas-liquid chromatography and appear to be free of the usually occurring "critical pairs". The mechanism of retention of the fatty acid methyl esters on low-loaded silver nitrate-impregnated silicas is discussed. It is shown that argentation HPLC is a rapid semi-preparative pre-fractionation method for highly unsaturated fatty esters with 3-6 double bonds.

Animals↗

DDT residues in cod livers from the Maritime Provinces of Canada.

The residues of DDT and its analogs were estimated in a number of cod liver samples (Gadus morhua) collected at six sampling sites off the Atlantic Coast of the Maritime Provinces of Canada during the summer of 1971. The residue levels indicated widespread distribution of DDT over the entire region sampled. Variations in the relative contributions of p,p'-DDE and p,p'-DDT to the total DDT residues (xiDDT) were noted and the tendency for these residues to preferentially accumulate in lipid rich tissues was demonstrated. The residue levels found in cod livers were compared to the residue levels determined in cod flesh taken from some of the same specimens and also to residue levels in commercially refined cod liver oils. Samples of each of these types were stored at -29 degrees C and analyzed in 1973 for DDT and PCB simultaneously. The xiDDT residues determined by both methods of analysis were compared.

Animals↗

Rapid preparation of micro sample and gas-liquid chromatographic determination of methoxychlor residues in animal tissues and water.

Methods are described for analyses of methoxychlor (2,2-bis(p-methoxyphenyl)-1,1,1-trichloroethane) in tissues of fish and invertebrate animals and in water samples. These micro methods offer economy in solvents and materials, and reduce health and fire hazards. Furthermore, the methods sufficiently reduce sample weights to allow analyses of small animals (mayflies) or animal organs. Samples are extracted in a ball-mill with n-hexane, and lipids are removed from fatty samples by freezeout. Small Florisil columns give final lipid cleanup and partial separation of methoxychlor from interfering materials. Methoxychlor is determined on a short gas-liquid chromatographic column (retention time 2.2 min). Animal tissue samples as small as 0.1 g and water samples of 5 ml can be analyzed at sensitivities of 10 and 1 ppb, respectively. Methoxychlor recoveries were 84-101% (fish and crab tissue), 92-100% (cod liver oil), and 98-107% (water) over a concentration range of 0.-10 ppm. Results obtained for fish tissue by using micro procedures agreed to within 20% of results reported by an independent laboratory using an established method.

Animals↗

The influence of fish oil diet and norepinephrine treatment on fatty acid composition of rat heart phospholipids and the positional fatty acid distribution in phosphatidylethanolamine.

The effect of chronic norepinephrine (NE) administration with increasing dosage from 1-4 mg/kg over a period of 2 weeks was studied on cardiac phospholipids and their fatty acid distribution in rats. Animals were fed a control diet or a 10% cod liver oil (CLO)-enriched diet. The relative distribution of various polyunsaturated fatty acids esterified to the 1- and 2-position of the phosphatidylethanolamine fraction was estimated. NE stress during control feeding significantly reduced the total phospholipid content in rat heart. No differences in the phospholipid class distribution were found. However, CLO feeding as well as chronic NE administration resulted in a decrease of omega 6 fatty acids, mainly C 18:2 omega 6 and C 20:4 omega 6, which was compensated with an increase in omega 3 fatty acids, mainly C 20:5 omega 3 and C 22:6 omega 3. The changes in fatty acid composition qualitatively agree with those reported by Gudbjarnason et al. (23), except that the mortality in our NE-treated control or CLO-fed groups was considerably lower. It can probably be attributed to a different mode of NE administration. On the other hand, at the end of the CLO feeding period in rats treated with NE or not, comparing with control fed rats without NE treatment, the incidence rate of ST segment elevation in electrocardiogram (ECG) recorded under light diethylether-induced anesthesia was higher. Independent of whether the fatty acid composition of myocardial phospholipids was dietary or pharmacologically manipulated, most of the polyunsaturated fatty acids were found at the 2-position of the phosphatidylethanolamine molecules. The polyunsaturated fatty acids account for 45-50% of the fatty acyl residues and preferentially occupy the 2-position, where they can exchange for each other.

Animals↗

Docosahexaenoic acid in cardiac metabolism and function.

The polyene fatty acid compostition of cardiac phospholipids is modified by a) dietary cod liver oil, b) norepinephrine, c) chronic administration of nicotine to animals fed a high cholesterol diet. Polyene fatty acids stimulate microsomal oxydation of epinephrine to cardiotoxic adrenochrome. Adrenochrome stimulates microsomal peroxydation or oxygenation of polyene fatty acids. There is an exponential relationship between docosahexaenoic acid of cardiac phospholipids and the heart rate.

Animals↗

Complimentary and alternative medicine use among patients starting warfarin.

The use of complimentary and alternative medicines (CAM) among the UK population is on the increase. For patients requiring warfarin therapy, it is important to maintain an adequate and safe level of anticoagulation. As some forms of CAM can interact with warfarin, it is imperative that any patient considered for warfarin, is asked about their use of CAM. Our report describes the incidence and type of CAM usage among patients about to start or recently commenced on warfarin therapy attending our outpatient anticoagulant clinic. All patients attending clinic for the first time were seen by a pharmacist. A retrospective analysis of the pharmaceutical care plans for all patients seen during 2003 were analysed regarding the degree of CAM usage. Of 631 care plans reviewed, 170 (26.9%) patients were taking some form of CAM. Ninety-nine (58% of all CAM users) were taking a CAM that could interact with warfarin; the commonest forms were cod-liver oil capsules and garlic capsules. We conclude that many patients new to warfarin therapy were significant users of CAM, many of which had the potential to interact with warfarin. By taking a full drug history, potential CAM/warfarin interactions could be avoided.

Adolescent↗

A comparison of pyrrolidide and picolinyl ester derivatives for the identification of fatty acids in natural samples by gas chromatography-mass spectrometry.

The pyrrolidide and picolinyl ester derivatives of the fatty acids in two natural lipid samples rich in unsaturated fatty acids, pig testis lipids and cod liver oil were satisfactorily resolved on capillary columns of fused silica coated with stationary phases of varying polarity. The picolinyl esters, in particular, when subjected to gas chromatography-mass spectrometry on a column containing a cross-linked methyl silicone, gave distinctive mass spectra, which could be interpreted in terms of both the numbers and positions of the double bonds.

Animals↗

Platelet - vessel wall interaction: role of blood clotting.

Vascular damage initiates not only the adhesion and aggregation of blood platelets but also coagulation, which is of mixed (intrinsic and extrinsic) origin. Evidence is presented that thrombin, generated as a result of the injury, is a prerequisite for platelet aggregation. Platelets, after activation, in their turn promote coagulation. Prostaglandin I2 (PGI2 or prostacyclin) inhibits coagulation induced by damaged vascular tissue. This effect of PGI2 is mediated by the inhibition of platelets in their participation in the generation of factor Xa and thrombin. Dietary cod liver oil, by changing plasma coagulability, decreases the procoagulation activity of vessel walls, and arterial thrombosis. Another fish oil with similar effects on plasma coagulability and some other haemostatic parameters does not modify vessel wall-induced clotting, nor does it significantly lower arterial thrombosis tendency; this indicates the physiological relevance of vessel wall-induced clotting in arterial thrombus formation. Some evidence is also given for the importance of vessel wall-induced clotting in primary haemostasis.

Adenosine Diphosphate↗

The effect of dietary alteration of prostaglandin synthesis on blood pressure and the reversal of hypertension in the one-kidney, one-clip rat.

This study examined the effect of diet-induced changes in prostaglandin synthesis on systolic blood pressure in one-kidney, one clip (1k, 1C) hypertensive rats and on the fall in blood pressure after unclipping. It tested the hypothesis that inhibition of prostaglandin synthesis exacerbates hypertension in this model and prevents complete reversal after unclipping. Rats with sustained hypertension within 8 weeks of renal artery clipping were fed synthetic diets supplemented to 20% of total energy with either safflower oil (linoleic acid) or a mixture of cod liver oil (90%) (containing eicosapentaenoic acid) and linseed oil (10%) (containing linolenic acid) for 4 weeks. The latter oil mixture resulted in a predictable reduction in kidney PGE2 and 6-keto PGF1 alpha (hydrolysis product of PGI2), aortic 6-keto PGF1 alpha and serum TXB2. However, at the end of 4 weeks dietary treatment there were no differences in systolic blood pressure between the two diet groups, and the blood pressure fall 24 hours after unclipping was similar. These findings therefore do not support a role for prostanoids in the maintenance or reversal of 1K, 1C hypertension.

6-Ketoprostaglandin F1 alpha↗

Effect of diets containing different oils on brain fatty acid composition in sea bass (Dicentrarchus labrax L.).

Four groups of adult sea bass were given diets containing about 8% of one of four different oils having a different fatty acid composition: linseed oil, grape-seed oil, containing high amounts of linolenic and linoleic acids respectively, hydrogenated coconut oil, mainly containing saturated fatty acids, and cod liver oil which was considered as reference. Total lipid, phospholipid and polar lipid contents of the brain of the different groups of sea bass were unaffected. The fatty acid composition of the brain agreed with the dietary history of sea bass: thus adult sea bass brain is capable of incorporating dietary fatty acids. Sea bass brain and structural lipids of the liver appeared to be similarly sensitive to the dietary input in contrast with mammalian brain which was reported to be more resistant than other tissues. The more striking dietary effect on liver total lipid fatty acid composition is ascribed to the incorporation of dietary fatty acids in depot fats.

Animals↗

Effects of dietary fat on benz-a-pyrene-induced forestomach tumorigenesis in mice chronically exposed to arsenic.

The modulating effect of As (As2O3) in drinking water plus dietary fatty acids on benzo-a-pyrene (BP) induced forestomach tumorigenesis was assayed in mice fed with corn oil (CO), Olein (O), palmstearin (PS) and cod liver oil (CLO) enriched formulae, and a mixed-fat (S-stock) diet during 28 weeks. Neither pre-or neoplastic lesions were recorded in esophagus or forestomach of mice treated with As alone. CLO diet showed a protective effect. Indeed, multiple epidermal hyperplasia were significantly reduced compared to S group and a reduction in papillomas multiplicity with respect to S, CO and O groups was also observed. Mice fed on PS formula showed minor papillomas per mouse with respect to CO and O. These results indicate that As plus BP did not exhibit any synergistic effect on the development of epidermoid tumor lesions, whereas CLO diet exerted antipromoting activity . In spite of their common essential fatty acid deficiency (EFAD) condition, PS and O lots showed contradictory results. Hence, the dietary enrichment in 18:1 n-9 (O diet) per se, and not the EFAD state, may play a deleterious role. Corn diet showed no promoting activity on the epidermoid tumor development in this model.

Animals↗

The oxidation of benzo[a]pyrene-7,8-dihydrodiol mediated by lipid peroxidation in the rat intestine and the effect of dietary lipids.

This study has demonstrated that the microsomal fraction of the rat small intestinal mucosa has the capacity to catalyse the oxidation of benzo[a]pyrene(BP)-7,8-diol to BP-diol-epoxides (BPDEs) both by a mechanism involving the mixed-function oxidase system (NADPH-dependent) and as a result of the initiation of peroxidation of the membrane phospholipids by ferrous ions, ascorbate and ADP. The NADPH-dependent reaction was fastest in the proximal part of the intestine and resulted in the formation of approximately equal amounts of BPDE I and BPDE II. The lipid peroxidation-catalysed reaction favoured the production of BPDE I and was maximal in the middle region of the intestine, closely paralleling the rate of lipid peroxidation in the intestinal sections. Feeding rats on a cod liver oil diet, rich in C20:5 and C22:6, significantly increased the incorporation of these fatty acids into the microsomal fractions. This resulted in a greatly increased rate of lipid peroxidation in vitro and a significantly higher rate of lipid peroxidation-catalysed BP-7,8-diol oxidation compared to rats fed fat-free, mono-unsaturated lard or corn oil (58% C18:2) diets. Thus the rate of conversion of BP-7,8-diol to its ultimate carcinogenic forms during lipid peroxidation in the intestinal fractions of rats fed a polyunsaturated fat was quantitatively more important than the NADPH-catalysed reaction as measured in vitro.

Animals↗