Essentiality of dietary ω3 fatty acids for premature infants: Plasma and red blood cell fatty acid composition

Pre-term infants, that are not breast-fed, are deprived of vital intrauterine fat accretion during late pregnancy and must rely on formula to obtain fatty acids essential for normal development, particularly of the visual system. Preterm infants (30 wk postconception) receiving human milk were compared to infants given one of the following formulae: Formula A was a commercial preterm formula with predominantly 18∶2ω6 (24.2%) and low (0.5%) 18∶3ω3; Formula B was based on soy oil and contained similar 18∶2ω6 levels (20%) and high 18∶3ω3 (2.7%); Formula C was also a soy oil-based formula (20% 18∶2, 1.4% 18∶3) but was supplemented with marine oil to provide ω3 long-chain polyunsaturated fatty acids (LCP) at a level (docosahexaenoic acid, DHA, 0.35%) equivalent to human milk. At entry (10 days of age), the fatty acid composition of plasma and red blood cell (RBC) membrane lipids of the formula groups were identical. By 36 wk postconception, the DHA content in lipids of group A was significantly reduced compared to that in the human milk and marine oil formula groups. Omega-3 LCP results were further amplified by 57 wk with compensatory increases in 22∶5ω6 in both plasma and RBC lipids. Provision of 2.7% α-linolenic acid in formula group B was sufficient to maintain 22∶6ω3 levels equivalent to those in human milk-fed infants at 36 wk but not at 57 wk. Effects on the production of thiobarbituric acid reactive substances and fragility of RBC attributable to the marine oil supplementation were negligible. The results support the essentiality of ω3 fatty acids for preterm infants to obtain fatty acid profiles comparable to infants receiving human milk. Formula for preterm infants should be supplemented with ω3 fatty acids including LCP. Based on a paper presented at the Symposium on Milk Lipids held at the AOCS Annual Meeting, Baltimore, MD, April 1990.     

Alteration and recovery of bleeding times,platelet aggregation and fatty acid composition of individual phospholipids in platelets of human subjects receiving a supplement of cod-liver oil

The effect of supplementation with cod-liver oil containing eicosapentaenoic acid (EPA), 20∶5ω3, on bleeding times, thrombin-induced platelet aggregation, platelet protein, platelet cholesterol, and the level and fatty acid composition of individual phospholipids in the platelets of human subjects was determined. Measurement of these parameters was conducted before the subjects received the supplement (day 0), after they received the supplement for 14 days (day 14), and 14 days after the supplement was terminated (day 28) so as to monitor recovery. The mean bleeding times exhibited a marked increase (by 81%) with supplementation and returned to near basal (day 0) values within 14 days after the supplement was terminated. Cod-liver oil supplementation significantly reduced thrombin-induced platelet aggregation with a partial recovery being exhibited by day 28. The content of phospholipid, cholesterol and protein (μg/10⁹ platelets) was not significantly different (P>0.05) when isolated from the subjects at day 0, 14 and 28, as neither were the composition of individual phospholipids [phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI) and sphingomyelin (SPH)] given as % of total phospholipid. However, the fatty acid compositions of all platelet phospholipids were altered significantly by the fish oil supplement. In PC, EPA rose from 0.3 to 2.9% of total fatty acids and docosahexaenoate from 0.7 to 1.8% concomitant with a drop in arachidonate (from 14.1 to 9.6%) and linoleate (from 10.2 to 7.9%); these levels approached basal levels 14 days after supplementation was terminated. The highest percentage of EPA with supplementation was found in PE (4.3%), while the arachidonate fell from 38.8 to 30.5%, with low percentages of EPA occurring in PS (0.7%) and PI (0.5%). The level of 24∶1 in SPH increased significantly (from 17.8 to 24.8) with supplementation and reverted to basal values by day 28. These results suggest a close relationship of the observed fatty acid changes in individual platelet phospholipids to the altered hematological parameters and platelet-vessel wall interactions produced by cod-liver oil supplementation.     

Effects of diets high in saturated fat and cholesterol on the lipid composition of canine platelets

The phospholipid composition of platelets from dogs on various experimental diets was determined. Thyroidectomized foxhounds were fed a control diet or the control diet supplemented with (1) beef tallow, (2) beef tallow and cholesterol, or (3) beef tallow, cholesterol, and safflower oil for 23 weeks prior to isolation of platelets. Platelets from animals fed the control diet contained 36.7% phosphatidylcholine (PC), 22.8% phosphatidylethanolamine (PE), 18.4% sphingomyelin (Sph), 11.8% phosphatidylserine (PS), 6.3% phosphatidylinositol (PI), and 2.2% lysophosphatidylcholine. The PE was 77.6% in the plasmalogen form. No highly significant changes in the phospholipid class composition resulted from the experimental diets. Cholesterol supplementation of the diets, however, caused consistent alterations in the fatty acid compositions of the platelet phospholipids including increases in the percentages of 18∶1ω9 (oleic acid), 18∶2ω6 (linoleic acid), and 20∶3ω6 (homo-gamma linolenic acid) and a decrease in the percentage of 20∶4ω6 (arachidonic acid). Addition of safflower oil to the tallow-cholesterol diet partially reversed these effects. These cholesterol-induced alterations in fatty acid composition could be due to exchange with plasma lipids, de novo synthesis, or altered platelet metabolism. The mechanism remains to be determined. Der. Nelson’s current affiliation is the Lipid Metabolism Branch, Division of Heart and Vascular Diseases, National Heart, Lung, and Blood Institute.     

The effect of borage oil consumption on the composition of individual phospholipids in human platelets

The effect of supplementation with borage oil containing γ-linolenic acid (GLA, 18∶3n−6) on the levels and fattya cid compositions of individual human platelet phospholipids was evaluated. For this purpose, male volunteers were given an average daily intake of 5.23 g of GLA (as borage oil) for 42 days, after which the supplement was withdrawn for an additional 42-day period. No significant differences were found in the relative amounts of the choline phospholipids (PC), ethanolamine phospholipids (PE), phosphatidylserine (PS), phosphatidylinositol (PI), and sphingomyelin (SPH) at days 0, 22, 43, 64, and 85. However, marked differences were observed in the fatty acid compositions of all the phospholipids including a marked, and reversible, rise in the level of dihomo-γ-linolenic acid (DGLA, 20∶3n−6), without a significant elevation in arachidonic acid (AA, 20∶4n−6) and decreases in n−3 polyunsaturated fatty acids. In the case of PC, a net rise in DGLA of 1.8 mol% was observed by day 22 (from 2.1 to 3.9 mol%). The DGLA/AA ratios at day 43 exhibited considerable variability across phospholipids with PC>PS>PE=PI; the PC, PE, PS, and PI accounted for 67.6, 16.7, 12.9, and 2.6%, respectively, of the total DGLA in platelet phospholipids. Interestingly, despite the lack of DGLA in SPH, this phospholipid exhibited a marked enrichment in nervonic acid (NA, 24∶1n−9) from 16.2 to 24.7 mol% upon borage oil consumption. The observed alterations may represent biochemical strategies for adaptation to dietary fatty acid modifications and the regulation of platelet membrane functioning.     

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.     

Diets rich in lean beef increase arachidonic acid and long-chain ω3 polyunsaturated fatty acid levels in plasma phospholipids

Diets rich in meat are claimed to contribute to the high tissue arachidonic acid (20∶4ω6) content in people in Westernized societies, but there are very few direct data to substantiate this assertion. Because meat contains a variety of long-chain polyunsaturated fatty acids (PUFA) that are susceptible to oxidation, we initially examined the effect of cooking on the long-chain PUFA content of beef, and then determined the effect of ingestion of lean beef on the concentration of long-chain PUFA in plasma phospholipids (PL). First, we examined the effect of grilling (5–15 min) and frying (10 min) different cuts of fat-trimmed lean beef on the long-chain PUFA content. Second, we investigated the effect of including 500 g lean beef daily (raw weight) for 4 wk on the fatty acid content and composition of plasma PL in 33 healthy volunteers. This study was part of a larger trial investigating the effect of lean beef on plasma cholesterol levels. In the first two weeks, the subjects ate a very low-fat diet (10% energy) followed by an increase in the dietary fat by 10% each week for the next 2 wk. The added fat consisted of beef fat, or olive oil (as the oil or a margarine) or safflower oil (as the oil or a margarine). This quantity of beef provided 60, 230, 125, 140 and 20 mg/d, respectively, of eicosatrienoic acid (20∶3ω6), 20∶4ω6, eicosapentaenoic acid (20∶5ω3), docosapentaenoic acid (22∶5ω3) and docosahexaenoic acid (22∶6ω3). Grilling for 10–15 min, but not frying, of the fat-trimmed lean beef resulted in 20–30% losses of the 20 and 22 carbon PUFA. The consumption of the lean beef during the first two-week period, when there was a very low level of dietary fat, was associated with significant increases in the proportion and concentration of 20∶3ω6, 20∶4ω6, 20∶5ω3 and 22∶5ω3 in the plasma PL and a significant decrease in the proportion and content of 18∶2ω6. The addition of beef fat or olive oil to the diets containing lean beef did not alter the plasma PL fatty acid profile compared with the very low-fat diet, whereas the addition of safflower oil maintained the significant increases in 20∶4ω6 and 22∶5ω3 but led to decreases in 18∶3ω3 and 20∶5ω3 compared with the very lowfat diet. The results showed that diets rich in lean beef increased the 20∶3ω6, 20∶4ω6 and the long-chain ω3 PUFA levels in the plasma PL. A high level of linoleic acid in diets rich in lean beef prevented the rise in the plasma level of 20∶3ω6 and 20∶5ω3, two fatty acids known to antagonize the effects of 20∶4ω6 on platelet aggregation.     

Dietary manipulation of macrophage phospholipid classes: Selective increase of dihomogammalinolenic acid

Because alterations in the dietary content of fatty acids are an important method for modulating macrophage eicosanoid production, we have quantitated the levels of n−6 and n−3 polyunsaturated fatty acids in peritoneal macrophage individual phospholipids from mice fed diets (3 wk) with either safflower oil (SAF), predominantly containing 18∶2n−6, borage (BOR) containing 18∶2n−6 and 18∶3n−6, fish (MFO) containing 20∶5n−3 and 22∶6n−3, and borage/fish mixture (MIX) containing 18∶2n−6, 18∶3n−6, 20∶5n−3 and 22∶6n−3. Dietary n−3 fattya cids were readily incorporated into macrophage phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI). The increase in n−3 fatty acid levels was accompanied by a decrease in the absolute levels of 18∶2n−6, 20∶4n−6 and 22∶4n−6 in PC, PE and PS. Interestingly, PI 20∶4n−6 levels were not significantly lowered (P>0.05) in MIX and MFO macrophages relative to SAF and BOR. These data demonstrate the unique ability of this phospholipid to selectively maintain its 20∶4n−6 levels. In BOR and MIX animals, 20∶3n−6 levels were significantly increased (P<0.05) in all phospholipids relative to SAF and MFO. The combination of borage and fish oils (MIX diet) produced the highest 20∶3n−6/20∶4n−6 ratio in all phospholipids. These data show that the macrophage eicosanoid precursor levels of 20∶3n−6, 20∶4n−6 and n−3 acids can be selectively manipulated through the use of specific dietary regimens. This is noteworthy because an increase in phospholipid levels of 20∶3n−6 and 20∶5n−3, while concomitantly reducing 20∶4n−6, may have therapeutic potential in treating inflammatory disorders.     

Molecular species composition of the major diacyl glycerophospholipids from muscle,liver, retina and brain of cod (Gadus morhua)

The molecular species composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS) from white muscle, liver, retina and brain of cod (Gadus morhua) were determined by high-performance liquid chromatography of the respective 1,2-diacylglycerol 3,5-dinitrobenzoyl derivatives. A minimum of 69 diacyl species was identified. In muscle and liver saturated fatty acid/polyunsaturated fatty acid (PUFA) and monounsaturated fatty acid/PUFA molecular species were predominant, particularly 16∶0/20∶5 and 16∶0/22∶6 in PC, 16∶0/22∶6 and 18∶1/22∶6 in PE and 18∶0/22∶6 and 18∶1/22∶6 in PS. Didocosahexaenoyl species were major components of PC, PE and PS from retina, comprising 29.3, 71.8 and 59.7% of the respective totals. Didocosahexaenoyl species were also abundant in PE and PS from brain, accounting for 13.8 and 24.0% of the totals, respectively. DiPUFA species were important in muscle, totalling 21.2% in PC and 38.3% in PE. PC from all tissues had the largest amounts of species containing only saturated or monounsaturated fatty acids, accounting for 59.8% of PC from brain, including 12.8% of 18∶1/24∶1 plus 24∶1/18∶1.     

Influence of dietary fat composition on intestinal absorption in the rat

Omega-3 fatty acids influence the function of the intestinal brush border membrane. For example, the omega-3 fatty acid eicosapentaenoic acid (20∶5ω3) has an antiabsorptive effect on jejunal uptake of glucose. This study was undertaken to determine whether the effect of feeding α-linolenic acid (18∶3ω3) or EPA plus docosahexaenoic acid (22∶6ω3) on intestinal absorption of nutrients was influenced by the major source of dietary lipid, hydrogenated beef tallow or safflower oil. Thein vitro intestinal uptake of glucose, fatty acids and cholesterol was examined in rats fed isocaloric diets for 2 weeks: beef tallow, beef tallow + linolenic acid, beef tallow + eicosapentaenoic acid/docosahexaenoic acid, safflower oil, safflower oil + linolenic acid, or safflower oil + eicosapentaenic acid/docosahexaenoic acid. Eicosapentaenoic acid/docosahexaenoic acid reduced jejunal uptake of 10 and 20 mM glucose only when fed with beef tallow, and not when fed with safflower oil. Linolenic acid had no effect on glucose uptake, regardless of whether it was fed with beef tallow or safflower oil. The jejunal uptake a long-chain fatty acids (18∶0, 18∶2ω6, 18∶3ω3, 20∶4ω6, 20∶5ω3 and 22∶6ω3) and cholesterol was lower in salfflower oil than with beef tallow. When eicosapentaenoic acid/docosahexaenoic acid was given with beef tallow (but not with safflower oil), there was lower uptake of 18∶0, 20∶5ω3 and cholesterol. The demonstration of the inhibitory effect of linolenic acid or eicosapentaenoic acid/docosahexaenoic acid on cholesterol uptake required the feeding of a saturated fatty acid diet (beef tallow). These changes in uptake were not explained by differences in the animals’ food intake, body weight gain or intestinal weight. Feeding safflower oil was associated with an approximately 25% increase in the jejunal and ileal mucosal surface area, but this increase was prevented by combining linolenic acid or eicosapentaenoic acid/docosahexaenoic acid with safflower oil. Different inhibitory patterns were observed when mixtures of fatty acids were present together in the incubation medium, rather than in the diet: for example, when 18∶0 was in the incubation medium with 20∶4ω6, the uptake of 20∶4ω6 was reduced, whereas the uptake was unaffected by 18∶2ω6 or 20∶5ω3. Thus, (1) the inhibitory effect of eicosapentaenoic acid/docosahexaenoic acid on jejunal uptake of glucose, fatty acids and cholesterol was influenced by the major dietary lipid, saturated (beef tallow) or polyunsaturated fatty acid (safflower oil); and (2) different omega-3 fatty acids (linolenic acid versus eicosapentaenoic acid/docosahexaenoic acid) have a variable influence on the intestinal absorption of nutrients.     

Dietary cod liver oil decreases arachidonic acid in rat gastric mucosa and increases stress-induced gastric erosions

The purpose of this study was to examine the influence of long-term feeding of dietary fat rich in either n−3 or n−6 fatty acids on the availability of arachidonic acid (20∶4n−6) in major phospholipids of gastric mucosa in rats. Three groups of male Wistar rats were fed either a standard diet, a cod liver oil-enriched diet (10% by weight), or a corn oil-enriched diet (10% by weight) for 8 mon. Dietary cod liver oil significantly reduced the level of 20∶4n−6 in phosphatidylcholine (PC) and in phosphatidylethanolamine (PE) of gastric mucosa. The loss of 20∶4n−6 was compensated for by eicosapentaenoic acid (20∶5n−3) in PC, whereas the decrease in 20∶4n−6 in PE corresponded to the increase in three n−3 fatty acids: 20∶5n−3, docosapentaenoic acid (22∶5n−3), and docosahexaenoic acid (22∶6n−3). The level of 20∶5n−3 was higher than the level of 22∶6n−3 both in PC and PE of mucosa in rats fed cod liver oil. Diets supplemented with corn oil increased the level of 18∶2n−6 but decreased the monoene fatty acids 16∶1 and 18∶1n−7 in PC but not in PE of gastric mucosa. The 20∶4n−6 levels of both PC and PE were markedly reduced by dietary cod liver oil, to about one-third of control levels. Similar changes were also observed in the stomach wall. Gastric erosions were observed in all rats exposed to restriction stress, but this form of stress induced twice the number of erosions in rats fed fish oil compared to control rats or rats fed corn oil. We conclude that a diet rich in fish oil altered the balance between n−6 and n−3 fatty acids in major gastric mucosal phospholipids, markedly reduced the availability of 20∶4n−6, and increased the incidence of gastric erosions induced by restriction or emotional stress.     

Dietary fats containing concentrates ofcis ortrans octadecenoates and the patterns of polyunsaturated fatty acids of liver phosphatidylcholine and phosphatidylethanolamine

The effects of the mixedcis- 18∶1 isomers and mixedtrans-18∶1 isomers present in partially hydrogenated soybean oil (PHSO) upon the patterns of polyunsaturated fatty acids (PUFA) in liver phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were studied in rats fed concentrates ofcis- 18∶1 ortrans- 18∶1 isomers isolated as triacylglycerides from PHSO. Thecis- 18∶1 andtrans- 18∶1 concentrates were fed at levels equal to those present in PHSO fed at 17.9% of the diet. All diets contained the required amounts of both linoleic and linolenic acids. Thetrans- 18∶1 concentrate was found to suppress the levels of 20∶4ω6 and 20∶3ω9, and to increase the levels of 18∶2ω6 and 20∶5ω3 in PC and PE. Thecis- 18∶1 concentrate suppressed 20∶4ω6 in PC, 20∶5ω3 in PC and PE, and 18∶2ω6 was more effective than thetrans concentrate in suppressing 22∶6ω3. Thetrans- 18∶1 concentrate was more effective in suppressing 20∶4ω6. Thetrans-18∶ isomers appear to modify PUFA metabolism by inhibition of PUFA synthesis, whereas thecis- 18∶1isomers appear to compete with 2-position fatty acyl transfer and to inhibit ω3 PUFA acylation.     

Fatty acid content of marine oil capsules

The use of dietary ω3 fatty acid capsules has been associated with a decrease in plasma triglyceride levels. In addition, populations consuming diets rich in fish appear to have a decreased incidence of cardiovascular disease. Eicosapentaenoic acid (EPA, 20∶5ω3) and docosahexaenoic acid (DHA, 22∶6ω3) are major fatty acids in fish oils. It is believed that fish oils exert their biolotic effect through these fatty acids. Many individuals are currently taking fish oil capsules to lower lipids, increase bleeding time, and possibly decrease cardiovascular risk. These capsules also have been classified as food additives with less stringent controls on content. We assessed the fatty acid, cholesterol, and vitamin A and E content of eight commercially available capcules along with cod liver oil. The content of EPA was found to range from 8.7–26.4% (wt%) with a mean of 17.3% (82.4% of labeled content), and that of DHA from 8.9–17.4% with a mean of 11.5% (90.0% of labeled content) as assessed by capillary column gas-liquid chromatography. The mean content of the polyunsaturated ω3 fatty acids was 31.9%, and that of the ω6 fatty acids was 1.4%. The content of saturated fatty acids was 32,0%, and that of monounsaturated fatty acids was 25.1%. Cholesterol content was low, with a range of 0.7–8.3 mg/g, the α-tocopherol range was 0.62–2.24 mg/g, and the range of retinyl esters was 0.4–298.4 μg/g. Cod liver oil had substantially more retinyl esters (2450.1 μg/g) than did fish oil capsules. Our data serve as an independent guide to fish oil capsule fatty acid content upon single lot analysis, and indicate that these capsules contain as much saturated fat as they contain ω3 fatty acids.     

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.     

Relationship between dietary supply of long-chain fatty acids and membrane composition of long- and very long chain essential fatty acids in developing rat photoreceptors

The present study was designed to determine if dietary supply of long-chain fatty acid (LCFA, C20∶4n-6, and/or C22∶6n-3), reflecting levels that might be incorporated into infant formulas, influences the fatty acid composition of the visual cell membrane. The rod outer segment (ROS) of the retina was analyzed from rats fed diets varying in the ratio of 18∶2n-6 to 18∶3n-3 with or without 20∶4n-6 [arachidonic acid (AA)] and 22∶6n-3 (docosahexaenoic acid) from birth to six weeks of age. The level of very long chain fatty acids (VLCFA, C₂₄−C₃₆) was identified using gas chromatography and gas chromatography-mass spectrometry. In the ROS, the highest relative percent of AA was attained in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of animals fed 1% AA diet, whereas feeding 0.7% docosahexaenoic acid (DHA) diet significantly increased the DHA level in PC, phosphatidylserine, and phosphatidylinositol compared to feeding diets containing AA. VLCFA of n-6 and n-3 up to C₃₆ were found in PC, with the most abundant fatty acids being C₃₂ and C₃₄. In PC, phosphatidylserine and PE, the n-6 tetraenoic VLCFA level was highly increased in animals fed 1% AA compared to other dietary groups. This study suggests that dietary fat containing small amounts of AA or DHA is an important factor influencing membrane fatty acid composition of the visual cell during development. Based on a presentation at the AOCS Annual Meeting & Expo in San Antonio, Texas, May 7–11, 1995.     

Blood lipid docosahexaenoic and arachidonic acid in term gestation infants fed formulas with high docosahexaenoic acid,low eicosapentaenoic acid fish oil

The effect of fish oil high in docosahexaenoic acid (22∶6n−3) and low in eicosapentaenoic acid (20∶5n−3) in formula on blood lipids and growth of full-term infants was studied. Infants were fed formula with about 15% oleic acid (18∶1), 32% linoleic acid (18∶2n−6), 4.9% linolenic acid (18∶3n−3) and 0, 0.10 or 0.22% 22∶6n−3, or 35% 18∶1, 20% 18∶2n−6, 2.1% 18∶3n−3 and 0, 0.11 or 0.24% 22∶6n−3 from 3 d to 16 wk of age (n=16, 18, 17, 21, 17, 16, respectively). The formulae had <0.1% 20∶5n−3 and no arachidonic acid (20∶4n−6). Breast-fed infants (n=26) were also studied. Plasma phospholipid and red blood cell (RBC) phosphatidylcholine (PC) and phosphatidylethanolamine (PE) fatty acids were determined at 3 d and 4, 8, and 16 wk of age. These longitudinal analyses showed differences in blood lipid 22∶6n−3 between breast-fed and formula-fed infants depending on the feeding duration. At 16 wk, infants fed formula with 0.10, 0.11% 22∶6n−3, or 0.22% 22∶6n−3 had similar 22∶6n−3 levels in the plasma phospholipid and RBC PC and PE compared with breast-fed infants and higher 22∶6n−3 than infants fed formula without 22∶6n−3. Formula with 0.24% 22∶6n−3, however, resulted in higher plasma phospholipid 22∶6n−3 than in breast-fed infants at 16, but not 4 or 8 wk of age. Plasma and RBC phospholipid 20∶4n−6 was lower in formula-fed than breast-fed infants, but no differences in growth were found. Higher blood lipid C₂₀ and C₂₂ n−6 and n−3 fatty acids in infants fed formula with 20% 18∶2n−6 and 2.4% 18∶3n−3 compared with 32% 18∶2n−6 and 4.9% 18∶3n−3 show the increase in blood lipid 22∶6n−3 in response to dietary 22∶6n−3 depending on other fatty acids in the formula.     

Dietary saturated fat level alters the competition between α-linolenic and linoleic acid

Male weanling rats were fed semi-synthetic diets high in saturated fat (beef tallow) vs high in linoleic acid (safflower oil) with or without high levels of α-linolenic acid (linseed oil) for a period of 28 days. The effect of feeding these diets on cholesterol content and fatty acid composition of serum and liver lipids was examined. Feeding linseed oil with beef tallow or safflower oil had no significant effect on serum levels of cholesterol. Serum cholesterol concentration was higher in animals fed the safflower oil diet than in animals fed the beef tallow diet without linseed oil. Feeding linseed oil lowered the cholesterol content in liver tissue for all dietary treatments tested. Consumption of linseed oil reduced the arachidonic acid content with concomitant increase in linoleic acid in serum and liver lipid fractions only when fed in combination with beef tallow, but not when fed with safflower oil. Similarly, ω3 fatty acids (18∶3ω3, 20∶5ω3, 22∶5ω3, 22∶6ω3) replaced ω6 fatty acids (20∶4ω6, 22∶4ω6) in serum and liver lipid fractions to a greater extent when linseed oil was fed with beef tallow than with safflower oil. The results suggest that the dietary ratio of linoleic acid to saturated fatty acids or of 18∶3ω3 to 18∶2ω6 may be important to determine the cholesterol and arachidonic acid lowering effect of dietary α-linolenic acid.     

Dietary docosahexaenoic acid affects stearic acid desaturation in spontaneously hypertensive rats

Docosahexaenoic acid (DHA, 22∶6n−3) is an n−3 polyunsaturated fatty acid which attenuates the development of hypertension in spontaneously hypertensive rats (SHR). The effects of DHA on delta-9-desaturase activity in hepatic microsomes and fatty acid composition were examined in young SHR. Two groups of SHR were fed either a DHA-enriched diet or a control diet for 6 wk. Desaturase activity and fatty acid composition were determined in hepatic microsomes following the dietary treatments. Delta-9-desaturase activity was decreased by 53% in DHA-fed SHR and was accompanied by an increase in 16∶0 and a reduction in 16∶1n−7 content in hepatic microsomes. The DHA diet also increased the levels of eicosapentaenoic acid (20∶5n−3) and DHA. The n−6 fatty acid content was also affected in DHA-fed SHR as reflected by a decrease in gamma-linolenic acid (18∶3n−6), arachidonic acid (20∶5n−6), adrenic acid (22∶4n−6), and docosapentaenoic acid (22∶5n−6). A higher proportion of dihomo-gamma-linolenic acid (20∶3n−6) and a lower proportion of 20∶4n−6 is indicative of impaired delta-5-desaturase activity. The alterations in fatty acid composition and metabolism may contribute to the antihypertensive effect of DHA previously reported.     

Lymphatic fatty acids from rats fed human milk and formula supplemented with fish oil

Absorption of long-chain polyunsaturated fatty acids from human milk and formula supplemented with fish oil was studied to determine if the distribution route into lymphatic triacylglycerol (TAG) and phospholipid (PL) varies with the dietary source. Rats were intraduodenally infused with human milk or formula containing graded amounts of fish oil (0, 0.5, or 1.0 g/100 mL), and the mesenteric lymph was collected. Arachidonic acid (20∶4n−6) levels in lymphatic TAG and PL were highest from animals fed human milk. In the animals infused with formula containing fish oil, as the amount of eicosapentaenoic acid (EPA, 20∶5n−3) infused increased, there was essentially an equal increase of EPA associated with both lymphatic TAG and PL. Animals intraduodenally infused with human milk or formula without fish oil had only minor levels (less than 1%) of EPA in the lymph. In the fish oil-treated animals, as the amount of docosahexaenoic acid (DHA, 22∶6n−3) infused increased, there was a 16-fold increase in DHA associated with lymphatic TAG, but only a 3-fold increase in DHA associated with lymphatic PL. The highest level of DHA in rats infused with human milk was observed in lymphatic PL. Hence, fish oil can be added to formula as a source of long-chain polyunsaturated fatty acids, but the distribution of fatty acids into lymphatic TAG and PL is not the same as that observed with human milk.     

Does a threshold for the effect of dietary omega-3 fatty acids on the fatty acid composition of nuclear envelope phospholipids exist?

Existence of a dietary maximal level or threshold for incorporation of ω3 fatty acids into membrane phospholipids is of interest as it may further define understanding of the dietary requirement for ω3 fatty acids. To test whether feeding increasing levels of dietary ω3 fatty acids continues to increase membrane ω3 fatty acid content, weanling rats were fed a nutritionally adequate semipurified diet which provided increasing amounts of C₂₀ and C₂₂ ω3 fatty acids, such as 20∶5ω3 and 22∶6ω3. Dietary 20∶5ω3 and 22∶6ω3 were provided by substituting a purified shark oil concentrate of high 22∶6ω3 content for safflower oil high in 18∶2ω6. After four weeks of feeding, nuclear envelopes from four animals in each diet group were prepared, lipid was extracted and phospholipids separated. Arachidonic acid content in membrane phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylserine was significantly reduced by feeding increased dietary levels of ω3 fatty acids. Decline of 20∶4ω6 level in phospholipid tended to stabilize when the dietary content of total ω3 fatty acids reached 4–5% of total fatty acids. Above this level, dietary ω3 fatty acids did not result in a further decrease in membrane content of 20∶4nω6. Increase in membrane phospholipid content of 20∶5ω3 occurred as the dietary intake of ω3 fatty acids increased from 1.1% to 5% of total fatty acids. A dietary ω3 fatty acid level of 2.2–3% was sufficient to result in maximum incorporation of 22∶6ω3 into membrane phosphatidylcholine and phosphatidylethanolamine, but not into phosphatidylinositol or phosphatidylserine.     

Eicosapentaenoic acid and docosahexaenoic acid selectively attenuate U46619-induced smooth muscle cell proliferation

It is well known that vascular smooth muscle cell (SMC) proliferation is a key step in atheromatous plaque formation. Thromboxane A₂ (T×A₂), released from aggregating platelets and an injured vessel wall, may play an important role in the development of atheromatous plaque. Many animal studies have suggested that n−3 polyunsaturated fatty acids eicosapentaenoic acid (EPA, 20∶5n−3) and docosahexaenoic acid (DHA, 22∶6n−3) present in the fish oils have antiatherosclerotic effects. In the present study, we investigated the effect of EPA and DHA on I×A₂-induced SMC proliferation. To determine the functional selectivity of n−3 fatty acids, we also tested the effect of arachidonic acid (AA, 20∶4n−6), γ-linolenic acid) (INA, 18∶3n−6), and oleic acid (OA, 18∶1n−9) on T×A₂-induced SMC proliferation. Only LPA and DHA prevented the SMC proliferation induced by the T×A₂ mimetic U46619. When EPA and DHA were added together in the ratio in which they are present in menhaden oil, EPA and DHA acted synergistically to block the SMC proliferation induced by the TXA₂-mimetic. These findings suggest that the n−3 polyunsaturated fatty acids in fish oils may exert antiatherosclerotic effects by blocking the mitogenstimulated proliferation of SMC.