Distribution of polyunsaturated fatty acids including conjugated linoleic acids in total and subcellular fractions from healthy and cancerous parts of human kidneys

Differences in the FA composition of subcellular fractions from healthy and cancerous kidney tissues from the same patients were examined. Only minor differences in CLA content were found between the healthy and the cancerous tissue portions. Regarding the distribution pattern, CLA incorporation into nuclei and cytosol was significantly higher than incorporation into plasma membranes and mitochondria, which could be correlated to the neutral lipid content of these fractions. The subcellular distribution pattern of CLA was similar to that observed with monounsaturated FA but unlike that found with 18∶2n−6, which underlines the different physiological properties of CLA and 18∶2n−6. Because PUFA have been suggested to have an effect on cancer risk, the contents of n−3 and n−6 PUFA were determined in kidney and renal cell carcinoma (RCC). The 18∶2n−6 content and Δ⁵ desaturase activity were significantly lower, and the 18∶3n−6, 20∶3n−6, and 20∶5n−3 contents and Δ⁶ desaturase activity were significantly higher in RCC than in healthy renal tissue, indicating a changed PUFA metabolism in RCC. Previous research has suggested that CLA inhibits the elongation and desaturation of 18∶2n−6 into 20∶4n−6. In that case, one might speculate that a diet enriched in CLA would be a useful tool in preventing RCC. However, the involvement of CLA in preventing renal cancer could not be demonstrated definitively from the design of this experiment. Further understanding of the cause and/or consequence of the difference in FA metabolism may lead to a better understanding of RCC.     

Gas chromatography-chemical ionization-mass spectrometric fatty acid analysis of a commercial supercritical carbon dioxide lipid extract from New Zealand green-lipped mussel (Perna canaliculus)

Supercritical fluid extracts of New Zealand green-lipped mussels (NZGLM) have been suggested to have therapeutic properties related to their oil components. The large number of minor FA in NZGLM extract was characterized by a GC-CIMS/MS method that excels at identification of double-bond positions in FAME. The extract contained five major lipid classes: sterol esters, TAG, FFA, sterols, and polar lipids. The total FA content of the lipid extract was 0.664 g/mL. Fifty-three unsaturated FA (UFA) were fully identified, of which 37 were PUFA, and a further 21 UFA were detected for which concentrations were too low for assignment of double-bond positions. There were 17 saturated FA, with 14∶0, 16∶0, and 18∶0 present in the greatest concentration. The 10 n−3 PUFA detected included 20∶5n−3 and 22∶6n−3, the two main n−3 FA; n−3 PUFA at low concentrations were 18∶3, 18∶4, 20∶3, 20∶4, 21∶5, 22∶5, 24∶6, and 28∶8. There were 43 UFA from the n−4, n−5, n−6, n−7, n−8, n−9, n−10, n−11 families, with 16∶2n−4, 16∶1n−5, 18∶1n−5, 18∶2n−6, 20∶4n−6, 16∶1n−7, 20∶1n−7, 16∶1n−9, 18∶1n−9, and 20∶1n−9 being the most abundant. In general, we estimated that FAME concentrations greater than 0.05% (w/w) were sufficient to assign double-bond positions. In total, 91 FA were detected in an extract of the NZGLM, whereas previous studies of fresh flesh from the NZGLM had reported identification of 42 FA. These data demonstrate a remarkable diversity of NZGLM FA.     

Positive association between plasma antioxidant capacity and n−3 PUFA in red blood cells from women

PUFA are susceptible to oxidation. However, the chain-reaction of lipid peroxidation can be interrupted by antioxidants. Whether an increased concentration of PUFA in the body leads to decreased antioxidant capacity and/or increased consumption of antioxidants is not known. To elucidate the relationship between plasma total antioxidant capacity (TAC), the concentration of antioxidant vitamins, and the proportion of PUFA in red blood cells (RBC), plasma TAC was measured by a Trolox equivalent antioxidant capacity assay in blood samples from 99 Icelandic women. Concentrations of tocopherols and carotenoids in the plasma were determined by HPLC, and the FA composition of RBC total lipids was analyzed by GC. Plasma TAC and the plasma concentration of α-tocopherol correlated positively with the proportion of total n−3 PUFA, 20∶5n−3, and 22∶6n−3 in RBC, whereas the plasma lycopene concentration correlated negatively with the proportion of total n−3 PUFA and 20∶5n−3. On the other hand, plasma TAC correlated negatively with the proportion of n−6 PUFA in RBC. Plasma TAC also correlated positively with the plasma concentration of α-tocopherol, alcohol consumption, and age. Both the plasma concentration of α-tocopherol and age correlated positively with the proportion of n−3 PUFA in RBC; however, n−3 PUFA contributed independently to the correlation with plasma TAC. Because the proportion of n−3 PUFA in RBC reflects the consumption of n−3 PUFA, these results suggest that dietary n−3 PUFA do not have adverse effects on plasma TAC or the plasma concentration of most antioxidant vitamins.     

Lipid class and fatty acid composition of the boreal soft coral Gersemia rubiformis

Total lipid, phospholipid, and FA composition and distribution of FA between polar lipids (PL) and neutral lipids (NL) were investigated in the boreal soft coral Gersemia rubiformis from the Bering Sea. The total lipids were mostly hydrocarbons and waxes (33.7%) and PL (33.1%). The content of monoalkyldiacylglycerols (9.7%) exceeded the content of TAG (6.7%). PC and PE constituted 31.4% and 25.6% of total phospholipids, respectively. Principal FA were 16∶0, 16∶1n−7, 18∶0, 18∶1n−9, 18∶1n−7, 20∶1n−7, 20∶4n−6, 20∶4n−3, 20∶5n−3 22∶5n−3, 22∶6n−3, 24∶5n−6, and 24∶6n−3. Most n−6 PUFA (52% of total FA) were associated with the PL fraction; this was especially true for arachidonic and tetracosapentaenoic acids. The NL were enriched with mono-, di-, trienoic, and n−3 PUFA. The variation in EPA levels in both NL and PL suggests an origin of this acid from lipids of diatoms consumed by the corals.     

Fatty acids bound to <Emphasis Type="Italic">Fasciola hepatica</Emphasis> 12 kDa fatty acid-binding protein,a candidate vaccine,differ from fatty acids in extracts of adult flukes

The FA composition of Fasciola hepatica 12 kDA purified native FA-binding protein (nFh12), a candidate vaccine against fascioliasis, is described. The FA chain lengths ranged between 12 and 24 carbons. The principal FA were 16∶0 18∶1n−9, 18∶0, 20∶4n−6, and 20∶1n−9. The acids 16∶0, 18∶1n−9, and 18∶0 comprised over half the FA that were bound to the whole FA-binding protein. Small amounts (1.0–2.8%) of isoanteiso methyl-branched FA also were characterized. Forty-one different FA were identified in extracts of the adult flukes, with the three most abundant FA also being 16∶0, 18∶1n−9, and 18∶0. A similar proportion of saturated vs. unsaturated FA was observed between the whole extract from F. hepatica and the nFh12 protein. However, the n−3/n−6 ratio of PUFA was significantly different, being 1.2 in the whole extract vs. 9.6 in the nFh12 protein complex. The nFh12 protein binds more n−5, n−6, and n−7 PUFA and less n−3 and n−9 PUFA than the whole extract. In addition, cholesterol (56%), sitosterol (36%), and fucosterol (8%) also were bound to the nFh12 protein complex.     

Effect of tetracosahexaenoic acid on the content and release of histamine, and eicosanoid production in MC/9 mouse mast cell

6,9,12,15,18,21-Tetracosahexaenoic acid (24∶6n−3) was isolated from a brittle star, Ophiura sarsi Lütken, at>95% purity to evaluate its physiological functions. The effects of 24∶6n−3 on the production of leukotriene (LT)-related compounds such as LTB₄, LTC₄ and 5-hydroxyeicosatetraenoic acid, and the accumulation and release of histamine in an MC/9 mouse mast cell line were studied. We found that 24∶6n−3 could inhibit the antigen-stimulated production of LT-related compounds as well as other n−3 polyunsaturated fatty acids (PUFA) such as eicosapentaenoic acid (20∶5n−3) and docosahexaenoic acid (22∶6n−3), which are major n−3 PUFA in fish oils; 24∶6n−3 was also shown to reduce the histamine content in MC/9 cells at 25 μM (27% reduction from the control), and the effect was diminished with increase of the fatty acid concentration (up to 100 μM). These two n−3 PUFA, 20∶5n−3 and 22∶6n−3, also reduced the histamine content (16 and 20% reduction at 25 μM, respectively), whereas arachidonic acid (20∶4n−6) increased it (18% increase at 25 μM). Spontaneous- and antigen-induced release of histamine was not influenced with these PUFA (at 25 μM). Ionophore-stimulated release of histamine was suppressed by the PUFA (13,9,15, and 11% reduction with 20∶4n−6, 20∶5n−3, 22∶6n−3, and 24∶6n−3, respectively). The patterns of the effects of 24∶6n−3 on the synthesis of eicosanoids and histamine content were more similar to those of 22∶6n−3 than 20∶5n−3. From these results, 24∶6n−3 can be expected to have anti-inflammatory activity and antiallergic activities similar to those of 22∶6n−3.     

Composition and seasonal variation of fatty acids of <Emphasis Type="Italic">Diplodus vulgaris</Emphasis> L. from the Adriatic Sea

Muscle tissue from the common two-banded sea bream Diplodus vulgaris L. originating from the Adriatic Sea, Croatia, was analyzed. The FA composition of neutral (TAG) and polar (PE, PC, PI/PS) lipid classes was determined, as well as the lipid and water contents during winter and summer periods. Both the total lipid and water contents were higher in the winter period. We identified 16 different FA. The major constituents of the total FA in both seasons were saturates: palmitic (16∶0) and stearic acids (18∶0); monoenes: oleic (18∶1n−9) and palmitoleic acids (16∶1n−7); and polyunsaturates: arachidonic acid (20∶4n−6), EPA (20∶5n−3), and DHA (22∶6n−3), but their amounts and ratios differed significantly between the two seasons and between lipid fractions. The FA composition showed a noticeable pattern of seasonality that reflected fluctuations mainly in TAG. The diminution of the monounsaturated FA content in the summer was clearly followed by an increase in PUFA content. Diplodus vulgaris is a good source of natural n−3 PUFA and would therefore be suitable for inclusion in highly unsaturated low-fat diets.     

Incorporation of n−3 fatty acids into plasma and liver lipids of rats: Importance of background dietary fat

The health benefits of long-chain n−3 PUFA (20∶5n−3 and 22∶6n−3) depend on the extent of incorporation of these FA into plasma and tissue lipids. This study aimed to investigate the effect of the background dietary fat (saturated, monounsaturated, or n−6 polyunsaturated) on the quantitative incorporation of dietary 18∶3n−3 and its elongated and desaturated products into the plasma and the liver lipids of rats. Female weanling Wistar rats (n=54) were randomly assigned to six diet groups (n=9). The fat added to the semipurified diets was tallow (SFA), tallow plus linseed oil (SFA-LNA), sunola oil (MUFA), sunola oil plus linseed oil (MUFA-LNA), sunflower oil (PUFA), or sunflower oil plus linseed oil (PUFA-LNA). At the completion of the 4-wk feeding period, quantitative FA analysis of the liver and plasma was undertaken by GC. The inclusion of linseed oil in the rat diets increased the level of 18∶3n−3, 20∶5n−3, and, to a smaller degree, 22∶6n−3 in plasma and liver lipids regardless of the background dietary fat. The extent of incorporation of 18∶3n−3, 20∶5n−3, and 22∶5n−3 followed the order SFA-LNA>MUFA-LNA>PUFA-LNA. Levels of 22∶6n−3 were increased to a similar extent regardless of the type of major fat in the rat diets. This indicates that the background diet affects the incorporation in liver and plasma FA pools of the n−3 PUFA with the exception of 22∶6n−3 and therefore the background diet has the potential to influence the already established health benefits of long-chain n−3 fatty acids.     

Long-chain n−3 polyunsaturated fatty acid incorporation into human atrium following fish oil supplementation

Recent studies have demonstrated that long-chain n−3 PUFA (LCn-3PUFA) are beneficial in reducing the risk of cardiac arrhythmias. This study was conducted to determine the extent of incorporation of LCn-3PUFA into human atrium following supplementation with a fish oil concentrate high in LCn-3PUFA. Volunteers preparing for coronary bypass surgery were randomized either to the treatment group (n=8), receiving 6 g/d of fish oil concentrate (4.4 g of LCn-3PUFA), or the placebo group (n=9), receiving 6 g/d of olive oil for a minimum period of 6 wk. Blood samples were collected prior to commencement of treatment, and preoperatively before bypass surgery. Atrial biopsies were obtained during surgery. The plasma and atrium samples were analyzed by GC following trans-methylation to determine FA profile. Post-supplementation, the treatment group had significantly higher plasma levels of 20∶5n−3, 22∶5n−3, and 22∶6n−3 than the placebo group. Analysis of the atrium total lipids revealed a significant increase in the proportion of 20∶5n−3 following fish oil supplementation. There was no significant difference in the concentration of 22∶5n−3 and 22∶6n−3 in the atrium total lipids; however, an upward trend was observed in subjects receiving fish oil supplementation. In the phospholipid fraction of the atrium, both 20∶5n−3 and 22∶6n−3 increased, whereas 20∶4n−6 levels decreased. This study demonstrates for the first time that short-term supplementation with fish oil concentrate results in significant incorporation of LNc-3PUFA with a concomitant depletion of the eicosanoid substrate (20∶4n−6) in the human atrium.     

Dietary modulation of fatty acid profiles and oxidative status of rat hepatocyte nodules: Effect of different n−6/n−3 fatty acid ratios

Male Fischer rats were fed the AIN76A diet containing varying n−6/n−3 FA ratios using sunflower oil (SFO), soybean oil (SOY), and SFO supplemented with EPA-50 and GLA-80 (GLA) as fat sources. Hepatocyte nodules, induced using diethylnitrosamine followed by 2-acetylaminofluorene/partial hepatoctomy promotion, were harvested, with surrounding and respective dietary control tissues, 3 mon after partial hepatectomy. The altered growth pattern of hepatocyte nodules in rats fed SFO is associated with a distinct lipid pattern entailing an increased concentration of PE, resulting in increased levels of 20∶4n−6. In addition, there is an accumulation of 18∶1n−9 and 18∶2n−6 and a decrease in the end products of the n−3 metabolic pathway in PC, suggesting a dysfunctional Δ-6-desaturase enzyme. The hepatocyte nodules of the SFO-fed rats exhibited a significantly reduced lipid peroxidation level that was associated with an increaser in the glutathione (GSH) concentration. The low n−6/n−3 FA ratio diets significantly decreased 20∶4n−6 in PC and PE phospholipid fractions with a concomitant increase in 20∶5n−3, 22∶5n−3, and 22∶6n−3. The resultant changes in the 20∶4/20∶5 FA ratio and the 20∶3n−6 FA level in the case of the GLA diet suggest a reduction of prostaglandin synthesis of the 2-series. The GLA diet also counteracted the increased level of 20∶4n−6 in PE by equalizing the nodule/surrounding ratio. The low n−6/n−3 ratio diets significantly increased lipid peroxidation levels in hepatocyte nodules, mimicking the level in the surrounding and control tissue while GSH was decreased. An increase in n−3 FA levels and oxidative status resulted in a reduction in the number of glutathione-S-transferase positive foci in the liver of the GLA-fed rats. Modulation of cancer development with low n−6/n−3 ratio diets containing specific dietary FA could be a promising tool in cancer intervention in the liver.     

Comparison of the effects of dietary alpha-linolenic,stearidonic, and eicosapentaenoic acids on production of inflammatory mediators in mice

The effects of dietary stearidonic acid (18∶4n−3) on inflammatory mediator release in whole blood and splenocytes was investigated in Balb/c mice, and the effects were compared with those of two other n−3 PUFA: α-linolenic acid (18∶3n−3) and EPA (20∶5n−3). TAG mixtures containing 10% of 18∶4n−3, 18∶3n−3, or 20∶5n−3 as the respective sole n−3 PUFA were enzymatically synthesized. Diets containing synthesized TAG mixtures were fed to Balb/c mice for 3 wk. The release of prostaglandin E₂ (PGE₂) and tumor necrosis factor (TNF) were measured in whole blood and splenocytes stimulated with lipopolysaccharide. In whole blood, the production of INF was suppressed by all dietary n−3 PUFA (18∶3n−3, 18∶4n−3, and 20∶5n−3) as compared with the control diet, which contained TAG prepared from safflower oil. PGE₂ production was not significantly changed. Differences among the n−3 PUFA (18∶3n−3), 18∶4n−3, and 20∶5n−3) were not observed. In splenocytes, PGE₂ production was suppressed by dietary n−3 PUFA, but TNF production was not. GC analysis of plasma and splenocyte FA profiles showed an increase in the levels of 20∶4n−3, 20∶5n−3, and 22∶6n−3 in mice fed the diet containing 18∶4n−3.     

Lipid and FA composition of the pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis>: Influence of season and maturation

The lipid and FA composition of the total lipids of the pearl oyster Pinctada fucata martensii, in different seasons and in different areas, were analyzed to clarify its lipid physiology and to estimate the possible influence of its prey phytoplankton. TAG and sterols were the major components in the neutral lipids in all conditions, whereas high levels of phospholipids (PE and PC) were found in the polar lipids. The major FA in the TAG in all samples were 14∶0, 16∶0, and 18∶0 as saturated FA (saturates); 16∶1n−7, 18∶1n−9, and 18∶1n−7 as monoenoic FA (monoenes); and 20∶4n−6 (arachidonic acid: AA), 20∶5n−3 (EPA), and 22∶6n−3 (DHA) as PUFA. The major components found in the polar lipids were 16∶0 and 18∶0 as saturates; 22∶2n−9, 15 and 22∶2n−7, 15 as non-methylene-interrupted dienes (NMID), and AA, 22∶3n−6, 9, 15, EPA, and DHA as PUFA. Although it is a marine animal, characteristically high levels of AA were found in both the TAG and phospholipids. This result suggests that lipids of P. fucata may be influenced by those of its phytoplanktonic prey. The increase in levels of NMID from TAG to PE with a decrease in those of monoenes suggests that the tissues of this species are able to biosynthesize only the less unsaturated PUFA, such as NMID. In particular, NMID derivatives are considered to be biosynthesized in the PE; thus, they might play a particular role in the membrane, because NMID were characteristically localized only in the PE.     

Fatty acid profiles and relative mobilization during fasting in adipose tissue depots of the American marten (Martes americana)

The American marten (Martes americana) is a boreal forest marten with low body adiposity but high metabolic rate. The study describes the FA composition in white adipose tissue depots of the species and the influence of food deprivation on them. American marten (n=8) were fasted for 2 d with 7 control animals. Fasting resulted in a 13.4% weight loss, while the relative fat mass was >25% lower in the fasted animals. The FA composition of the fat depots of the trunk was quite similar to other previously studied mustelids with 14∶0, 16∶0, 18∶0, 16∶1n−7, 18∶1n−9, and 18∶2n−6 as the most abundant FA. In the extremities, there were higher proportions of monounsaturated FA (MUFA) and PUFA. Food deprivation decreased the proportions of 16∶0 and 16∶1n−7, while the proportion of long-chain MUFA increased in the trunk. The mobilization of FA was selective, as 16∶1n−7, 18∶1n−9, and particular n−3 PUFA were preferentially mobilized. Relative mobilization correlated negatively with the carbon chain length in saturated FA (SFA) and n−9 MUFA. The Δ9 desaturation of SFA enhanced the mobilization of the corresponding MUFA, but the positional isomerism of the first double bond did not correlate consistently with relative mobilization in MUFA or PUFA. In the marten, the FA composition of the extremities was highly resistant to fasting, and the tail tip and the paws contained more long-chain PUFA to prevent the solidification of lipids and to maintain cell membrane fluidity during cooling.     

Tissue phospholipid fatty acid composition in genetically lean (Fa/−) or obese (fa/fa) zucker female rats on the same diet

The fatty acid composition of serum total lipids, of phospholipids of various organs (liver, heart, kidney), and of nervous structures (brain, retina, sciatic nerve, myelin, synaptosomes) have been compared in lean (Fa/−) and genetically obese (fa/fa) Zucker female rats. Both received a standard commercial diet including 37% of 18∶2n−6 and 5% of n−3 polyunsaturated fatty acids (PUFA), 1.7% of which were in the form of 20∶5n−3 and 22∶6n−3. In comparison with lean rats, the results for the obese rats pointed out (i) no difference in the fatty acid composition of nervous structures: (ii) a decrease of 18∶2n−6 (from −8% to −35%) and of 20∶4n−6 (from −9% to −49%) in serum, liver and in kidney; this was compensated for by an increase in 20∶3n−6 (from +30% to +320%) and in total n−3 PUFA (from +68% to +76%); (iii) a decrease of 20∶4n−6 (−18%) and of 22∶6n−3 (−24%) in heart compensated for by an increase in 18∶2n−6 (+39%) and in 20∶3n−6 (+233%); and (iv) constant levels of total PUFA (n−6 and n−3) in the various fractions studied, except in serum where this level decreased (−23%). Finally, except for the nervous structures, tissue phospholipids of obese rats included a lower proportion of 20∶4n−6 and a higher proportion of 20∶3n−6. This resulted in a significant reduction in the 20∶4n−6/20∶3n−6 ratio; by contrast, the 20∶3n−6/18∶2n−6 ratio increased. The results suggest that in Zucker rats, the obese character (fa/fa) affects the desaturation-elongation process of 18∶2n−6 to 20∶4n−6 by specifically decreasing Δ5-desaturase activity.     

Mice raised on milk transgenically enriched with n−3 PUFA have increased brain docosahexaenoic acid

The brain contains high levels of the long-chain n−3 FA DHA(22∶6n−3), mainly in the gray matter and synaptosomes. Adequate intake of DHA is crucial for optimal nervous system function, particularly in infants. Supplementation of infant formulas with DHA at levels similar to human breast milk is recommended for biochemical and functional benefits to neonates. We generated transgenic mice that produce elevated levels of n−3 PUFA in their milk by expressing the Caenorhabditis elegans n−3 FA desaturase under the control of a lactation-induced goat beta-casein promoter. To examine the postnatal effects of consuming the n−3-enriched milk, we compared the growth and brain and plasma FA composition of mouse pups raised on milk from transgenic dams with those observed for pups raised on milk from nontransgenic dams. A significant decrease in arachidonic acid (ARA, 20∶4n−6) and concomitant increases in n−3 PUFA were observed in the phospholipid fraction of transgenic mouse milk. The n−6∶n−3 FA ratios were 4.7 and 34.5 for the transgenic and control milk phospholipid fractions, respectively. DHA and DPA (22∶5n−6) comprised 15.1% and 2.8% of brain FA from weanling mice nursed on transgenic dams, as compared with 6.9% and 9.2% for weanling mice nursed on control dams, respectively. This transgenic mouse model offers a unique approach to disassociate the effects and fetal programming resulting from a high n−6∶n−3 FA ratio gestational environment from the postnatal nutritional effects of providing milk with differing n−6∶n−3 FA ratios.     

The Effects of dietary n−3/n−6 ratio on brain development in the mouse: a dose response study with long-chain n−3 fatty acids

This study examines the effects of the ratio of n−3/n−6 fatty acids (FA) on brain development in mice when longchain n−3 FA are supplied in the diet. From conception until 12 days after birth, B6D2F₁ mice were fed liquid diets, each providing 10% of energy from olive oil, and a further 10% from different combinations of free FA concentrates derived from safflower oil (18∶2n−6), and fish oil (20∶5n−3 and 22∶6n−3). The range of dietary n−3/n−6 ratios was 0,025, 0.5, 1.0, 2.0, and 4.0, with an n−6 content of greater than 1.5% of energy in all diets, and similar levels of total polyunsaturated fatty acids (PUFA). In an additional group of ratio 0.5, 18∶2n−6 was partially replaced by its δ6 desaturation product, 18∶3n−6. Biochemical analyses were conducted on 12-day-old pup brains, as well as on samples of maternal milk. No obvious effects on overall pup growth and development were observed, apart from a smaller litter size at ratio 1. Co-variance analysis indicated that increasing the n−3/n−6 ratio was associated with slightly smaller brains, relative to body weight. We found that 18∶2n−6 and 20∶5n−3 were the predominant n−6 and n−3 FA in the milk; in the brain these were 20∶4n−6 and 22∶6n−3, respectively. Increasing dietary n−3/n−6 ratios generally resulted in an increase in n−3 FA, with a corresponding decrease in n−6 FA. The n−3/n−6 ratio of the milk lipids showed a strong linear relationship with the diet, but in the brain the rate of increase tended to decrease beyond 0.5 (phosphatidylcholine, PC) and 0.25 (phosphatidylethanolamine, PE), such that there was a significant quadratic contribution to the relationship. The partial replacement of dietary 18∶2n−6 with 18∶3n−6 raised levels of 20∶4n−6 in milk, brain PC, and brain PE. These results indicate that the n−3/n−6 ratio of the phospholipids in the developing mouse brain responds maximally to maternal dietary long-chain n−3/n−6 ratios of between 0.25 and 0.5.     

Effects of geographic source, rearing system, and season on the nutritional quality of wild and farmed Perca fluviatilis

The effects of season, geographic source (Lake Geneva, Rhine River), and rearing system (extensive, semiextensive, and intensive systems) on the lipid content and FA composition of fillets of Perca fluviatilis were studied. Significant differences in the total lipid content were found between fish coming from the Rhine River and Lake Geneva (1.21 and 1.48%, respectively). Seasonal effects were investigated quarterly for perch sampled in the Rhine River. Intensively reared perch displayed a higher lipid content (1.48%) than the other farmed perch, i.e., 1.26% for a semiextensive system and 1.16% for an extensive system. No significant difference in lipid content was found (i) between lacustrine fish and intensively reared fish or (ii) among fish from the Rhine River and the semiextensive or extensive rearing systems. The main FA were 22∶6n−3 (DHA, 21.3–37.1% of total FA), 16∶0 (17.7–20.2%), 20∶5n−3 (EPA, 9.2–13.2%), 18∶1 (8.0–11.5%), 20∶4n−6 [arachidonic acid (ARA), 1.9–10.7%], 16∶1 (4.3–6.0%), and 18∶2n−6 (2.1–6.0%). In comparison with perch coming from the Rhine River, the lacustrine fish were characterized by higher total n−6 PUFA and a lower proportion of both total monounsaturated FA (MUFA) and total n−3 PUFA. Among rearing systems, extensively farmed fish had higher n−6 PUFA and lower n−3 PUFA contents. Wild fish showed higher ARA and 18∶2n−6 than farmed fish. They also had significantly more EPA (12.5–13.2%) than farmed perch (9.2–10.9%). For DHA no difference existed between (i) the lacustrine fish (31.9% of total FA) and the intensively reared fish (33.0%) and (ii) the Rhine (37.1%) and semiextensively reared fish (36%). Effects of size, diet composition, and environmental conditions on the total lipid contents and FA composition are discussed.     

Headspace gas chromatography to determine human low density lipoprotein oxidation

We previously described a rapid headspace gas chromatographic method for the determination of hexanal, an important decomposition product of n−6 polyunsaturated fatty acid (PUFA) peroxidation in rat liver samples and human red blood cell membranes. This method was applied to the measurement of Cu²⁺ catalyzed-oxidation of freshly prepared human low density lipoproteins (LDL) from 10 healthy adult volunteers. A twofold variation in oxidative susceptibility was found by this assay for hexanal and other volatiles. Hexanal values correlated significantly (P<0.05) with total polyunsaturated fatty acid (PUFA), 18∶2 and n−6 PUFA contents of LDL; but poorly with 20∶4 and with vitamin E. Therefore, in addition to α-tocopherol, other endogenous antioxidants and factors may contribute, to LDL's resistance to oxidation. This simple, rapid and sensitive method for oxidative susceptibility provides a useful component in the analysis of the prooxidant/antioxidant status of biological samples. The method is used routinely in our laboratories to determine specific peroxidation products of n−6 and n−3 PUFA.     

A significant inverse relationship between concentrations of plasma homocysteine and phospholipid docosahexaenoic acid in healthy male subjects

The aim of this study was to investigate the possibility of a relationship between plasma homocysteine (Hcy) and phospholipid FA (PUFA) in healthy Australian males. One hundred thirty six healthy male subjects aged 20–55 yr were recruited from the Melbourne metropolitan area. Each volunteer completed a semiquantitative food frequency questionnaire and gave a blood sample. Plasma Hcy concentrations were determined by an established HPLC method; the plasma phospholipid FA were determined by standard methods. Plasma Hcy concentration was significantly negatively correlated with plasma phospholipid concentration of the PUFA 20∶5n−3 (r=−0.226, P=0.009), 22∶5n−3 (r=−0.182, P=0.036), 22∶6n−3 (r=−0.286, P=0.001), total n−3 (r=−0.270, P=0.002) and the ratio n−3/n−6 PUFA (r=−0.265, P=0.002), and significantly positively correlated with 20∶4n−6 (r=0.180, P=0.037). In the partial correlation analysis, after controlling for serum vitamin B₁₂ and folate concentration, plasma Hcy was significantly negatively correlated with the plasma phospholipid concentration of 22∶6n−3 (r=−0.205, P=0.019), total n−3 (r=−0.182, P=0.038) and the ratio n−3/n−6 PUFA (r=−0.174, P=0.048). Evidence indicates that an increased concentration of n−3 PUFA in tissues has a beneficial effect on cardiovascular health. Our findings provide further evidence that increased consumption of dietary n−3 PUFA increases the concentration of n−3 PUFA in plasma phospholipid, which is associated with a protective effect on cardiovascular diseases and lower plasma Hcy levels. The mechanism that might explain the association between plasma 22∶6n−3 and Hcy levels is not clear.     

Supplementation and delivery of n−3 fatty acids through spray-dried milk reduce serum and liver lipids in rats

Indian diets comprising staples such as cereals, millets, and pulses provide 4.8 energy % from linoleic acid (18∶2n−6) but fail to deliver adequate amounts of n−3 FA. Consumption of long-chain n−3 PUFA such as EPA (20∶5n−3) and DHA (22∶6n−3) is restricted to those who consume fish. The majority of the Indian population, however, are vegetarians needing additional dietary sources of n−3 PUFA. The present work was designed to use n−3 FA-enriched spray-dired milk powder to provide n−3 FA. Whole milk was supplemented with linseed oil to provide α-linolenic acid (LNA, 18∶3n−3), with fish oil to provide EPA and DHA, or with groundnut oil (GNO), which is devoid of n−3 PUFA, and then spray-dired. Male Wistar rats were fed the spray-dired milk formulations for 60 d. The rats given formulations containing n−3 FA showed significant increases (P<0.001) in the levels of LNA or EPA/DHA in the serum and in tissue as compared with those fed the GNO control formulation. Rats fed formulations containing n−3 FA had 30–35% lower levels of serum total cholesterol and 25–30% lower levels of serum TAG than control animals. Total cholesterol and TAG in the livers of rats fed the formulations containing n−3 FA were lower by 18–30% and 11–18%, respectively, compared with control animals. This study showed that spray-dried milk formulations supplemented with n−3 FA are an effective means of improving dietary n−3 FA intake, which may decrease the risk factors associated with cardiovascular disease.