Modification of liver fatty acid metabolism in mice by n-3 and n-6 delta 6-desaturase substrates and products

The effects of dietary supplementation of either alpha-linolenic acid (18:3(n-3)) or stearidonic acid (18:4(n-3)) in combination with either linoleic acid (18:2(n-6)) or gamma-linolenic acid (18:3(n-6)) on liver fatty acid composition in mice were examined. Essential fatty acid deficient male C57BL/6 mice were separated into four groups of seven each and were fed a fat-free semi-purified diet supplemented with 1% (w/w) fatty acid methyl ester mixture (1:1), 18:2(n-6)/18:3(n-3), 18:2(n-6)/18:4(n-3), 18:3(n-6)/18:3(n-3), or 18:3(n-6)/18:4(n-3). After 7 days on the diets, fatty acid compositions in liver phosphatidylcholine and phosphatidylethanolamine fractions were analyzed. In groups fed 18:4(n-3) (18:2(n-6)/18:4(n-3) or 18:3(n-6)/18:4(n-3)) as compared to those fed 18:3(n-3) (18:2(n-6)/18:3(n-3) or 18:3(n-6)/18:3(n-3)), the levels of 20:4(n-3), 20:5(n-3) and 22:5(n-3) were increased, whereas those of 20:3(n-6) and 20:4(n-6) were decreased. When 18:3(n-6) replaced 18:2(n-6) as the source of n-6 acids, the levels of 18:3(n-6), 20:3(n-6), 20:4(n-6) and 22:5(n-6) were increased, whereas those of 20:4(n-3) and 20:5(n-3) were reduced. Replacing 18:3(n-3) by 18:4(n-3) reduced the (n-6)/(n-3) ratio by approx. 30%, whereas replacing 18:2(n-6) by 18:3(n-6) increased the (n-6)/(n-3) ratio by approx. 2-fold. These findings indicated that delta 6-desaturase products were metabolized more readily than their precursors. Both products also competed for the subsequent metabolic enzymes. However, the n-6 fatty acids derived from 18:3(n-6) were incorporated more favourably into liver phospholipids than n-3 fatty acids derived from 18:4(n-3).     

Brain astrocyte synthesis of docosahexaenoic acid from n-3 fatty acids is limited at the elongation of docosapentaenoic acid

The phospholipids, particularly phosphatidylethanolamine, of brain gray matter are enriched with docosahexaenoic acid (22:6n-3). The importance of uptake of preformed 22:6n-3 from plasma compared with synthesis from the alpha-linolenic acid (18:3n-3) precursor in brain is not known. Deficiency of 18:3n-3 results in a compensatory increase in the n-6 docosapentaenoic acid (22:5n-6) in brain, which could be formed from the precursor linoleic acid (18:2n-6) in liver or brain. We studied n-3 and n-6 fatty acid incorporation in brain astrocytes cultured in chemically defined medium using delipidated serum supplemented with specific fatty acids. High performance liquid chromatography with evaporative light scattering detection and gas liquid chromatography were used to separate and quantify cell and media lipids and fatty acids. Although astrocytes are able to form 22:6n-3, incubation with 18:3n-3 or eicosapentaenoic acid (20:5n-3) resulted in a time and concentration dependent accumulation of 22:5n-3 and decrease in 22:6n-3 g/g cell fatty acids. Astrocytes cultured with 18:2n-6 failed to accumulate 22:5n-6. Astrocytes secreted cholesterol esters (CE) and phosphatidylethanolamine containing saturated and monounsaturated fatty acids, and arachidonic acid (20:4n-6) and 22:6n-3. These studies suggest conversion of 22:5n-3 limits 22:6n-3 synthesis, and show astrocytes release fatty acids in CE.     

On the singular, dual, and multiple positional specificity of manganese lipoxygenase and its G316A mutant

Abstract manganese lipoxygenase (Mn-LO) oxygenates 18:3n-3 and 18:2n-6 to bis-allylic 11S-hydroperoxy fatty acids, which are converted to 13R-hydroperoxy fatty acids. Other unsaturated C(16)-C(22) fatty acids, except 17:3n-3, are poor substrates, possibly because of ineffective enzyme activation (Mn(II)-->Mn(III)) by the produced hydroperoxides. Our aim was to determine whether unsaturated C(16)-C(22) fatty acids were oxidized by Mn(III)-LO. Mn(III)-LO oxidized C(16), C(19), C(20), and C(22) n-3 and n-6 fatty acids. The carbon chain length influenced the position of hydrogen abstraction (n-8, n-5) and oxygen insertion at the terminal or the penultimate 1Z,4Z-pentadienes. Dilinoleoyl-glycerophosphatidylcholine was oxidized by Mn-LO, in agreement with a "tail-first" model. 16:3n-3 was oxidized at the bis-allylic n-5 carbon and at positions n-3, n-7, and n-6. Long fatty acids, 19:3n-3, 20:3n-3, 20:4n-6, 22:5n-3, and 22:5n-6, were oxidized mainly at the n-6 and the bis-allylic n-8 positions (in ratios of approximately 3:2). The bis-allylic hydroperoxides accumulated with one exception, 13-hydroperoxyeicosatetraenoic acid (13-HPETE). Mn(III)-LO oxidized 20:4n-6 to 15R-HPETE ( approximately 60%) and 13-HPETE ( approximately 37%) and converted 13-HPETE to 15R-HPETE. Mn(III)-LO G316A oxygenated mainly 16:3n-3 at positions n-7 and n-6, 19:3n-3 at n-10, n-8, and n-6, and 20:3n-3 at n-10 and n-8. We conclude that Mn-LO likely binds fatty acids tail-first and oxygenates many C(16), C(18), C(20), and C(22) fatty acids to significant amounts of bis-allylic hydroperoxides.     

Fatty acids of some algae from the Bohai Sea.

The fatty acid compositions of 22 species of marine macrophytes, belonging to the Ceramiales, Cryptonemiales, Nemalionales, Laminariales, Chordariales, Scytosiphonales, Desmarestiales, Dictyosiphonales, Fucales, Dictyotales and Ulvales and collected from the Bohai Sea, were determined by capillary gas chromatography. The contents of polyunsaturated fatty acids (FAs) in the Bohai Sea algae, in comparison with the same species from the Yellow Sea were found to be lower. Red algae had relatively high levels of the acids 16:0, 18:1(n-7), 18:1(n-9), 20:5(n-3) and 20:4(n-6), and those examined were rich in C(20) PUFAs, these chiefly being arachidonic and eicosapentaenoic acids. The major FAs encountered in the Phaeophyta were 14:0, 16:0, 18:1(n-9), 18:2(n-6), 18:3(n-3), 18:4(n-3), 20:4(n-6) and 20:5(n-3). C(18)PUFAs are of greater abundance in the brown algae than in the red algae examined. All three green algae from the Ulvales had similar fatty acid patterns with major components, 16:0, 16:4(n-3), 18:1(n-7), 18:2(n-6), 18:3(n-3), and 18:4(n-3). They contained 16:3(n-3) and more 16:4(n-3), were rich in C(18)PUFAs, chiefly 18:3(n-3) and 18:4(n-3) and had 18:1(n-7)/18:1(n-9) ratios higher than 1.     

Effect of different contents of extruded linseed in the sow diet on piglet fatty acid composition and hepatic desaturase expression during the post-natal period

n-3 polyunsaturated fatty acids (n-3 PUFA) contribute to the normal growth and development of numerous organs in the piglet. The fatty acid composition of piglet tissues is linked to the fatty acid composition of sow milk and, consequently, to the composition of sow diet during the gestation and lactation period. In this study, we investigated the impact of different contents of extruded linseed in the sow diet on the fatty acid composition and desaturase gene expression of piglets. Sows received a diet containing either sunflower oil (low 18:3n-3 with 18:3n-3 representing 3% of total fatty acids) or a mixture of extruded linseed and sunflower oil (medium 18:3n-3 with 9% of 18:3n-3) or extruded linseed (high 18:3n-3 with 27% of 18:3n-3) during gestation and lactation. Fatty acid composition was evaluated on sow milk and on different piglet tissues at days 0, 7, 14, 21 and 28. The postnatal evolution of delta5 (D5D) and delta6 (D6D) desaturase mRNA expression was also measured in the liver of low 18:3n-3 and high 18:3n-3 piglets. The milk of high 18:3n-3 sows had higher proportions of n-3PUFA than that of low 18:3n-3 and medium 18:3n-3 sows. Piglets suckling the high 18:3n-3 sows had greater proportions of 18:3n-3, 20:5n-3, 22:5n-3 and 22:6n-3 in the liver, and of 22:5n-3 and 22:6n-3 in the brain than low 18:3n-3 and medium 18:3n-3 piglets. D5D and D6D mRNA expressions in piglet liver were not affected by the maternal diet at any age. In conclusion, extruded linseed in the sow diet modifies the n-3PUFA status of piglets during the postnatal period. However, a minimal content of 18:3n-3 in the sow diet is necessary to increase the n-3PUFA level in piglet liver and brain. Moreover, modifications in the n-3PUFA fatty acid composition of piglet tissue seem linked to the availability of 18:3n-3 in maternal milk and not to desaturase enzyme expression.     

Metabolism of alpha-linolenic acid in humans

Alpha-linolenic acid (18:3n-3) is essential in the human diet, probably because it is the substrate for the synthesis of longer-chain, more unsaturated n-3 fatty acids eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3) which are required for tissue function. This article reviews the recent literature on 18:3n-3 metabolism in humans, including fatty acid beta-oxidation, recycling of carbon by fatty acid synthesis de novo and conversion to longer-chain polyunsaturated fatty acids (PUFA). In men, stable isotope tracer studies and studies in which volunteers increased their consumption of 18:3n-3 show conversion to 20:5n-3 and 22:5n-3, but limited conversion to 22:6n-3. However, conversion to 18:3n-3 to 20:5n-3 and 22:6n-3 is greater in women compared to men, due possibly to a regulatory effect of oestrogen, while partitioning of 18:3n-3 towards beta-oxidation and carbon recycling was lower than in men. These gender differences may be an important consideration in making dietary recommendations for n-3 PUFA intake.     

Biosynthesis of polyunsaturated fatty acids in zooxanthellae and polyps of corals

The fatty acid (FA) composition of zooxanthellae, polyp tissue, and intact colonies was determined in soft coral Sinularia sp. and hard coral Acropora sp. Analysis of the distribution of polyunsaturated fatty acids (PUFAs) among the zooxanthellae and the host organism showed that 18: 3n-6 and C_18–22 PUFAs of the n-3 series (18: 4n-3, 20: 5n-3, 22: 5n-3, and 22: 6n-3) were mainly synthesized by the zooxanthellae and that C_20–22 PUFAs of the n-6 series (20: 3n-6, 20: 4n-6, and 22: 4n-6) were synthesized in the polyp tissue. Soft coral polyps were able to synthesize tetracosapolyenoic FAs (24: 5n-6 and 24: 6n-3) and 18: 2n-7, their zooxanthellae synthesized C₁₆ PUFAs (16: 2n-7, 16: 3n-4, and 16: 4n-1). It is supposed that the biosynthesis of 16: 2n-7 in Sinularia sp. and 18: 3n-6 in Acropora sp. is catalyzed by Δ6 desaturase. The relatively even distribution of three FAs (18: 2n-6, 18: 3n-6, and 16: 2n-7) among lipids of zooxanthellae and coral polyps indicates the possible transport of these FAs between symbionts and the host organism.     

Differential effects of n-3 fatty acid deficiency on phospholipid molecular species composition in the rat hippocampus

In this study, we have examined the effects of n-3 fatty acid deficient diets on the phospholipids (PL) molecular species composition in the hippocampus. Female rats were raised for two generations on diets containing linoleic acid (18:2n-6), with or without supplementation of alpha-linolenic acid (18:3n-3) or 18:3n-3 plus docosahexaenoic acid (22:6n-3). At 84 days of age, the hippocampal phospholipids were analyzed by reversed phase HPLC-electrospray ionization mass spectrometry. Depleting n-3 fatty acids from the diet led to a reduction of 22:6n-3 molecular species in phosphatidylcholine (PC), phosphatidylethanolamine (PE), PE-plasmalogens (PLE), and phosphatidylserine (PS) by 70-80%. In general, 22:6n-3 was replaced with 22:5n-6 but the replacement at the molecular species level did not always occur in a reciprocal manner, especially in PC and PLE. In PC, the 16:0,22:6n-3 species was replaced by 16:0,22:5n-6 and 18:0,22:5n-6. In PLE, substantial increases of both 22:5n-6 and 22:4n-6 species compensated for the decreases in 22:6n-3 species in n-3 fatty acid deficient groups. While the total PL content was not affected by n-3 deficiency, the relative distribution of PS decreased by 28% with a concomitant increase in PC.The observed decrease of 22:6n-3 species along with PS reduction may represent key biochemical changes underlying losses in brain-hippocampal function associated with n-3 deficiency.     

Effects of two Avtemia diets with different contents of polyunsaturated fatty acids on the lipid composition of larvae of Atlantic herring (Clupea harengus

Atlantic herring larvae ( Clupea harengus ) were fed two enriched Artemia diets with different contents of (n-3) highly unsaturated fatty acids (HUFA), one containing low levels of 20: 5(n-3) and no 22: 6(n-3), the other containing substantial levels of both 20: 5(n-3) and 22: 6(n-3). After 30 days of culture, fatty acid compositions of lipid classes in the heads, bodies and eyes of the larvae were analysed. Fish fed Artemia with the low (n-3) HUFA diet lacking 22: 6(n-3) had lower amounts of total (n-3)HUFA and, in particular, of 22:6(n-3) in individual phospholipids and total neutral lipids of heads, bodies and eyes as compared to fish fed Artemia with high levels of (n-3)HUFA. The amount of 22: 6(n-3) in the fatty acids of phosphatidyl-ethanolamine of eyes was particularly susceptible to dietary depletion. The implications of these findings are discussed, particularly in relation to dietary requirements for 22: 6(n-3) during development of neural tissue in predatory fish iarvae.     

Mitochondrial and peroxisomal oxidation of arachidonic and eicosapentaenoic acid studied in isolated liver cells

The partitioning between peroxisomal and mitochondrial beta-oxidation of [1-14C]eicosapentaenoic acid (20:5(n-3] and [1-14C]arachidonic acid (20:4(n-6)) was studied. In hepatocytes from fasted rats approximately 70% of the fatty acid substrate was oxidized with oleic, linoleic, eicosapentaenoic and docosahexaenoic (22:6(n-3)) acid, even more with adrenic (22:4(n-6)) and less with arachidonic acid. When the mitochondrial oxidation was suppressed by fructose refeeding and by (+)-decanoylcarnitine, the fatty acid oxidation in per cent of that in cells from fasted rats was with 18:1(n-9) 7%, 18:2(n-6) 8%, 20:4(n-6) 12%, 20:5(n-3) 20%, 22:4(n-6) 57% and for 22:6(n-3) 29%. The fraction of 14C recovered in palmitate and other newly synthesized fatty acids after fructose refeeding decreased in the order 22:4(n-6) greater than 22:6(n-3) greater than 20:5(n-3) greater than 20:4(n-6) and was very small with 18:1(n-9) and 18:2(n-6). In cells from both fed and fructose-refed animals 20:5(n-3) was efficiently elongated to 22:5(n-3) and 22:6(n-3). 20:5(n-3) and 20:4(n-6) were not elongated after fasting. The phospholipid incorporation with [1-14C]20:5(n-3) decreased during prolonged incubations while it remained stable with [1-14C]arachidonic acid. The results suggest that peroxisomes contribute more to the oxidation of 20:5(n-3) than with 20:4(n-6) although both substrates are probably oxidized mainly in the mitochondria.     

In vivo conversion of 18- and 20-C essential fatty acids in rats using the multiple simultaneous stable isotope method

An important question for mammalian nutrition is the relative efficiency of C18 versus C20 essential fatty acids (EFAs) for supporting the tissue composition of n-3 and n-6 pathway end products. One specific question is whether C22 EFAs are made available to tissues more effectively by dietary alpha-linolenic acid (18:3n-3) and linoleic acid (18:2n-6) or by dietary eicosapentaenoic acid (20:5n-3) and dihomo-gamma-linolenic acid (20:3n-6). To address this question in a direct manner, four stable isotope compounds were given simultaneously in a novel paradigm. A single oral dose of a mixture of 2H5-18:3n-3, 13C-U-20:5n-3, 13C-U-18:2n-6, and 2H5-20:3n-6 was administered to rats given a defined diet. There was a preferential in vivo conversion of arachidonic acid (20:4n-6) to docosatetraenoic acid (22:4n-6) and of 22:4n-6 to n-6 docosapentaenoic acid (22:5n-6) when the substrates originated from the C18 precursors. However, when the end products docosahexaenoic acid (22:6n-3) or 22:5n-6 were expressed as the total amount in the plasma compartment divided by the dosage, this parameter was 11-fold greater for 20:5n-3 than for 18:3n-3 and 14-fold greater for 20:3n-6 than for 18:2n-6. Thus, on a per dosage basis, the total amounts of n-3 and n-6 end products accreted in plasma were considerably greater for C20 EFA precursors relative to C18.     

Chain elongation of eicosapentaenoic acid in the macrophage

In order to elucidate the metabolic fate of eicosapentaenoic acid (20:5 (n-3], a major n-3 fatty acid constituent of fish oil, resident and casein-elicited mouse peritoneal macrophages were incubated with [3H]20:5 (n-3). Comparative experiments with arachidonic acid (20:4 (n-6] were also conducted. After 4, 8 and 18 h incubation, [3H]20:5 (n-3) was extensively elongated into [3H]22:5(n-3) while [3H]20:4(n-6) was only moderately elongated into [3H]22:4(n-6) in both resident and elicited macrophages. No measurable conversion of [3H]22:5(n-3) into [3H]22:6(n-3) (delta 4 desaturation) could be demonstrated. These data demonstrate that the highly active chain elongation of 20:5(n-3) by macrophage elongase, as well as the lack of detectable delta 4 desaturase activity, are responsible for the accumulation of 22:5(n-3) in this cell.     

The effect of altering the 20:5n-3 and 22:6n-3 content of a meal on the postprandial incorporation of n-3 polyunsaturated fatty acids into plasma triacylglycerol and non-esterified fatty acids in humans

Previous studies suggest that consuming meals containing large amounts of fish oil is associated with selective postprandial incorporation of 20:5n-3 and 22:6n-3 into plasma non-esterified fatty acids (NEFA). We investigated the effect of consuming meals containing different amounts of 20:5n-3 and 22:6n-3 comparable to dietary habits of western populations on the postprandial incorporation of 18:3n-3, 20:5n-3 and 22:6n-3 into plasma triacylglycerol (TAG) and NEFA over 6h in middle aged subjects. 20:5n-3 incorporation into plasma TAG was greater than 22:6n-3 irrespective of the test meal. Conversely, 22:6n-3 incorporation into plasma NEFA was greater than 20:5n-3, irrespective of the test meal. There was no effect of the amount of 20:5n-3+22:6n-3 in the test meal on the 18:3n-3 incorporation into plasma TAG or NEFA. These findings suggest differential metabolism of 20:5n-3 and 22:6n-3 in the postprandial period when consumed in amounts typical of western dietary habits.     

The lipid composition of two species of serrasalmid fish in relation to dietary polyunsaturated fatty acids

The lipid composition of two species of Serrasalmid fish with different natural feeding habits were compared in relation to the polyunsaturated fatty acids (PUFA) supplied in their diets. Mylossoma aureum , a herbivorous piranha, was maintained on oatmeal flakes in which : 2(n-6) and : 3(n-3) were the only PUFA and accounted for 40–8 and 1.2%, respectively of dietary fatty acids. Serrasalmus nattereri , the carnivorous red piranha, was fed mosquito larvae containing .0-33.4% of their total fatty acids as : 2(n-6)+18 : 3(n-3) and 4.9-8.5% as 20 : 4(n-6)+20 : 5(n-3). The two species had similar lipid class compositions in liver, brain, viscera and carcass, except that lipids from M. aureum were generally richer in triacylglycerols. In both species, visceral and carcass lipid contained high levels of triacylglycerols whose principal PUFA was : 2(n-6). In M. aureum the major PUFA in liver total lipid and triacylglycerols was : 2(n-6) whilst the major PUFA in liver phospholipids were : 4(n-6) and : 5(n-6), with : 6(n-3) being a minor component. The level of : 6(n-3) in ethanolamine glycerophospholipids was significantly greater in brain than liver of M. aureum. Although absent from dietary lipid, : 6(n-3) was the major PUFA in phosphatidylcholine and ethanolamine glycerophospholipids from both the liver and brain of S, nattereri . In both species, the ratio of (n-6)/(n-3)PUFA was consistently lower in tissue lipids than in dietary lipids. The results are consistent with (i) the herbivorous M. aureum converting dietary C18 PUFA to their C20 and C22 homologues, (ii) the carnivorous S, nattereri forming : 6(n-3) from either 18:3(n-3) or 20: 5(n-3) and (iii) both species selectively desaturating and elongating (n-3) rather than (n-6) PUFA.     

Comparison of 20-, 22-, and 24-carbon n-3 and n-6 polyunsaturated fatty acid utilization in differentiated rat brain astrocytes

Astrocytes convert n-6 fatty acids primarily to arachidonic acid (20:4n-6), whereas n-3 fatty acids are converted to docosapentaenoic (22:5n-3) and docosahexaenoic (22:6n-3) acids. The utilization of 20-, 22- and 24-carbon n-3 and n-6 fatty acids was compared in differentiated rat astrocytes to determine the metabolic basis for this difference. The astrocytes retained 81% of the arachidonic acid ([(3)H]20:4n-6) uptake and retroconverted 57% of the docosatetraenoic acid ([3-(14)C]22:4n-6) uptake to 20:4n-6. By contrast, 68% of the eicosapentaenoic acid ([(3)H]20:5n-3) uptake was elongated, and only 9% of the [3-(14)C]22:5n-3 uptake was retroconverted to 20:5n-3. Both tetracosapentaenoic acid ([3-(14)C]24:5n-3) and tetracosatetraenoic acid ([3-(14)C]24:4n-6) were converted to docosahexaenoic acid (22:6n-3) and 22:5n-6, respectively. Therefore, the difference in the n-3 and n-6 fatty acid products formed is due primarily to differences in the utilization of their 20- and 22-carbon intermediates. This metabolic difference probably contributes to the preferential accumulation of docosahexaenoic acid in the brain.     

Leptin levels in rat offspring are modified by the ratio of linoleic to alpha-linolenic acid in the maternal diet

The supply of polyunsaturated fatty acids (PUFA) is important for optimal fetal and postnatal development. We have previously shown that leptin levels in suckling rats are reduced by maternal PUFA deficiency. In the present study, we evaluated the effect of maternal dietary intake of (n-3) and (n-6) PUFA on the leptin content in rat milk and serum leptin levels in suckling pups. For the last 10 days of gestation and throughout lactation, the rats were fed an isocaloric diet containing 7% linseed oil (n-3 diet), sunflower oil (n-6 diet), or soybean oil (n-6/n-3 diet). Body weight, body length, inguinal fat pad weight, and adipocyte size of the pups receiving the n-3 diet were significantly lower during the whole suckling period compared with n-6/n-3 fed pups. Body and fat pad weights of the n-6 fed pups were in between the other two groups at week one, but not different from the n-6/n-3 group at week 3. Feeding dams the n-3 diet resulted in decreased serum leptin levels in the suckling pups compared with pups in the n-6/n-3 group. The mean serum leptin levels of the n-6 pups were between the other two groups but not different from either group. There were no differences in the milk leptin content between the groups. These results show that the balance between the n-6 and n-3 PUFA in the maternal diet rather than amount of n-6 or n-3 PUFA per se could be important for adipose tissue growth and for maintaining adequate serum leptin levels in the offspring.     

Alternate pathways in the desaturation and chain elongation of linolenic acid, 18:3(n-3), in cultured glioma cells.

Cultured C6 glioma cells rapidly incorporate and metabolize the essential fatty acids, 18:2(n-6) and 18:3(n-3), to 20- and 22-carbon polyunsaturated fatty acids. Using several deuterated fatty acid substrates we have obtained data that suggest alternate pathways, one possibly involving delta 8-desaturation, may exist in glioma cells for formation of 20:5(n-3) and 22:6(n-3) from 18:3(n-3). With 18:3(n-3)-6,6,7,7-d4 practically no 18:4(n-3)-6,7-d2 or 20:4(n-3)-8,9-d2 was detected whereas 20:3(n-3)-8,8,9,9-d4 accounted for 3.4% and delta 5,11,14,17-20:4-8,8,9,9-d4 for 21.1% of the total deuterated fatty acids recovered in phospholipids after a 16 h incubation; 20:5(n-3)-8,9-d2, 22:5(n-3)-10,11-d2, and 22:6(n-3)-10,11-d2 accounted for 42.4%, 13.2%, and 2.8% of deuterated acyl chains, respectively. When added exogneously, 20:3-8,8,9,9,-d4 was extensively converted to delta 5,11,14,17-20:4(n-3)-8,8,9,9-d4 (45%) and 20:5(n-3)-8,9-d2 (24%); a small amount (4%) of 18:3(n-3)-d4 also was detected. Both 20:4(n-3)-8,9-d2 and 18:4(n-3)-12,13,15,16-d4 were also converted to 20:5(n-3) and 22:6(n-3) with 8 and 0% of the respective original deuterated substrate remaining after 16 h. A possible pathway for 18:3(n-3) metabolism in glioma cells is described whereby an initial chain elongation step is followed by successive delta 5 and delta 8 desaturation reactions resulting in 20:5(n-3) formation and accounting for the ordered removal of deuterium atoms. Alternatively, extremely effective retroconversion may occur to chain shorten 20:3(n-3)-d4 to 18:3(n-3)-d4 followed by rapid conversion through the classical desaturation and chain elongation sequence.(ABSTRACT TRUNCATED AT 250 WORDS)     

The levels of essential unsaturated fatty acids in human milk on the 3rd, 4th, 5th, and 6th days after labour

The composition of fatty acids in human milk lipids was determined in 41 women on the 3rd, 4th, 5th and 6th days after labour by the method of gas chromatography. In these investigations no significant differences were demonstrated in the fatty acids in the lipid fractions between these consecutive days. The level of polyunsaturated fatty acids of the n-6 and n-3 groups was about 11.9-13.6%, including linoleic acid (18:2, n-6) about 7.7-9.8%, and alpha-linolenic acid (18:3, n-3) about 0.7-1%. In the analysis group of n-6 fatty acids the determined acids were: linoleic acid (18:2, n-6), gamma-linolenic acid (18:3, n-6), eicosadienoic acid (20:2, n-6), eicosatrienoic acid (20:3, n-6), arachidonic acid (20:4, n-6), docosahexaenoic acid (22:6, n-6). From the group of n-3 acids the identified ones were: alpha-linolenic acid (18:3, n-3), eicosapentaenoic acid (20:5, n-3), docosapentaenoic acid (22:5, n-3) and docosahexaenoic acid (22:6, n-3). The obtained quotients of fatty acids n-6 through n-3 on the consecutive days were: 7.2:1-7.8:1, indicating a too low level of the n-3 acids in the investigated milk. The acids prevailing in human milk lipids were: oleic (18:1, n-9) and palmitic (16:0) which accounted for 37-39% and 25-26% respectively. The polyunsaturated to saturated fatty acid ratio (P:S) ranged from 0.28 to 0.33.     

The esterification and modification of n-3 and n-6 polyunsaturated fatty acids by hepatocytes and liver microsomes of turbot (Scophthalmus maximus)

The present study was undertaken to establish whether the formation of 22:6n-3 from 18:3n-3 and/or 20:5n-3 can occur in turbot liver and if this conversion is consistent with the operation of a Delta4 desaturase-independent pathway. At the same, time the effects of feeding a diet devoid of long chain polyunsaturated fatty acids (PUFA) on the patterns of esterification and modification of 18:3n-3, 20:5n-3 and 18:2n-6 by turbot hepatocytes and liver microsomes were examined. For this purpose, two groups of fish (25-30 g) were employed: one was fed a commercial diet containing fish oil (FO) and thus rich in long chain n-3 PUFA and the other was fed an experimental diet based on olive oil (OO). After 5 months of feeding, hepatocytes and liver microsomes isolated from individuals in the two groups of fish were incubated with [1-(14)C]-PUFA [either 18:3n-3, 20:5n-3 or 18:2n-6]. After 3 h of incubation, most radioactivity from all three radiolabelled substrates incorporated into lipids by hepatocytes and microsomes was recovered in the original substrate. The formation of desaturation products of n-3 radiolabelled substrates was higher in hepatocytes isolated from OO-fed than FO-fed fish. Small amounts of radiolabelled 22:6n-3 were formed from [1-(14)C]18:3n-3 and [1-(14)C]20:5n-3, but only by hepatocytes from fish fed OO, which also synthesised a small amount of radiolabelled 24:6n-3 from 14C-20:5n-3. Elongation products predominated over desaturation products in hepatic microsomes from both groups of fish studied, particularly in microsomes from fish fed FO. The results confirm that regardless of the long chain PUFA content of the diet, the production of 22:6n-3 in turbot liver from 18:3n-3 and/or 20:5n-3, and of 20:4n-6 from 18:2n-6, is very limited. The presence of radiolabelled 24:6n-3 in microsomes coupled with the absence of radiolabelled 22:6n-3 suggests that the formation of 22:6n-3 that does occur in turbot liver cells, may involve C24 intermediates and peroxisomal beta-oxidation.     

Inhibition of Icosanoid Production in MC/9 Mouse Mast Cells by n-3 Polyunsaturated Fatty Acids Isolated from Edible Marine Algae

The effects of two polyunsaturated fatty acids, 18:4n-3 and 16:4n-3 purified from the marine algae, Undaria pinnatifida and Ulva pertusa, on icosanoid production in MC/9 mouse mast cells were assessed. Both fatty acids suppressed the production of leukotriene B₄ (LTB₄), leukotriene C₄ (LTC₄), and 5-hydroxyeicosatetraenoic acid (5-HETE). The order of the suppressive activity for the two marine algae-derived fatty acids and three other common polyunsaturated fatty acids was as follows; 22:6n-3=18:4n-3=18:3n-3>20:5n-3=16:4n-3 for LTB₄; 22:6n-3=18:4n-3=18:3n-3>16:4n-3>20:5n-3 (no suppression) for LTC₄; 22:6n-3=18:4n-3>18:3n-3>20:5n-3=16:4n-3 for 5-HETE.