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.     

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.     

Identification and expression of mammalian long-chain PUFA elongation enzymes

In mammalian cells, Sprecher has proposed that the synthesis of long-chain PUFA from the 20-carbon substrates involves two consecutive elongation steps, a Δ6-desaturation step followed by retroconversion (Sprecher, H., Biochim. Biophys. Acta 1486, 219–231, 2000). We searched the database using the translated sequence of human elongase ELOVL5, whose encoded enzyme elongates monounsaturated and polyunsaturated FA, as a query to identify the enzyme(s) involved in elongation of very long chain PUFA. The database search led to the isolation of two cDNA clones from human and mouse. These clones displayed deduced amino acid sequences that had 56.4 and 58% identity, respectively, to that of ELOVL5. The open reading frame of the human clone (ELOVL2) encodes a 296-amino acid peptide, whereas the mouse clone (Elovl2) encodes a 292-amino acid peptide. Expression of these open reading frames in baker's yeast, Saccharomyces cerevisiae, demonstrated that the encoded proteins were involved in the elongation of both 20-and 22-carbon long-chain PUFA, as determined by the conversion of 20∶4n−6 to 22∶4n−6, 22∶4n−6 to 24∶4n−6, 20∶5n−3 to 22∶5n−3, and 22∶5n−3 to 24∶5n−3. The elongation activity of the mouse Elovl2 was further demonstrated in the transformed mouse L cells incubated with long-chain (C₂₀-and C₂₂-carbon) n−6 and n−3 PUFA substrates by the significant increase in the levels of 24∶4n−6 and 24∶5n−3, respectively. This report demonstrates the isolation and identification of two mammalian genes that encode very long chain PUFA specific elongation enzymes in the Sprecher pathway for DHA synthesis.     

Lipids and delta6-desaturase activity alterations in rat liver microsomal membranes induced by fumonisin B1

Alterations in the membrane structure and function of hepatocyte membranes by fumonisin B₁ (FB₁) have been proposed to play an important role in the disruption of growth regulatory effects and hence in the cancer-promoting ability of the mycotoxin. Detailed analyses of lipids in liver microsomal fractions of rats exposed to different dietary levels of FB₁ over a period of 21 d indicated an increase in PC, PE, PI, and cholesterol (Chol). These changes decreased the PC/PE and increased the total phospholipid/Chol ratios. When considering FA content, the quantities of total FA increased (P<0.05) in the major phospholipid fractions as a result of the increased phospholipid levels. However, when considering the relative levels (mg/100 mg of the total FA) of specific FA, the monounsaturated FA (16∶1n−7 and 18∶1n−9) and 18∶2n−6 increased (P<0.05), whereas the long-chain PUFA decreased (P<0.05) in the main phospholipid fractions. Enzyme analyses indicated that the activity of the Δ6-desaturase was significantly reduced in liver microsomal preparations in a dose-dependent manner. An increase in the 20∶3n−6/20∶4n−6 ratio also suggested a decrease in the activity of the Δ5-desaturase. Disruption of microsomal lipid metabolism at different levels by FB₁ could play an important role in the alteration of growth regulatory effects in the liver.     

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.     

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.     

FA composition of plasma,red blood cell,and liver phospholipids of newborn piglets fed trans isomers of alpha-linolenic acid

The effects of dietary cis and trans α-linolenic acid (18∶3n−3) on the FA composition of plasma, red blood cell, and liver phospholipids were studied in newborn piglets. Animals were fed for 14 d with one of three diets: a control diet (group A) containing cis 18∶3n−3 at a level of 2.0% of total FA, a diet (group B) in which a part of the 18∶3n−3 acid was isomerized (1.3% of cis 18∶3n−3 and 0.7% of trans 18∶3n−3), or a diet (group C) with 2.0% cis 18∶3n−3 and 0.7% trans 18∶3n−3. Feeding animals with diets containing trans 18∶3n−3 resulted in the presence of trans isomers of 18∶3n−3, trans isomers of EPA, and trans isomers of DHA in phospholipids; however, the level of total trans n−3 PUFA in tissues was less than 0.3% of total tissue FA. In group B, the reduction of dietary amounts of cis 18∶3n−3 was associated with a decrease in individual and total cis n−3 PUFA. In contrast, in group C there was no decrease in tissue n−3 PUFA despite the increased dietary level of trans 18∶3n−3. These results suggest that the isomerization of a part of dietary n−3 PUFA, leading to the reduction of their levels in the diet, could induce a decrease in n−3 PUFA in phospholipids. The physiological effects of trans PUFA are not known and should be considered in future studies.     

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.     

Liver desaturase activities and FA composition in monkeys. Effect of a low-protein diet

The aim of the present study was to measure Δ9-, Δ6-, and Δ5-desaturase activities in liver microsomes, as well as phospholipid FA composition of liver and erythrocytes in monkeys fed a control or low-protein diet during the postweaning period. Ten Saimiri sciureus boliviensis (Cebidae) of both sexes were employed; at 12 mon of age they were separated into two groups fed ad libitum on a control or a low-protein diet for 24 mon. Saimiri sciureus had active Δ9, Δ6, and Δ5 liver desaturase enzymes, and these activities were influenced by the diet. A low-protein diet produced a significant reduction in Δ5-desaturation capacity, an increase in Δ9-desaturase activity, and no change in Δ6-desaturase activity (P<0.05). These changes, evoked by protein deprivation, were reflected in the liver phospholipid FA composition. Increases in the proportion of saturated FA and in monounsaturated oleic acid (18∶1n−9) and a decrease in the proportion of PUFA of the n−6 and n−3 series were produced in the animals fed a low-protein diet (P<0.0001). Differences between the two dietary groups were less pronounced in the FA composition of erythrocyte phospholipids. The authors are members of the Carrera del Investigador del Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.     

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.     

Tissue deposition of n−3 FA pathway intermediates in the synthesis of DHA in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

The tissue distribution of newly synthesized 22∶6n-3 and intermediate PUFA was examined in rainbow trout to further our understanding of the metabolism of this EFA in fish. Rainbow trout were fed a pulse of deuterated linoleic acid (D₅−17,17,18,18,18−18∶3n-3), and the tissue distribution of deuterated anabolites was determined at interval up to 35 d post-dose by GC-negative chemical ionization MS of the pentafluorobenzyl derivatives. D₅−22∶6n-3 was the major deuterated FA in liver and cecal mucosa 2 and 5 d post-dose. All the n−3 FA pathway intermediates were found in liver, cecal mucosa, and blood including D₅−24∶5n-3 and D₅−24∶6n-3. Brain and eyes also contained the full suite of intermediate deuterated FA, but with a different profile from liver when analyzed over a longer time course up to 35 d. D₅−20∶5n-3 was the major component in brain up to 7 d, after which D₅−22∶6n-3 became predominant, but D₅−22∶5n-3 constituted ca. 20% of FA throughout the time period. The pattern in eyes was similar but less pronounced. In visceral adipose tissue there was a much greater accumulation of the initial substrate, D₅−18∶3n-3, with D₅−18∶4n-3 and D₅−22∶6n-3 the predominant deuterated FA at all time points. There was a similar though less pronounced trend in eye socket adipose tissue. The C₂₄ PUFA were not detected in visceral fat and barely detected in eye socket fat. The results show that the kinetics of accumulation and depletion of the various n−3 PUFA differ between tissues. The presence of pathway intermediate FA provides evidence that liver and ceca possess the full metabolic pathway for synthesis of 22∶6n-3, whereas brain and eyes are less active, with an accumulation of pentaene intermediate FA, and adipose tissue is inactive.     

Dietary fish oil dose-response effects on ileal phospholipid fatty acids and contractility

We have reported that dietary fish oil (FO) leads to the incorporation of long-chain n−3 PUFA into the gut tissue of small animal models, affecting contractility, particularly of rat ileum. This study examined the FO dose response for the incorporation of n−3 PUFA into ileal tissue and how this correlated with in vitro contractility. Groups of ten to twelve 13-wk-old Wistar-Kyoto rats were fed 0, 1, 2.5, and 5% FO-supplemented diets balanced with sunflower seed oil for 4 wk, after which ileal total phospholipid FA were determined and in vitro contractility assessed. For the total phospholipid fraction, increasing the dietary FO levels led to a significant increase first evident at 1% FO, with a stepwise, nonsaturating six-fold increase in n−3 PUFA as EPA (20∶5n−3), DPA (docosapentaenoic acid, 22∶5n−3), and DHA, but mainly as DHA (22∶6n−3), replacing the n−6 PUFA linoleic acid (18∶2n−6) and arachidonic acid (20∶4n−6) over the dosage range. There was no difference in KCl-induced depolarization-driven contractility. However, a significant increase in receptor-dependent maximal contractility occurred at 1% FO for carbachol and at 2.5% FO for prostaglandin E₂, with a concomitant increase in sensitivity for prostaglandin E₂ at 2.5 and 5% FO. These results demonstrate that significant increases in ileal membrane n−3 PUFA occurred at relatively low doses of dietary FO, with differential receptor-dependent increases in contractility observed for muscarinic and prostanoid agonists.     

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.     

Isomers of hexadecenoic and hexadecadienoic acids in <Emphasis Type="Italic">Androsace septentrionalis</Emphasis> (Primulaceae) seed oil

Seeds of Androsace septentrionalis of the genus Androsace (tribus Primuleae) from the plant family Primulaceae were studied for their oil content and FA composition. The seed oil of A. septentrionalis was found to contain two unusual FA rarely occurring in plants: 11-cis-hexadecenoic acid (16∶1Δ11c or 16∶1n−5) and 9-cis,12-cis-hexadecadienoic acid (16∶2Δ9c,12c or 16∶2n−4). It also contained an unusually high amount (21.4%) of 9-cis-hexadecenoic acid (palmitoleic acid; 16∶1Δ9c or 16∶1n−7), i.e., at a level higher than that of oleic acid, in addition to common FA. Compared with most plant seed oils, at 3.8% the level of 18∶1Δ11c (or 18∶1n−7) also was elevated. The nonidentity of the Androsace 16∶2-acid with the 16∶2-acid, which is very typical for Ranunculus spp., as well as its identity with the 16∶2-acid typically found in Asclepiadaceae was established by co-chromatography. The structure and composition of the constituent FA of A. septentrionalis were also determined by various chromatographic methods (TLC, Ag⁺-TLC, capillary GLC) and spectroscopic methods (IR, GC-MS). The significant deviation of the Androsace FA pattern from that of other Primuleae, indicating a separate phylogenetic position of Androsace, is discussed.     

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.     

Influence of diet on fatty acids of three subtropical fish, subfamily caesioninae (Caesio diagramma and C. tile) and family siganidae (Siganus canaliculatus)

The total lipid and fatty acid compositions of tissues and the stomach contents of three subtropical marine fish species, subfamily Caesioninae, Caesio diagramma and C. tile, and family Siganidae Siganus canaliculatus, were investigated to clarify the differences between these species. Triacylglycerols (TAG) were the dominant depot lipids of the three species, whereas wax esters were found as a minor component. In particular, muscle lipids were found to contain mainly glycerol derivatives such as TAG and phospholipids. The major fatty acids identified in the three species were 16∶0, 18∶0, 18∶1n−9, and 22∶6n−3 (docosahexaenoic acid, DHA). In addition, noticeable levels of 16∶1n−7, 18∶1n−7, 20∶4n−6 (arachidonic acid, AA), and 20∶5n−3 (eicosapentaenoic acid) were found. DHA was the most abundant polyunsaturated fatty acid (PUFA) in the muscle and viscera lipids of the three species. The high DHA levels in the lipids of all the organs were found to be higher than those of the lipid extracted from the stomach contents of the three fishes. In addition, the specimens of S. canaliculatus contained significantly higher levels of AA in its tissues than did the other two species. A high AA content is unusual since such high levels of n−6 PUFA are rarely found in higher marine organisms. These levels may be due to its characteristic feeding pattern, because S. canaliculatus prefer and mainly feed on seaweed, which often contains high amounts of n−6 PUFA, such as linoleic acid (18∶2n−6) and AA.     

Effect of the Δ<Superscript>6</Superscript>-desaturase inhibitor SC-26196 on PUFA metabolism in human cellsinhibitor SC-26196 on PUFA metabolism in human cells

The objective of this study was to determine the effect of 2,2-diphenyl-5-(4-{[(1E)-pyridin-3-yl-methylidene]-amino}piperazin-1-yl)pentanenitrile (SC-26196), a Δ⁶-desaturase inhibitor, on PUFA metabolism in human cells. SC-26196 inhibited the desaturation of 2 μM [1-¹⁴C] 18∶2n−6 by 87–95% in cultured human skin fibroblasts, coronary artery smooth muscle cells, and astrocytes. By contrast, SC-26196 did not affect the conversion of [1-¹⁴C]20∶3n−6 to 20∶4 in the fibroblasts, demonstrating that it is selective for Δ⁶-desaturase. The IC₅₀ values for inhibition of the desaturation of 2 μM [1-¹⁴C] 18∶3n−3 and [3-¹⁴C]24∶5n−3 in the fibroblasts, 0.2–0.4 μM, were similar to those for the inhibition of [1-¹⁴C] 18∶2n−6 desaturation, and the rates of recovery of [1-¹⁴C] 18∶2n−6 and [3-¹⁴C] 24∶5n−3 desaturation after removal of SC-26196 from the culture medium also were similar. SC-26196 reduced the conversion of [3-¹⁴C] 22∶5n−3 and [3-¹⁴C] 24∶5n−3 to DHA by 75 and 84%, respectively, but it had no effect on the retroconversion of [3-¹⁴C] 24∶6n−3 to DHA. These results demonstrate that SC-26196 effectively inhibits the desaturation of 18- and 24-carbon PUFA and, therefore, decreases the synthesis of arachidonic acid, EPA, and DHA in human cells. Furthermore, they provide additional evidence that the conversion of 22∶5n−3 to DHA involves Δ⁶-desaturation.     

Molecular evidence that the rate-limiting step for the biosynthesis of arachidonic acid in <Emphasis Type="Italic">Mortierella alpina</Emphasis> is at the level of an elongase

The oil-producing fungus Mortierella alpina 1S-4 is an industrial strain for arachidonic acid (AA) production. To determine its physiological properties and to clarify the biosynthetic pathways for PUFA, heterologous and homologous gene expression systems were established in this fungus. The first trial was performed with an enhanced green fluorescent protein gene to assess the transformation efficiency for heterologous gene expression. As a result, strong fluorescence was observed in the spores of the obtained transformant, suggesting that the foreign gene was inherited by the spores. The next trial was performed with a homologous PUFA elongase (GLELOp) gene, this enzyme having been reported to catalyze the elongation of GLA (18∶3n−6) to dihomo-γ-linolenic acid (20∶−6), and to be the rate-limiting step of AA production. The FA composition of the transformant was different from that of the host strain: The GLA content was decreased whereas that of AA was increased. These data support the hypothesis that the GLELOp enzyme plays an important role in PUFA synthesis, and may indicate how to control PUFA biosynthesis.     

Very long chain PUFA in murine testicular triglycerides and cholesterol esters

Very long chain (VLC) PUFA of the n−6 and n−3 series are known to occur in mammalian testis. The aim of this work was to characterize further two testicular lipid classes with VLCPUFA, cholesterol esters (CE) and total triglycerides (TG) in rat and mouse testis. The VLCPUFA predominating in these lipids were a series of n−6 pentaenes and tetraenes with 24 to 32 carbons, including small amounts of odd-chain PUFA, 28∶5n−6 and 24∶5n−6 prevailing in CE and TG, respectively. Most of the VLCPUFA of TG were concentrated in a small fraction of TG, made up by 1-O-alkyl-2,3-DAG. This TG subclass was absent altogether from the TG of sexually immature testis. The TG and the CE with VLCPUFA only occurred in testis of adult fertile animals. The proportion of VLCPUFA in total TG and CE was higher in rodents than in other mammals. In the n−6 PUFA-rich adult mouse testis, the amounts of testicular triacylglycerols decreased significantly after consumption of fish oil for 2 wk. Whereas 18∶2n−6 was significantly reduced, the amounts of 22∶5n−6 and longer n−6 PUFA were less affected in all mamor testicular lipids including PC and PE, where they were unchanged. The 1-O-alkyl-2,3-DAG and their n−6 VLCPUFA were virtually unaffected by the diet. The VLCPUFA-containing molecular species of CE and TG may represent a form of storage of cholesterol and polyenoic FA required to sustain spermatogenesis. Via chain-shortening, VLCPUFA stored in the neutral lipids may serve as precursors of the major C₂₂ PUFA typical of cell membrane glycerophospholipids, protecting testicular cells against shifts in FA composition induced by dietary changes.