Effect of n−3 fatty acid deficiency on fatty acid composition and metabolism of aminophospholipids in rat brain synaptosomes

Docosahexaenoic acid (DHA, 22∶6n−3) is one of the major polyunsaturated fatty acids esterified predominantly in aminophospholipids such as ethanolamine glycerophospholipid (EtnGpl) and serine glycerophospholipid (SerGpl) in the brain. Synaptosomes prepared from rats fed an n−3 fatty acid-deficient safflower oil (Saf) diet had significantly decreased 22∶6n−3 content with a compensatory increased 22∶5n−6 content when compared with rats fed an n−3 fatty acid-sufficient perilla oil (Per) diet. When the Saf group was shifted to a diet supplemented with safflower oil plus 22∶6n−3 (Saf+DHA) after weaning, 22∶6n−3 content was found to be restored to the level of the Per group. The uptake of [³H]ethanolamine and its conversion to [³H]EtnGpl did not differ significantly among the three dietary groups, whereas the formation of [³H]lysoEtnGpl from [³H]ethanolamine was significantly lower in the Saf group than in the other groups. The uptake of [³H]serine, its incorporation into [³H]SerGpl, and the conversion into [³H]EtnGpl by decarboxylation of [³H]SerGpl did not differ among the three dietary groups. The observed decrease in lysoEtnGpl formation associated with a reduction of 22∶6n−3 content in rat brain synaptosomes by n−3 fatty acid deprivation may provide a clue to reveal biochemical bases for the dietary fatty acids-behavior link.     

Effects of postnatal ethanol exposure on brain growth and lipid composition in n−3 fatty acid-deficient and-adequate rats

The artificial rearing model was used to investigate the effects of short-term exposure to ethanol on growth and fatty acid composition of forebrain (FB) and cerebellum (CB) during the brain growth spurt in either n−3 fatty acid-adequate (AD) or n−3 deficient (DEF) rat pups. On postnatal day 5, offspring of female rats that had been fed AD or DEF diets from day 5 of life were assigned to three groups: members of two groups were gastrostomized and artificially fed formulas appropriate for their maternal history, and the third group (suckled control) was fostered to lactating dams of a similar dietary history. Half of the artificially reared pups in each dietary condition were fed ethanol in their formula (7% vol/vol) in one-quarter of their daily feedings, while the others received maltose-dextrin substituted isocalorically for ethanol. Blood alcohol concentrations did not differ betwen the dietary groups. FB weight on postnatal day 9 was lower in ethanol-exposed offspring in both dietary conditions. Brain fatty acid composition reflected dietary history in that, compared with AD pups, DEF pups had lower percentages of docosahexaenoic acid, higher percentages of 22∶5n−6, and a higher n−6/n−3 fatty acid ratio. However, the effects of ethanol exposure were inconsistent, lowering the n−6/n−3 ratio in the phosphatidylethanolamine (PF) fraction in FB but not in CB, while increasing this ratio in the phosphatidylcholine (PC) fraction in FB of the DEF pups only. Thus, while ethanol had some effects on lipid composition, there was no difference between the dietary groups in their vulnerability to the effects of early short-term ethanol exposure on brain growth.     

Dietary α-linolenic acid increases brain but not heart and liver docosahexaenoic acid levels

Fish oil-enriched diets increase n−3 FA in tissue phospholipids; however, a similar effect by plant-derived n−3 FA is poorly defined. To address this question, we determined mass changes in phospholipid FA, individual phospholipid classes, and cholesterol in the liver, heart, and brain of rats fed diets enriched in flax oil (rich in 18∶3n−3), fish oil (rich in 22∶6n−3 and 20∶5n−3), or safflower oil (rich in 18∶2n−6) for 8 wk. In the heart and liver phospholipids, 22∶6n−3 levels increased only in the fish oil group, although rats fed flax oil accumulated 20∶5n−3 and 22∶5n−3. However, in the brain, the flax and fish oil diets increased the phospholipid 22∶6n−3 mass. In all tissues, these diets decreased the 20∶4n−6 mass, although the effect was more marked in the fish oil than in the flax oil group. Although these data do not provide direct evidence for 18∶3n−3 elongation and desaturation by the brain, they demonstrate that 18∶3n−3-enriched diets reduced tissue 20∶4n−6 levels and increased cellular n−3 levels in a tissuedependent manner. We hypothesize, based on the lack of increased 22∶6n−3 but increased 18∶3n−3 in the liver and heart, that the flax oil diet increased circulating 18∶3n−3, thereby presenting tissue with this EFA for further elongation and desaturation.     

Effects of dietary α-linolenic acid on the conversion and oxidation of13C-α-linolenic acid

The effects of a diet rich in α-linolenic acid vs. one rich in oleic acid on the oxidation of uniformly labeled¹³C-α-linolenic acid and its conversion into longer-chain polyunsaturates (LCP) were investigatedin vivo in healthy human subjects. Volunteers received a diet rich in oleic acid (n=5) or a diet rich in α-linolenic acid (n=7; 8.3 g/d) for 6 wk before and during the study. After 6 wk, subjects were given 45 mg of¹³C-α-linolenic acid dissolved in olive oil. Blood samples were collected att=0, 5, 11, 24, 96, and 336 h. Breath was sampled and CO₂ production was measured each hour for the first 12 h. The mean (±SEM) maximal absolute amount of¹³C-eicosapentaenoic acid (EPA) in plasma total lipids was 0.04 ±0.01 mg in the α-linolenic acid group, which was significantly lower (P=0.01) than the amount of 0.12±0.03 mg¹³C-EPA in the oleic acid group. Amounts of¹³C-docosapentaenoic acid (DPA) and¹³C-docosahexaenoic acid (DHA) tended to be lower as well. The mean proportion of labeled α-linolenic acid (ALA) recovered as¹³CO₂ in breath after 12 h was 20.4% in the ALA and 15.7% in the oleic acid group, which was not significantly different (P=0.12). The cumulative recovery of¹³C from¹³C-ALA in breath during the first 12 h was negatively correlated with the maximal amounts of plasma¹³C-EPA (r=−0.58,P=0.047) and¹³C-DPA (r=−0.63,P=0.027), but not of¹³C-DHA (r=−0.49,P=0.108). In conclusion, conversion of¹³C-ALA into its LCP may be decreased on diets rich in ALA, while oxidation of¹³C-ALA is negatively correlated with its conversion into LCP. In a few pilot samples, low¹³C enrichments of n−3 LCP were observed in a diet rich in EPA/DHA as compared to oleic acid.     

The effects of dietary α-linolenic acid compared with docosahexaenoic acid on brain, retina, liver, and heart in the guinea pig

The aim of this study was to compare two different strategies to elevate brain, retina, liver, and heart docosahexaenoic acid (DHA) levels in guinea pigs. Fist, we used an increasing dose of α-linolenic acid (AIA) relative to a constant linoleic acid (LA) intake, and second, we used two levels of dietary DHA provided in conjunction with dietary arachidonic acid (AA). The percentage DHA and AA of total phospholipids in retina, liver, and heart, and in the brain phosphotidylethanolamine and phosphatidylcholine was studied in female pigmented guinea pigs (3 wk old) fed one of five semisynthetic diets containing 10% (w/w) lipid for 12 wk. The LA content in the diets was constant (17% of total fatty acids), with the ALA content varying from 0.05% (diet SFO), to 1% (diet Mix), and to 7% (diet CNO). Two other diets LCP) and LCP3) had a constant LA/ALA ratio (17.5∶1) but varied in the levels of dietary AA and DHA supplementation. Diet LCP1 was structured to closely replicate the principal long chain polyunsaturated fatty acids (PUFA) found in human breast milk and contained 0.9% AA and 0.6% DHA (% of total fatty acids) whereas diet LCP3 contained 2.7% AA and 1.8% DHA. At the end of the study, animals were sacrificed and tissues taken for fatty acid analyses. We found no significant effects of diets on the growth of guinea pigs. Diets containing ALA has profoundly different effects on tissue fatty acid compositions compared with diets which contained the long chain PUFA (DHA and AA). In the retina and brain phospholipids, high-ALA diets or dietary DHA supplementation produced moderate relative increases in DHA levels. There was no change in retinal or brain AA proportions following dietary AA supplementation, even at the highest level. This was in contrast to liver and heart where tissue DHA proportions were low and AA predominated. In these latter tissues, dietary ALA had little effect on tissue DHA proportions although the proportion of AA was slightly depressed at the highest dietary ALA intake, but dietary DHA and AA supplements led to large increases (up to 10-fold) in the proportions of these PUFA. Tissue uptake of dietary AA and DHA appeared maximal for the LCP1 diet (replicate of breast milk) in the heart. There were no significant changes in the plasma levels of 11-dehydrothromboxane B₂ (a thromboxane A₂ metabolite), for any diet. The data confirm that dietary ALA is less effective than dietary DHA supplementation (on a gram/gram basis) in increasing tissue DHA levels and that tissues vary greatly in their response to exogenous AA and DHA, with the levels of these long chain metabolites being most resistant to change in the retina and brain compared with liver and heart. Dietary DHA markedly increased tissue DHA proportions in both liver and heart, whereas the major effect of dietary AA was in the liver. Future studies of the effects of dietary DHA and AA supplementation should examine a variety of tissues rather than focusing only on neural tissue.     

Comparison of growth and fatty acid metabolism in rats fed diets containing equal levels of γ-linolenic acid from high γ-linolenic acid canola oil or borage oil

We have utilized transgenic technology to develop a new source of γ-linolenic acid (GLA) using the canola plant as a host. The aim of the present study was to compare the growth and fatty acid metabolism in rats fed equal amounts of GLA obtained from the transgenic canola plant relative to GLA from the borage plant. Young male Sprague-Dawley rats (n=10/group) were randomized and fed a purified AIN93G diet (10% lipid by weight) containing either a mixture of high GLA canola oil (HGCO) and corn oil or a control diet containing borage oil (BO) for 6 wk. GLA accounted for 23% of the triglyceride fatty acids in both diets. Growth and diet consumption were monitored every 2–3 d throughout the study. At study termination, the fatty acid composition of the liver and plasma phospholipids was analyzed by gas chromatography. The growth and diet consumption of the HGCO group were similar to the BO group. There were no adverse effects of either diet on the general health or appearance of the rats, or on the morphology of the major organs. There was no significant difference between the diet groups for total percentage of n−6 polyunsaturated fatty acids present in either the total or individual phospholipid fractions of liver or plasma. The relative percentage of GLA and its main metabolite, arachidonic acid, in each phospholipid fraction of liver or plasma were also similar between groups. The percentage of 18∶2n−6 in liver phosphatidylethanolamine and phosphatidylinositol/serine was higher (P<0.05) and 22∶5n−6 was lower in the HGCO group than the BO group. This finding could be attributed to the higher 18∶3n−3 content in the HGCO diet than the BO diet. Results from this long-term feeding study of rats show for the first time that a diet containing transgenically modified canola oil was well-tolerated, and had similar biological effects, i.e., growth characteristics and hepatic metabolism of n−6 fatty acids, as a diet containing borage oil.     

Increased α-linolenic acid intake increases tissue α-linolenic acid content and apparent oxidation with little effect on tissue docosahexaenoic acid in the guinea pig

The essential fatty acids do not have identical roles in nutrition. Linoleic acid (LA) accumulates throughout the body of most mammals, whereas α-linolenic acid (ALA) is rarely found in tissue lipids to the same extent as LA. It has been argued that this is the result of metabolism of ALA to docosahexaenoic acid (DHA) or that ALA is rapidly β-oxidized to acetyl CoA and CO₂. In this study, we consider the effect of high and low ALA levels on the tissue distribution of ALA and other n-3 polyunsaturated fatty acids (PUFA) in all tissues. Guinea pigs were fed one of two defined diets for 3 wk from wearning with both diets containing 1.8% (by weight) of LA and either 1.7% ALA or 0.03% ALA. The high ALA diet was associated with significantly increased ALA levels in all tissues except the brain and significantly increased levels of long-chain n-3 PUFA in all tissues except intestines, brain, carcass, and skin. The long-chain n-3 PUFA content of the whole body was less than 5% of that of the ALA content in both diet groups, and the major long-chain n-3 PUFA (>66% of total) in the body was 22∶5n−3. The brain was the only tissue where the DHA content exceeded that of 22∶5n−3. On the low ALA diet, there appeared to be conservation of ALA based on a comparison of the ratio of LA to ALA in the tissues compared with that in the diet. On the high ALA diet there was a loss of ALA relative to LA in the tissues compared with the diet. These studies suggest that the low levels of tissue ALA in the guinea pig are likely the result of β-oxidation or excretion via the skin and fur rather than metabolism to DHA.     

Modification of milk formula to enhance accretion of long-chain n−6 and n−3 polyunsaturated fatty acids in artificially reared infant rats

Artificially reared infant rats were used to determine the effects of long-chain polyunsaturated fatty acid (LCP-UFA) supplementation on blood and tissue concentrations of arachidonic acid (AA) and docosahexaenoic acid (DHA). Beginning at 7 d of age, infant rats were fed for 10 d with rat milk formulas supplemented with AA at 0,0.5 and 1.0%, or supplemented with DHA at 0,0.5 and 1.0% of total fatty acid. The supplementation of AA increased accretion of the fatty acid in tissue and blood phospholipids with a maximum increase of 9% in brain, 15% in liver, 25% in erythrocytes, and 43% in plasma above the values of unsupplemented infant rats. Rat milk formula containing 1.0% of AA had no added benefits over that containing 0.5% of AA. The supplementation of DHA increased phospholipid DHA by a maximum of 24% in brain, 87% in liver, 54% in erythrocytes, and 360% in plasma above the unsupplemented control. The increase in tissue and blood DHA was concentration-dependent on formula fatty acid. Brain phosphatidylcholine and phosphatidylethanolamine were similarly enriched with AA and DHA by supplementation of the corresponding fatty acids. In general the observed increase of AA was accompanied by a decrease in 16:0, 18:1n−9, and/or 18:2n−6, whereas the increased DHA was associated with a reduction of 18:1n−9, 18:2n−6, and/or 20:4n−6. Clearly, infant rats were more responsive to DHA than AA supplementation, suggesting a great potential of dietary manipulation to alter tissue DHA concentrations. However, the supplementation of DHA significantly decreased tissue and blood AA/DHA ratios (wt%/wt%), whereas there was little or no change in the ratio by AA supplementation. Although the physiological implications of the levels of AA and DHA, and AA/DHA ratios achieved under the present experimental conditions are not readily known, the findings suggest that artificial rearing could provide a suitable model to investigate LCPUFA requirements using various sources of AA and DHA in rats.     

Effects of high-γ-linolenic acid canola oil compared with borage oil on reproduction,growth, and brain and behavioral development in mice

Previous research in rats and mice has suggested that γ-linolenic acid (GLA) derived from borage oil (BO: 23% GLA) may be an appropriate source for increasing levels of long-chain n−6 FA in the developing brain. Recently, transgenic technology has made available a highly enriched GLA seed oil from the canola plant (HGCO: 36% GLA). The first objective of this study was to compare the effects of diets containing equal levels of GLA (23%) from either BO or HGCO on reproduction, pup development, and pup brain FA composition in mice. The second objective was to compare the effects of the HGCO diluted to 23% GLA (GLA-23) with those of undiluted HGCO containing 36% GLA (GLA-36). The diets were fed to the dams prior to conception and throughout pregnancy and lactation, as well as to the pups after weaning. The behavioral development of the pups was measured 12 d after birth, and anxiety in the adult male offspring was assessed using the plus maze. The findings show that despite equivalent levels of GLA, GLA-23 differed from BO in that it reduced pup body weight and was associated with a slight increase in neonatal pup attrition. However, there were no significant effects on pup behavioral development or on performance in the plus maze. An increase in dietary GLA resulted in an increase in brain 20∶4n−6 and 22∶4n−6, with a corresponding decrease in 22∶6n−3. Again, despite their similar levels of GLA, these effects tended to be larger in GLA-23 than in BO. In comparison with GLA-23, GLA-36 had larger effects on growth and brain FA composition but no differences with respect to effects on reproduction and behavioral development. These findings suggest that the HGCO can be used as an alternative source of GLA.     

Comparative effects of α- and γ-linolenic acids on rat liver fatty acid oxidation

It has been reported that both n−3 and n−6 octadecatrienoic acids can increase hepatic fatty acid oxidation activity. It remains unclear, however, whether different enzymes in fatty acid oxidation show a similar response to n−3 and n−6 octadecatrienoic acids. The activity of hepatic fatty acid oxidation enzymes in rats fed an oil mixture rich in α-linolenic acid (18:3n−3) and borage oil rich in γ-linolenic acid (18:3n−6) was therefore compared to that in rats fed an oil mixture rich in linoleic acid (18:2n−6) and a saturated fat (palm oil) in this study. Linseed oil served as the source of 18:3n−3 for the oil mixture rich in this octadecatrienoic acid and contained 30.6% 18:3n−3 but not 18:3n−6. Borage oil contained 25.7% 18:3n−6 and 4.5% 18:3n−3. Groups of seven rats each were fed diets containing 15% various fats for 15 d. The oxidation rate of palmitoyl-CoA in the peroxisomes was higher in rats fed a fat mixture rich in 18:3n−3 (3.03 nmol/min/mg protein) and borage oil (2.89 nmol/min/mg protein) than in rats fed palm oil (2.08 nmol/min/mg protein) and a fat mixture rich in 18:2n−6 (2.15 nmol/min/mg protein). The mitochondrial palmitoyl-CoA oxidation rate was highest in rats fed a fat mixture rich in 18:3n−3 (1.93 nmol/min/mg protein), but no significant differences in this parameter were seen among the other groups (1.25–1.46 nmol/min/mg protein). Compared to palm oil and fat mixtures rich in 18:2n−6, a fat mixture rich in 18:3n−3 and borage oil significantly increased the hepatic activity of carnitine palmitoyl-transferase and acyl-CoA oxidase. Compared to palm oil and a fat mixture rich in 18:2n−6, a fat mixture rich in 18:3n−3, but not fats rich in 18:3n−6, significantly decreased 3-hydroxyacyl-CoA dehydrogenase activity. Compared to palm oil and a fat mixture rich in 18:2n−6, borage oil profoundly decreased mitochondrial acyl-CoA dehydrogenase activity, but a fat mixture rich in 18:3n−3 increased it. 2,4-Dienoyl-CoA reductase activity was significantly lower in rats fed palm oil than in other groups. Compared to other fats, borage oil significantly increased Δ³, Δ²-enoyl-CoA isomerase activity. Activity was also significantly higher in rats fed 18:2n−6 oil than in those fed palm oil. It was confirmed that both dietary 18:3n−6 and 18:3n−3 increased fatty acid oxidation activity in the liver. These two dietary octadecatrienoic acids differ considerably, however, in how they affect individual fatty acid oxidation enzymes.     

Effects of storage time and added antioxidant on fatty acid composition of red blood cells at −20°C

The stability of PUFA in venous red blood cells (RBC) of women aged 25 to 55 years (n=12) was investigated during storage at −20°C. The RBC sample from each participant was divided into seven portions: one baseline with the antioxidant BHT, another without BHT, samples without BHT stored for 2, 4, 9, or 17 wk, and samples with BHT stored for 17 wk. No difference was found in proportions of PUFA at baseline and after storage for 2 and 4 wk without BHT, and 17 wk with BHT. After 9 wk without BHT the proportion of 22∶6n−3 in RBC was lower, and after 17 wk without BHT proportions of all PUFA were lower than at baseline. High proportion of 22∶6n−3 in RBC at baseline was associated with more stable concentration of total FA in RBC without BHT during 17 wk. The findings indicate that PUFA in RBC from healthy women are stable at −20°C for 4 wk, without BHT and for at least 17 wk with BHT.     

Dietary α-linolenic acid lowers postprandial lipid levels with increase of eicosapentaenoic and docosahexaenoic acid contents in rat hepatic membrane

This study was designed to examine the effects of dietary n−3 and n−6 polyunsaturated fatty acids (PUFA) on postprandial lipid levels and fatty acid composition of hepatic membranes. Male Sprague-Dawley rats were trained for a 3−h feeding protocol and fed one of five semipurified diets: one fat-free diet or one of four diets supplemented with 10% (by weight) each of corn oil, beef tallow, perilla oil, and fish oil. Two separate experiments were performed, 4-wk long-term and 4-d short-term feeding models, to compare the effects of feeding periods. Postprandial plasma lipid was affected by dietary fats. Triacylglycerol (TG) and total cholesterol levels were decreased in rats fed perilla oil and fish oil diets compared with corn oil and beef tallow diets. Hepatic TG and total cholesterol levels were also reduced by fish oil and perilla oil diets. Fatty acid composition of hepatic microsomal fraction reflected dietary fatty acids and their metabolic conversion. The major fatty acids of rats fed the beef tallow diet were palmitic, stearic, and oleic. Similarly, linoleic acid (LA) and arachidonic acid in the corn oil group, α-linolenic acid (ALA) and eicosapentaenoic acid (EPA) in the perilla oil group, and palmitic acid and docosahexaenoic acid (DHA) in the fish oil group were detected in high proportions. Both long- and short-term feeding experiments showed similar results. In addition, microsomal DHA content was negatively correlated with plasma lipid levels. Hepatic lipid levels were also negatively correlated with EPA and DHA contents. These results suggest that n−3 ALA has more of a hypolipidemic effect than n−6 LA and that the hypolipidemic effect of n−3 PUFA may be partly related to the increase of EPA and DHA in hepatic membrane.     

Lipid and γ-linolenic acid accumulation in strains of zygomycetes growing on glucose

Strains of Zygomycetes, belonging to the genera Zygorhynchus, Mortierella, Rhizopus, Mucor, and Cunninghamella, when cultivated on glucose produced significant quantities of γ-linolenic acid (GLA). After exhaustion of the nitrogen source from the culture medium, all strains accumulated cellular lipids in concentrations ranging from 10 to 28% (oil/dry mycelium). However, in some strains after the depletion of the carbon source (glucose) from the culture medium, a reconsumption of the accumulated oil and synthesis of fat-free cell material was observed. Accumulation of large amounts of oil in the mycelium resulted in the production of oil with low GLA content. Rhizopus stolonifer strain LGAM (9)1, and Cunninghamella sp. strain LGAM (9)2 produced more than 30 mg GLA/g of dry cellular mass. Cunninghamella sp. accumulated 28.1% oil/dry cellular mass, which contained 11.9% GLA. The production of GLA was 260 mg/L of culture medium.     

The effect of low α-linolenic acid diet on glycerophospholipid molecular species in guinea pig brain

The changes in guinea pig brain (cerebrum) glycerophospholipid molecular species resulting from a low α-linolenic acid (ALA) diet are described. Two groups of six guinea pigs were raised from birth to 16 wk of age on either an n-3 deficient diet containing 0.01 g ALA/100 g diet or n-3 sufficient diet containing 0.71 g AlA/100 g diet. Molecular species of diradyl glycerophosphoethanolamine. (GroPEtn), glycerophosphocholine, glycerophosphoserine, and glycerophosphoinositol were analyzed by high-performance liquid chromatography with on-line electrospray ionization mass spectrometry (HPLC/ESI/MS). Alkenylacyl GroPEtn species were determined by comparing spectra before and after mild acid treatment while diacyl- and alkylacyl species were distinguished by HPLC/ ESI/MS. The proportions of phospholipid classes and of the diradyl GroPEtn subclasses were not altered by diet changes. The main polyunsaturated molecular species of diradyl GroPEtn subclasses and of phosphatidylcholine and phosphatidylserine (PtdSer) contained 16∶0, 18∶0, or 18∶1 in combination with docosahexaenoic acid (DHA, 22∶6n-3), docosapentaenoic (DPA, 22∶5n-6), or arachidonic acid (ARA, 20∶4n-6). A significant proportion of DPA containing species were present in both diet groups, but in n-3 fatty acid deficiency, the proportion of DPA increased and DHA was primarily replaced by DPA. The combined value of main DHA and DPA containing species in the n-3 deficient group ranged from 91-111% when compared with the n-3 sufficient group, indicating a nearly quantitative replacement. The n-3 fatty acid deficiency did not lower the content of ARA containing molecular species of PtdSer of the guinea pig brain as reported previously for the rat brain. The molecular species of phosphatidylinositol were not altered by n-3 fatty acid deficiency. The present data show that the main consequence of a low ALA diet is the preferential replacement of DHA-containing molecular species by DPA-containing molecular species in alkenylacyl- and diacyl GroPEtn and PtdSer of guinea pig brain.     

Effects of dietary n−3 polyunsaturated fatty acids on T-Cell membrane composition and function

Dietary n−3 PUFA have been shown to attenuate T-cell-mediated inflammation, in part, by suppressing T-cell activation and proliferation. n−3 PUFA have also been shown to promote apoptosis, another important mechanism for the prevention of chronic inflammation by maintaining T-cell homeostasis through the contraction of populations of activated T cells. Recent studies have specifically examined Fas death receptor-mediated activation-induced cell death (AICD), since it is the form of apoptosis associated with peripheral T-cell deletion involved in immunological tolerance and T-cell homeostasis. Data from our laboratory indicate that n−3 PUFA promote AICD in T helper 1 polarized cells, which are the mediators of chronic inflammation. Since Fas and components of the deathinducing signaling complex are recruited to plasma membrane microdomains (rafts), the effect of dietary n−3 PUFA on raft composition and resident protein localization has been the focus of recent investigations. Indeed, there is now compelling evidence that dietary n−3 PUFA are capable of modifying the composition of T-cell membrane microdomains (rafts). Because the lipids found in membrane microdomains actively participate in signal transduction pathways, these results support the hypothesis that dietary n−3 PUFA influence signaling complexes and modulate T-cell cytokinetics in vivo by altering T-cell raft composition.     

Effect of fatty acid positional distribution and triacylglycerol composition on lipid by-products formation during heat treatment: III—Cyclic fatty acid monomers study

The influence of isomeric triacylglycerols (TG) containing 18:3n-3 and 18:2n-6 on the formation of cyclic fatty acid monomers (CFAM) after heat treatment was assessed. Diacid TG, containing linoleic acid (L) or linolenic acid (α-Ln) along with palmitic acid (P), and positioned either in the central position (PLP and PLnP, respectively), or in one of the two outer positions (PPL and PPLn, respectively) were synthesized. Monoacid TG of trilinolein and trilinolenin mixed with tripalmitin were also prepared. The CFAM formed after heating were analyzed after total hydrogenation. The results obtained with the model TG were compared to another model consisting of a canola oil and its randomized counterpart. In diacid TG, the location of α-Ln in the central position of the TG molecule (PLnP) greatly enhanced the formation of the CFAM upon heating at temperatures below 240°C. On the other hand, 18:3n-3 in monoacid TG (LnLnLn) was highly resistant to CFAM formation within the same range of temperatures (180–220°C). The TG structure, more than the TG composition, seemed to explain the differences in the CFAM formation between the original canola oil and its interesterified counterpart. Like α-Ln, 18:2n-6 was more prone to cyclization when attached at the central position of the model TG. Conversely, the influence of the TG composition on the cyclization rate was less important for L than for α-Ln. It was concluded that positioning the C18 polyunsaturated fatty acid in the central position of TG rendered the oils much more sensitive to the cyclization reaction upon heat treatment.     

Effects of aging and dietary n−3 fatty acids on rat brain phospholipids: Focus on plasmalogens

The aging brain undergoes modifications in the lipid composition of cell membranes and especially in plasmalogens. These phospholipids represent between one-half and twothirds of the ethanolamine phospholipids in the brain. They are known to facilitate membrane fusion and act as endogenous antioxidants. During normal aging and in some pathological conditions, plasmalogen and DHA levels fall. In this context, we aimed to evaluate the influence of n−3 FA intake on plasmalogens in the brain during aging. Littermates from two generations of n−3-deficient rats were fed an n−3-deficient diet or an equilibrated diet containing either α-linolenic acid alone (α-LNA) or with two doses of DHA (0.3 or 0.6% w/w). After weaning, 9 mon of diet, or 21 mon of diet, plasmalogen levels were assessed, and the sn-2 substitutions of plasmenylethanolamines were analyzed in the cortex, striatum, and hippocampus. Our results showed that plasmalogen contents were not influenced by the diet. Plasmalogen levels were significantly decreased in aged rats compared with adults, whereas DHA levels increased in the hippocampus and remained stable in the cortex and striatum. DHA levels were significantly and similarly increased in total phospholipids and especially in plasmenylethanolamines after 9 mon of diet containing α-LNA alone or combined with DHA. This study showed that each structure sustained specific age-induced modifications. Dietary n−3 FA may not oppose the physiological decrease in brain plasmalogen levels during aging. Moreover, α-LNA appears to be equally as potent as preformed DHA at replacing DHA in the brain of our rat model.     

Effects of hydrocarbon structure on fatty acid,fatty alcohol,and β-hydroxy acid composition in the hydrocarbon-degrading bacterium <Emphasis Type="Italic">Marinobacter hydrocarbonoclasticus</Emphasis>

The lipids of the gram-negative bacterium Marinobacter hydrocarbonoclasticus grown in a synthetic seawater medium supplemented with various hydrocarbons as the sole carbon source were isolated, purified, and their structures determined. The hydrocarbons were normal, iso, anteiso, and mid-chain branched alkanes, phenylalkanes, cyclohexylalkanes, and a terminal olefin. According to the sequential procedure used for lipid extraction, three pools were isolated: unbound lipids extracted with organic solvents (corresponding to metabolic lipids and to the main part of membrane lipids), OH⁻ labile lipids [mainly ester-bound in the lipopolysaccharides (LPS)], and H⁺ labile lipids (mainly amide-bound in the LPS). Each pool contained FA, fatty alcohols, and β-hydroxy acids. The proportions of these lipids in the unbound lipid pools were 84–98%, 1.1–11.6%, and 0.1–3.6% (w/w), respectively. The chemical structures of the lipids were strongly correlated with those of the hydrocarbons fed; analytical data suggested a metabolism essentially through oxidation into primary alcohol, then into FA and degradation via the β-oxidation pathway. Subterminal oxidation of the hydrocarbon chains, α-oxidation of FA or double-bond oxidation in the case of the terminal olefin, were minor, although sometimes substantial, routes of hydrocarbon degradation. Cyclohexyldodecanedid not support growth, likely because of the toxicity of cyclohexylacetic acid formed in the oxidation of the alkyl side chain. In the OH⁻ and H⁺ labile lipid pools, β-hydroxy acids, the lipophilic moiety of LPS, generally dominated (28–72% and 64–98%, w/w, respectively). The most remarkable feature of these cultures on hydrocarbons was the incorporation in LPS of β-hydroxy acids with C_odd, ω-unsaturated, iso, or anteiso alkyl chains in addition to the specific β-hydroxy acid of M. hydrocarbonoclasticus, 3-OH-n-12∶0. These β-hydroxy acids were tolerated insofar as their geometry and steric hindrance were close to those of the 3-OH-n-12∶0 acid.     

Effects of margarine and butter consumption on distribution of trans-18∶1 fatty acid isomers and conjugated linoleic acid in major serum lipid classes in lactating women

Trans FA (TFA) have at least one trans double bond and comprise several isomers and types, including many of the CLA (e.g., c9, t11–18∶2 CLA). Some TFA may have adverse effects (e.g., cardiovascular disease), whereas some are though to have beneficial effects (e.g., anticarcinogenicity). The presence of TFA in human tissues and fluids is related to dietary intake, although this relationship is not completely understood—especially in regard to serum lipid fractions. This study was conducted as part of an investigation designed to test the influence of butter (B), “low TFA” margarine (LT), and regular margarine (RM) on milk fat content. Here we tested the secondary hypothesis that consumption of B, LT, and RM by lactating women would result in differential distribution of TFA and CLA in major serum lipid classes. Breastfeeding women (n=11) participated in this randomized Latinsquare study consisting of five periods: intervention I (5 d), washout I (7 d), intervention II (5 d), washout II (7 d), and intervention III (5 d). Extracted serum lipid was separated into cholesterol ester (CE), TAG, and phospholipid (PL) fractions and analyzed for total and isomeric TFA and CLA concentrations. Data indicate that TAG consistently contained the highest concentration of total t-18∶1. No interaction between treatment and fraction was found for any of the t-18∶1 isomers identified. Absolute concentration of each t-18∶1 isomer was greatest during the RM period, regardless of fraction. On a relative basis, concentrations of t10–18∶1 and t12–18∶1 were most responsive to treatment in the CE fraction. The concentration of c9, t11–18∶2 CLA was highest in the TAG fraction and lowest in the PL fraction, regardless of treatment. In summary, these results indicate (i) that there is a differential distribution of some isomeric TFA and CLA among human serum lipid fractions and (ii) that dietary TFA intake influences absolute and relative concentrations of some of the isomers in selected fractions.     

Effect of nitrogen sources on γ-linolenic acid accumulation in Spirulina platensis

The effect of ammonium phosphate on the growth, lipid content, and γ-linolenic acid accumulation was determined in the cyanobacterium Spirulina platensis. After 14 d on media containing 0.041 g N/L, γ-linolenic acid concentration of cultured cells increased up to 35.3±0.13%, w/w. In treatments containing NaNO₃, NH₄NO₃, and NH₄Cl, γ-linolenic acid concentration increased up to 31.2±0.23%, w/w. After the same period, lipid content in the dry biomass was 12.2±0.03%, w/w, with (NH₄)₂HPO₄ compared to about 14.1±0.12%, w/w, in treatments with NaNO₃. When (NH₄)₂HPO₄ concentration in the medium was increased to 0.082 g N/L, 30.8±0.28%, w/w, γ-linolenic acid had formed after 10 d and the lipid percentage in the dry cell mass was 16.7±0.16%, w/w. However, in treatments with NaNO₃, NH₄NO₃, or NH₄Cl, γ-linolenic acid concentration increased up to 30.6±0.23%, w/w, and the lipid content was found to be 18.0±0.17 to 18.9±0.03%, w/w. These data showed that (NH₄)₂HPO₄ is a suitable source of nitrogen for growth of S. platensis, with increased accumulation of γ-linolenic acid at lower N concentration.