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Background: Fatty acids (FAs) in human milk are important nutrients for infants. They play important roles in energy supply, nervous system development, and metabolic function maintenance. However, how the composition of major milk FAs change with lactation stages remains controversial., Objectives: To systematically review the concentration range of major FAs in human milk at various lactation stages., Methods: A total of 12 papers involving 50 sets of data with 3507 participants were reviewed according to the PRISMA checklist and flow diagram. The inclusion criteria was the literatures had the FAs contents in breast milk of healthy lactation mothers at three lactation stages and the dietary patterns could be calculated. The exclusion criteria were: the studies were duplicates, were unrelated to dietary patterns or breast milk composition, and/or the study populations were unhealthy. We searched PubMed, the China National Knowledge Infrastructure, WanFang, and Web of science. Agency for Health Care Research and Quality (AHRQ) was used to assess the bias of studies. The mean values of polyunsaturated fatty acids (PUFAs) including docosahexaenoic acid (DHA), arachidonic acid (AA), eicosapentaenoic acid (EPA), α-linolenic acid (ALA), linoleic acid (LA), monounsaturated fatty acids (MUFAs), and saturated fatty acids (SFAs, including lauric acid and palmitic acid), in human milk at three lactation stages (colostrum 1-7 d, transitional milk 8-14 d, mature milk 15 d-3 mo) of healthy lactating women were investigated in terms of the high protein dietary pattern. Publication biases were evaluated by Egger's test., Results: According to the percentage in total fat of colostrum, transitional milk, and mature milk (% wt/wt), respectively, the results showed that PUFA (25.72%, 24.92%, and 22.69%), AA (0.85%, 0.76%, and 0.59%), DHA (0.53%, 0.47%, and 0.39%), EPA (0.15%, 0.10%, and 0.10%), and MUFA (37.39%, 37.21%, and 36.14%) contents in breast milk decreased with lactation, while another two PUFA forms, LA (17.47%, 17.82%, and 17.48%), and ALA (1.09%, 1.39%, and 1.24%) arrived at a peak in the transitional milk and then decreased in the mature milk, SFA (37.46%, 38.64%, and 40.52%), and lauric acid contents (2.78%, 4.91%, and 4.97%) increased with the lactation stages., Conclusion: These findings could shed light on the dynamic change progress of major FA metabolism, potentially enhancing the knowledge of lactation biology, and improving infant feeding practices to meet their needs., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Oxylipins are a family of saturated and unsaturated fatty acids peroxidation products with bioactive properties. We have developed an improved method for the measurement of ex vivo oxylipin production by peripheral blood mononuclear cells (PBMCs) and neutrophils. We aimed to develop an analytical method to determine the production rates of polyunsaturated fatty acids (PUFAs), PUFA-oxylipin, and saturated-oxylipins by stimulated PBMCs and neutrophils based on solid phase extraction and HPLC-MS/MS technology. A UHPLC system coupled to a Q-Exactive Hybrid Quadrupole-Orbitrap mass spectrometer was used to identify and quantify oxylipin production. For each oxylipin and PUFA their differential response was calculated with respect to a deuterated internal standard factor (ISF). To calculate oxylipin and PUFAs in the culture samples, the individual ISF was used for each oxylipin and PUFA with respect to the deuterated internal standard. PBMCs and neutrophils showed a different pattern of oxylipin production and fatty acid secretion. Lipopolysaccharide (LPS) did not stimulate oxylipin production or fatty acids secretion in PBMCs, whereas phorbol myristate acetate (PMA) stimulation increased the production rate of 5-HETE, 15-HETE, 15-HEPE, 17-DoHE, PGE2, AA, and DHA. LPS stimulation decreased 16-hydroxyl-palmitatte (16-OHPAL) production and DHA secretion in neutrophils, while PMA stimulation increased the production rate of AA and its derivate oxylipins, 5-HETE, 15-HETE, and PGE2. In conclusion, we have developed a new method to determine oxylipins derived from both saturated and unsaturated fatty acids in culture cell media. This method has enough sensitivity, and accuracy, to determine oxylipin production and fatty acid secretion by PBMCs and neutrophils., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Oxygenated polyunsaturated fatty acids (PUFAs)play an outstanding role in the physiological and pathological regulation of several biological processes. These oxygenated metabolites can be produced both enzimatically, yielding almost pure enantiomers, and non-enzymatically. The free radical-mediated non-enzymatic oxidation commonly produces racemic mixtures which are used as biomarkers of oxidative stress and tissue damage. The biological activity of oxygenated PUFAs is often associated with only one enantiomer, making it necessary of availing of lipidomics platforms allowing to disclose the role of single enantiomers in health and disease. Polysaccharide-based chiral stationary phases (CSPs) play a dominating part in this setting. As for the cellulose backbone, 4-methylbenzoate derivatives exhibit very high chiral recognition ability towards this class of compounds. Concerning the phenylcarbamate derivatives of cellulose and amylose, the tris(3,5-dimethylphenylcarbamate) variants show the best enantioresolving ability for a variety of oxygenated PUFAs. Moreover, also the amylose tris(5-chloro-2-methylphenylcarbamate)-based selector produces relevant chromatographic performances. The extreme versatility of those CSPs mostly depends on their compatibility with the most relevant elution modes: normal- and reversed-phase, as well as polar organic/ionic-mode. In this review article, a selection of enantioseparation studies of different oxygenated PUFAs is reported, with both tris(benzoates) and tris(phenylcarbamates) of cellulose and amylose., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Polyunsaturated fatty acids (PUFAs) in the cellular membrane can be oxidized by various enzymes or reactive oxygen species (ROS) to form many oxidized lipids. These metabolites are highly bioactive, participating in a variety of physiological and pathophysiological processes. Mass spectrometry (MS), coupled with Liquid Chromatography, has been increasingly recognized as an indispensable tool for the analysis of oxidized lipids due to its excellent sensitivity and selectivity. We will give an update on the understanding of the molecular mechanisms related to generation of various oxidized lipids and recent progress on the development of LC-MS in the detection of these bioactive lipids derived from fatty acids, cholesterol esters, and phospholipids. The purpose of this review is to provide an overview of the formation mechanisms and technological advances in LC-MS for the study of oxidized lipids in human diseases, and to shed new light on the potential of using oxidized lipids as biomarkers and mechanistic clues of pathogenesis related to lipid metabolism. The key technical problems associated with analysis of oxidized lipids and challenges in the field will also discussed., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Glutathione (GSH) conjugates of oxygenated polyunsaturated fatty acids comprise a group of pro-inflammatory and pro-resolving lipid mediators formed in immunocompetent cells. While the pro-inflammatory conjugates such as the cysteinyl leukotrienes (cys-LTs), eoxins (EXs) and five-oxo-GSH conjugate (FOG 7 ) derive from arachidonic acid (AA), the group of conjugates in tissue regeneration (CTRs) such as maresin CTRs (MCTRs), protectin CTRs (PCTRs) and resolvin CTRs (RCTRs) are biosynthesized from docosahexaenoic acid (DHA). Here, we present a gradient UPLC-MS/MS method for the analysis of pro-inflammatory and pro-resolving GSH conjugates using positive electrospray ionization (ESI(+)) and collision-induced fragmentation for unambiguous identification and structural information, and a negative ionization (ESI(-)) mode for quantification of the GSH conjugates. The method was employed to detect GSH conjugates in human platelets and macrophages. MCTRs were detected in platelets upon addition of exogenous docosahexaenoic acid (DHA) and the biosynthesis was independent on leukotriene C 4 (LTC 4 ) synthase activity. Pathogenic bacteria stimulated the formation of EXs and PCTRs in M2 macrophages, whereas Ca 2+ -ionophore activated the biosynthesis of LTC 4 in M1 and M2 macrophage phenotypes. Together, our methodology covers the qualitative and quantitative analysis of GSH conjugates and gives an analytical basis for the detection and structural elucidation of cysteinyl-containing lipid mediators., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Emu (Dromaius novaehallandiae), the flightless bird native to Australia and found in many countries, is receiving much attention for its nutritional benefits as well as its medicinal value. Emu oil contains high amounts of polyunsaturated fatty acids and antioxidants. It has potent anti-inflammatory actions and thus can be used topically and orally to treat conditions such as mucositis, inflammatory bowel syndrome, and auricular inflammation, and to prevent chemotherapy-induced bone loss. Emu oil also has a hypocholesterolemic effect, transdermal penetration-enhancing activity, cosmetic and insect repellent activity, and so on. However, its mechanism(s) of actions are unclear and have not, to our knowledge, been studied to date. Previous studies suggest that the fatty acids of the ω-9, ω-6, and ω-3 series, which are present in emu oil, may act on cyclooxygenase, lipoxygenase, and lipoxin pathways to bring about its anti-inflammatory and other beneficial actions. The aim of this review was to provide a brief summary of the current knowledge of research on emu products, mainly emu oil, for the possible use as a complementary and alternative natural medicine for various chronic diseases. In this review we also highlighted the future research scope of emu oil for its possible antidiabetic activity. Thus, emu oil is an attractive pharmacologic agent to further explore for its therapeutic activity to treat various ailments., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Male infertility is a relatively common condition affecting 1 in 20 men of reproductive age. The etiology of this condition is thought to involve the excessive generation of reactive oxygen species by human spermatozoa; however, the cause of this aberrant activity is unknown. In this study we demonstrate that defective human sperm populations are characterized by high cellular contents of both esterified and unesterified fatty acids and a decrease in the proportion of the total fatty acid pool made up by docosahexaenoic acid. The free unsaturated fatty acid content of these cells was positively correlated with the induction of mitochondrial superoxide generation (P<0.001). This relationship was causal and mediated by the range of unesterified, unsaturated fatty acids that are present in human spermatozoa. Thus direct exposure of these cells to free unsaturated fatty acids stimulated mitochondrial superoxide generation and precipitated a loss of motility and an increase in oxidative DNA damage, two key attributes of male infertility. We conclude that defective human spermatozoa are characterized by an abnormally high content of fatty acids that, in their unesterified, unsaturated form, promote ROS generation by sperm mitochondria, creating a state of oxidative stress and a concomitant loss of functional competence., (Copyright 2009 Elsevier Inc. All rights reserved.)

Osteoporosis is associated with an increase in marrow fat. Fats, particularly polyunsaturated fats, either in co-cultures or diet, have been shown to significantly influence bone remodeling. Whether the increase in marrow fat seen in osteoporosis is also associated with a change in fatty acid composition is not known. This study was undertaken to investigate the fatty acid composition in subjects of varying bone mineral density (BMD). Samples of marrow fat and subcutaneous fat from 126 subjects (98 females, 34 males, mean age 69.7+/-10.5 years) undergoing orthopedic surgery were analyzed for fatty acid composition by gas chromatography. These results were correlated with BMD assessed by DXA. A total of 22 fatty acids were identified in marrow and subcutaneous fat. Significant differences in fatty acid composition existed between marrow and subcutaneous fat as well as between marrow fat samples obtained from the proximal femur and proximal tibia. Other than cis-7-hexadecenoic acid [C16:1 (n=9)] and docosanoic acid [C22:0], no difference in marrow fatty acid composition was evident between subject groups of varying BMD (normal, low bone mass, and osteoporosis). In conclusion, there exists a wide range of individual fatty acids in marrow fat. Marrow fatty acid composition differs from that of subcutaneous fat and varies between predominantly erythropoetic and fatty marrow sites. Other than cis-7-hexadecenoic acid [C16:1 (n=9)] and docosanoic acid [C22:0], no difference in marrow fatty acid composition was evident between subjects of varying BMD.

Fish consumption during gestation can provide the fetus with long-chain polyunsaturated fatty acids (LCPUFA) and other nutrients essential for growth and development of the brain. However, fish consumption also exposes the fetus to the neurotoxicant, methyl mercury (MeHg). We studied the association between these fetal exposures and early child development in the Seychelles Child Development Nutrition Study (SCDNS). Specifically, we examined a priori models of Omega-3 and Omega-6 LCPUFA measures in maternal serum to test the hypothesis that these LCPUFA families before or after adjusting for prenatal MeHg exposure would reveal associations with child development assessed by the BSID-II at ages 9 and 30 months. There were 229 children with complete outcome and covariate data available for analysis. At 9 months, the PDI was positively associated with total Omega-3 LCPUFA and negatively associated with the ratio of Omega-6/Omega-3 LCPUFA. These associations were stronger in models adjusted for prenatal MeHg exposure. Secondary models suggested that the MeHg effect at 9 months varied by the ratio of Omega-6/Omega-3 LCPUFA. There were no significant associations between LCPUFA measures and the PDI at 30 months. There were significant adverse associations, however, between prenatal MeHg and the 30-month PDI when the LCPUFA measures were included in the regression analysis. The BSID-II mental developmental index (MDI) was not associated with any exposure variable. These data support the potential importance to child development of prenatal availability of Omega-3 LCPUFA present in fish and of LCPUFA in the overall diet. Furthermore, they indicate that the beneficial effects of LCPUFA can obscure the determination of adverse effects of prenatal MeHg exposure in longitudinal observational studies.

We have assessed the activity of Na(+)/K(+)-ATPase, cAMP, free fatty acids (FFA) and metallothionein (MT) in the posterior gills of the brackish water shore crab Carcinus aestuarii during acclimation to 10 ppt dilute seawater (DSW). Following 3-18 days acclimation in DSW specific activity of Na(+)/K(+)-ATPase in native gill homogenates and partially purified membrane vesicles was progressively increased, from 1.7- to 3.9-fold. After short-term acclimation of crabs in DSW with added sucrose to make media isosmotic with the haemolymph the specific Na(+)/K(+)-ATPase activity in homogenates was not increased, relative to SW enzyme activity. Moreover, hyposmotic conditions led to depletion of cAMP in gills. In partially purified membrane vesicles isolated from posterior gills, fatty acids with compositions 16:0, 18:0, 18:1, 20:4 and 20:5 dominated in both SW- and DSW-acclimated Carcinus. During a year in which the metabolic activity of crabs was increased, the arachidonic/linoleic acids ratio (ARA/LA) for DSW-acclimated crabs was markedly increased relative to that in SW. Increased Na(+)+K(+)-ATPase activity under hyposmotic stress may be modulated at least partially by the changed proportion of fatty acids in the purified membranes of posterior gills. Long-term acclimation of shore crabs to DSW resulted in a 2.6-fold increase in cytosolic metallothionein (MT) content in posterior gills over those in SW crabs. Assuming an antioxidant role of MT associated with intracellular zinc partitioning, the observed MT induction in posterior gills may be considered an adaptive response of C. aestuarii to hyposmotic stress.

Fatty acid (FA) compositions were determined for the invader amphipod Dikerogammarus villosus collected from July to September 2002, in an overheated, high-conductivity dammed reservoir in north-eastern France. Predominant fatty acids were the polyunsaturated fatty acids (PUFA): eicosapentaenoic acid (EPA), linoleic acid (LA), arachidonic acid (ARA), linolenic acid (LNA) together with the monounsaturated fatty acid 18:1omega9 and the saturated fatty acid 16:0. FA markers indicated that available food was constituted of incompletely degraded phytodetritus and terrestrial inputs, as well as animal remains. PUFA contents depended on the diet and the capacity of animals to desaturate and elongate LNA and LA in long chain PUFA as EPA and ARA respectively. Based on their FA compositions, we showed that gammarids represent naturally-occurring freshwater sources of essential PUFA, and could play a fundamental role in pelagic-benthic coupling and energy recycling in the ecosystem. The complexity of the feeding strategies of D. villosus--detritivorous, omnivorous, carnivorous--makes this species efficient at exploiting different components of the available food and may be a key factor in its high invasive success.

Fatty acid (FA) composition in the intraabdominal (IAB), subcutaneous (SC) and peripheral adipose tissues of the semiaquatic American mink (Mustela vison) was examined in comparison to the diet by gas-liquid chromatography. There was a clear compositional gradient from the IAB via SC to peripheral adipose tissues and the anatomically different adipose tissues accumulated or metabolized FA selectively. The total lipids of the body appendages had smaller proportions of saturated (SFA) and larger proportions of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) than the lipids of the trunk adipose tissues. Especially n-3 PUFA were enriched in the periphery. The appendages were also characterized with a high ratio of unsaturated FA to SFA, an increased Delta9-desaturation index and increased mean numbers of double bonds and carbon atoms in a FA molecule. The proportions of SFA and MUFA of the diet resembled the trunk adipose tissues while the dietary percentage of n-3 PUFA surpassed those of the trunk fat depots but was lower than those of the peripheral fats. These data confirm that the FA signatures of mammals reflect not only their dietary history but also metabolic modifications of ingested FA.

Reduced plasma retinol concentrations occur in human malaria but the benefits of supplementation remain uncertain. We assessed the in vivo efficacy of retinol administration, and its effect on lipid peroxidation, in a Plasmodium berghei murine model. Animals received vehicle (n=17) or retinol (i) before P. berghei inoculation (four doses), (ii) at parasitaemia 10-15% (three to four doses) or (iii) before and after inoculation (six to seven doses; n=15 in each group), with euthanasia on day 8 post-inoculation or when the parasitaemia exceeded 50%. Multiple-dose pre-inoculation retinol reduced endpoint parasitaemia by 24% (P=0.001 versus controls). A reduction of 18% (P=0.042) was observed when retinol was given to parasitaemic animals. Retinol was ineffective when given both before and after infection (11% reduction; P=0.47). Although retinol supplementation did not change plasma retinol concentrations, liver retinol content increased and correlated inversely with endpoint parasitaemia (r=-0.45, P=0.001). Malaria infection augmented concentrations of the free radical lipid peroxidation end-product F(2)-isoprostanes in plasma, erythrocytes and liver by 1.8-, 2.8- and 4.9-fold, respectively, but retinol supplementation had no effect on these increases. Consistent with some human malaria studies, prophylactic retinol reduces P. berghei parasitaemia. This effect relates to augmentation of tissue retinol stores rather than to retinol-associated changes in oxidant status.

The aim of this study was to examine the fatty acid composition and non-enzymatic lipid peroxidation (LP) of mitochondria and microsomes obtained from liver, heart and brain of Lonchura striata. The percentage of total unsaturated fatty acid was approximately 30-60% in the organelles from all tissues studied. Brain mitochondria and both organelles of liver exhibited the highest percentage of polyunsaturated fatty acid (PUFA) (30 and 18%, respectively). The arachidonic acid (AA) content was 7% in mitochondria of liver and brain and 3% in heart mitochondria. The percentage of docosahexanoic acid (DHA) was 8% in brain mitochondria and approximately 2-3% in heart and liver mitochondria. The peroxidizability index (PI) of brain mitochondria and both organelles from liver was higher than that of organelles from heart and brain microsomes. Liver organelles and brain mitochondria were affected by LP, as indicated by the increase in chemiluminescence and a decrease of AA and DHA. These changes were not observed during LP of brain microsomes and both organelles from heart. These results indicate: 1) PI positively correlates with PUFA percentage and LP; 2) The resistance to LP detected in heart organelles would contribute to the cardiac protection against oxidative damage.

Objectives: Insulin resistance in diabetes mellitus type 2 (DM2) can result from membrane lipid alterations. Blacks are at a higher risk of developing DM2; therefore, we investigated whether membrane lipid differences exist between blacks and whites and if differences contribute to impaired insulin binding in diabetes., Methods: Subjects were recruited from four groups: white control (n = 10), black control (n = 10), white diabetic (n = 5), and black diabetic (n = 10). Diabetic subjects who had DM2 with insulin resistance on insulin monotherapy were matched by age and sex. The following determinations were made: fasting serum glucose, fasting serum insulin, plasma lipid profile, red blood cell (RBC) membrane lipids and cholesterol, and RBC insulin binding., Results: The membrane lipid analysis showed racial differences in phosphatidyl ethanolamine (PE) and phosphatidyl choline (PC). The plasma membrane of whites showed higher PE and lower PC levels than that in blacks. The RBC rheologic (PE/phosphatidyl serine) properties (deformability) were lower in diabetics and black subjects. The saturated nature of RBC ([sphingomyelin + PC)/(PE + phosphatidyl serine]) was the lowest in white control subjects (P < 0.056)., Conclusion: The combination of increased saturated/polyunsaturated fatty acids, increased saturated nature, and increased cholesterol/phospholipid can contribute to decreased membrane fluidity, resulting in insulin resistance. Also, decreased RBC deformability can make oxygen delivery through the capillaries difficult, create tissue hypoxia, and contribute to some of the known complications of diabetes. Membrane lipid alteration may be one of the reasons for a higher incidence of diabetes among blacks.

Clinical trials have revealed that progression of immunoglobulin A nephropathy (IgAN), the most common form of human glomerulonephritis, is inhibited by dietary (n-3) polyunsaturated fatty acid (PUFA) supplementation. The early stages of IgAN can be mimicked by feeding mice the mycotoxin deoxynivalenol (DON). Here, the effects of consuming the (n-3) PUFA eicosapentaenoic acid (EPA) on DON-induced IgAN were assessed relative to dose dependency and to expression of interleukin (IL-6). In the dose-response study, weight gain and feed intake did not differ among mice consuming 20 ppm DON supplemented with 0%, 0.1%, 0.5% and 3% EPA for 16 weeks. Mice fed the two highest EPA concentrations exhibited markedly increased splenic EPA, docosapentaenoic acid and docosahexaenoic acid, whereas arachidonic acid was decreased in all three EPA fed groups. Deoxynivalenol consumption significantly increased serum IgA and IgA immune complexes as well as kidney mesangial IgA deposition. All three IgAN markers were attenuated in mice fed 3% EPA diet but not in those fed 0.1% or 0.5% EPA. Elevated IgA production was observed in spleen and Peyer's patch (PP) cell cultures derived from mice fed DON in control diets, but this was reduced in cultures from mice fed 0.1%, 0.5% and 3% EPA. Acute DON exposure increased serum levels of IL-6, a cytokine that drives differentiation of IgA-committed B cells to IgA secretion. Relatedly, expression of IL-6 mRNA and IL-6 heteronuclear RNA, a marker of IL-6 transcription, was increased in spleen and PP. All three indicators of IL-6 expression were suppressed in mice consuming 3% EPA. Suppressed IL-6 corresponded to decreased binding activity of two factors that regulate transcription of this cytokine, cyclic AMP response element-binding protein and activator protein-1. The results indicate that a threshold existed for EPA relative to suppression of experimental IgAN and that the threshold dose was effective at inhibiting IL-6 transcription.

Insufficient availability of n-3 polyunsaturated fatty acids (PUFAs) during pre- and neonatal development decreases accretion of docosahexaenoic acid (DHA, 22:6n-3) in the developing brain and is associated with sub-optimal sensory and cognitive function in humans, altered behavior in animals, and may contribute to neurodevelopmental disorders such as attention deficit hyperactivity disorder and schizophrenia. This study examined the effects of variation in dietary availability of n-3 PUFAs on brain fatty acid composition and the consequent effects on locomotor activity in male and female Long-Evans rats. Rats were raised from conception using purified diets and breeding protocols designed to produce four groups with distinct brain phospholipid compositions varying in DHA content and/or the proportion of n-3 and n-6 PUFAs. Locomotor behavior was measured for a 2-h period on postnatal days 28, 42, 56, and 70. In males, decreased brain DHA produced alterations in activity that were most pronounced post-adolescence and with the greatest decrease in DHA. However, the behavioral effects in males were not linearly related to brain DHA level. In contrast, no significant effects of variation in brain fatty acid composition were observed in females. This suggests that variation in brain DHA content produces sex-specific alterations in locomotor activity and that the neurochemical alterations underlying the observed behavioral changes vary depending on the degree of DHA depletion.

The present research is focussed on the optimization of a fast and economically convenient method for the sample preparation and gas chromatographic separation of fatty acids contained in lipidic samples. This, as part of a wider project that has, as ultimate goal, the automation of this specific analytical procedure. The developed approach was applied to the analysis of cod liver oil, a highly complex lipid characterized by a wide nutritional interest. Derivatization was carried out by using low quantities (microL amounts) of a reagent and solvent, while the derived fatty acid methyl esters (FAMEs) were separated in 120 s on a 10 m x 0.1 mm i.d. polar micro-bore column. The total analysis time required for six samples was approximately 45 min (7.5 min/sample), considering a simultaneous process of methylation and GC separation of previously prepared samples. The results obtained were compared to those derived from conventional applications on the same sample. With regard to the validation of the rapid method, peak area/retention time repeatability, linear range, limit of detection (LOD) and quantitation (LOQ) were determined. Peak assignment was carried out by exploiting bidimensional group-type mapping information obtained in a comprehensive gas chromatographic application.

This study was conducted to determine whether dietary ganglioside (GG) increases the content of ether phospholipids (EPL) in intestinal mucosa. Weanling Sprague-Dawley rats were fed a semipurified diet consisting of 20% fat as a control diet. Two experimental diets were formulated by adding either 0.1% (w/w fat) GGs (GG diet) or 1.0% (w/w fat) sphingomyelin (SM diet) to the control diet. Fatty acid methyl esters from the alkenylacyl, alkylacyl and diacyl subclasses of phospholipids were measured to determine total and molecular percentage of EPL comprising the choline phosphoglyceride (CPG) and ethanolamine phosphoglyceride (EPG) fraction. Animals fed the GG diet significantly increased total EPL content both in CPG (by 36%) and in EPG (by 66%), and the molecular percentage of EPL in CPG (by 76%) and in EPG (by 59%) compared to animals fed the control diet. Dietary GG-induced increase in EPL resulted in a higher level of polyunsaturated fatty acids (PUFA) specifically in 20:4n-6 and 22:6n-3 compared to control animals, leading to a decrease in the ratio of saturated fatty acids (SFA) to PUFA both in CPG and in EPG. Feeding animals the SM diet showed a higher level of EPL than control animals with a concomitant increase in 22:6n-3 in EPL. The present data demonstrate that dietary GG increases the content and composition of EPL containing PUFA in the weanling rat intestine.

Objective: The relevance of oxidative stress in mice fed a choline-deficient diet (CDD) was investigated in relation to the oxidative stress marker, hydroxyoctadecadienoic acid (HODE) in comparison with F2-isoprostanes. Further, the protective effects of antioxidants against oxidative damage were assessed by using HODE., Methods: We recently proposed total HODE as a biomarker for oxidative stress in vivo. Biological samples such as plasma, urine, and tissues were first reduced and then saponified to convert various oxidation products of linoleates to HODE. In the present study, this method was applied to measure oxidative damage in mice induced by CDD for 1 mo., Results: CDD, when compared with choline-controlled diet (CCD), increased liver weight and fatty acid accumulation but the increase in body weight was less significant. Remarkable increases in HODE and 8-iso-prostaglandin F(2alpha) in liver and plasma were observed when mice were fed with the CDD for 1 mo compared with the CCD. The HODE level was about two to three orders higher than the F2-isoprostane level. This increase was decreased to the level of the CCD when alpha-tocopherol or 2,3-dihydro-5-hydroxy-4,6-di-tert-butyl-2,2-dipentylbenzofuran, a potent synthetic antioxidant, was mixed with the CDD. The stereoisomer ratio of HODE (9-and-13 (Z,E)-HODE/9-and-13 (E,E)-HODE) was decreased by CDD compared with CCD, which was spared by the addition of alpha-tocopherol and 2,3-dihydro-5-hydroxy-4,6-di-tert-butyl-2,2-dipentylbenzofuran. However, the increase in plasma glutamic-pyruvic transaminase and fatty acids in liver induced by the CDD was not recovered by any antioxidant., Conclusions: This study clearly demonstrated that oxidative stress was involved in fatty liver formation induced by the CDD and that HODE was a good biomarker for an oxidative stress in vivo.

Objective: We investigated the effects of vitamin E and C supplementation on the fatty acid composition of mononuclear cells and on the clinical observations in patients who had chronic hepatitis C and received interferon-alpha-2b (IFN-alpha-2b) and ribavirin combination therapy., Methods: Patients were randomly allocated to receive daily 500 mg of vitamin E and 750 mg of vitamin C (vitamin group, n = 14) or no supplement (non-vitamin group, n = 16) in addition to IFN-alpha-2b and ribavirin therapy. The fatty acid composition of mononuclear cell phospholipids was analyzed before and at 2, 4, and 8 wk after treatment., Results: After vitamin supplementation, plasma and red blood cell alpha-tocopherol and plasma ascorbic acid levels increased in the vitamin group. Serum levels of alanine aminotransferase decreased significantly after 2 wk of treatment in both groups. At the start of treatment, a lower level of eicosapentaenoic acid (EPA) and a higher level of the molar ratio of arachidonic acid to EPA in mononuclear cells were observed in the present patients compared with healthy volunteers, and a significant correlation between the molar ratio and serum alanine aminotransferase level was found. The EPA level of mononuclear cells was maintained in the vitamin group during treatment, whereas a significant decrease was observed in the non-vitamin group at 4 and 8 wk after treatment., Conclusions: Antioxidant vitamin supplementation during IFN-alpha-2b and ribavirin therapy prevented a decrease in EPA of mononuclear cell phospholipids. If a further decrease in the ratio of arachidonic acid to EPA can be achieved by using oral EPA supplementation, the efficacy of IFN-alpha-2b and ribavirin therapy may be improved.

Objective: We determined whether hyperglycemia-induced oxidative burst is augmented in the presence of saturated, unsaturated, and trans-fatty acids (FAs)., Methods: The time course of superoxide (O2-) production in human umbilical vein endothelial cells treated with 5.5 mM (D100) or 27.5 mM (D500) of dextrose and select FAs were measured using the hydroethidene fluorescence method., Results: The rate of O2- production in cells treated with D500 was significantly higher (55% to 125% increase) than the rate observed in control cells treated with D100 (P < 0.001). The rate of O2- production (mean +/- standard deviation, in arbitrary units) was increased when cells were treated with 250, 500, and 1000 muM of myristic acid (C14; 0.189 +/- 0.04, 0.240 +/- 0.04, 0.234 +/- 0.02, respectively; P > 0.1, P < 0.05, P < 0.05, respectively), palmitate (C16; 0.151 +/- 0.03, 0.218 +/- 0.01, 0.289 +/- 0.07; P > 0.1, P < 0.05, P < 0.01), and stearic acid (C18; 0.321 +/- 0.03, 0.259 +/- 0.02, 0.341 +/- 0.03; P < 0.01 for all comparisons) compared with control cells treated with D100 only (0.184 +/- 0.01). In the presence of D500, myristic acid did not significantly augment O2- production, whereas stearic acid caused a significant further increase in O2- production. The cis unsaturated FA (oleic, C18:1; linoleic, C18:2; or linolenic acid, C18:3) stimulated O2- production significantly more than did saturated FA or trans-unsaturated FA such as elaidic, linolelaidic, or linolenelaidic acid., Conclusions: Fatty acids have the ability to modulate oxidative load in hyperglycemia and this effect may be related to the length, saturation, and possibly cis/trans orientation of the FA.

Objective: We examined the relative contributions of meat and fish to the dietary intakes of long-chain omega-3 polyunsaturated fatty acids (LCn3PUFAs)., Methods: A database detailing 4550 foods and 4298 recipes recorded in the 1995 Australian National Nutrition Survey (NNS95) was updated with new fatty acid compositional data then used to determine intakes from 24-h dietary recalls of 13858 individuals. This approach was validated with food frequency questionnaires from 8321 of these individuals., Results: Fatty acid intakes were comparable to our previous estimates from NNS95 except for LCn3PUFAs, which were considerably higher. Mean intakes in adults estimated from the 24-h recalls were 75, 71, and 100 mg/d for eicosapentaenoic acid, docosapentaenoic acid (DPA), and docosahexaenoic acid, respectively, giving a total of 246 mg/d. This is 30% greater than our previously published estimate of LCn3PUFA intake, the difference being attributable to inaccuracies in pre-existing data on the fatty acid composition of certain foods, particularly the DPA content of meats. We estimate that 43% of the LCn3PUFAs consumed by adults in the NNS95 survey originated from meat, poultry, and game compared with 48% from fish and seafood. Steak and kidney pies and other meat-containing cereal-based products accounted for an additional 4%. Beef and lamb contributed 28% of the total LCn3PUFA intake, whereas pork and poultry contributed 4% and 10%, respectively. Food frequency questionnaires produced similar results., Conclusion: Meat is a major source of LCn3PUFA, particularly DPA, for most Australians. When DPA is included in the definition of LCn3PUFAs, almost half the average adult intake of LCn3PUFA appears to originate from meat sources.

Throughout the maturation of germ cells, a morphological, biochemical and functional differentiation of mitochondria has been shown to occur. Ageing is known to cause changes involved in energy metabolism. These changes have been related to molecular and functional alterations in the properties of biological membranes. Variations in membrane lipid composition and lipid-protein interactions occur with ageing in several tissues. The present paper describes the relationship between these membrane alterations and the activities of lipid-dependent enzymes of isolated testis mitochondria in rats of from 10 days of age to 24 months. The specific activities of these enzymes are lower in preparations from adult and aged rats as compared to those from young rats. Temperature breaks of Arrhenius plots show age-dependent shifts to higher temperatures for the NADH-dehydrogenase, succinate-dehydrogenase, cytochrome c oxidase, and ATPase in senescent animals. Analysis of the membrane fatty acid composition reveals a distinct age-dependent fall in the content of polyunsaturated fatty acids accompanied by an increase in the proportion of saturated fatty acids and a decrease in polyunsaturated fatty acid percentage. The results suggest that during spermatogenesis and the ageing process some changes in the composition of the fatty acids in the surrounding membrane affect the protein-lipid interactions, producing a decrease in mitochondrial enzyme activities.

The uropygial gland is a holocrine secretory gland of birds. The lipid and the waxy sebum that the gland secretes is coated on the beak and transferred to the plumage in preening. The composition of the gland secretions from birds of different species has been determined, but little is known about the lipids of the secretion of the gland of the rock dove Columba livia. The amount of secretion, the total lipid content and the fatty acids composition of the secretion of C. livia captured in the nonbreeding season was reported. The mean amount of the secretion within the gland was 30 mg; the mean lipid content of the secretion was 0.385 mg/mg of secretion, which was equivalent to approximately 38% of the secretion. The weight of the secretion relative to gland weight was 32%. If we assume that the amount of the gland secretion constitutes a valid parameter to determine the degree of the gland development, our results indicate that the physiological role of the gland does not depend upon gland mass (GW); the rock dove in particular has a small gland, but its secretion represented 32% of the gland's mass. The composition of the lipids extracted from the gland secretion consisted of C14 to C20 fatty acids, most of them were unsaturated. The secretion of the gland contained approximately 59% of unsaturated fatty acids with a prevalence of oleic acid (37%) and a low content of linoleic (6%) and arachidonic acids (7%). The saturated long chain fatty acids were mainly 14:0, 16:0 and 18:0 in a percentage of approximately 34%. No sexual differences were found in any of the evaluated parameters.

The effects of high power ultrasound treatment (20 kHz) on some components of refined sunflower oil were studied in order to verify if and to what extent modifications in the lipidic fraction can occur. Traditional parameters including free acidity, total polar compounds, peroxide value, and conjugated dienes concentration were determined in refined sunflower oil samples before, immediately after the ultrasonic treatment and during storage. Differences between treated and untreated samples were detected only for peroxide values (PV). The PV increased from 5.38 meq. O2/kg oil for untreated oil to 6.33 meq. O2/kg oil for sunflower oil sonicated (20 kHz; 150 W; 2 min). The fatty acid composition was obtained by high resolution gas chromatography. No significant changes were observed regarding the decrease in fatty acid composition before and immediately after the treatment. Regarding the volatile fraction, some off-flavour compounds for example hexanal and limonene resulting from the ultrasonic degradation of sunflower oil were identified by gas chromatography coupled to mass spectrometry (GC-MS).

Total lipid, fatty acids and prostaglandins (PGF(2 alpha) and PGE(2)) in the ovary of kuruma prawns (Marsupenaeus japonicus) were measured during ovarian development. The level of ovarian total lipid increased with an increase in the gonad-somatic index (GSI). No significant difference was found in fatty acid composition among different stages of ovarian development. However, the content of arachidonic acid (precursor of PG(2)), but not eicosapentanoic acid (precursor of PG(3)), was significantly lower at stages I and II than at stage V (P<0.01). When ovarian PGF(2 alpha) and PGE(2) levels were plotted against GSI, no correlation was found in either PG. However, in terms of ovarian developmental stages, the level of ovarian PGs was high (approx. 20 pg/mg) at stage I, followed by marked decreases at stages IV and V (PGF(2 alpha), P<0.01) and stage IV (PGE(2), P<0.01). These results suggest that ovarian PGs and arachidonic acid are deeply involved in ovarian maturation in kuruma prawns.

I investigated whether metabolism of essential fatty acids and the concentrations of their long-chain metabolites (long-chain polyunsaturated fatty acids [LCPUFAs]) are altered in fetal or perinatal growth retardation, maternal hypercholesterolemia, low-grade systemic inflammation, insulin resistance, and atherosclerosis, conditions that predispose to the development of coronary heart disease (CHD).I critically reviewed the literature pertaining to the metabolism of essential fatty acids in CHD and conditions that predispose to it.LCPUFAs enhance endothelial nitric oxide synthesis, suppress the production of the proinflammatory cytokines tumor necrosis factor and interleukin-6, attenuate insulin resistance, and have antiatherosclerotic properties. Low-birthweight infants have decreased concentrations of LCPUFAs, especially arachidonic acid. Neonatal arachidonic acid status is related to intrauterine growth, and LCPUFAs improve fetal and postnatal growth. LCPUFAs are useful in the management of hyperlipidemia, inhibit 3-hydroxy-3-methylglutaryl coenzyme A reductase activity, and may mediate the beneficial actions of statins. Plasma concentrations of various LCPUFAs are low in diabetes mellitus, hypertension, and CHD and in populations at high risk of CHD. Breast milk is rich in LCPUFAs, and this may explain why and how adequate (6 mo to 1 y) breast feeding protects against the development of obesity, hypertension, insulin resistance, and CHD.LCPUFAs are essential for the growth and development of the fetus and infant. LCPUFAs can prevent various conditions that predispose to the development of CHD. The low incidence of CHD seen in adequately breast-fed infants can be linked to the LCPUFA content of breast milk. Based on this evidence, I suggest that provision of LCPUFAs during critical periods of growth, especially from the second trimester of pregnancy to age 5 y, prevents CHD in adult life.

Recent studies have shown that long-term in vivo exposure of dogs to neutral sulfur(IV)/sulfite aerosols induces mild inflammatory reactions, whereas the combination of neutral sulfite with acidic sulfur(VI)/sulfate aerosols evokes less pronounced effects. To understand underlying mechanisms, we studied in vitro the role of lipid mediators in the responses of alveolar macrophages (AMs) to sulfur-related compounds under neutral (pH 7) or moderate acidic (pH 6) conditions. Canine AMs incubated with sulfite at pH 7 released threefold higher amounts of platelet-activating factor than control (P < 0.005). Generation of arachidonic acid, leukotriene B4, 5-hydroxy-eicosatetraenoic acid, prostaglandin E2, thromboxane B2 and 12-hydroxyheptadecatrienoic acid increased twofold (P < 0.0005). However, these metabolites remained unchanged following incubation of AMs with sulfite at pH 6 or with sulfate at pH 7 or pH 6. Mediator release by sulfite-treated AMs at pH 7 stimulated respiratory burst activity of neutrophils. Inhibition of MAPK pathway by PD 98059, of cytosolic (cPLA2) and secretory phospholipases A2 by AACOCF3 and thioetheramide-PC, respectively, reduced sulfite-induced eicosanoid formation in AMs. Sulfite activated cPLA2 activity twofold at pH 7. This mechanism of sulfite-stimulated responses in phospholipid metabolism predicts that chronic exposure to sulfur(IV)/sulfite is associated with a considerable health risk.

In order to study the effects of exogenous EtOH and/or Fe(+2) on membrane lipid peroxidation, exogenous EtOH, FeCl(2), FeCl(2) & EtOH, NaCl and NaCl & EtOH were injected into fertile chicken eggs. Controls were either shams or injected with saline. These injections were made at 0 days or 0-2 days of development and tissue removed at stage 37 (11 days of development). Embryonic exposure to exogenous EtOH and/or Fe(+2) promoted decreased brain mass, decreased levels of brain membrane polyunsaturated fatty acids, elevated levels of brain lipid hydroperoxides, and elevated levels of Fe(+2) within embryonic brain and liver. These alterations were more severe in triple-injected embryos (E0-2/E11) as compared to single-injected embryos (E0/E11). While exogenous treatments of either EtOH and/or FeCl(2) promoted increased levels of endogenous brain Fe(+2), the effects were not additive. These observations are consistent with the hypothesis that embryonic exposure to exogenous EtOH and/or Fe(+2) promotes brain membrane lipid peroxidation.

An analytical GC-MS method based on nonpolar fused silica capillary column was developed to analyze the lipophilic constituents, mainly alkamides, from the root extracts of Echinacea purpurea (L.) Moench. In particular, the proposed method was applied to evaluate the phytochemical impacts of cucumber mosaic cucumovirus (CMV) infection on the plant's lipophilic marker phytochemicals. Methanolic (70% v/v) extracts, obtained from root materials by ultrasonic treatments, were subjected to liquid-liquid extraction with n-hexane-ethyl acetate (1:1 v/v) to recover the lipophilic, volatile to semivolatile, principles. Seventeen components, including the 11 alkamides known to E. purpurea roots, were identified in the GC-MS traces of the analyzed fractions and efficiently separated in a turnaround time of 25 min. CMV infection was found to be responsible for significant variations in the relative compositions of the major constituents, in particular germacrene D, Dodeca-2E, 4E, 8Z, 10Z(E)-tetraenoic acid isobutylamide cis/trans isomers, Undeca-2Z, 4E-diene-8, 10-diynoic acid isobutylamide and Dodeca-2E, 4Z-diene-8, 10-diynoic acid isobutylamide.

In this study total lipid (TL) content and the fatty acid composition changes in the liver of red deer stags (n=35) and fallow deer bucks (n=19) were examined at various reproductive stages. Samples were taken from mature red deer stags in the rut (September), in the post-rut (October), at the end of the rut (November) and after sexual activity had ceased (January). Sampling from mature fallow bucks was performed in the pre-rut (early October), in the rut (November) and after the reproduction period (January). The results obtained indicated significant (P<0.001) fatty degeneration of the liver in the males at the rutting season. At that time the TL concentration (x+/-S.D. in wet weight) was 156+/-40 g/kg in the red deer stags and 405+/-46 g/kg in the fallow bucks. Subsequent to the rutting season this value decreased to 47+/-15 g/kg by November in the red deer stags and to 51+/-3 g/kg by January in the fallow bucks. Significant changes were also observed in the fatty acid composition of the liver lipids, determined by gas chromatography. Liver samples collected during the rutting season from both red deer stags and fallow bucks showed higher (P<0.001) total proportions of monounsaturated fatty acids and lower (P<0.001) weight percentages of polyunsaturated fatty acids than those taken in January. In comparison with January the liver samples taken in September from red deer stags showed significantly higher (P<0.001) proportions of fatty acids C14:0, C15:0 and C:16:1, and significantly lower (P<0. 001) proportions of C18:0, C20:4 (n-6) and C22:5 (n-3). The proportions of C14:0, C15:1, C17:1 and C18:3 (n-3) in the liver samples taken in November from fallow bucks were higher (P<0.001), while the proportions of C18:0 and C20:4 (n-6) were lower (P<0.001) than those measured prior to rutting (in October) or after the rut (in January).

We hypothesized that the cytotoxic effect of GLA observed in glioma but not normal glial cells reflects differences in GLA metabolism and/or antioxidant enzyme levels between these cells. The PUFA content of unsupplemented glioma cells was approximately 50% of that seen in unsupplemented astrocytes. Supplementation with 20 microM GLA for 24 h led to a 230 and 22% increase in glioma and astrocyte PUFA content, respectively, such that both supplemented cell types contained similar levels of PUFA. No major differences were seen in terms of GLA metabolites retained in the cells or secreted into the media following incubation with [(3)H]-GLA. No significant differences were observed in activity of MnSOD or CuZn-SOD between the cells. However, CAT and GPx activity in the glioma cells was significantly higher and lower, respectively, than observed in normal astrocytes. GLA supplementation resulted in a significant increase in CAT activity in normal astrocytes; glioma CAT activity was unchanged. No significant change was seen in the other antioxidant enzymes following GLA supplementation. These results suggest that the cytotoxic effect of GLA on glioma cells reflects both increased PUFA content and an inability to upregulate CAT.

The effects of the marine fatty acid 20:4n-3, an isomer of arachidonic acid (20:4n-6), have been compared to that of 20:5n-3 on 20:4n-6 oxygenation in human platelets and endothelial cells. In platelets, 20:4n-3 added along with 20:4n-6 was as potent as 20:5n-3 in inhibiting prostaglandin H synthase (PGH synthase) activity. From 2.5- to 10 microM of 20:4n-6, the synthesis of thromboxane B2 and 12-hydroxy-5,8,10-heptadecatrienoic acid, reflecting the PGH/thromboxane synthase activity, was lowered by 5 and 10 microM of both fatty acids. In contrast, 20:4n-3, but not 20:5n-3, strongly stimulated the lipoxygenase activity at each concentration of 20:4n-6 used whatever the amount of 20:4n-3 added. The effects of both n-3 polyunsaturated fatty acids on endothelial cell PGH/prostacyclin synthases were compared after 2- and 24-hr incubation with the cells, leading to moderate (2 hr) and high (24 hr) concentrations of these fatty acids in membrane phospholipids. The incorporation of 20:4n-3 and 20:5n-3 occurred mostly in phosphatidylcholine and phosphatidylethanolamine and did not alter the 20:4n-6 level of phospholipid classes after 2-hr supplementation, whereas it was drastically decreased after 24 hr. The synthesis of prostacyclin obtained after cell stimulation by 0.1 U/mL thrombin was unaffected by the fatty acid modifications induced after 2-hr supplementation, whereas it was strongly depressed after 24 hr. It was concluded that 20:4n-3 is not an agonist for platelet activation, despite its close structural analogy with 20:4n-6, and is as potent as 20:5n-3 in inhibiting PGH synthase activities, showing that the double bond at C5 is not necessary for inhibition. In contrast, the oxygenation of 20:4n-6 by 12-lipoxygenase was stimulated by 20:4n-3 but not by 20:5n-3, which might be related to the efficient oxygenation of 20:4n-3 by this enzyme compared with 20:5n-3.

For more than 25 years 12-S-hydroxyheptadecatrienoic acid (HHT) has been known to be a product of thromboxanesynthase (TX-Syn) when synthesized with thromboxane A2 (TXA2). Although there are some hints that HHT has anti-aggregatory effects, to date, it has neither been shown to have any specific pathological relevance nor is there much information about its physiological role. This review presents a summary of the physicochemical properties of HHT, its chemical synthesis, the impact of various biological systems on its enzymatic and non-enzymatic production and its physiological function and metabolization, as well as a survey of the most important methods for analyzing this unsaturated hydroxy-fatty acid. Due to the low antibody-raising potency expected in HHT, no immunological system for HHT quantification has been developed so far. In our report we present the development and validation of a sensitive and reliable, competitive radioimmunoassay (RIA) suitable for the quantitative determination of HHT. HHT was produced by an enhanced enzymatic method using platelet-rich plasma (PRP). With an effective and modified liquid-liquid and solid-phase extraction method we were able to produce highly purified HHT (97% purity by GC/MS) in sub-milligram ranges. These fractions were used for the synthesis of BSA-antigen-conjugates and for immunization of rabbits. The tritiated tracer was synthesized using prostaglandin H synthase for the production of prostaglandin H2 (PGH2) followed by an aqueous reaction with Fe(2+)-solution to rear-range PGH2 to HHT. The dynamic range of the assay was from 30-400 pg/tube, with a sensitivity of approximately 40 pg/tube. The evaluation of the assay was performed by a HPLC-RIA method as well as by correlation with a quantitative HPLC method and correlation with TXB2 concentrations in a blood coagulation study. The assay may be useful for the quantification of HHT in several tissues and body fluids under various physiological conditions and may also help to understand the possible physiological role of HHT in biological processes.

For years, intravenous lipid supply has been considered as a means of providing an efficient fuel to many tissues of the body and of preventing or correcting essential fatty acid deficiency. The potential for lipid emulsions to modulate cell function via their content of specific fatty acids and of liposoluble vitamins has not received much attention yet. Soybean [long-chain triglycerides (LCT)] emulsions provide a valuable source of energy, but they are excessively rich in omega-6 essential fatty acids (FAs). Their infusion is associated with an accumulation of linoleate and a reduction of long-chain (> or = C20) omega-6 and omega-3 fatty acids in cell membrane phospholipids, as well as with a depletion of antioxidant status, associated with a reduction of alpha-tocopherol in plasma lipoproteins. Infusions of the mixed medium-chain triglycerides (MCT)/LCT (50%:50%; wt:wt) largely bypass these disadvantages. In addition, plasma elimination of MCT/LCT is faster than that of LCT. Recent advances indicate a great potential for omega-3 FAs incorporated into membrane phospholipids to modulate cell response to various stimuli and to influence several intracellular metabolic processes. Furthermore, some of these FA directly influence the production and the action of important mediators, the eicosanoids. In practical terms, an increased intake of omega-3 FAs may reduce inflammatory and thrombotic responses while protecting tissue microperfusion and immune defenses. Such properties may find interesting applications in several types of intensive care unit patients, provided that omega-3 FA incorporation takes place promptly. We recently had the opportunity to study in vitro and in vivo the metabolism of emulsions made of a mixture of MCT, soybean LCT, and fish oil triglycerides. Plasma elimination of such preparations appeared to be very fast, and their infusion was not associated with a prolonged residence of emulsion particles. In addition, uptake of remnants enriched with omega-3 FAs and liposoluble vitamins was fairly fast and occurred in several types of cells, leading to an efficient incorporation of omega-3 FAs in cell membranes within a few hours. The understanding that remnant uptake plays a significant role in the delivery of components included in lipid emulsions opens new areas of investigation and is likely to find several conditions of applications for new types of preparations.

A new interface coupled to a mass spectrometer was developed for the direct analysis of volatile organic compounds from small volumes of aqueous samples, including blood or tissue homogenates (St-Germain et al. 1995, Anal. Chem. 67:4536-4541). The greatest advantages of our system are minimal sample treatment, an instantaneous response time coupled with detection limits in the range of < 1 ppb for most compounds. For the analysis of low-molecular weight aldehydes, such as formaldehyde, acetaldehyde, propanal, and hexanal, lower detection limits were obtained when samples were converted to methoxime derivatives prior to injection. The detection limit for hexanal in water or Krebs-Ringer solution was 0.01 microM (10 pmol injected). The reproducibility of replicate injections was 4.4%. The usefulness of our system was illustrated by measuring aldehyde accumulation in peroxidized solutions of polyunsaturated fatty acids and rat tissue homogenates. Data confirmed that peroxidation of omega-3 fatty acids produces propanal, whereas omega-6 fatty acids form hexanal. Peroxidation of heart and brain homogenates formed predominantly propanal. However, the recovery of hexanal after sample treatment with methoxylamine depended on the derivatization time and temperature, suggesting that this aldehyde may form Schiff base linkages. These results show that spray extraction coupled to mass spectrometry provides a quick (< 1 min), clean and reproducible way to detect aldehydes produced from lipid peroxidation in aqueous samples.

A highly sensitive LC method has been developed and validated for quantitation of Ro 24-0238 in human plasma using Ro 24-2446 as an internal standard. With 1 ml of plasma, the limit of quantitation of the method was 50 pg ml-1 of Ro 24-0238. After solid-phase extraction with C18 reversed-phase cartridges, the samples were reconstituted in an acidic buffer solution; under these conditions, Ro 24-0238 and Ro 24-2446 (IS) were converted to their cationic forms. The LC system employed a strong cation exchange column and a narrow bore reversed-phase column, connected via a column switching valve. The cationic analyte and internal standard were separated from most of the endogenous components of plasma on the cation exchange column. A small fraction containing the analyte and the internal standard was transferred by automated valve switching to the narrow bore reversed-phase column, which further resolved the individual components. The chromatography was monitored by UV absorption at 322 nm. The overall intra-assay precision was 3.6% (RSD) and the per cent error was less than +/- 11%. The overall inter-assay precision was 3.9% (RSD). Linearity was demonstrated in a concentration range of 50-5000 pg ml-1. This method has been applied to pharmacokinetic studies of Ro 24-0238 in man.

1. Adipose tissues associated with the mystacial and superciliary vibrissae of the Saimaa seal (Phoca hispida saimensis) contained 31% of hexadecenoic acid (16:1 omega 7), which was almost twice the already high amount in blubber. The amount of tetradecenoic acid (14:1 omega 5) was also double that in blubber. 2. At the same time the levels of long-chain highly unsaturated docosapentaenoic (22:5 omega 3) and docosahexaenoic acids (22:6 omega 3) were only two-thirds of those found in blubber. 3. It is suggested that the excess of low-melting monoenoic fatty acids (14:1 and 16:1) in adipose tissues near the vibrissae might not be compensation for the low supply of dietary polyunsaturated fatty acids in these tissues, but that the self-induced excess of monoenoic fatty acids ensured the proper functioning of these well-developed sense organs.

1. The main differences in fatty acid composition of blubber between the Saimaa seal (Phoca hispida saimensis) and the Baltic seals (Phoca hispida botnica and Halichoerus grypus) were, that in fresh water the proportions of polyunsaturated C18 acids and arachidonic acid (20:4 omega 6) were high. 2. The trend became more evident when data on ringed seals from northern oceans were compared with the present data. 3. The proportions of exogenous 20:1 and 22:1 were extremely low in ringed seals from Lake Saimaa and in Baltic seals compared with seals living in oceans.

The pyridoindole derivative stobadine [(-)-cis-2,8-dimethyl-2,3,4,4a,5,9b-hexahydro-1H-pyrido(4,3b)indole] has been described as a drug with antihypoxic and antiarrhythmic cardioprotective properties. Here its reactivity with peroxyl radicals in liposomes using a lipid-soluble azo-initiator of peroxyl radicals, 2,2'-azo-bis(2,4-dimethyl-valeronitrile) (AMVN), was examined. Stobadine exerted scavenging as evidenced by the inhibition of: (i) cis-parinaric acid fluorescence decay (half-maximal effect at 20 microM), or (ii) luminol-sensitized chemiluminescence (half-maximal effect at 33 microM). In rat liver microsomes, stobadine was equally efficient in inhibiting lipid peroxidation induced by lipid-soluble (AMVN) or water-soluble 2,2'-azo-bis(2-aminopropane)-HCl (AAPH), azo-initiators of peroxyl radicals with half-maximal effect at 17 microM. Stobadine partitions in a two-phase system (octanol-water) with the coefficient log P = 0.57 +/- 0.03, explaining its ability to quench peroxyl radicals in both lipid and aqueous phases. Stobadine is not an efficient scavenger of superoxide radicals. The second order rate constant for the reaction of stobadine with superoxide was estimated to be 7.5 x 10(2) M-1 sec-1 as measured by superoxide-induced lucigenin-amplified chemiluminescence. ESR measurements showed that stobadine in liposomes does not reduce the chromanoxyl radical of a vitamin E homologue with a 6-carbon side-chain, 2,5,7,8-tetramethyl-2-(4'-methylpentyl)chroman-6-ol(chromanol++ +-alpha-C6), in agreement with pulse-radiolysis results obtained using Trolox in homogeneous solution (Steenken et al., Chem Res Toxicol 5: 355-360, 1992). Stobadine increased the magnitude of the chromanoxyl and ascorbyl radical ESR signal generated by lipoxygenase+arachidonate. This was interpreted to be due to the interaction of stobadinyl radicals with the chromanol ring and ascorbate, respectively. It is suggested that high reactivity of stobadine radicals requires the presence of reducing antioxidants (vitamin E, vitamin C) to exhibit its antioxidant effects in physiological systems.

1. The biosynthesis of polyunsaturated fatty acids (PUFA) of the n-6 and n-3 series was investigated in cultured Sertoli cells. 18:2n-6, 18:3n-6, 20:2n-6, 18:3n-3 and 20:3n-3 were added individually at a concentration of 20 mumol to culture media. 2. Maximum incorporation of 20- and 22-carbon PUFA into membrane lipids was observed after 72 hr of incubation with all the exogenous substrates used. 3. As reported in other cell systems, the delta 6 desaturation was the first rate-limiting step; the major factor regulating this activity was the concentration of linoleic acid or alpha-linolenic acid in the medium. 4. Our data show that the delta 5-desaturation represents a second regulatory step in PUFA biosynthesis. 5. The sum of n-6 and n-3 PUFA of the 22 carbon chain length constantly represented between 11 and 12% of total fatty acids, regardless of the exogenous substrate used. 6. Our kinetic studies of the incorporation of PUFA of the n-6 and n-3 series did not permit detection of a delta 8 desaturase activity.

In our on-going studies of experimental uveitis, we previously obtained a preliminary indication of phagocyte-mediated retinal lipid peroxidation by measuring conjugated dienes (CD), thiobarbituric acid reactive substances (TBARS) and fluorescent chromolipids. Using gas chromatography/mass spectrometry (GC/MS), the current study detected hydroperoxide-derived 10-, 11-, 13-, 14-, and 17-hydroxydocosahexaenoic acid (HDHE) in retinal membranes. Docosahexaenoic acid (22:6) is the major polyunsaturated fatty acid (PUFA) in photoreceptor membranes. Hydroperoxides from other retinal PUFA were found also. Arachidonic acid (20:4) yielded 8-, 9-, 11-, 12-hydroxyeicosatetraenoic acid (HETE) as major products. Since 12-HETE could also arise from lipoxygenase catalyzed oxygenation of free 20:4, the source of 12-HETE could be both peroxidative and lipoxygenase pathways. Concomitantly, peroxidative loss of 22:6 and accumulation of 20:4 were also noted. At the peak of inflammation, loss of 22:6 was close to 50% of the original amount in the control retinas. In the same time period, 20:4 increased more than two-fold. The present data suggest that the oxygen radicals derived from phagocytes initiate the retinal lipid peroxidation, and the resultant formation of hydroperoxides, oxidative loss of 22:6 and accumulation of 20:4 appear to serve as amplification factors in subsequent biochemical events, such as chemotaxis of PMNs and activation of cyclooxygenase.

1. Absolute and relative amounts of eicosapentaneoic acid (EPA) and docohexaenoic acid (DHA) in muscle of eels from four different fish farms were compared with samples from wild eels from two different areas of northern Italy. 2. Farmed eels were richer in DHA and EPA than wild animals. 3. The addition of cod liver oil to the diet of farmed eels led to a significant accumulation of EPA and DHA, but no change in total lipid content. 4. The calculation of two indices related to highly unsaturated fatty acid (HUFA) content (FLQ = fish lipid quality; AI = aterogenic index), indicated the higher nutritional value of farmed vs wild fish. 5. We conclude that farmed eels are an adequate source of fish products for human nutrition and propose use of the above-mentioned indices as an effective means for assessing fish nutritional quality for populations at high risk of chronic degenerative disease.

1. The phospholipid fatty acid compositions of the sponges Ircinia strobilina, Ircinia felix, Ircinia campana, Ircinia sp., Spongia tubulifera and Dysidea etherea were studied, revealing the presence, besides other common fatty acids, of considerable amounts (2-5%) of the novel 23-methyl-5,9-tetracosadienoic acid (1). 2. The demospongic acids 5,9-tetracosadienoic acid, 23-methyl-5,9-tetracosadienoic acid (1), and 5,9-pentacosadienoic acid, were particularly abundant in sponges of the genus Ircinia, in contrast to the most common 5,9-hexacosadienoic acid found in other species. These findings are discussed in terms of the taxonomy of the Dictyoceratida. 3. The complete characterization of the novel phospholipid fatty acid 23-methyl-5,9-tetracosadienoic acid (1) is presented.

Our laboratory recently identified trans,trans-muconaldehyde (MUC), a six-carbon diene dialdehyde, as a hematotoxic microsomal metabolite of benzene (Latriano et al., Proc Natl Acad Sci USA 83: 8356-8360, 1986). We also showed that MUC is metabolized in vitro to trans,trans-muconic acid (MA), a six-carbon diene dicarboxylic acid and known urinary metabolite of benzene. To elucidate further the role of ring-opened metabolites in benzene toxicity, the metabolism of benzene and MUC was examined in the benzene sensitive DBA/2N mouse strain and the less benzene sensitive C57BL/6 strain. A sensitive assay for urinary MA analysis was developed. The percent of benzene dose excreted as urinary MA within the first 24 hr after treatment decreased with an increase in benzene dose, i.e. from 9.8 to 0.4% in DBA/2N mice and from 17.6 to 0.2% in C57BL/6 mice treated with 0.5 to 880 mg/kg benzene. DBA/2N mice excreted significantly (P less than or equal to 0.05) more MA compared with C57BL/6 mice after treatment with hematotoxic benzene doses (220-880 mg/kg). At low benzene doses (0.5 to 2.5 mg/kg), C57BL/6 mice excreted significantly (P less than or equal to 0.05) more MA compared with DBA/2N mice. There were no significant differences in the metabolism of MUC to MA between the two strains after treatment with 0.5 to 3.0 mg/kg. Furthermore, mice from both strains excreted similar amounts of muconic acid when treated with 0.7 to 7.1 mg/kg MA. These results are consistent with the hypothesis that reactive ring-opened metabolites such as trans,trans-muconaldehyde play a role in benzene hematotoxicity. Sensitivity towards benzene may be due, in part, to increased metabolism to ring-opened compounds.

Prostaglandins PGE1 and PGE2 extracted from bovine semen, purified via silicic column chromatography were quantified by gas chromatography as their methoxime methyl ester trimethylsilyl ether derivatives. The PGE2 and PGE2 concentrations of 19 bovine semen samples ranged from 395 +/- 225 and 487 +/- 407 ng/ml, respectively. A constant 1:1 ratio between PGE1 and PGE2 was observed. There was not relationship between PGE and sperm motility, but high sperm counts were generally associated with decreased PGE levels. The direct precursors of PGE1 and PGE2, i.e. 20:3n6 and 20:4n6, occurred in low concentrations compared to other related unsaturated fatty acids, i.e. 18:2n6 and 22:5n6 of the n-6 family.

Fatty acids from the several lipid classes of selected steelhead trout (Salmo gairdnerii) parr and smolt tissues, previously separated by thin-layer chromatography, were analyzed by gas-liquid chromatography. The fatty acid composition of the parr was markedly different from that of the smolt; the former being characterized by relatively low amounts of polyunsaturated fatty acids and relatively high amounts of linoleic acid, much like the typical freshwater lipid pattern. The fatty acid composition of the smolt was characterized by large proportions of long-chain polyunsaturated fatty acids. Generally, the fatty acid composition of the smolt resembled the typical seawater lipid pattern. The change in fatty acid composition of the smolt is anticipatory to seawater entry and is independent of diet and water temperature. These alterations suggest that the assumption of a polyunsaturated lipid pattern during parr-smolt transformation (smoltification) is preadaptive to seawater entry.

Following administration of the LD50 or the LD100 of Gramoxon (PQ), the phospholipids (PL) of the lung, liver and kidney were separated and both the PLs and the fatty acids isolable from them were examined quantitatively. The different doses of PQ caused different changes of the saturated and unsaturated fatty acids in the various organs. The changes in the PL unsaturated fatty acids point to PQ-induced lipid peroxidation enhancement and membrane damage.

There is growing evidence from both experimental and epidemiologic studies to suggest that regular fish consumption reduces the risk of cardiovascular disease in humans. However, a crude analysis of Canadian descriptive data does not support this hypothesis. Age-adjusted mortality from ischemic heart disease and cerebrovascular disease is actually higher in the Atlantic Provinces than in the Prairies where fish consumption is much lower. Possible reasons for this, including the importance of the type of fish consumed, and the effects of methods of preparation are discussed.