Increase In Eicosapentaenoic Acid Research Articles

Metabolomics Reveals the Impact of Overexpression of Cytosolic Fructose-1,6-Bisphosphatase on Photosynthesis and Growth in Nannochloropsis gaditana.

Nannochloropsis gaditana, a microalga known for its photosynthetic efficiency, serves as a cell factory, producing valuable biomolecules such as proteins, lipids, and pigments. These components make it an ideal candidate for biofuel production and pharmaceutical applications. In this study, we genetically engineered N. gaditana to overexpress the enzyme fructose-1,6-bisphosphatase (cyFBPase) using the Hsp promoter, aiming to enhance sugar metabolism and biomass accumulation. The modified algal strain, termed NgFBP, exhibited a 1.34-fold increase in cyFBPase activity under photoautotrophic conditions. This modification led to a doubling of biomass production and an increase in eicosapentaenoic acid (EPA) content in fatty acids to 20.78-23.08%. Additionally, the genetic alteration activated the pathways related to glycine, protoporphyrin, thioglucosides, pantothenic acid, CoA, and glycerophospholipids. This shift in carbon allocation towards chloroplast development significantly enhanced photosynthesis and growth. The outcomes of this study not only improve our understanding of photosynthesis and carbon allocation in N. gaditana but also suggest new biotechnological methods to optimize biomass yield and compound production in microalgae.

Lipidomic and transcriptomic analysis of the increase in eicosapentaenoic acid under cobalamin deficiency of Schizochytrium sp.

Schizochytrium sp. is a heterotrophic microorganism capable of accumulating polyunsaturated fatty acids and has achieved industrial production of docosahexaenoic acid (DHA). It also has the potential for eicosapentaenoic acid (EPA) production. In this study, it was found that the cell growth, lipid synthesis and fatty acid composition of Schizochytrium sp. were significantly affected by the level of cobalamin in the medium, especially with regard to the content of EPA in the fatty acids. The content of EPA in the fatty acids increased 17.91 times, reaching 12.00%, but cell growth and lipid synthesis were significantly inhibited under cobalamin deficiency. The response mechanism for this phenomenon was revealed through combined lipidomic and transcriptomic analysis. Although cell growth was inhibited under cobalamin deficiency, the genes encoding key enzymes in central carbon metabolism were still up-regulated to provide precursors (Acetyl-CoA) and reducing power (NADPH) for the synthesis and accumulation of fatty acids. Moreover, the main lipid subclasses observed during cobalamin deficiency were glycerolipids (including glycerophospholipids), with EPA primarily distributed in them. The genes involved in the biosynthesis of these lipid subclasses were significantly up-regulated, such as the key enzymes in the Kennedy pathway for the synthesis of triglycerides. Thus, this study provided insights into the specific response of Schizochytrium sp. to cobalamin deficiency and identified a subset of new genes that can be engineered for modification.

P068 Polyunsaturated fatty acid excess fuels pro-inflammatory transcriptional networks in GPX4 deficient intestinal epithelial cells

Abstract Background There is increasing evidence that the rise in Crohn’s disease (CD) incidence is partially driven by constituents of a Western-style diet such as polyunsaturated fatty acids (PUFA). Recently we could demonstrate that PUFA excess induces a CD-like enteritis in mice lacking antioxidant GPX4 in their intestinal epithelial cells (IECs) and that CD patients clinical disease activity positively correlates with PUFA intake. In this project we investigated the role of epithelial lipid composition and lipid mediators during PUFA induced metabolic gut inflammation. Furthermore, we transcriptionally profiled GPX4 deficient small intestinal cells after stimulation with ω3- and ω6-PUFA. Methods Mice lacking one allele of Gpx4 in their IECs (GPX4+/-IEC) and wild-type controls (WT) were fed a PUFA (10% fish oil) enriched Western diet (PUFA-WD) for 12 weeks. Intestinal inflammation was assessed by histopathology. IECs were harvested by scraping of small intestinal sections and lipid profiles and mediators were analysed with LC/MS. GPX4 in murine small intestinal cells (MODE-K) was knocked down by siRNA (siGpx4) scrambled siRNA served as control (siCtrl). MODE-K were stimulated with ω3-, ω6-PUFA, 5,6- and 8,9-DHET or vehicle. Cytokine expression in cell culture supernatant was analysed by ELISA and transcriptome was analysed by bulk RNA sequencing. Results Histopathology revealed gut inflammation in GPX4+/-IEC but not WT mice after 12 weeks PUFA-WD feeding. LC/MS analysis of IECs derived from GPX4+/-IEC and WT mice fed the PUFA-WD depicted a significant increase in eicosapentaenoic acid (20:5), docosapentaenoic acid (22:5) and docosahexaenoic acid (22:6) containing phosphatidylethanolamine (PE) and phosphatidylcholine (PC) species (Fig 1a). All other PE and PC species showed a reduced amount, confirming the enrichment of the specific PUFAs within the IECs by the PUFA-WD. Lipid mediator analysis further revealed an increase in 5,6-DHET and 8,9-DHET only in IECs derived from GPX4+/-IEC but not WT mice (Fig 1b). When stimulated with PUFA, siGpx4 MODE-K produced high amounts of neutrophil attracting chemokine CXCL-1 which is attenuated by the supplementation of anti-inflammatory lipid mediators 5,6-DHET and 8,9-DHET (Fig 2). Transcriptome analysis revealed the upregulation of pro-inflammatory pathways including TNFa, MAPK and NF-κB in siGpx4 vs. siCtrl MODE-K after PUFA stimulation (Fig 3). Conclusion By integrating lipidomic and transcriptomic data of a reductionist cell culture system and a model organism, we could show that PUFA excess in a GPX4 deficient individual fuels a pro-inflammatory transcriptional network which outweighs anti-inflammatory ω3-PUFA metabolites influence culminating in gut inflammation.

Parenteral Fish-Oil Containing Lipid Emulsions Limit Initial Lipopolysaccharide-Induced Host Immune Responses in Preterm Pigs.

Multicomponent lipid emulsions are available for critical care of preterm infants. We sought to determine the impact of different lipid emulsions on early priming of the host and its response to an acute stimulus. Pigs delivered 7d preterm (n = 59) were randomized to receive different lipid emulsions for 11 days: 100% soybean oil (SO), mixed oil emulsion (SO, medium chain olive oil and fish oil) including 15% fish oil (MO15), or 100% fish oil (FO100). On day 11, pigs received an 8-h continuous intravenous infusion of either lipopolysaccharide (LPS—lyophilized Escherichia coli) or saline. Plasma was collected for fatty acid, oxylipin, metabolomic, and cytokine analyses. At day 11, plasma omega-3 fatty acid levels in the FO100 groups showed the highest increase in eicosapentaenoic acid, EPA (0.1 ± 0.0 to 9.7 ± 1.9, p < 0.001), docosahexaenoic acid, DHA (day 0 = 2.5 ± 0.7 to 13.6 ± 2.9, p < 0.001), EPA and DHA-derived oxylipins, and sphingomyelin metabolites. In the SO group, levels of cytokine IL1β increased at the first hour of LPS infusion (296.6 ± 308 pg/mL) but was undetectable in MO15, FO100, or in the animals receiving saline instead of LPS. Pigs in the SO group showed a significant increase in arachidonic acid (AA)-derived prostaglandins and thromboxanes in the first hour (p < 0.05). No significant changes in oxylipins were observed with either fish-oil containing group during LPS infusion. Host priming with soybean oil in the early postnatal period preserves a higher AA:DHA ratio and the ability to acutely respond to an external stimulus. In contrast, fish-oil containing lipid emulsions increase DHA, exacerbate a deficit in AA, and limit the initial LPS-induced inflammatory responses in preterm pigs.

Effect of Supplemental Enteral Fish Oil on the Development of Psychological Complications in Critically Ill Multiple-Trauma Patients: 6 Months' Follow-Up.

Posttraumatic stress disorder (PTSD) is common in intensive care unit (ICU) patients. Defic it intake of ω-3 polyuns aturated fatty ac ids (PUFAs) may be as sociated with developm ent of PTSD. This study randomized mechanically ventilated patients suffering from multiple trauma (n = 150) into 2 groups: a study and a control group that received enteral feeding with or without fish oil. Fifty-one patients were interviewed 6 months after discharge from the ICU. Psychometric parameters of PTSD were assessed by questionnaires. The fatty acid composition of erythrocyte membranes was performed by homogenization of the cells in hexane-isopropanol. No differences were found in baseline characteristics between the groups. Erythrocyte membrane composition showed significantly higher concentrations of ω-3 fatty acids in the study group. Anxiety and depression symptoms were correlated with an increase in eicosapentaenoic acid (EPA) on days 4 and 8 for depression and EPA on day 4 and docosahexaenoic acid (DHA) on day 8 for anxiety. Total ω-3 content was positively correlated with anxiety and depression as well. An inverse correlation was found between DHA and EPA in the treatment group and with the total ω-3 and DHA in the control group for the Brief Illness Perceptions Questionnaire. Administration of an ω-3 PUFA-enriched diet during the ICU stay did not prevent development of PTSD in trauma patients 6 months after discharge from ICU.

Resveratrol Modifies Lipid Composition of Two Cancer Cell Lines.

Resveratrol (Resv) offers health benefits in cancer and has been reported to modulate important enzymes of lipid metabolism. Studies of its effects on lipid composition in different subtypes of breast-cancer cells are scarce. Thus, we investigated the alterations in phospholipids (PL), fatty acids (FA), and lipid metabolism enzymes in two breast-cancer cell lines after Resv treatment. MCF-7 and MDA-MB-231 cells were treated with 80 and 200 μM of Resv, respectively, for 24 hours. We analyzed PL with radiolabeled inorganic phosphate (32Pi) by thin-layer chromatography, FA by gas chromatography-mass spectrometry, and lipid metabolism enzymes (DGAT2, FAS, ρACCβ, pAMPKα, and AMPK) by Western blot. Resv treated MDA-MB-231 phospholipids showed a reduction in phosphatidylcholine (63%) and phosphatidylethanolamine (35%). We observed an increase in eicosapentaenoic acid (EPA) (73%) and docosahexaenoic acid (DHA) (65%) in MCF-7 cells after Resv treatment. Interestingly, the same treatment caused 50% and 90% increases in EPA and DHA, respectively, in MDA-MB-231 cells. In MCF-7 cells, Resv increased the expression of ρACCβ (3.3-fold) and AMPKα/ρAMPKα (1.5-fold) and in MDA-MB-231 cells it inhibited the expression of ρACCβ (111.8-fold) and AMPKα/ρAMPKα (1.2 fold). Our results show that Resv modified PL and saturated and unsaturated FA especially in MDA-MB-231 cells, and open new perspectives to the understanding of the reported anticancer effect of Resv on these cells.

Medium-Chain Triglycerides Attenuate Liver Injury in Lipopolysaccharide-Challenged Pigs by Inhibiting Necroptotic and Inflammatory Signaling Pathways.

This study was conducted to investigate whether medium-chain triglycerides (MCTs) attenuated lipopolysaccharide (LPS)-induced liver injury by down-regulating necroptotic and inflammatory signaling pathways. A total of 24 pigs were randomly allotted to four treatments in a 2 × 2 factorial design including diet (0 and 4% MCTs) and immunological challenge (saline and LPS). After three weeks of feeding with or without 4% MCTs, pigs were challenged with saline or LPS. MCTs led to a significant increase in eicosapentaenoic acid, docosahexaenoic acid and total (n-3) polyunsaturated fatty acid concentrations. MCTs attenuated LPS-induced liver injury as indicated by an improvement in liver histomorphology and ultrastructural morphology of hepatocytes, a reduction in serum alanine aminotransferase and alkaline phosphatase activities as well as an increase in claudin-1 protein expression. In addition, MCTs also reduced serum tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 concentrations, liver TNF-α and IL-1β mRNA expression and protein concentrations and enhanced liver heat shock protein 70 protein expression in LPS-challenged pigs. Moreover, MCTs decreased mRNA expression of receptor-interacting serine/threonine-protein kinase (RIP) 3, mixed-lineage kinase domain-like protein (MLKL) and phosphoglycerate mutase 5 and inhibited MLKL phosphorylation in the liver. Finally, MCTs decreased liver mRNA expression of toll-like receptor (TLR) 4, nucleotide-binding oligomerization domain protein (NOD) 1 and multiple downstream signaling molecules. MCTs also suppressed LPS-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation and increased extracellular signal-related kinase 1/2 phosphorylation in the liver. These results indicated that MCTs are capable of attenuating LPS-induced liver damage by suppressing hepatic necroptotic (RIP1/RIP3/MLKL) and inflammatory (TLR4/NOD1/p38 MAPK) signaling pathways.

Does administration of eicosapentaenoic acid increase soluble thrombomodulin level in statin-treated patients with stable coronary artery disease?

Interventions targeting the serum eicosapentaenoic acid (EPA)/arachidonic acid (AA) ratio could be useful for the prevention of coronary artery disease (CAD). Few data exist regarding the effects of administration of EPA on the serum levels of soluble thrombomodulin (sTM) as a marker of endothelial damage, or on the relationship between the sTM and EPA/AA ratio in patients with CAD receiving statin treatment. We assigned stable CAD patients already receiving statin therapy to an EPA group (1800mg/day: n = 50) or control group (n = 50). A significant increase of the sTM level was observed in the EPA group as compared to that in the control group 0.40 (0.10/0.70) FU/mL vs. 0.20 (0/0.40) FU/mL, p = 0.004 at the 6-month follow-up examination. Multivariate regression analysis after adjustments for coronary risk factors and changes of the serum lipid levels identified an increased EPA/AA ratio as an independent predictor of increased serum sTM level (β = 0.244, p = 0.02). The results suggest that an increased sTM level caused by additional administration of EPA to statin might be associated with an increased EPA/AA ratio. The increase of the serum sTM after administration of EPA might reflect an increase of the TM expression on the endothelial surface rather than endothelial damage in CAD patients under statin treatment.Clinical Trial Registration Information UMIN ( http://www.umin.ac.jp/ ), Study ID: UMIN000010452.

Changes in serum fatty acid and lipoprotein subclass concentrations from prepuberty to adulthood and during aging.

Concentrations in serum were determined for 18 fatty acids (FAs) and 21 lipoprotein main and subclasses by chromatographic analyses and the average size was calculated for very low density (VLDL), low density (LDL) and high density (HDL) particles. 283 ethnic Norwegian children and adults from the rural Fjord region of Western Norway were compared with the objectives to reveal patterns and gender differences during the development from prepuberty to adulthood and during aging in adults. Both genders showed a large increase in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from child to adult. Males, but not females, show a significant increase in most C16–C18 FAs from prepuberty to adulthood. These changes in males correlate to a pattern of increased concentrations of triglycerides, VLDL and LDL particles, especially the atherogenic subclasses of small and very small LDL particles. Furthermore, concentrations of medium, large and very large HDL particles decrease, while concentration of very small HDL particles increase leading to reduced average size of HDL particles. Females only showed significant increase in concentrations of small and very small LDL particles, very small HDL particles and apolipoprotein B. While EPA and DHA continued to increase during aging in women, no validated model for connecting age to FA profile was obtained for men. Women showed significant increase in concentrations of all subclasses of LDL particles during aging, while men exhibited a more complex pattern with increase also in apolipoprotein A1 and HDL particles.Electronic supplementary materialThe online version of this article (doi:10.1007/s11306-016-0968-y) contains supplementary material, which is available to authorized users.

Cardiometabolic Profiles and Gene Expression Following Fish Oil Supplementation: Is There a Role for FADS Genotypes?

Consumption of omega‐3 fatty acids (FAs) modifies cardiometabolic risk factors in hyperlipidemic and diabetic individuals; however, their effects in young healthy adults are unclear. We examined the impact of fish oil supplements on blood cardiometabolic markers, FA composition, eicosanoid profiles and gene expression. We then examined whether these endpoints varied with an individual's fatty acid desaturase (FADS) genotype. Young male adults (18 – 25 yrs) consumed daily fish oil supplements (up to 3g omega‐3 FAs) for 12 weeks. Significant reductions in blood triglycerides were seen by week 12. Paradoxically, two omega‐6 derived eicosanoids (PGF2α and TXB2) were increased with supplementation. Whole blood gene expression revealed no differences in key genes regulating eicosanoid production, i.e., ALOX5, ALOX12, ALOX15, COX‐2 and iPLA2. FA analysis showed increases in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) content in serum and red blood cells (RBCs), and decreases in arachidonic acid (AA). Compared to major allele carriers, individuals with FADS minor allele variants experienced greater increases in EPA levels in serum and RBCs during supplementation. There were no genotype differences for DHA or AA levels. Collectively, our results showed that fish oil supplements alter the eicosanoid profile independent of changes in gene expression. Additionally, we demonstrated that changes in FA levels after supplementation are influenced by an individual's FADS genotype. Further investigations regarding the relationship between fish oil supplements, eicosanoid profiles, and FADS genotype are warranted.Grant Funding Source: Supported by NSERC

MODIFICATION OF SERUM AND TISSUE LIPIDS IN RATS FED WITH BLENDED AND INTERESTERIFIED OILS CONTAINING GROUNDNUT OIL WITH LINSEED OIL

Groundnut oil (GNO) was modified to contain equal proportions of omega-6/ omega-3 fatty acids by blending with linseed oil. Blended oil (BLE) was subjected to interesterification reaction using immobilized lipase to prepare transesterified oil (TRA).A structured lipid (STR) was also prepared by acidolysis reaction usingGNO and omega-3 fatty acid concentrates from linseed oil. These oils were then fed to rats for a period of 60 days.A reduction in low density lipoprotein cholesterol level by 26 and 23% and a reduction in serum triglyceride concentration by 18.9 and 17.6% were observed in rats given TRA and STR, respectively, as compared with rats fed with GNO. Rats fed with BLE and interesterified oils showed incorporation of a-linolenic acid and an increase in eicosapentaenoic acid and docosahexaenoic acid in serum and liver lipids. These modified oils could be used for enriching Indian diets with omega-3 fatty acids. PRACTICAL APPLICATIONS There is a realization that dietary fat should contain balanced amount of omega-6 and omega-3 fatty acids. The oils used in Indian cooking generally contains high amount of omega-6 but not omega-3 fatty acids. The present investigation was undertaken to develop oil that can provide equal amounts of omega-6 and omega-3 fatty acids by the blending process. The fatty acids in blended oils were randomized through lipase catalyzed acidolysis and transesterification reaction. These modified oils significantly reduced the serum and liver lipids when fed to rats. The modified oils also showed altered physical properties. This study therefore showed that it is possible to restructure the edible oils through biotechnological approaches to get the desired fatty acid composition needed for better nutrition and required physical properties in oils.

NGF blocks polyunsaturated fatty acids biosynthesis in n − 3 fatty acid-supplemented PC12 cells

Regulation of polyunsaturated fatty acid (PUFA) biosynthesis in proliferating and NGF-differentiated PC12 pheochromocytoma cells deficient in n−3 docosahexaenoic acid (DHA 22:6n−3) was studied. A dose- and time-dependent increase in eicosapentaenoic acid (EPA, 20:5n−3), docosapentaenoic acid (DPA, 22:5n−3) and DHA in phosphatidylethanolamine (PtdEtn) and phosphatidylserine (PtdSer) glycerophospholipids (GPL) via the elongation/desaturation pathway following alpha-linolenic acid (ALA, 18:3n−3) supplements was observed. That was accompanied by a marked reduction of eicosatrienoic acid (Mead acid 20:3n−9), an index of PUFA deficiency. EPA supplements were equally effective converted to 22:5n−3 and 22:6n−3. On the other hand, supplements of linoleic acid (LNA, 18:2n−6) were not effectively converted into higher n−6 PUFA intermediates nor did they impair elongation/desaturation of ALA. Co-supplements of DHA along with ALA did not interfere with 20:5n−3 biosynthesis but reduced further elongation to 22-hydrocarbon PUFA intermediates. A marked decrease in the newly synthesized 22:5n−3 and 22:6n−3 following ALA or EPA supplements was observed after nerve growth factor (NGF)-induced differentiation. NGF also inhibited the last step in 22:5n−6 formation from LNA. These results emphasize the importance of overcoming n−3 PUFA deficiency and raise the possibility that growth factor regulation of the last step in PUFA biosynthesis may constitute an important feature of neuronal phenotype acquisition.

Plasma Fatty Acid Profile of Gestating Ewes Supplemented with Fishmeal

Problem statement: The very long chain n-3 polyunsaturated fatty acids (>18C) cannot be adequately synthesized by ruminant tissues to meet their requirements; therefore, their concentration in body depends on the supply through feed. It may be possible to improve the essential fatty acid status of ruminant animals, during gestation by manipulating the maternal diet with Fishmeal (FM). The objectives of this research were to (1) determine the effect of fishmeal supplementation on the plasma fatty acid profile of ewes during late gestation and (2) determine the status of the plasma docosahexaenoic acid (22:6n3) of lambs born to these ewes. Approach: Eight gestating ewes [Rideau-Arcott, 97±5 kg initial body weight, 100 days of gestation] were used in a completely randomized design. Ewes were individually-housed and fed either a control diet (supplemented with soybean meal) or a fishmeal supplemented diet. Blood samples were collected via jugular venipuncture for plasma fatty acids analysis on 100, 114, 128 and 142 days of gestation after morning feeding. Blood samples from the lambs were also collected via jugular venipuncture immediately after birth and before receiving their mothers’ colostrum. Plasma fatty acids were analyzed by gas-liquid chromatography. Results: The ewes from both groups, i.e., control and fishmeal supplemented, had a similar fatty acid profile prior to supplementation (at 100 days, p>0.05). Thereafter, there was an increase in eicosapentaenoic acid (20:5n3), docosahexaenoic acid, total n3-PUFA and total very long chain n3-PUFA (>C18) contents in plasma for the fishmeal supplemented ewes compared to the control (p 0.05) in total saturated fatty acids, total monounsaturated fatty acids, total conjugated linoleic acid, total trans-18:1, total cis-18:1, or total n6-PUFA contents in ewe plasma between control and fishmeal supplemented groups. Lambs born to ewes fed the fishmeal supplemented diet had greater (p<0.05) plasma concentrations of eicosapentaenoic acid (0.7 Vs 0.4, g/100 g FA), docosahexaenoic acid (1.6 Vs 0.9, g/100 g FA) and total very long chain n3-PUFA (3.3 vs. 2.0, g/100 g FA) than lambs born to ewes fed the control diet. Conclusion: The ewes supplemented with fishmeal supplementation showed a positive response with the enrichment of docosahexaenoic acid, eicosapentaenoic acid and total very long chain n3-PUFA in plasma during gestation and these fatty acids were transferred to the fetus as well.

Effect of dietary fish oil on n-3 fatty acid content of meat from broiler chickens fed low protein diets

An experiment was carried out to evaluate the effects of dietary fish oil and crude protein (CP) levels on fatty acid composition of the thigh and breast tissues of broiler chickens. Four hundred and fifty, 1-day-old mixed-sex broiler chicks were used in a completely randomised design with factorial arrangement consisting of two dietary CP levels (21 v. 18.0% and 19 v. 17.1% for grower and finisher diets, respectively), and three levels of fish oil inclusion (0, 2 or 4%). Specific increases in eicosapentaenoic acid (EPA, 20 : 5) and docosahexaenoic acid (DHA, 22 : 6) were observed in breast and thigh tissues as a response to increased fish oil supplementation. The n-6 : n-3 fatty acid ratio in breast and thigh meat samples decreased (P &lt; 0.05) in birds fed low protein diets, but dietary protein level led to no alteration in the total n-3 fatty acids of the tissues (P &gt; 0.05). Thigh tissue of chickens fed low protein diets had a higher concentration of DHA (P &lt; 0.05), while the DHA and EPA concentrations in breast tissue were not affected by dietary protein level (P &gt; 0.05). Results of the present study showed that fish oil can be used to fortify EPA and DHA levels in chicken meat and there is an interaction between dietary fatty acids and protein level on meat oxidative stability, and a reduction in dietary protein level may lead to a better oxidative stability of chicken meat.

Highly Unsaturated Fatty Acid (HUFA) Retention in the Freshwater Cladoceran, Moina macrocopa, Enriched With Lipid Emulsions

Fatty acid profiles of Moina macrocopa enriched with lipids from inexpensive squid or canola oil were compared with profiles of Moina enriched with a commercial enrichment diet, A1 DHA Selco® (ADS). Arachidonic acid (AA) significantly increased from &lt;0.01 to 0.28 mg/g body weight in Moina enriched with ADS at 1 g/l for 12 h and to 0.38 mg/g in Moina enriched in 2 g/l canola oil for 12 h. The highest increase in eicosapentaenoic acid (EPA; from 0.15 to 0.38 mg/g) was obtained when Moina were enriched in 1 g/l ADS for 12 h. The highest increases in docosahexaenoic acid (DHA; originally &lt;0.01) were obtained in Moina enriched in 1 g/l ADS for 12 h (increased to 0.09) or in 2 g/l ADS for 24 h (rose to 0.8 mg/g). The AA:EPA ratio was highest in Moina treated with 2 g/l canola oil for 12 h or 2 g/l squid oil for 24 h. The AA:DHA ratio was highest in Moina enriched with 2 g/l canola oil for 12 h while the DHA:EPA ratio was highest in Moina enriched with 2 g/l squid oil for 24 h.

A Mediterranean-style, low–glycemic-load diet decreases atherogenic lipoproteins and reduces lipoprotein (a) and oxidized low-density lipoprotein in women with metabolic syndrome

The objective was to assess the impact of a Mediterranean-style, low–glycemic-load diet (control group, n = 41) and the same diet plus a medical food (MF) containing phytosterols, soy protein, and extracts from hops and Acacia (MF group, n = 42) on lipoprotein atherogenicity in women with metabolic syndrome. Plasma lipids, apolipoproteins (apos), lipoprotein subfractions and particle size, low-density lipoprotein (LDL) oxidation, and lipoprotein (a) were measured at baseline, week 8, and week 12 of the intervention. Three-day dietary records were collected at the same time points to assess compliance. Compared with baseline, women decreased energy intake from carbohydrate (P < .001) and fat (P < .001), whereas they increased energy intake from protein (P < .001). A significant increase in energy from monounsaturated fatty acids was also observed as well as increases in eicosapentaenoic acid and docosahexaenoic acid, whereas trans-fatty acid intake was reduced (P < .00001). The atherogenic lipoproteins, large very low-density lipoprotein (P < .0001) and small LDL (P < .0001), were reduced, whereas the ratio of large high-density lipoprotein to smaller high-density lipoprotein particles was increased (P < .0001). Apolipoprotein B was reduced for all women (P < .0001), with a greater reduction in the MF group (P < .025). Oxidized LDL (P < .05) and lipoprotein (a) (P < .001) were reduced in both groups at the end of the intervention. Consumption of a Mediterranean-style diet reduces the risk for cardiovascular disease by decreasing atherogenic lipoproteins, oxidized LDL, and apo B. Inclusion of an MF may have an additional effect in reducing apo B.

Whole Grain Products, Fish and Bilberries Alter Glucose and Lipid Metabolism in a Randomized, Controlled Trial: The Sysdimet Study

Background Due to the growing prevalence of type 2 diabetes, new dietary solutions are needed to help improve glucose and lipid metabolism in persons at high risk of developing the disease. Herein we investigated the effects of low-insulin-response grain products, fatty fish, and berries on glucose metabolism and plasma lipidomic profiles in persons with impaired glucose metabolism.Methodology/Principal Findings Altogether 106 men and women with impaired glucose metabolism and with at least two other features of the metabolic syndrome were included in a 12-week parallel dietary intervention. The participants were randomized into three diet intervention groups: (1) whole grain and low postprandial insulin response grain products, fatty fish three times a week, and bilberries three portions per day (HealthyDiet group), (2) Whole grain enriched diet (WGED) group, which includes principally the same grain products as group (1), but with no change in fish or berry consumption, and (3) refined wheat breads (Control). Oral glucose tolerance, plasma fatty acids and lipidomic profiles were measured before and after the intervention. Self-reported compliance with the diets was good and the body weight remained constant. Within the HealthyDiet group two hour glucose concentration and area-under-the-curve for glucose decreased and plasma proportion of (n-3) long-chain PUFAs increased (False Discovery Rate p-values <0.05). Increases in eicosapentaenoic acid and docosahexaenoic acid associated curvilinearly with the improved insulin secretion and glucose disposal. Among the 364 characterized lipids, 25 changed significantly in the HealthyDiet group, including multiple triglycerides incorporating the long chain (n-3) PUFA.Conclusions/SignificanceThe results suggest that the diet rich in whole grain and low insulin response grain products, bilberries, and fatty fish improve glucose metabolism and alter the lipidomic profile. Therefore, such a diet may have a beneficial effect in the efforts to prevent type 2 diabetes in high risk persons.Trial RegistrationClinicalTrials.gov NCT00573781

The Effects of an Oral Supplement Enriched With Fish Oil, Prebiotics, and Antioxidants on Nutrition Status in Crohn's Disease Patients

Research in the treatment of Crohn's disease (CD) supports anti-inflammatory benefits of n-3 fatty acids from fish oil, prebiotics, and antioxidants. A nutritionally balanced inflammatory bowel disease nutrition formula (IBDNF) enriched with these compounds has the potential to improve nutrition status and disease activity in CD. This is an open-label pilot study investigating the effects of IBDNF on nutrition status in CD patients. Twenty-eight patients with active CD on stable medication were asked to consume 16 oz of IBDNF/d for 4 months. Nutrition status was assessed with dual-energy X-ray absorptiometry scans and serum micronutrient levels. Disease activity and quality of life were measured using the Crohn's Disease Activity Index (CDAI) and the Inflammatory Bowel Disease Questionnaire (IBDQ). Twenty patients completed the final visit. After 4 months, there was a significant decrease in plasma phospholipid levels of arachidonic acid with increases in eicosapentaenoic acid (EPA) and docosahexaenoic acid. Ten patients had a final EPA concentration of >2%. There was improvement in fat-free and fat mass in patients with final EPA >2% (P = .014 and P = .05). Vitamin D (25-OH) levels improved in all patients (18.5-25.9 ng/mL, P < .001). Those with EPA >2% had significantly lower CDAI (116 ± 94.5 vs 261.8 ± 86.5; P = .005) and higher IBDQ (179.1 ± 26.6 vs 114.6 ± 35.9, P < .001) compared to those with EPA <2%. IBDNF has the potential to deposit fat-free and fat mass, improve vitamin D status, and improve quality of life in CD patients.

Effect of omega-3 fatty acids alone and in combination with raloxifene on biomarkers of breast cancer risk in postmenopausal healthy women at high risk.

e11036 Background: Prevention represents the most effective approach to reducing mortality from breast cancer. To translate our preclinical findings in this regard we are conducting a 2-year randomized clinical trial in healthy postmenopausal women with increased breast cancer risk based on breast density (>25%). Methods: We are studying the effects of omega-3 fatty acids (FA) alone and in combination with raloxifene on several biomarkers including those of inflammation (CRP, IL-6), estrogen metabolism (2-hydroxyestrone, 16-α-hydroxyestrone), oxidative status (glutathione [GSH]), IGF-1 signaling (IGF-1, IGFBP-3) and plasma FA. Our study consists of 5 groups: (1) no intervention; (2) raloxifene 60mg; (3) raloxifene 30mg; (4) omega-3-acid ethyl esters (Lovaza) 4g; (5) raloxifene 30mg + Lovaza 4g. We report here preliminary findings from the first 46 women after 1 year. Results: Raloxifene reduced serum IGF-1 (12% in Group 3 and 20% in Group 2) compared to control (Group 1) in a dose-dependent fashion (p=0.005) while Lovaza alone had no effect (Group 4). The effect of Lovaza in combination with raloxifene (Group 5) was comparable to raloxifene alone (Group 3). Serum CRP, IL-6, IGFBP-3, 2-hydroxyestrone and 16-α-hydroxyestrone were unchanged in all groups. Blood GSH showed an increasing trend with Lovaza, but was not significant (p=0.07). Plasma FA changes by Lovaza included 2.5-fold increases in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), an increase in the omega-3/omega-6 ratio (0.12 + 0.01 at baseline to 0.30 + 0.06 at 1 year) and decreased arachidonic acid (AA) (0.54 + 0.05 μmol/mL at baseline to 0.35 + 0.02 at 1 year) (p<0.05). Conclusions: Our results support the role of decreased IGF-1 levels as a potential mechanism of breast cancer risk reduction by raloxifene which may not be driven by effects on IGFBP-3. The Lovaza-induced increase in omega-3 FA levels and decrease in AA are consistent with our preclinical findings. An increased trend of blood GSH by Lovaza may indicate enhanced antioxidant activity. Collectively, these results suggest that the combination of omega-3 FA and raloxifene may be a promising approach for impacting critical targets for breast cancer prevention.

Increase of Eicosapentaenoic Acid in Thraustochytrids through Thraustochytrid Ubiquitin Promoter-Driven Expression of a Fatty Acid Δ5 Desaturase Gene

Thraustochytrids, marine protists known to accumulate polyunsaturated fatty acids (PUFAs) in lipid droplets, are considered an alternative to fish oils as a source of PUFAs. The major fatty acids produced in thraustochytrids are palmitic acid (C(16:0)), n - 6 docosapentaenoic acid (DPA) (C(22:5)(n) (- 6)), and docosahexaenoic acid (DHA) (C(22:6)(n) (- 3)), with eicosapentaenoic acid (EPA) (C(20:5)(n) (- 3)) and arachidonic acid (AA) (C(20:4)(n) (- 6)) as minor constituents. We attempted here to alter the fatty acid composition of thraustochytrids through the expression of a fatty acid Δ5 desaturase gene driven by the thraustochytrid ubiquitin promoter. The gene was functionally expressed in Aurantiochytrium limacinum mh0186, increasing the amount of EPA converted from eicosatetraenoic acid (ETA) (C(20:4)(n) (- 3)) by the Δ5 desaturase. The levels of EPA and AA were also increased by 4.6- and 13.2-fold in the transgenic thraustochytrids compared to levels in the mock transfectants when ETA and dihomo-γ-linolenic acid (DGLA) (C(20:3)(n) (- 6)) were added to the culture at 0.1 mM. Interestingly, the amount of EPA in the transgenic thraustochytrids increased in proportion to the amount of ETA added to the culture up to 0.4 mM. The rates of conversion and accumulation of EPA were much higher in the thraustochytrids than in baker's yeasts when the desaturase gene was expressed with the respective promoters. This report describes for the first time the finding that an increase of EPA could be accomplished by introducing the Δ5 desaturase gene into thraustochytrids and indicates that molecular breeding of thraustochytrids is a promising strategy for generating beneficial PUFAs.