Presence Of Eicosapentaenoic Acid Research Articles

Differential Effect of Omega-3 Fatty Acids on Platelet Inhibition by Antiplatelet Drugs In Vitro.

The omega-3 polyunsaturated fatty acids (PUFAs) Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) exert multiple cardioprotective effects, influencing inflammation, platelet activation, endothelial function and lipid metabolism, besides their well-established triglyceride lowering properties. It is not uncommon for omega-3 PUFAs to be prescribed for hypertriglyceridemia, alongside antiplatelet therapy in cardiovascular disease (CVD) patients. In this regard, we studied the effect of EPA and DHA, in combination with antiplatelet drugs, in platelet aggregation and P-selectin and αIIbβ3 membrane expression. The antiplatelet drugs aspirin and triflusal, inhibitors of cyclooxygenase-1 (COX-1); ticagrelor, an inhibitor of the receptor P2Y12; vorapaxar, an inhibitor of the PAR-1 receptor, were combined with DHA or EPA and evaluated against in vitro platelet aggregation induced by agonists arachidonic acid (AA), adenosine diphosphate (ADP) and TRAP-6. We further investigated procaspase-activating compound 1 (PAC-1) binding and P-selectin membrane expression in platelets stimulated with ADP and TRAP-6. Both DHA and EPA displayed a dose-dependent inhibitory effect on platelet aggregation induced by AA, ADP and TRAP-6. In platelet aggregation induced by AA, DHA significantly improved acetylsalicylic acid (ASA) and triflusal's inhibitory activity, while EPA enhanced the inhibitory effect of ASA. In combination with EPA, ASA and ticagrelor expressed an increased inhibitory effect towards ADP-induced platelet activation. Both fatty acids could not improve the inhibitory effect of vorapaxar on AA- and ADP-induced platelet aggregation. In the presence of EPA, all antiplatelet drugs displayed a stronger inhibitory effect towards TRAP-6-induced platelet activation. Both omega-3 PUFAs inhibited the membrane expression of αIIbβ3, though they had no effect on P-selectin expression induced by ADP or TRAP-6. The antiplatelet drugs exhibited heterogeneity regarding their effect on P-selectin and αIIbβ3 membrane expression, while both omega-3 PUFAs inhibited the membrane expression of αIIbβ3, though had no effect on P-selectin expression induced by ADP or TRAP-6. The combinatory effect of DHA and EPA with the antiplatelet drugs did not result in enhanced inhibitory activity compared to the sum of the individual effects of each component.

Effect of Polyunsaturated Fatty Acids on Temozolomide Drug-Sensitive and Drug-Resistant Glioblastoma Cells.

Glioblastomas (GBMs) are notoriously difficult to treat, and the development of multiple drug resistance (MDR) is common during the course of the disease. The polyunsaturated fatty acids (PUFAs) gamma-linolenic acid (GLA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) have been reported to improve MDR in several tumors including breast, bladder, and leukaemia. However, the effects of PUFAs on GBM cell MDR are poorly understood. The present study investigated the effects of PUFAs on cellular responses to temozolomide (TMZ) in U87MG cells and the TMZ-resistant (TMZR) cells derived from U87MG. Cells were treated with PUFAs in the absence or presence of TMZ and dose-response, viable cell counting, gene expression, Western blotting, flow cytometry, gas chromatography-mass spectrometry (GCMS), and drug efflux studies were performed. The development of TMZ resistance caused an increase in ABC transporter ABCB1 and ABCC1 expression. GLA-, EPA-, and DHA-treated cells had altered fatty acid composition and accumulated lipid droplets in the cytoplasm. The most significant reduction in cell growth was seen for the U87MG and TMZR cells in the presence of EPA. GLA and EPA caused more significant effects on ABC transporter expression than DHA. GLA and EPA in combination with TMZ caused significant reductions in rhodamine 123 efflux from U87MG cells but not from TMZR cells. Overall, these findings support the notion that PUFAs can modulate ABC transporters in GBM cells.

Valorization of microalgal biomass to value-added products using integrated supercritical CO2 extraction and sub-critical hydrothermal liquefaction

The study depicts the use of different extraction techniques to utilize each fraction of microalgal consortium biomass (Chlorella sp. and Phormidium sp.) for its conversion to value-added products (VAPs). The algal biomass was rich in proteins (30.8%), lipids (28.1%), carbohydrates and essential amino acids (36.29% of total protein). The lipid yield obtained after supercritical CO2 extraction (SFE) was found to be 3.0 ± 0.4% compared to the conventional soxhlet extraction (28.2 ± 2.3%) with chloroform: methanol (1:1 v/v). However, SFE extracted lipid showed higher Polyunsaturated fatty acids (PUFAs) content (20.68%) with the presence of eicosapentaenoic acid, docosahexaenoic acid, α-linolenic acid, linoleic acid and arachidonic acid. The PUFA content in lipid obtained after soxhlet extraction was only 3.48%. The residual biomass after SFE showed recovery of 17.1 ± 0.7% proteins and 27.1 ± 0.9 mg g−1 of total carotenoids. Subcritical-hydrothermal liquefaction (HTL) of the microalgae at 230 °C and 2.7 MPa produced 30.7% biocrude (dried biomass basis) containing aldehyde/ketone (37.14%), aromatics (27.63%) and alkanes/alkenes/alkynes (15.3%) along with other compounds. This fraction consisted of VAPs targeting multiple industries-food (11%), fuel (22%), pharmaceutical (56%) and cosmeceutical (12%). A novel approach including sequential SFE-HTL process showed a lower biocrude yield (8.6%) but showed upgradation in biocrude quality having 50% aromatic compound of which >48% were pyrazines derivatives (flavoring agents for the food industry) and 20.8% pharmaceutically valuable compounds, besides extracting PUFAs from SFE. Hence, a new microalgal valorization approach has been developed by integrating two extraction techniques to produce multiple VAPs to target a minimum waste generation approach.

Using field evaluation and systematic iteration to rationalize the accumulation of omega-3 long-chain polyunsaturated fatty acids in transgenic Camelina sativa.

SummaryThe Brassicaceae Camelina sativa (gold of pleasure) is now an established niche crop and being used as a transgenic host for a range of novel seed traits. Most notable of these is the accumulation of omega‐3 long‐chain polyunsaturates such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), fatty acids normally only found in marine organisms. As part of continued efforts to optimize the accumulation of these non‐native fatty acids via seed‐specific expression of algal genes, a new series of iterative constructs was built and introduced into Camelina. Seed fatty acid composition was determined, and the presence of EPA and DHA was confirmed. To provide an additional level of evaluation, full environmental release was carried out on selected events, providing a real‐world gauntlet against which to assess the performance of these novel lines. Composition of the seed oil triacylglycerol was determined by mass spectrometry, allowing for conclusions as to the contribution of different activities to the final accumulation of EPA and DHA. Since these data were derived from field‐grown material, they also represent a robust demonstration of the stability of the omega‐3 LC‐PUFA trait in Camelina. We propose that field trialling should be routinely incorporated in the plant synthetic biology ‘design–build–test–learn’ cycle.

Dietary composition and fatty acid content of giant salmonflies ( Pteronarcys californica ) in two Rocky Mountain rivers

AbstractMany aquatic invertebrates are declining or facing extinction from stressors that compromise physiology, resource consumption, reproduction, and phenology. However, the influence of these common stressors specifically on consumer–resource interactions for aquatic invertebrate consumers is only beginning to be understood. We conducted a field study to investigate Pteronarcys californica (i.e., the “giant salmonfly”), a large‐bodied insect that is ecologically and culturally significant to rivers throughout the western United States. We sampled gut contents and polyunsaturated fatty acid composition of salmonflies to compare resource consumption across river (Madison or Gallatin, Montana), sex (male or female), and habitat (rock or woody debris). We found that allochthonous detritus comprised the majority of salmonfly diets in the Gallatin and Madison Rivers, making up 68% of the gut contents on average, followed by amorphous detritus, diatoms, and filamentous algae. Diets showed little variation across river, sex, or length. Minor differences in diets were detected by habitat type, with a higher proportion of diatoms in the diets of salmonflies collected from rocky habitat compared to woody debris. Fatty acid composition generally supported the results of gut content analysis but highlighted the importance of primary producers. The presence of eicosapentaenoic acid (20:5n‐3) and alpha linolenic acid (18:3n‐3) indicated consumption of diatoms and filamentous green algae, respectively. Our research underscores the importance of a healthy riparian zone that provides allochthonous detritus for invertebrate nutrition as well as the role of algae as an important source of fatty acids.

Scope for biodiesel and bioactive compounds production in the diatom Nitzschia punctata

The isolated diatom Nitzschia punctata was subjected to different trophic modes - autotrophy, heterotrophy, and mixotrophy and evaluated variations in growth rate, morphology, biochemical composition, and fatty acid profiles of diatoms in response to different nutritional modes. Glucose was used as a nutrient source in hetero and mixotrophic cultivation. Significant morphological variations in mucilage (jelly) stalks were found in hetero- and mixotrophically grown diatoms. The biomass yield was highest in mixotrophic diatoms (139.16 ± 3 mg/L), followed by autotrophic (127.04 ± 4 mg/L) and heterotrophically grown diatoms (95.22 ± 3 mg/L). Confocal microscopy studies using Nile red revealed significant bright yellow-gold fluorescence dispersed intermittently along with the mucilage stalks of mixotrophic and heterotrophic diatom cultures. Fourier transform infra-red (FTIR) spectroscopic studies showed a sharp absorption peak in hetero- and mixotrophic cultures at 1741 cm-1, indicating the presence of esters of fatty acids (R-C(O)-O-R). Elemental composition analysis using scanning electron microscopy revealed higher C (~48%) in mixotrophic cultivation. Fatty acid methyl esters (FAME) assessment through Gas chromatography and mass spectroscopy (GC–MS) showed higher proportions of palmitoleic acid (C16:1; ~18 – 29%) and oleic acid (C18:1; ~26 – 35%) with saturates and monounsaturates of C16 – C18 up to 90%, confirming of superior biodiesel quality property, which is desirable. The presence of Eicosapentaenoic acid (EPA) in hetero- and mixotrophic cultivation proves the scope for its potential utilisation as a source of bioactive compounds for industrial application.

Quality evaluation of Artemia cysts from three Algerian populations

The aim of this study was to determine the nutritional value of three populations of Artemia, one from the Bethioua Sebkha (Oran) and two new biotopes at El Melah (Bechar) and Timimoune (Adrar), by evaluating the cyst and nauplius biometry, hatching parameters and fatty acid profiles. This information is of relevance to aquaculture initiatives, and fills in knowledge gaps from previous research at Algerian sites. The size of nauplii varied from 453.26 ± 0.3 µm and 478.73 ± 0.2 µm. The chorion thickness was between 13.28 µm and 10.50 µm. The highest hatching percentage and hatching efficiency were obtained after decapsulation of Bethioua cysts. The shortest hatching synchrony time was also found for the Bethioua cysts. All cyst samples from the the Bethioua population analysed in this study showed a freshwater-type fatty acid profile, rich in linolenic acid (LNA, 18:3n.3), opposed to a marine-type fatty acid profile rich in presence of eicosapentaenoic acid (EPA, 20:5n-3). The Bethioua population exhibited the best hatching performance and are ideally suited for use in aquaculture.

Abstract 15015: Eicosapentaenoic Acid, but Not Docosahexaenoic Acid or a Mixed Omega-3 Fatty Acid Supplement, Inhibits Low-density Lipoprotein Oxidation in a Time-dependent Manner

Background: Eicosapentaenoic acid (EPA), an omega-3 fatty acid (O3FA), reduces oxidation of low-density lipoproteins (LDL) in patients with hypertriglyceridemia, an effect that may contribute to lower cardiovascular (CV) events as reported in the REDUCE-IT trial. By contrast, DHA-containing products have failed to show a reduction in CV events, which may be due, in part, to differences in antioxidant activity. We compared the effects of EPA versus DHA and a mixed O3FA (EPA/DHA) supplement on oxidation of human LDL in vitro . Methods: LDL was isolated from human plasma by isopycnic centrifugation, separated into test samples of 100 μg/mL, and incubated at 37°C for 30 min in the absence (vehicle) or presence of EPA, DHA, or mixed O3FA supplement at equimolar levels (2.5 μM). All samples were then subjected to copper-induced oxidation (20 μM) as measured by formation of malondialdehyde (MDA). The FA content of the O3FA supplement was measured using gas chromatography/mass spectrometry. Results: EPA significantly inhibited LDL oxidation in a time-dependent manner compared with vehicle; after 4 hours, EPA inhibited MDA levels by 96% compared with the vehicle oxidation level (0.51 ± 0.01 vs 11.4 ± 0.4 μM; p <0.001). While DHA exhibited antioxidant activity at 2 hours at a level below EPA (2.5 ± 0.1 vs 11.4 ± 0.4; p <0.001), even this level of activity was lost by 4 hours. The mixed O3FA supplement failed to show any antioxidant activity through 4 hours (11.4 ± 0.5 μM). Fatty acid analysis showed that the O3FA supplement, in addition to EPA and DHA, contained more than 30 other fatty acids, including saturated fats, that may have nullified any potential benefits. Conclusions: These data support potent LDL antioxidant effects of EPA that were sustained over time compared with DHA, which had a weaker, transient effect, or a mixed O3FA supplement, which had no beneficial effect at all. This potent antioxidant mechanism of EPA may contribute to reduced CV risks seen in REDUCE-IT compared with negative findings from trials using DHA-containing formulations.

Comparison of fatty acids profile of the gonads and eggs of Lutjanus guttatus (Perciformes: Lutjanidae) obtained from wild and captive broodstock

The proportions of fatty acids present in eggs and gonads of wild and captive Lutjanus guttatus (Steindachner, 1869) were evaluated to assist in obtaining nutritional information necessary to improve the diets provided in aquaculture production centers of this species. Samples of eggs and gonads were obtained from broodstock (1.0±0.1 kg) kept in the Pacific Marine Park (PMP), Costa Rica, and from individuals caught by fishermen in their natural environment. The samples were taken in triplicate during the period spanning from August to September 2015. Captive snappers were fed a fresh diet. Spawns and gonads were obtained from two groups of 16 specimens each of captive and wild fish, both groups having a sex ratio of 1:1 male to female. Gonads from females were selected according to their maturity stage, and male gonads were sampled based on the quantity, viscosity and color of semen. There were no statistically significant differences (p≥0.05) between egg diameters and oil droplets size; viability rates, expressed as percentages, were considered to be high and appropriate for comparison (91.2% captive, 86.1% wild). There were also no statistically significant differences in the fatty acid composition of eggs obtained from wild and captive females (p=0.2188), or in gonads from males (p=0.6179) and wild and captive females (p=0.1153). The presence of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) was observed in all of the samples analyzed, while arachidonic acid (ARA) was found in amounts ranging from 0.38 to 5.07% and was not present in the eggs of wild females or in the gonads of captive females.

Divergent Regulation of Myotube Formation and Gene Expression by E2 and EPA during In-Vitro Differentiation of C2C12 Myoblasts.

Estrogen (E2) and polyunsaturated fatty acids (n-3PUFA) supplements independently support general wellbeing and enhance muscle regeneration in-vivo and myotube formation in-vitro. However, the combined effect of E2 and n-3PUFA on myoblast differentiation is not known. The purpose of the study was to identify whether E2 and n-3PUFA possess a synergistic effect on in-vitro myogenesis. Mouse C2C12 myoblasts, a reliable model to reiterate myogenic events in-vitro, were treated with 10nM E2 and 50μM eicosapentaenoic acid (EPA) independently or combined, for 0–24 h or 0–120 h during differentiation. Immunofluorescence, targeted qPCR and next generation sequencing (NGS) were used to characterize morphological changes and differential expression of key genes involved in the regulation of myogenesis and muscle function pathways. E2 increased estrogen receptor α (Erα) and the expression of the mitogen-activated protein kinase 11 (Mapk11) within 1 h of treatment and improved myoblast differentiation and myotube formation. A significant reduction (p < 0.001) in myotube formation and in the expression of myogenic regulatory factors Mrfs (MyoD, Myog and Myh1) and the myoblast fusion related gene, Tmem8c, was observed in the presence of EPA and the combined E2/EPA treatment. Additionally, EPA treatment at 48 h of differentiation inhibited the majority of genes associated with the myogenic and striated muscle contraction pathways. In conclusion, EPA and E2 had no synergistic effect on myotube formation in-vitro. Independently, EPA inhibited myoblast differentiation and overrides the stimulatory effect of E2 when used in combination with E2.

Eicosapentaenoic acid inhibits oxidation of high density lipoprotein particles in a manner distinct from docosahexaenoic acid

The omega-3 fatty acid eicosapentaenoic acid (EPA) reduces oxidation of ApoB-containing particles in vitro and in patients with hypertriglyceridemia. EPA may produce these effects through a potent antioxidant mechanism, which may facilitate LDL clearance and slow plaque progression. We hypothesize that EPA antioxidant effects may extend to ApoA-containing particles like HDL, potentially preserving certain atheroprotective functions. HDL was isolated from human plasma and incubated at 37 °C in the absence (vehicle) or presence of EPA and/or DHA; 5.0 or 10.0 μM each. Samples were then subjected to copper-induced oxidation (10 μM). HDL oxidation was inhibited similarly by EPA and DHA up to 1 h. EPA (10 μM) maintained significant HDL oxidation inhibition of 89% (0.622 ± 0.066 μM MDA; p < .001) at 4 h, with continued inhibition of 64% at 14 h, vs. vehicle (5.65 ± 0.06 to 2.01 ± 0.10 μM MDA; p < .001). Conversely, DHA (10 μM) antioxidant benefit was lost by 4 h. At a lower concentration (5 μM), EPA antioxidant activity remained at 81% (5.53 ± 0.15 to 1.03 ± 0.10 μM MDA; p < .001) at 6 h, while DHA lost all antioxidant activity by 4 h. The antioxidant activity of EPA was preserved when combined with an equimolar concentration of DHA (5 μM each). EPA pretreatment prevented HDL oxidation in a dose-dependent manner that was preserved over time. These results suggest unique lipophilic and electron stabilization properties for EPA as compared to DHA with respect to inhibition of HDL oxidation. These antioxidant effects of EPA may enhance certain atheroprotective functions for HDL.

Influence of omega-3 fatty acids on bovine luteal cell plasma membrane dynamics

Fish oil is a rich source of omega-3 fatty acids which disrupt lipid microdomain structure and affect mobility of the prostaglandin F2α (FP) receptor in bovine luteal cells. The objectives of this study were to determine the effects of individual omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on 1) membrane fatty acid composition, 2) lipid microdomain structure, and 3) lateral mobility of the FP receptor in bovine luteal cells. Ovaries were collected from a local abattoir (n=5/experiment). The corpus luteum was resected and enzymatically digested using collagenase to generate a mixed luteal cell population. In all experiments, luteal cells were treated with 0, 1, 10 or 100μM EPA or DHA for 72h to allow incorporation of fatty acids into membrane lipids. Results from experiment 1 show that culturing luteal cells in the presence of EPA or DHA increased these luteal fatty acids. In experiment 2, both EPA and DHA increased spatial distribution of lipid microdomains in a dose-dependent manner. Single particle tracking results from experiment 3 show that increasing both EPA and DHA concentrations increased micro- and macro-diffusion coefficients, increased domain size, and decreased residence time of FP receptors. Collectively, results from this study demonstrate similar effects of EPA and DHA on lipid microdomain structure and lateral mobility of FP receptors in cultured bovine luteal cells. Moreover, only 10μM of either fatty acid was needed to mimic the effects of fish oil.

Promoting apoptosis effect of polyunsaturated fatty acids in prostate adenocarcinoma cells

Objective To detect whether deep sea fish oil, eicosapentaenoic acid (EPA) and docosahex-aenoic acid (DHA) in n-3 polyunsaturated fatty acids (PUFAs) and arachidonic acid (AA) in n-6 PUFAs play an important role in benign prostatic hyperplasia (BPH) cells and different prostate cancer (PCa) cells. Methods Using methyl thiazol tetrazolium (MTT) assay, the effects of Fish oil, EPA, DHA and AA on the survival rate of PCa cells and prostate cells were tested. Western blotting and real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) were used to detect the gene expression level of cysteinyl aspartate-specific protease (Caspase)-3. Results After the PCS-440-010 cells were incubated with different concentrations of Fish oil for 24 h, survival rate had no statistically significant difference as compared with control group. After PC-3 and DU145 cells were incubated with different concentrations of Fish oil for 24 h, the survival rate decreased by 27% and 40% (P<0.05) as compared with control group. After LNCaP cells were incubated with different concentrations of Fish oil for 24 h, survival rate had no significant change. After PC-3 (decreased by 13% and 26%) and DU145 (decreased by 16% and 30%) cells were incubated cells were incubated in the presence of EPA or DHA for 24 h, the survival rate was decreased obviously as compared with control group (P<0.05). After DU145 cells were incubated with AA for 24 h, the survival rate had no significantl changes. After DU145 cells were incubated with DHA 50 μmol/L for 12, 24, 48 and 72 h, there was time-dependent between the survival rate of DU145 cells and DHA stimulus. DHA could increase the expression of Caspase-3 mRNA in a concentration-dependent manner in DU145 cells. Conclusion DHA can reduce the survival rate of PCa cells concentration- and time-dependently, but had no effects on benign prostate cells. Key words: Prostatic adenocarcinoma; Androgen resistance prostate cancer; Polyunsaturated fatty acids; Docosahex-aenoic acid

The influence of eicosapentaenoic acid and docosahexaenoic acid on expression of genes connected with metabolism and secretory functions of ageing 3T3-L1 adipocytes

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are n-3 long chain polyunsaturated fatty acids. The purpose of our study was to evaluate the influence of EPA and DHA on expression of genes connected with metabolism and secretory functions of ageing adipocytes. Young, mature and old differentiated 3T3-L1 adipocytes were cultured for 48h in the presence of EPA, or DHA. Both fatty acids increased the expression of Pparg, FATP1, FATP4 and ATGL genes, but only in young 3T3-L1 adipocytes. Moreover, in young, mature and old cells DHA elevated the expression of CPT1 gene. In addition, EPA and DHA enhanced the expression of leptin, adiponectin and apelin genes only in young cells. Investigated fatty acids changed mRNA levels of IL6 and MCP1 in young, mature and old cells. EPA increased the expression of these two genes, whereas DHA decreased it. Furthermore, EPA and DHA treatment changed the expression of IRS1 and GLUT4 genes involved in insulin signalling pathway, but their effects were opposite. Expression of these genes was decreased by EPA and increased by DHA in young, mature and old cells. In summary, the investigated fatty acids are able to affect the expression of genes associated with lipid metabolism, secretory functions and insulin resistance in ageing 3T3-L1 adipocytes, but their impact is age-dependant. Young cells seem to be more sensitive to EPA and DHA than mature and old ones. Furthermore, the effect of these two fatty acids is not always identical, and therefore requires further investigation.

Long-Chain Omega-3 Polyunsaturated Fatty Acids Have Developmental Effects on the Crop Pest, the Cabbage White Butterfly Pieris rapae.

Nutritional enhancement of crops using genetic engineering can potentially affect herbivorous pests. Recently, oilseed crops have been genetically engineered to produce the long-chain omega-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) at levels similar to that found in fish oil; to provide a more sustainable source of these compounds than is currently available from wild fish capture. We examined some of the growth and development impacts of adding EPA and DHA to an artificial diet of Pieris rapae, a common pest of Brassicaceae plants. We replaced 1% canola oil with EPA: DHA (11:7 ratio) in larval diets, and examined morphological traits and growth of larvae and ensuing adults across 5 dietary treatments. Diets containing increasing amounts of EPA and DHA did not affect developmental phenology, larval or pupal weight, food consumption, nor larval mortality. However, the addition of EPA and DHA in larval diets resulted in progressively heavier adults (F 4, 108 = 6.78; p = 0.011), with smaller wings (p < 0.05) and a higher frequency of wing deformities (R = 0.988; p = 0.001). We conclude that the presence of EPA and DHA in diets of larval P. rapae may alter adult mass and wing morphology; therefore, further research on the environmental impacts of EPA and DHA production on terrestrial biota is advisable.

Influence of an n-3 long-chain polyunsaturated fatty acid-enriched diet on experimentally induced synovitis in horses.

Dietary n-3 long-chain polyunsaturated fatty acid (LCPUFA) supplementation has previously been shown to modify joint-related inflammation in several species, although information in the horse is lacking. We investigated whether dietary supplementation with n-3 LCPUFA would modify experimentally induced synovitis in horses. Twelve, skeletally mature, non-pregnant mares were randomly assigned to either a control diet (CONT) or an n-3 long-chain fatty acid-enriched treatment diet (N3FA) containing 40g/day of n-3 LCPUFA for 91days. Blood samples taken on days 0, 30, 60 and 90, and synovial fluid collected on days 0 and 90 were processed for lipid composition. On day 91, joint inflammation was stimulated using an intra-articular (IA) injection of 100ng of recombinant equine IL-1beta (reIL-1β). Synovial fluid samples taken at post-injection hours (PIH) 0, 4, 8 and 24 were analysed for prostaglandin E2 (PGE2 ), matrix metalloproteinase (MMP) activity and routine cytology. Synovium and articular cartilage samples collected at PIH 8 were analysed for gene expression of MMP 1 and MMP 13, interleukin-1beta (IL-1β), cyclooxygenase 2 (COX-2), tumour necrosis factor-alpha and the aggrecanases, a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5. A 90-day feeding period of n-3 LCPUFA increased serum phospholipid and synovial fluid lipid compositions of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) compared to CONT horses. The reIL-1β injection caused an inflammatory response; however, there was no effect of dietary treatment on synovial fluid PGE2 content and MMP activity. Synovial tissue collected from N3FA horses exhibited lower expression of ADAMTS-4 compared to CONT horses. Despite the presence of EPA and DHA in the synovial fluid of N3FA horses, dietary n-3 LCPUFA supplementation did not modify synovial fluid biomarkers compared to CONT horses; however, the lower ADAMTS-4 mRNA expression in N3FA synovium warrants further investigation of n-3 LCPUFA as a joint therapy.

The metabolism of 8-heptadecene in Pyropia (Bangiaceae, Rhodophyta)

The metabolic pathway of 8-heptadecene in red algae was investigated. The results showed that the amounts of 8-heptadecene in the primitive bangiophycidean red algae Pyropia and Bangia were 30–50 % of the volatile compounds, much higher than that in the green alga Ulva pertusa, the brown alga Sargassum thunbergii and the florideophycidean red alga Gracilaria lemaneiformis. Studies on the metabolism of 8-heptadecene in Pyropia found that its enzymatic system has no significant catalytic activity on palmitic acid, oleic acid, linoleic acid, linolenic acid, and arachidonic acid. However, the isolated enzymatic solution showed activity in the presence of eicosapentaenoic acid. This activity produced about four times the amount of 8-heptadecene compared with other substrates and the control, indicating the solution had a specific catalytic function for eicosapentaenoic acid. Furthermore, the enzyme solution was strongly inhibited by NaN3 but not by phenidone and phenanthroline suggesting that the enzyme is structurally related to heme protein. Thus, it is believed that a constant amount of 8-heptadecene is maintained in the primitive red alga Pyropia and the 8-heptadecene is a metabolite of eicosapentaenoic acid and may catalyzed by enzymes including a heme lipoxygenase-like enzyme.

Abstract 585: Eicosapentaenoic acid increases mammary tumor inhibition of intermittent calorie restriction and regulates adipokines

Abstract It has become clear that the use of combination therapies for prevention of breast cancer may be more efficacious than individual therapies. Therefore, we combined two different interventions which had each been shown alone to inhibit mammary tumorigenesis but had not been tested in combination. The first intervention was consumption of the omega-3 fatty acid eicosapentaenoic acid (EPA) which is found in fish oil and the second intervention was calorie restriction. We utilized six groups of MMTV-Her2/neu mice as a model for human breast cancers which over express Her2/neu. Starting at 10 weeks of age half of the mice were fed a control diet composed of a modified AIN-93M formula with fat derived from soy oil. The other half consumed a diet with 71.75% of fat calories from EPA. Mice were further divided into ad libitum (AL), chronic calorie restricted (CCR) or intermittent caloric restricted (ICR) groups. AL groups (Control and EPA) received unrestricted access to their diets. CCR groups were given 75% of the total calories that the AL age-matched groups consumed. ICR groups were fed calories equal to 100% of the AL age-matched groups for three weeks followed by three weeks of calories equal to 50% of the AL age-matched groups. The 6 week cycles of restriction and refeeding were maintained until the mice were 60 weeks of age or were euthanized due to mammary tumors (MTs). MT incidence was lowest in the ICR-EPA group (15%) and highest in the AL-Con group (87%) while the AL-EPA, CCR-Con, CCR-EPA and ICR-Con had MT incidence rates of 63%, 47%, 40% and 59%, respectively The average survival until terminal end point was also longest in the ICR-EPA group at 59.2 weeks and shortest in the AL-Con group at 52.6 (P&amp;lt;0.0002). The average tumor burden was 0.98 grams for ICR-EPA mice and 1.78 grams for AL-Con mice but due to the very small number of MTs in the ICR-EPA group this was not a significant difference. Histopathology showed no significant difference in tumor grades between the groups. To better understand the effects of the interventions we examined serum for alterations in adipokines and growth factors. We found that EPA consumption dramatically reduced the levels of leptin (P&amp;lt;0.02) and increased the levels of adiponectin in the AL mice (P&amp;lt;0.001) but did not significantly alter animal weights at 25 weeks of age. Calorie restriction was also able to significantly decrease serum leptin levels and increase serum adiponectin levels. Serum IGF-I was significantly reduced by calorie restriction with the ICR-Con having the lowest level. The presence of EPA in the diet did not significantly change the levels of IGF-I. These results illustrate that MT inhibition is significantly increased when ICR and EPA are combined as compared to either intervention alone or no intervention. This effect may be related to alterations in serum growth factors and adipokines. Support: Susan G.Komen for the Cure and The Hormel Foundation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 585. doi:1538-7445.AM2012-585

Eicosapentaenoic acid protects against 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced hepatic toxicity in cultured rat hepatocytes

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a persistent and ubiquitous environmental contaminant. The health impact of TCDD exposure is of great concern to the general public. Recent reports have implied that eicosapentaenoic acid (EPA) might be a potential chemopreventive agent and influence hepatotoxicity. The aim of the current study was to explore the effectiveness of EPA in alleviating the toxicity of TCDD on primary cultured rat hepatocytes. EPA (5, 10 and 20μM) was added to cultures alone or simultaneously with TCDD (5 and 10μM). Rat hepatocytes were treated with TCDD and EPA for 48h, and then cytotoxicity was detected by [3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide] (MTT) assay and lactate dehydrogenase (LDH) release, while total antioxidant capacity (TAC) and total oxidative stress (TOS) levels were determined to evaluate the oxidative injury. The DNA damage was also analyzed by liver micronucleus assay (LMN) and 8-oxo-2-deoxyguanosine (8-OH-dG). The results of MTT and LDH assays showed that TCDD but not EPA decreased cell viability. TCDD also increased TOS level and significantly decreased TAC level in rat hepatocytes in a clear dose dependent manner. On the basis of increasing doses, the dioxin caused significant increases of micronucleated hepatocytes (MNHEPs) and 8-OH-dG as compared to control culture. Whereas, in cultures treated with EPA alone, TOS level did not change and the level of TAC significantly increased. The presence of EPA with TCDD minimized the toxic effects of the dioxin on primary hepatocytes cultures. Noteworthy, EPA has a protective effect against TCDD-mediated DNA damages.

Erratum: Lipid synthesized by micro‐algae grown in laboratory‐ and industrial‐scale bioreactors

In the above-mentioned paper the strain referred to as Nannochloropsis oculata was a not further described Chlorella strain. Therefore, the data presented in Figures 1, 2 and 3, as well as in Table 3, refer to Chlorella (?). and not N. oculata. The Abstract of the paper now reads as follows: Tetraselmis sp. and Chlorella (?), cultivated in industrial-scale bioreactors, produced 2.33 and 2.44% w/w lipid (calculated as the sum of fatty acid methyl esters) in dry biomass, respectively. These lipids contained higher amounts of neutral lipids and glycolipids plus sphingolipids, than phospholipids. Lipids of Tetraselmis sp. were characterized by the presence of eicosapentaenoic acid (that was located mainly in phospholipids), and octadecatetraenoic acid (that was equally distributed among lipid fractions), while these fatty acids were completely absent in Chlorella (?) lipids. Additionally, lipids produced by 16 newly isolated strains from Greek aquatic environments (cultivated in flask reactors) were studied. The highest percentage of lipids was found in Prorocentrum triestinum (3.69% w/w) while the lowest in Prymnesium parvum (0.47% w/w). Several strains produced lipids rich in eicosapentaenoic and docosahexaenoic acids. For instance, docosahexaenoic acid was found in high percentages in lipids of Amphidinium sp. S1, P. parvum, Prorocentrum minimum and P. triestinum, while lipids produced by Asterionella sp. (?) S2 contained eicosapentaenoic acid in high concentration. These lipids, containing ω-3-long-chain polyunsaturated fatty acids, have important applications in the food and pharmaceutical industries and in aquaculture. The authors apologize for this mistake and any inconvenience caused.