Metabolic Conversion of C20 Polymethylene-Interrupted Polyunsaturated Fatty Acids to Essential Fatty Acids

Polymethylene-interrupted (PMI)-polyunsaturated fatty acids (PUFA) are fatty acids present largely in gymnosperm. Sciadonic acid (SciA, 20:3 Δ-5,11,14) and juniperonic acid (JA, 20:4 Δ-5,11,14,17) are typical C20 PMI-PUFA with an isolated double bond at Δ5. Previously, we found that SciA and JA are converted to linoleic acid (LNA) and α-linolenic acid (ΑLA), respectively. The conversion process includes chain-shortening step by peroxisomal β-oxidation for elimination a double bond at Δ5, and subsequent chain-elongation step in microsomes. In this study, we examined the substrate specificity of this metabolism in rodent and human cells. Supplementation of SciA, eicosadienoic acid (EDA, 20:2 Δ-11,14) or JA to CHO-K1 cells (wild type) induced an accumulation of LNA, LNA or ALA, respectively, in cellular lipids. These changes were not observed in the peroxisomes-deficient CHO cells, indicating involvement of peroxisomes in the metabolism. Two types of human cells (MKN74 and HepG2) also converted the C20 PMI-PUFA and EDA to the respective essential fatty acids. In contrast, no chain-shortened metabolite of pinolenic acid (18:3 Δ-5,9,12) was detected in any cell lines tested. From these results, C20 PMI-PUFA and EDA, but not C18 PMI-PUFA, are suggested as being effectively converted to essential fatty acids by the fatty acid remodeling system in rodent and human cells.     

Gut Microbial Metabolites of Polyunsaturated Fatty Acids Correlate with Specific Fecal Bacteria and Serum Markers of Metabolic Syndrome in Obese Women

The aim of this human study was to assess the influence of prebiotic-induced gut microbiota modulation on PUFA-derived bacterial metabolites production. Therefore, we analyzed the circulating fatty acid profile including CLA/CLnA in obese women treated during 3 months with inulin-type fructan prebiotics. In these patients, we had already determined gut microbiota composition by phylogenetic microarray and qPCR analysis of 16S rDNA. Some PUFA-derived bacterial metabolites were detected in the serum of obese patients. Despite the prebiotic-induced modulation of gut microbiota, including changes in CLA/CLnA-producing bacteria, the treatment did not impact significantly on the circulating level of these metabolites. However, some PUFA-derived bacterial metabolites were positively correlated with specific fecal bacteria (Bifidobacterium spp., Eubacterium ventriosum and Lactobacillus spp.) and inversely correlated with serum cholesterol (total, LDL, HDL). These correlations suggest a potential beneficial effect of some of these metabolites but this remains to be confirmed by further investigation.     

Plasma Levels of 14:0, 16:0, 16:1n-7, and 20:3n-6 are Positively Associated,but 18:0 and 18:2n-6 are Inversely Associated with Markers of Inflammation in Young Healthy Adults

Inflammation is a recognized risk factor for the development of chronic diseases, such as type 2 diabetes and atherosclerosis. Evidence suggests that individual fatty acids (FA) may have distinct influences on inflammatory processes. The goal of this study was to conduct a cross-sectional analysis to examine the associations between circulating FA and markers of inflammation in a population of young healthy Canadian adults. FA, high-sensitivity C-reactive protein (hsCRP), and cytokines were measured in fasted plasma samples from 965 young adults (22.6 ± 0.1 years). Gas chromatography was used to measure FA. The following cytokines were analyzed with a multiplex assay: regulated upon activation normal T cell expressed and secreted (RANTES/CCL5), interleukin 1-receptor antagonist (IL-1Ra), interferon-γ (IFN-γ), interferon-γ inducible protein 10 (IP-10), and platelet-derived growth factor β (PDGF-ββ). Numerous statistically significant associations (p < 0.05, corrected for multiple testing) were identified between individual FA and markers of inflammation using linear regression. Myristic (14:0), palmitic (16:0), palmitoleic (16:1n-7), and dihomo-γ-linolenic (20:3n-6) acids were positively associated with all markers of inflammation. In contrast, stearic acid (18:0) was inversely associated with hsCRP and RANTES, and linoleic acid (18:2n-6) was inversely associated with hsCRP, RANTES and PDGF-ββ. In conclusion, our results indicate that specific FA are distinctly correlated with various markers of inflammation. Moreover, the findings of this study suggest that FA profiles in young adults may serve as an early indicator for the development of future complications comprising an inflammatory component.     

Higher Serum EPA or DHA, and Lower ARA Compositions with Age Independent Fatty Acid Intake in Japanese Aged 40 to 79

Docosahexaenoic acid (DHA) and arachidonic acid (ARA) are the predominant long-chain polyunsaturated fatty acids (PUFA) among membrane phospholipids in the mammalian brain and neural tissues. This cross-sectional study examined age effects on serum eicosapentaenoic acid (EPA), DHA, and ARA compositions assessed with reference to dietary intakes among 1,014 Japanese men and 1,028 Japanese women aged 40–79 years. Venous blood was collected early in the morning after at least 12-h fasting. Serum fatty acid (FA) compositions were expressed as molar percentages of the total FA (mol% of total). Diet was assessed using a 3-day dietary record that included photographs. Participants were categorized into groups by sex and age (40–49, 50–59, 60–69, and 70–79 years). Intakes of fish, EPA, and DHA tended to increase with age. Significant positive correlations between serum FA composition and the corresponding weight percentage of total FA intake were observed for EPA and DHA in all sex and age groups, and for ARA among females in their 40s. Serum EPA and DHA compositions were higher, while ARA decreased with age, and these associations remained consistent even after adjusting for corresponding FA intake. These results suggest potential effects of age on differences in blood EPA, DHA, and ARA compositions, independent of corresponding FA intake among community-dwelling Japanese men and women.     

Dietary Fatty‐Acid Compositions Are more Strongly Reflected in Fatty than Lean Dorsal Fillets of Common Carp (Cyprinus carpio L.)

In this fish‐feeding study, we tested similarity patterns between fatty acids (FA) in diets and common carp (Cyprinus carpio) of fish ponds used for semi‐intensive aquaculture, containing naturally occurring pond zooplankton and different feeds (marine or terrestrial feeds) until carp reached market size. We evaluated if and how total lipid contents in dorsal fillets can reflect dietary FA compositions in farm‐raised common carp and hypothesized that increasing total lipid contents in dorsal fillets significantly increase the similarity between dietary and dorsal fillets’ FA compositions. Results of this study showed that carps had higher total lipids when supplied with marine feeds and dietary FA compositions were indeed more strongly reflected in fatty (i.e. high total lipid contents) than in leaner dorsal fillets (low total lipid contents). Increasing total lipid contents in dorsal fillets significantly increased the similarity between the dietary and dorsal fillets’ FA compositions. In contrast, leaner dorsal fillets had FA patterns that were more distinct from dietary FA. Total lipid contents higher than ~60 mg/g dry weight in dorsal fillets had only limited effects on increasing the similarity between FA compositions of diets and dorsal fillets, and were independent of feed sources. It is thus suggested that higher total lipid contents in dorsal fillets can be used as a proxy to predict dietary FA profiles in common carps, or perhaps even in farm‐raised fish in general.     

Identification of n‐6 Monounsaturated Fatty Acids in Acer Seed Oils

Seed oils from Acer species are a potential source of the nutraceutical fatty acids, nervonic acid (cis‐15‐tetracosenoic acid, NA), and γ‐linolenic acid (cis‐6,9,12‐octadecatrienoic acid, GLA). To further characterize the genus, seed fatty acid content and composition were determined for 20 species of Acer. Fatty acid content ranged from 8.2% for Acer macrophyllum to over 36% for A. mono and A. negundo. The presence of very‐long‐chain fatty acids (VLCFA), with chain length of 20‐carbons or greater, and GLA were characteristic features of the seed oils. In all species, erucic acid (cis‐13‐docosenoic acid, EA) was the predominant VLCFA with the highest level of NA being only 8.6% in A. olivianum. Regioselective lipase digestion demonstrated that VLCFA are largely absent from the sn‐2 position of seed triacylglycerol, whereas GLA is primarily located at this position. Five Acer species contained low levels (<2%) of cis‐12‐octadecenoic acid and cis‐14‐eicosenoic acid, uncommon n‐6 fatty acids not previously reported from Acer.     

Short‐Chain Fatty Acids Enhance the Lipid Accumulation of 3T3‐L1 Cells by Modulating the Expression of Enzymes of Fatty Acid Metabolism

Short‐chain fatty acids (SCFA) such as acetic acid, propionic acid, and butyric acid are produced by fermentation by gut microbiota. In this paper, we investigate the effects of SCFA on 3T3‐L1 cells and the underlying molecular mechanisms. The cells were treated with acetic acid, propionic acid, or butyric acid when cells were induced to differentiate into adipocytes. MTT assay was employed to detect the viability of 3T3‐L1 cells. Oil Red O staining was used to visualize the lipid content in 3T3‐L1 cells. A triglyceride assay kit was used to detect the triacylglycerol content in 3T3‐L1 cells. qRT‐PCR and Western blot were used to evaluate the expression of metabolic enzymes. MTT results showed that safe concentrations of acetic acid, propionic acid, and butyric acid were less than 6.4, 3.2, and 0.8 mM, respectively. Oil Red O staining and triacylglycerols detection results showed that treatment with acetic acid, propionic acid, and butyric acid accelerated the 3T3‐L1 adipocyte differentiation. qRT‐PCR and Western blot results showed that the expressions of lipoprotein lipase (LPL), adipocyte fatty acid binding protein 4 (FABP4), fatty acid transporter protein 4 (FATP4), and fatty acid synthase (FAS) were significantly increased by acetic acid, propionic acid, and butyric acid treatment during adipose differentiation (p < 0.05). In conclusion, SCFA promoted lipid accumulation by modulating the expression of enzymes of fatty acid metabolism.     

SNPs of the FADS Gene Cluster are Associated with Polyunsaturated Fatty Acids in a Cohort of Patients with Cardiovascular Disease

Polymorphisms of the human Δ-5 (FADS1) and Δ-6 (FADS2) desaturase genes have been recently described to be associated with the level of several long-chain n-3 and n-6 polyunsaturated fatty acids (PUFAs) in serum phospholipids. We have genotyped 13 single nucleotide polymorphisms (SNPs) located on the FADS1–FADS2–FADS3 gene cluster (chromosome 11q12–13.1) in 658 Italian adults (78% males; mean age 59.7 ± 11.1 years) participating in the Verona Heart Project. Polymorphisms and statistically inferred haplotypes showed a strong association with arachidonic acid (C20:4n-6) levels in serum phospholipids and in erythrocyte cell membranes (rs174545 adjusted P value for multiple tests, P < 0.0001 and P < 0.0001, respectively). Other significant associations were observed for linoleic (C18:2n-6), alpha-linolenic (C18:3n-3) and eicosadienoic (C20:2n-6) acids. Minor allele homozygotes and heterozygotes were associated to higher levels of linoleic, alpha-linolenic, eicosadienoic and lower levels of arachidonic acid. No significant association was observed for stearidonic (C18:4n-3), eicosapentaenoic (C20:5n-3) and docosahexaenoic (C22:6n-3) acids levels. The observed strong association of FADS gene polymorphisms with the levels of arachidonic acid, which is a precursor of molecules involved in inflammation and immunity processes, suggests that SNPs of the FADS1 and FADS2 gene region are worth studying in diseases related to inflammatory conditions or alterations in the concentration of PUFAs. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Hypolipidemic Effects of Phospholipids (PL) Containing n-3 Polyunsaturated Fatty Acids (PUFA) Are Not Dependent on Esterification of n-3 PUFA to PL

Phospholipids (PL) containing n‐3 polyunsaturated fatty acids (PUFA) have beneficial effects of maintaining and promoting health compared with triacylglycerols (TAG) containing n‐3 PUFA or general PL. This study evaluated the effects of dietary PL containing n‐3 PUFA and elucidated the effects of the glycerophosphate structure and n‐3 PUFA on fatty acid (FA) metabolism in rats. Rats were fed a basal diet containing soybean oil alone, TAG containing n‐3 PUFA (1.8 %), soybean PL (2.7 %), PL containing n‐3 PUFA (2.7 %), or TAG containing n‐3 PUFA (1.8 %) + soybean PL (2.7 %). The present n‐3 PUFA‐supplemented diets had similar FA compositions, and the PL diets had similar PL compositions. TAG containing n‐3 PUFA reduced serum TAG contents, but did not affect serum cholesterol contents compared with soybean oil alone. PL diets containing n‐3 PUFA and the combination of TAG containing n‐3 PUFA and soybean PL resulted in decreased serum and liver TAG contents compared with the diet containing soybean oil alone, reflecting enhanced liver FA β‐oxidation. The results of this study show that TAG containing n‐3 PUFA with added soybean PL affects serum and liver TAG and cholesterol contents to a similar degree as PL containing n‐3 PUFA. TAG containing n‐3 PUFA and soybean PL are widely used as functional food ingredients and pharmaceutical constituents and are inexpensive compared with PL containing n‐3 PUFA. Therefore, the combination of TAG containing n‐3 PUFA and soybean PL has potential as a useful and inexpensive component of functional foods.     

Dietary Fatty Acids Differentially Regulate Secretion of Adiponectin and Interleukin‐6 in Primary Canine Adipose Tissue Culture

The aim of this study was to determine the effect of n3 polyunsaturated fatty acids (PUFA) on canine adipose tissue secretion of adiponectin, interleukin‐6 (IL6), and tumor necrosis factor‐α (TNFα). Subcutaneous and omental visceral adipose tissue samples were collected from 16 healthy intact female dogs. Concentrations of adiponectin were measured in mature adipocyte cultures, and concentrations of IL6 and TNFα were measured in undifferentiated stromovascular cell (SVC) cultures following treatment with eicosapentaenic acid (EPA, 20:5n‐3), arachidonic acid (ARA, 20:4n‐6), or palmitic acid (PAM, 16:0) at 25, 50, or 100 μM. Secretion of adiponectin from mature adipocytes was higher (p < 0.001) following EPA treatment at 50 μM compared to control in subcutaneous tissue, and higher following EPA treatment compared to PAM treatment at 25 μM in both subcutaneous (p < 0.001) and visceral tissues (p = 0.010). Secretion of IL6 from SVC derived from subcutaneous tissue was lower following EPA treatment and higher following PAM treatment compared to control both at 50 μM (p = 0.001 and p = 0.041, respectively) and 100 μM (p = 0.013 and p < 0.001, respectively). These findings of stimulation of adiponectin secretion and inhibition of IL6 secretion by EPA, and stimulation of IL6 secretion by PAM, are consistent with findings of increased circulating concentrations of adiponectin and decreased circulating concentration of IL6 in dogs supplemented with dietary fish oil, and show that the effect of fish oil on circulating concentrations of adiponectin and IL6 is, at least partially, the result of local effects of EPA and PAM on adipose tissue.     

Dietary trans α‐linolenic acid does not inhibit Δ5‐ and Δ6‐desaturation of linoleic acid in man

Dietary trans monoenes have been associated with an increased risk of heart disease in some studies and this has caused much concern. Trans polyenes are also present in the diet, for example, trans α‐linolenic acid is formed during the deodorisation of α‐linolenic acid‐rich oils such as rapeseed oil. One would expect the intake of trans α‐linolenic acid to be on the increase since the consumption of rapeseed oil in the western diet is increasing. There are no data on trans α‐linolenic acid consumption and its effects. We therefore carried out a comprehensive study to examine whether trans isomers of this polyunsaturated fatty acid increased the risk of coronary heart disease. Since inhibition of Δ6‐desaturase had also been linked to heart disease, the effect of trans α‐linolenic acid on the conversion of [U‐¹³C]‐labelled linoleic acid to dihomo‐γ‐linolenic and arachidonic acid was studied in 7 healthy men recruited from the staff and students of the University of Edinburgh. Thirty percent of the habitual fat was replaced using a trans ‘free’‐ or ‘high’ trans α‐linolenic acid fat. After at least 6 weeks on the experimental diets, the men received 3‐oleyl, 1,2‐[U‐¹³C]‐linoleyl glycerol (15 mg twice daily for ten days). The fatty acid composition of plasma phospholipids and the incorporation of ¹³C‐label into n‐6 fatty acids were determined at day 8, 9 and 10 and after a 6‐week washout period by gas chromatography‐combustion‐isotope ratio mass spectrometry. Trans α‐linolenic acid of plasma phospholipids increased from 0.04 ? 0.01 to 0.17 ? 0.02 and cis ? ‐linolenic acid decreased from 0.42 ? 0.07 to 0.29 ? 0.08 g/100 g of fatty acids on the high trans diet. The composition of the other plasma phospholipid fatty acids did not change. The enrichment of phosphatidyl ¹³C‐linoleic acid reached a plateau at day 10 and the average of the last 3 days did not differ between the low and high trans period. Both dihomo‐γ‐linolenic and arachidonic acid in phospholipids were enriched in ¹³C, both in absolute and relative terms (with respect to ¹³C‐linoleic acid). The enrichment was slightly and significantly higher during the high trans period (P<0.05). Our data suggest that a diet rich in trans α‐linolenic acid (0.6% of energy) does not inhibit the conversion of linoleic acid to dihomo‐γ‐linolenic and arachidonic acid in healthy middle‐aged men consuming a diet rich in linoleic acid.     

Profile of (n‐9) and (n‐7) Isomers of Monounsaturated Fatty Acids of Radish (Raphanus sativus L.) seeds

The present study was undertaken to determine the profile of the (n‐9) and (n‐7) isomers for C18:1, C20:1 and C22:1 fatty acids in radish seeds as well as their isomers. Radish (Raphanus sativus L. cvs. ‘Antep, Beyaz, Cherry Belle and Iri K?rm?z?’) seeds were produced in 2003–2005 from different sowing dates (SD). The n‐7 isomers of C18:1, C20:1 and C22:1 ranged from 0.7 to 1.3, 0.1 to 0.3 and 0.4 to 1.1 %, respectively. The average values of C18:1(n‐7) was highest (1 %) amongst the three acids. The ratios of (n‐7)/(n‐9) ranged from 4.5 % (‘Cherry Belle’, SD‐I) to 8.3 % (‘Antep’, SD‐III), 0.8 % (‘Iri K?rm?z?’, SD‐II) to 3 % (‘Iri K?rm?z?’, SD‐I) and 1.6 % (‘Cherry Belle’, SD‐I) to 3.7 % (‘Iri K?rm?z?’, SD‐I) for C18:1, C20:1 and C22:1. Erucic acid was the principal fatty acid with concentrations of nearly 34–39 % in ‘Antep’, 32–34 % in ‘Cherry Belle’, 30–33 % in ‘Beyaz’ and 21–22 % in ‘Ir? K?rm?z?’. The oleic acid content was higher in SD‐I and SD‐II than SD‐III in all cultivars. Correlation studies revealed that palmitoleic acid (C16:1) had a significant relationship between most of the fatty acids of the (n‐7)/(n‐9) family. The results indicated that palmitoleic acid is important in the synthesis of long‐chain fatty acids and that the data for the (n‐7)/(n‐9) ratios for C22:1 could be used as biochemical markers to determine the similarities or differences between radish cultivars.     

The Growth‐Inhibiting Effects of Beef Fatty Acids on MCF‐7 Cells Are Influenced Mostly by the Depot Location and Inconsistently by the Biohydrogenation Intermediate Content

Biohydrogenation intermediates (BHI) including conjugated linoleic acid (CLA) isomers are formed during ruminal biohydrogenation of polyunsaturated fatty acids (PUFA) in ruminants. Although many studies have examined the anticarcinogenic effects of CLA, few studies have reported the anticarcinogenic properties of BHI in their natural form found in dairy and beef fats. The present study compared the growth‐inhibitory effects of fatty acids from beef perirenal fat (PRF) or subcutaneous fat (SCF) with low or high levels of BHI in MCF‐7 human breast cancer cells. Cells were exposed for 72 h to media containing increasing doses (50 to 400 μM) of different beef fat treatments. Fatty‐acid analysis showed that BHI were readily incorporated into cell phospholipids (PL) in a treatment‐dependent manner, but higher BHI in PL did not consistently inhibit growth. Culturing with low‐BHI PRF or high‐BHI PRF did not lead to growth inhibition, but low‐BHI SCF inhibited growth, and inhibition was further increased by high‐BHI SCF. Other classes of fatty acids may, therefore, be interacting with BHI resulting in differential effects on growth inhibition in human breast cancer cells.     

Lower Concentration of n‐3 in the Red Blood Cells and Plasma of Lambs when their Dams were Fed a Diet High Compared with Low in n‐6 Fatty Acids at Joining

Feeding ewes a diet high in n‐6 in late gestation can affect fatty acid concentrations in the newborn lamb. The effect of feeding ewes a high n‐6 diet prior to conception and in early gestation on lamb n‐6 and n‐3 status has not previously been examined. The aim of the current study was to determine whether the concentration of n‐6 was higher and n‐3 was lower in lamb red blood cells (RBC) and plasma when Merino dams were fed a diet high in n‐6 either pre‐conception only or both pre‐conception and in early gestation. Dams were fed a diet low (silage) or high (oats/CSM) in n‐6 for either 6 weeks pre‐mating only or 6 weeks pre‐mating and 17 days post‐mating. The fatty acid status of lamb RBC and plasma was determined following birth and compared with dam fatty acids around parturition. The concentration of lamb RBC and plasma n‐3 was lower (p < 0.05) when dams received the high n‐6 compared with low‐n‐6 diet around mating, independent of the length of time of feeding. The concentration of n‐3 in lamb plasma was also higher when lambs were assessed as being likely rather than unlikely to have suckled prior to blood collection. Lamb RBC and plasma n‐3 fatty acids were lower when dams were fed the high compared with the low n‐6 diet for only a short time around mating. Transfer of fatty acids via the placenta and milk may account for the differences.     

Chemical Synthesis of Rare Natural Bile Acids: 11α‐Hydroxy Derivatives of Lithocholic and Chenodeoxycholic Acids

A method for the preparation of 11α‐hydroxy derivatives of lithocholic and chenodeoxycholic acids, recently discovered to be natural bile acids, is described. The principal reactions involved were (1) elimination of the 12α‐mesyloxy group of the methyl esters of 3α‐acetate‐12α‐mesylate and 3α,7α‐diacetate‐12α‐mesylate derivatives of deoxycholic acid and cholic acid with potassium acetate/hexamethylphosphoramide; (2) simultaneous reduction/hydrolysis of the resulting △¹¹‐3α‐acetoxy and △¹¹‐3α,7α‐diacetoxy methyl esters with lithium aluminum hydride; (3) stereoselective 11α‐hydroxylation of the △¹¹‐3α,24‐diol and △¹¹‐3α,7α,24‐triol intermediates with B₂H₆/tetrahydrofuran (THF); and (4) selective oxidation at C‐24 of the resulting 3α,11α,24‐triol and 3α,7α,11α,24‐tetrol to the corresponding C‐24 carboxylic acids with NaClO₂ catalyzed by 2,2,6,6‐tetramethylpiperidine 1‐oxyl free radical (TEMPO) and NaClO. In summary, 3α,11α‐dihydroxy‐5β‐cholan‐24‐oic acid and 3α,7α,11α‐trihydroxy‐5β‐cholan‐24‐oic acid have been synthesized and their nuclear magnetic resonance (NMR) spectra characterized. These compounds are now available as reference standards to be used in biliary bile acid analysis.     

Novel Odd-Chain Fatty Acids with a Terminal Double Bond in Ovaries of the Limpet <Emphasis Type="Italic">Cellana toreuma</Emphasis>

Our previous study characterized highly diverse dienoic fatty acids (FA), in particular an uncommon non‐methylene‐interrupted (NMI) FA, in the ovaries of the Japanese limpet Cellana toreuma belonging to the archaeogastropods, but many minor chemically unidentified FA remain. In this study, among previously unidentified FA (less than 0.1% of total FA), four novel NMI FA with a terminal double bond [7,18‐nonadecadienoic (19:2Δ7,18), 11,18‐nonadecadienoic (19:2Δ11,18), 7,20‐heneicosadienoic (21:2Δ7,20), and 11,20‐heneicosadienoic (21:2Δ11,20) acids] were found, along with known 14‐pentadecenoic (15:1Δ14), 16‐heptadecenoic (17:1Δ16), and 9,18‐nonadecadienoic (19:2Δ9,18) acids, based on capillary GC–MS of their methyl esters, 3‐pyridylcarbinol derivatives, and argentation thin‐layer chromatography. From our findings, possible biosynthetic pathways for the novel FA are discussed.