Spirulina Lipids Alleviate Oxidative Stress and Inflammation in Mice Fed a High-Fat and High-Sucrose Diet.

High-fat and high-sucrose diet (HFHSD)-induced obesity leads to oxidative stress and chronic inflammatory status. However, little is known about the beneficial effects of total lipids extracted from Spirulina. Hence, in the present study, Spirulina lipids were extracted with chloroform/methanol (SLC) or ethanol (SLE) and then their effects on oxidative stress and inflammation in the mice fed a HFHSD were investigated. The results show that the major lipid classes and fatty acid profiles of SLC and SLE were almost similar, but the gamma-linolenic acid (GLA) and carotenoid contents in SLE was a little higher than that in SLC. Dietary 4% SLC or SLE for 12 weeks effectively decreased the hepatic lipid hydroperoxide levels as well as increased the activities and mRNA levels of antioxidant enzymes in the mice fed a HFHSD. In addition, supplementation with SLC and SLE also markedly decreased the levels of serum pro-inflammatory cytokines and the mRNA expression of pro-inflammatory cytokines in the liver and epididymal white adipose tissue of mice fed a HFHSD, and the effects of SLC and SLE were comparable. These findings confirm for the first time that dietary Spirulina lipids could alleviate HFHSD-induced oxidative stress and inflammation.

Separation of γ-linolenic and other polyunsaturated fatty acids from Boraginaceae via supercritical CO2.

Essential fatty acids were extracted from Echium amoenum (Boraginaceae) seed oil via supercritical carbon dioxide and the results were compared with conventional Soxhlet method as the base case of 100% recovery. The response surface methodology was used to optimize the effective extraction parameters. The chemical composition of recovered oil was analyzed by polar and non-polar gas chromatograph-flame ionization detector columns. The experimental results indicated that echium seed contained 25 wt% oil and the maximum extraction oil recovery of 92% was obtained via supercritical CO(2) at optimal operating conditions (43°C, 280 bar, 1.5 mL/min, 25 min static time and 130 min dynamic time). At similar operating conditions, applying 2.5 and 5 mol% ethanol as a modifier enhanced the recovery to 96 and 112%, respectively. The results showed that supercritical fluid extraction is a viable technique for separation of constituents such as γ-linolenic acid (7-8%), palmitic acid (6-7%), stearic acid (3-4%), oleic acid (12-13%), linoleic acid (19-20%), α-linolenic acid (40-41%) and stearidonic acid (8-9%) from Boraginaceae.

Production of γ-linolenic acid using a novel heterologous expression system in the oleaginous yeast Lipomyces kononenkoae.

A novel expression system was established in the oleaginous yeast, Lipomyces kononenkoae. The expression vector pLK-rhPHG of L. kononenkoae was constructed and using the hygromycin phosphotransferase gene and green fluorescent protein gene as reporter genes. A delta 6-fatty acid desaturase gene (D6DM) from Cunninghamella echinulata MIAN6 was then expressed in this strain. The recombinant strain accumulated about 1.2% γ-linolenic acid in the total fatty acids.

Δ6-Desaturase from Mortierella alpina: cDNA cloning, expression, and phylogenetic analysis.

The open reading frame of the Δ(6)-desaturase gene was isolated from Mortierella alpina W15 and the gene was cloned into a pPIC3.5K vector. The vector was transformed into Pichia pastoris GS115 and expression was induced with methanol. The Δ(6)-desaturase expressed in P. pastoris GS115 catalyzed the conversion of linoleic acid to γ-linolenic acid but not the conversion of α-linolenic acid to octadecatetraenoic acid. The results indicate that the Δ(6)-desaturase gene from M. alpina W15 has substrate specificity in different organisms. Phylogenetic analysis revealed that Δ(6)-desaturase genes can be divided into four monophyletic groups. This work paves the way for further study of the functions of Δ(6)-desaturase in fatty acid metabolism and its three-dimensional structure.

Mertensia (Boraginaceae) seeds are new sources of γ-linolenic acid and minor functional compounds.

In this work, seeds from selected Mertensia species were analyzed for γ-linolenic acid-rich oils and minor functional compounds. Fatty acids (FA) were analyzed by GC-FID; tocopherols, sterols, squalene, and phenolics compounds by HPLC-DAD, and the structures of the latter were confirmed by LC-MS. M. maritima spp. asiatica and M. sibirica showed the highest amounts of γ-linolenic acid (22.8 and 18.7%, respectively) and total FA (15.9 and 10.9 g/100 g seeds, respectively). M. sibirica had the greatest levels of sterols, phenolics and tocopherols (244.8, 243.9 and 66.3 mg/100 g, respectively), in which stigmasterol, rosmarinic acid and α-tocopherol (237.7, 180.1 and 53.6 mg/100 g, respectively) were the most abundant components. M. maritima spp. asiatica and M. arizonica showed the highest amounts of squalene (2.5 and 1.1 mg /100 g seeds). Mertensia species constitute a new source of GLA-rich oils, suitable to be marketed by the pharmaceutical and food industries.

[Separation of gamma linolenic acid from evening primrose oil with urea inclusion--orthogonal experiment of optimizing technological parameters and observation of urea inclusion compound I].

OBJECTIVE: The influence on the urea inclusion compound under different conditions (allocated proportion, time of inclusion, temperature of inclusion) were studied through the orthogonal test, and theoretical reference of urea inclusion process for further optimization wound be offered. METHOD: The orthogonal experiment was adopted, and microscope was used to observe the shape, aperture size of the urea inclusion compound under different technological parameters, the GC was employed to inspect the purity of GLA. RESULT: The results indicated that the ratio of fatty acids and urea, inclusion of temperature, time of inclusion had great effect on urea inclusion compound. The three factors and its interactions significantly affected the purity of GLA. The results also showed that the best process was that the ratio of fatty acids and urea was 1 : 3, temperature of inclusion was--15 degrees C, time of inclusion was 24 h. CONCLUSION: Under the best condition, the purity of GLA reach up to 95.575 9%; and it is feasible to observe the shape and the amount of the urea inclusion compound to reflect and guide the urea inclusion technology.

Genetic variants in FADS1 and ELOVL2 increase level of arachidonic acid and the risk of Alzheimer's disease in the Tunisian population.

Polyunsaturated fatty acids (PUFAs) are closely related to various physiological conditions. In several age-related diseases including Alzheimer's disease (AD) altered PUFAs metabolism has been reported. However, the mechanism behind PUFAs impairment and AD developpement remains unclear. In humans, PUFAs biosynthesis requires delta-5 desaturase (D5D), delta-6 desaturase (D6D) and elongase 2 activities; which are encoded by fatty acid desaturase 1 (FADS1), fatty acid desaturase 2 (FADS2), and elongation of very-long-chain fatty acids-like 2 (ELOVL2) genes, respectively. In the present work, we aim to assess whether genetic variants in FADS1, FADS2 and ELOVL2 genes influence plasma and erythrocyte PUFA composition and AD risk. A case-control study was carried out in 113 AD patients and 161 healthy controls.Rs174556, rs174617, and rs3756963 of FADS1, FADS2, and ELOVL2 genes, respectively were genotyped using PCR-RFLP. PUFA levels were quantified using Gas Chromatography. Genotype distributions of rs174556 (FADS1) and rs3756963 (ELOVL2) were different between case and control groups. The genotype TT of rs174556 and rs3756963 single nucleotide polymorphism (SNP) increases significantly the risk of AD in our population. PUFA analysis showed higher plasma and erythrocyte arachidonic acid (AA) level in patients with AD, whereas only plasma docosahexaenoic acid (DHA) was significantly decreased in AD patients. The indexes AA/Dihomo-gamma-linolenic acid (DGLA) and C24:4n-6/Adrenic acid (AdA) were both higher in the AD group. Interestingly, patients with TT genotype of rs174556 presented higher AA level and AA/DGLA index in both plasma and erythrocyte. In addition, higher AA and AA/DGLA index were observed in erythrocyte of TT genotype ofrs3756963 carrier's patients. Along with, positive correlation between AA/DGLA index, age or Gamma-linolenic acid (GLA)/ Linoleic acid (LA) index was seen in erythrocyte and /or plasma of AD patients. After adjustment for confounding factors, the genotype TT of rs174556, erythrocyte AA and AA/DGLA index were found to be predictive risk factors for AD while plasma DHA was found associated with lower AD risk. Both rs174556 and rs3756963 influence AD risk in the Tunisian population and they are likely associated with high AA level. The combination of the two variants increases further the susceptibility to AD. We suggest that FADS1 and ELOVL2 variants could likely regulate the efficiency of AA biosynthesis which could be at the origin of inflammatory derivate.

Fish Oil and Primrose Oil Suppress the Progression of Alzheimer's Like Disease Induced by Aluminum in Rats.

The role of fish oil, primrose oil and their mixture in ameliorating the changes in Alzheimer's like model was evaluated in rats. Primrose oil and primrose/fish oil mixture fatty acids composition was assessed by gas chromatography. The rat experiment consisted of 5 groups; the first fed on balanced diet as control normal (CN); the other four groups treated with intraperitoneal aluminum lactate and consumed dyslipidemic diet; one group served as control Alzheimer's like disease (CA) while the other three groups (test groups) received daily oral dose from primrose oil, fish oil and primrose/fish oil mixture separately for 5 weeks. Results showed primrose oil and primrose/ fish oil mixture to contain gamma linolenic acid as 9.15 and 4.3% of total fatty acids, respectively. Eicosapentaenoic and docosahexaenoic were present as 10.9 and 6.5 %, respectively in the oil mixture. Dyslipidemia and increased erythrocyte sedimentation rate (ESR), plasma butyrylcholinesterase (BChE), brain malondialdehyde (MDA) and NO with decrease in plasma magnesium, brain catalase, reduced glutathione, body weight gain and brain weight were demonstrated in CA compared to CN. Brain histopathology and immuno-histochemistry showed neuronal degeneration and neurofibrillary tangles with elevated myeloperoxidase and nuclear factor-kappa B in CA compared to CN. The tested oils demonstrated neuro-protection reflected in the variable significant improvement of biochemical parameters, immuno-histochemistry and brain histopathology. Primrose/fish oil mixture was superior in reducing ESR, brain MDA, plasma activity of BChE and brain histopathological changes along with elevating plasma magnesium. Primrose/fish oil mixture and fish oil were more promising in improving plasma high density lipoprotein cholesterol (HDL-C) than primrose. Fish oil was the most efficient in improving plasma total cholesterol (T-C), low density lipoprotein cholesterol and T-C /HDL-C. Primrose/fish oil mixture and primrose oil were superior in elevating brain catalase compared to fish oil. Other parameters were equally improved by the different oil treatments. Primrose oil, fish oil and their mixture reduced the progression of Alzheimer's disease in rats with superiority to primrose/fish oil mixture.

Nutrition for the eye: different susceptibility of the retina and the lacrimal gland to dietary omega-6 and omega-3 polyunsaturated fatty acid incorporation.

The purpose of this study was to compare the susceptibility of the retina and the exorbital lacrimal gland to dietary supplies of long-chain omega-3 (omega3) and omega-6 (omega6) polyunsaturated fatty acids (LC-PUFAs). Male Wistar rats were fed a 5% lipid diet containing: (1) 10% eicosapentaenoic acid (EPA) and 7% docosahexaenoic acid (DHA), or (2) 10% gamma-linolenic acid (GLA), or (3) 10% EPA, 7% DHA and 10% GLA or (4) a balanced diet deprived of EPA, DHA and GLA for 3 months. Lipids were extracted from plasma phospholipids, retina and exorbital lacrimal gland, and fatty acid composition was determined by gas chromatography. Dietary supplementation with EPA and DHA increased omega3 PUFA levels in plasma phospholipids as well as in the retina and the exorbital lacrimal gland. By contrast, GLA supplementation favored omega6 PUFA incorporation, and particularly the incorporation of the end-chain omega6 product, docosapentaenoic acid (DPA), into all tissues. Supplementation with EPA, DHA and GLA increased the levels of DHA, EPA and dihomo-GLA (dGLA), whereas arachidonic acid (AA) was unchanged and DPA decreased in the retina and the lacrimal gland. The ability of both tissues to incorporate PUFAs from blood was evaluated. The results showed that the retina was more selective than the lacrimal gland for EPA. In spite of the different susceptibility of the retina and the lacrimal gland to dietary PUFAs, these results suggest that the concomitant use of dietary omega3 and omega6 PUFAs may be useful in modulating inflammation in both tissues.

Fatty Acid, Triacylglycerol and Unsaponifiable Matters Profiles and Physicochemical Properties of Chinese Evening Primrose Oil.

This work focused on physicochemical property assaying, fatty acid composition, triacylglycerol (TAG) profiles, and unsaponifiable matters composition of the Chinese evening primrose oil. The cold press oil possessed very low acid value and peroxide value, and relatively high iodine value. Fatty acid composition results indicated that this oil was especially high in linoleic acid and linolenic acid. Characterization of TAG composition was achieved by a two-dimensional HPLC coupling of nonaqueous reverse-phase and silver ion HPLC with atmospheric pressure chemical ionization MS method. There was a total of 38 TAGs including 27 regioisomers which had been determined. Unsaponifiable matters composition results revealed that this oil possessed a number of phytosterols, in which ß-sitosterol and stigmasterol were most predominant.

Ribes taxa: A promising source of γ-linolenic acid-rich functional oils.

Fifty Ribes species and R. nigrum-based cultivars from eight Ribes sections were surveyed for γ-linolenic acid (GLA, 18:3, n-6)- and stearidonic acid (SDA, 18:4, n-3)-rich oils. R. pallidiflorum, R. glabellum and R. pubescens seed oils contain noticeable GLA amounts: 13.3, 11.8, and 11.9% of total fatty acids (FA), respectively. However, the highest GLA contents were found in the seed oils of several blackcurrant cultivars, highlining Ribes 'Myuryucheene' with 20.2% GLA of total FA. Principal Component Analysis showed that similarities in FA profiles allow grouping species as botanical criteria for Ribes sections do. The main GLA-taxa detected in this study correspond to blackcurrant cultivars, all of them native to Siberia. Considering that such cultivars are notable fruit-producers, its cultivation in Siberia besides producing fruits in very difficult agronomic areas, could produce a valuable by-product, i.e. the seeds, which will add economic value to agricultural systems if devoted to GLA-rich oils extraction.

Regioisomeric structure determination of alpha- and gamma-linolenoyldilinoleoylglycerol in blackcurrant seed oil by silver ion high-performance liquid chromatography and mass spectrometry.

Gamma-linolenic acid (Gla) and oils containing Gla have evident positive effects on a variety of disorders, and therefore, the structure of triacylglycerols (TAGs) containing Gla is of special interest. The regioisomeric structures of TAGs 18:3(n-3)/18:2/18:2 (Ala/L/L) and 18:3(n-6)/18:2/18:2 (Gla/L/L) in blackcurrant seed oil were determined by Ag-HPLC/APCI-MS and Ag-HPLC/ESI-MS/MS techniques. In the latter, silver ion adducts were prepared by adding silver nitrate to the postcolumn flow. A new Ag-HPLC solvent system containing nonchlorinated solvents for the separation of Gla- and Ala-containing TAGs is introduced. Ag-HPLC separation of Ala/L/L and Gla/L/L was sufficient and regioisomers sn-LnLL + sn-LLLn and sn-LLnL (Ln, linolenic acid) could be differentiated well with the MS methods used. No discrimination was made between the sn-1 and sn-3 positions. The results show that the methods used are suitable to discriminate and determine the regioisomeric structure of TAGs. The regioisomeric structure of TAG with the fatty acid combination Gla/L/L in blackcurrant seed oil was considered to be practically random (32.7-37.8% of sn-LGlaL). In the fatty acid combination Ala/L/L, the regioisomeric structure is nonrandom (7.3-12.1% of sn-LAlaL) with Ala preferentially in a primary position. It can be concluded that the positional distribution of Ala and Gla is different in Ln/L/L TAGs of blackcurrant seed oil.

Purification and identification of bovine cheese whey fatty acids exhibiting in vitro antifungal activity.

Milk lipids contain several bioactive factors exhibiting antimicrobial activity against bacteria, viruses, and fungi. In the present study, we demonstrate that free fatty acids (FFA) derived from the saponification of bovine whey cream lipids are active in vitro at inhibiting the germination of Candida albicans, a morphological transition associated with pathogenicity. This activity was found to be significantly increased when bovine FFA were enriched in non-straight-chain FFA. At low cell density, this non-straight-chain FFA-enriched fraction was also found to inhibit in a dose-dependant manner the growth of both developmental forms of C. albicans as well as the growth of Aspergillus fumigatus. Using an assay-guided fractionation, the main components responsible for these activities were isolated. On the basis of mass spectroscopic and gas chromatographic analysis, antifungal compounds were identified as capric acid (C10:0), lauroleic acid (C12:1), 11-methyldodecanoic acid (iso-C13:0), myristoleic acid (C14:1n-5), and gamma-linolenic acid (C18:3n-6). The most potent compound was gamma-linolenic acid, with minimal inhibitory concentration values of 5.4 mg/L for C. albicans and 1.3 mg/L for A. fumigatus, in standardized conditions. The results of this study indicate that bovine whey contains bioactive fatty acids exhibiting antifungal activity in vitro against 2 important human fungal pathogens.

[Prepared evening primrose oil microspheres by spray drying].

OBJECTIVE: To explore the best technique parameters on preparing evening primrose oil microspheres by spray drying technique. METHOD: GC and internal standard method were used to determine the content of gamma-linolenic acid methyl esters, the technological parameters were investigated by orthogonal experimental design with the yield and entrapment rate as indexes. RESULT: The charging rate as 3 mL x min(-1), atomizing pressure as 100 kPa and the temperature of inlet as 105 degrees C were the best technique parameters. CONCLUSION: Evening primrose oil microspheres accorded with the expecting demand. The main influencing factor is the charging rate. Spray drying is reasonable and practical for preparing evening primrose oil microspheres.

Polycyclic aromatic hydrocarbons content and fatty acids profile in coconut, safflower, evening primrose and linseed oils.

This study aimed at evaluating the polycyclic aromatic hydrocarbons (PAHs) contamination of commercial vegetable oils and examined the identity through the fatty acids profiles. Coconut, safflower, evening primrose, and linseed oils marketed in São Paulo (Brazil) were investigated totaling 69 samples. Four PAHs, benzo[a]anthracene (BaA), chrysene (Chr), benzo[b]fluoranthene (BbF), and benzo[a]pyrene (BaP), were detected in 96% of the samples at individual levels ranging from not detected to 14.99 µg kg-1. Chrysene was the abundant hydrocarbon found among all types of oils, with the highest median values. The results of the fatty acid profiles revealed that 43% showed different profiles according to the ones on their labels, with a higher incidence of adulteration of evening primrose oils. The maximum tolerable limits by European Regulation No. 835/2011 were exceeded for BaP in 12%, and for total 4 PAHs in 28%, with a greater contribution of adulterated samples.

Long-chain n-6 polyunsaturated fatty acids in breast milk decrease the risk of HIV transmission through breastfeeding.

BACKGROUND: Breastfeeding accounts for a sizable proportion of infant HIV infections. Some fatty acids (FAs) are potent immunomodulators with virucidal activity, and their primary source in breastfed children is breast milk. OBJECTIVES: The aims of the study were to examine whether the percentage weight concentration of FAs in breast milk was associated with the risk of mother-to-child transmission (MTCT) of HIV by breastfeeding and with shedding of cell-free virus (CFV) or cell-associated virus (CAV) in breast milk. DESIGN: We conducted a case-control study nested within a cohort of HIV-infected Tanzanian women and children. We matched 59 incident breastfeeding MTCT cases to 59 nontransmitting controls based on the child's age at sample collection. We quantified FAs, CFV, and CAV in a breast milk sample collected before the infant's first positive HIV test. RESULTS: After adjustment for indicators of maternal HIV disease stage, the risk of MTCT was inversely related to 11c,14c-eicosadienoic acid [odds ratio (OR) for quartile 4 compared with quartile 1: 0.21; P for trend = 0.04], arachidonic acid (OR: 0.21; P for trend = 0.03), and dihomo-gamma-linolenic acid (OR: 0.24; P for trend = 0.03); the latter 2 were also linearly, inversely related to virus shedding in breast milk. Lauric acid and pentadecanoic acid were associated with increased MTCT, whereas trans FAs were related to higher CAV and CFV. CONCLUSION: Increasing concentrations of long-chain n-6 polyunsaturated FAs in breast milk might reduce the risk of MTCT.

Changes in the content of gamma-linolenic C18:3 (n-6) and stearidonic C18:4 (n-3) acids in developing seeds of viper's bugloss Echium vulgare L.

Changes in the composition of fatty acids (FA) were determined in lipid extracts isolated from developing ovaries containing ovules and developing seeds of Echium vulgare L. The samples were collected successively over 20 days beginning with the first day after flowering. The contents of the n-6 FA family members, i.e., gamma-linolenic (GLA) (C(18:3)) and linoleic (LA) (C(18:2)) acids changed in a parallel manner and reached the maximum of 13.9% and 24%, respectively, on the 12th day, after which they fell systematically down to 8.6% and 18.2%, respectively, on the 20th day after flowering. Starting with day 13, the content of alpha-linolenic acid (ALA) (C(18:3) n-3) begins to grow intensively, from 24.2% to 39.3% on the 20th day after flowering. The increase in the content of stearidonic acid (SDA) (C(18:4) n-3), up to 10.5% on the 20th day after flowering, occurred steadily as the seeds developed, and was independent of the changes in the content of GLA and LA. The pattern of changes in the content of SDA, GLA, LA and ALA during the development of seeds, and the occurrence of SDA in the seed oil of other plants, demonstrate that the biosynthesis of SDA in the seeds is critically dependent on the presence of ALA. The above condition indicates that SDA biosynthesis in the seeds of Echium vulgare follows the scheme LA --> simultaneous, competitive, action of Delta(6) and Delta(15) desaturases, leading to the formation of GLA and ALA, respectively, and then ALA (Delta(6) des) --> SDA. The biosynthesis according to the scheme: GLA (Delta(15) des) --> SDA is highly unlikely.

High-performance liquid chromatography with nuclear magnetic resonance detection--A method for quantification of alpha- and gamma-linolenic acids in their mixtures with free fatty acids.

While many naturally occurring mixtures of free fatty acids are conveniently analyzed by hyphenated technique of LC-NMR, a complete separation of alpha- and gamma-linolenic acids for their quantitative determination appears impossible at least by the methods of reversed phase HPLC. However, they can be differentiated and quantified from 1H NMR spectra measured in the course of isocratic acetonitrile-chloroform (90:10, with C8 and C18 columns in series) LC-NMR analysis without the need for any derivatization.

[Influence of methyl esterification on the ultraviolet absorption of gamma-linolenic acid].

OBJECTIVE: To study the influence of methyl esterification on the ultraviolet absorption of gamma-linolenic acid. METHODS: To measure the ultraviolet absorption value of some gamma-linolenic acid product by Ultraviolet Spectrophotometry before and after methyl esterification and to measure the fatty acids formation with gas chromatographic method. Then to do a correlation analysis of the results. RESULTS: The ultraviolet absorption value of gamma-linolenic acid before methyl esterification was very significantly correlated with that after methyl esterification. CONCLUSION: It is feasible to mesure the ultraviolet absorption value of polyunsaturated fatty acids (such as gamma-linolenic acid) without methyl esterification.

Sardinian Boraginaceae are new potential sources of gamma-linolenic acid.

The aim of this work was to establish the richness in γ-linolenic acid (GLA, 18:3n6) and stearidonic acid (SDA, 18:4n3) of several Sardinian Boraginaceae species. To this end, seeds of selected species were collected from their natural habitats and analysed. The highest GLA contents were found in the seed oils of two endemic Borago taxa, i.e. B. morisiana (24.4 and 24.6% GLA of total fatty acids for samples from San Pietro Island and Sardinia Island, respectively), and 22.9% GLA for B. pygmaea. Both Borago species contained more GLA than B. officinalis collected in the same ecosystems. SDA was found in significant amounts in Echium plantagineum seed oil from the Lattias Mountains (15% SDA of total fatty acids). It is notable that both Borago GLA-rich species are under threat of extinction, thus revealing the importance of the preservation of the natural Sardinian ecosystems for endangered species and human health.