Biomimetic Thiyl Radical Chemistry by γ-Irradiation of Micelles and Vesicles Containing Unsaturated Fatty Acids

Different types of lipid aggregations, such as micelles and liposomes, can be used as biomimetic models. The uses of γ-irradiation as a valid methodology for simulating the biological generation of thiyl radicals in these models are summarized and, in particular, thiyl radical catalyzed cis trans isomerization of unsaturated lipids is underlined. The efficiency of antioxidants against lipid isomerization assayed by biomimetic models and the importance of trans lipids as novel biomarkers of radical stress are also described.     

Production of C6–C14 Medium-Chain Fatty Acids in Seeds and Leaves via Overexpression of Single Hotdog-Fold Acyl-Lipid Thioesterases

ACYL-LIPID THIOESTERASES (ALT) are a type of plant acyl–acyl carrier protein thioesterase that generate a wide range of medium-chain fatty acids and methylketone (MK) precursors when expressed heterologously in Escherichia coli. While this makes ALT-type thioesterases attractive as metabolic engineering targets to increase production of high-value medium-chain fatty acids and MKs in plant systems, the behavior of ALT enzymes in planta was not well understood before this study. To profile the substrate specificities of ALT-type thioesterases in different plant tissue types, AtALT1-4 from Arabidopsis thaliana, which have widely varied chain length and oxidation state preferences in E. coli, were overexpressed in Arabidopsis seeds, Camelina sativa seeds, and Nicotiana benthamiana leaves. Seed-specific overexpression of ALT enzymes led to medium-chain fatty acid accumulation in Arabidopsis and Camelina seed triacylglycerols, and transient overexpression in N. benthamiana demonstrated that the substrate preferences of ALT-type thioesterases in planta generally agree with those previously determined in E. coli. AtALT1 and AtALT4 overexpression in leaves and seeds resulted in the accumulation of 12–14 carbon-length fatty acids and 6–8 carbon-length fatty acids, respectively. While it was difficult to completely profile the products of ALT-type thioesterases that generate MK precursors (i.e. β-keto fatty acids), our results nonetheless demonstrate that ALT enzymes are catalytically diverse in planta. The knowledge gained from this study is a significant step towards being able to use ALT-type thioesterases as metabolic engineering tools to modify the fatty acid profiles of oilseed crops, other plants, and microorganisms.     

Δ6 Desaturase mRNA Abundance in HepG2 Cells Is Suppressed by Unsaturated Fatty Acids

The effect of unsaturated fatty acids on the abundance of Δ6 desaturase (D6D) mRNA and the fatty acid composition of HepG2 cell membranes was examined. Supplementation of HepG2 cells with oleic acid (18:1n-9, OA), linoleic acid (18:2n-6, LA), α-linolenic acid (18:3n-3, ALA), arachidonic acid (20:4n-6, AA) or eicosapentaenoic acid (20:5n-3, EPA) reduced D6D mRNA abundance by 39 ± 6.6, 40 ± 2.2, 31 ± 5.2, 55 ± 4.8, and 52 ± 5.0%, respectively, compared with control cells. Despite the reduction in D6D mRNA abundance, the level of D6D conversion products (20:3n-9, EPA and AA) in OA, ALA and LA supplemented cells, respectively, was elevated above that in control cells. Our results suggest that although unsaturated fatty acids decrease the abundance of D6D mRNA by as much as 50%, the conversion of polyunsaturated fatty acids and accumulation of long chain polyunsaturated fatty acids (LCPUFA) in HepG2 cell phospholipids continues to occur.     

Heterogeneous Catalytic Esterification of ω-Sulfhydryl Fatty Acids: Avoidance of Thioethers,Thioesters, and Disulfides

Two mesoporous silicas functionalized with propylsulfonic (SBA-15-PSA) and arenesulfonic (SBA-15-ASA) acid groups, and a highly acidic, functionalized styrene divinylbenzene copolymer ion exchange resin (Amberlyst-15) were examined for their ability to catalyze the ethanolic esterification of the ω-sulfhydryl fatty acid, 11-mercaptoundecanoic acid (MUA), without catalyzing unwanted side reactions at the sulfhydryl group. All three solid acid catalysts catalyzed the MUA esterification in excess ethanol. The activation energy for the catalytic esterifications were determined from 50 to 75 °C, resulting in apparent E _a of 54, 71, and 59 kJ/mol for SBA-15-PSA, SBA-15-ASA, and Amberlyst-15, respectively. GC–MS analysis determined that all three catalysts produced near quantitative conversion of MUA to its ethyl ester with very little reactivity towards the sulfhydryl group. This was a marked improvement over the esterifications catalyzed by sulfuric and p-toluenesulfonic acids which produced thioethers and disulfide side products. The MUA ethyl ester synthesis was demonstrated on a gram scale at 70 °C catalyzed by Amberlyst-15, and the desired product was isolated in 80% yield at >95% purity with a minimum of purification.     

Pro- and Antioxidative Effect of α-Tocopherol on Edible Oils, Triglycerides and Fatty Acids

Using advanced electron paramagnetic resonance techniques (EPR), oxidation of crude vegetable oils and their components (fatty acids and triglycerides) by radicals generated from hydrogen peroxide was investigated. The correlation rotational times were determined allowing us to characterize radicals formed during edible oils oxidation. Additionally ¹H- and ¹⁴N-hyperfine coupling constants differentiate the fatty acids dependently on their unsaturation. The acids with a higher number of unsaturated bonds exhibit higher A_N values of PBN/·lipid adduct. The waste oil with high free fatty acids content underwent the oxidation reaction more efficiently, however due to saturation and the high content of the fatty acids the carbon-centered radicals formed (upon hydrogen peroxide radicals) and their PBN (N-tert-butyl-α-phenylnitrone) adducts were less stable. The antioxidant effect was dependent on the amount of α-tocopherol added. In small amounts of up to 0.35 mg/1 g of fatty acid or triglyceride, it inhibited the creation of PBN/·lipid adducts while with higher amounts it intensified adduct formation. The α-tocopherol (AT) addition influence was also studied as spin scavenging dependence and indicated that any addition of the antioxidant in the investigated samples led to free radical scavenging and the effect increased with the increase in AT content.     

ω-3 Fatty Acids and Cardiovascular Diseases: Effects,Mechanisms and Dietary Relevance

ω-3 fatty acids (n-3 FA) have, since the 1970s, been associated with beneficial health effects. They are, however, prone to lipid peroxidation due to their many double bonds. Lipid peroxidation is a process that may lead to increased oxidative stress, a condition associated with adverse health effects. Recently, conflicting evidence regarding the health benefits of intake of n-3 from seafood or n-3 supplements has emerged. The aim of this review was thus to examine recent literature regarding health aspects of n-3 FA intake from fish or n-3 supplements, and to discuss possible reasons for the conflicting findings. There is a broad consensus that fish and seafood are the optimal sources of n-3 FA and consumption of approximately 2–3 servings per week is recommended. The scientific evidence of benefits from n-3 supplementation has diminished over time, probably due to a general increase in seafood consumption and better pharmacological intervention and acute treatment of patients with cardiovascular diseases (CVD).     

Fatty Acids,CD36, Thrombospondin-1, and CD47 in Glioblastoma: Together and/or Separately?

Glioblastoma (GBM) is one of the most aggressive tumors of the central nervous system, characterized by a wide range of inter- and intratumor heterogeneity. Accumulation of fatty acids (FA) metabolites was associated with a low survival rate in high-grade glioma patients. The diversity of brain lipids, especially polyunsaturated fatty acids (PUFAs), is greater than in all other organs and several classes of proteins, such as FA transport proteins (FATPs), and FA translocases are considered principal candidates for PUFAs transport through BBB and delivery of PUFAs to brain cells. Among these, the CD36 FA translocase promotes long-chain FA uptake as well as oxidated lipoproteins. Moreover, CD36 binds and recognizes thrombospondin-1 (TSP-1), an extracellular matrix protein that was shown to play a multifaceted role in cancer as part of the tumor microenvironment. Effects on tumor cells are mediated by TSP-1 through the interaction with CD36 as well as CD47, a member of the immunoglobulin superfamily. TSP-1/CD47 interactions have an important role in the modulation of glioma cell invasion and angiogenesis in GBM. Separately, FA, the two membrane receptors CD36, CD47, and their joint ligand TSP-1 all play a part in GBM pathogenesis. The last research has put in light their interconnection/interrelationship in order to exert a cumulative effect in the modulation of the GBM molecular network.     

Human Amnion Epithelial Cells: A Potential Cell Source for Pulp Regeneration?

The aim of this study was to analyze the suitability of pluripotent stem cells derived from the amnion (hAECs) as a potential cell source for revitalization in vitro. hAECs were isolated from human placentas, and dental pulp stem cells (hDPSCs) and dentin matrix proteins (eDMPs) were obtained from human teeth. Both hAECs and hDPSCs were cultured with 10% FBS, eDMPs and an osteogenic differentiation medium (StemPro). Viability was assessed by MTT and cell adherence to dentin was evaluated by scanning electron microscopy. Furthermore, the expression of mineralization-, odontogenic differentiation- and epithelial–mesenchymal transition-associated genes was analyzed by quantitative real-time PCR, and mineralization was evaluated through Alizarin Red staining. The viability of hAECs was significantly lower compared with hDPSCs in all groups and at all time points. Both hAECs and hDPSCs adhered to dentin and were homogeneously distributed. The regulation of odontoblast differentiation- and mineralization-associated genes showed the lack of transition of hAECs into an odontoblastic phenotype; however, genes associated with epithelial–mesenchymal transition were significantly upregulated in hAECs. hAECs showed small amounts of calcium deposition after osteogenic differentiation with StemPro. Pluripotent hAECs adhere on dentin and possess the capacity to mineralize. However, they presented an unfavorable proliferation behavior and failed to undergo odontoblastic transition.     

NMR,GC–MS and ESI-FTICR-MS Profiling of Fatty Acids and Triacylglycerols in Some Botswana Seed Oils

In the search for non-traditional seed oils, physicochemical parameters, fatty acid (FA) and triacylglycerol (TAG) profiles for five Botswana seed oils, obtained by Soxhlet extraction, were determined. GC–MS and ¹H-NMR analyses showed the FA profiles for mkukubuyo, Sterculia africana, and manketti, Ricinodendron rautanenii, seed oils dominated by linoleic and oleic acids, 26.1, 16.7 and 51.9, 24.4%, respectively, with S. africana containing significant amounts of cyclic FAs (19.9%). Mokolwane, Hyphaene petersiana, seed oil was typically lauric; 12:0 and 14:0 acids were 25.9 and 13.4%, respectively. Morama, Tylosema esculentum, seed oil resembled olive oil; 18:1 (47.3%) and 18:2 (23.4%) acids dominated. Moretologa-kgomo, Ximenia caffra, seed oil had 45.8% of 18:1 FA, plus significant amounts of very long chain FAs: 26:1 (5.8%), 28:1 (13.9%), 30:1 (3.9%), and acetylenic acids, 9a-18:1 (1.5%) and 9a, 11t-18:2 (16.0%). TAG classes and regiochemistry were determined with ESI-FTICR-MS, and ¹³C-NMR spectra, respectively. Morama showed seven major TAG classes with C54:4 and C54:3 dominating; mokolwane had 16 major classes with C32:0, C38:0 and C42:2 dominating; manketti had 11 major classes with C54:7, C54:6 and C54:4 dominating; mkukubuyo had 12 major classes with C52:4, C52:3 and C54:4 dominating; moretologa-kgomo had 30 major TAG classes with C64:5, C64:3 and C62:3 dominating. Saturated FAs were generally distributed over the sn-1(3) position for morama, manketti, and moretologa-kgomo but at the sn-2 position for mokolwane and mkukubuyo. These findings indicate that morama and manketti seed oils can be developed for food uses, whilst moretologa-kgomo and mkukubuyo seed oils only for nonfood uses.     

Profiling of Fatty Acids Composition in Suet Oil Based on GC–EI-qMS and Chemometrics Analysis

Fatty acid (FA) composition of suet oil (SO) was measured by precolumn methylesterification (PME) optimized using a Box–Behnken design (BBD) and gas chromatography/electron ionization-quadrupole mass spectrometry (GC–EI-qMS). A spectral library (NIST 08) and standard compounds were used to identify FAs in SO representing 90.89% of the total peak area. The ten most abundant FAs were derivatized into FA methyl esters (FAMEs) and quantified by GC–EI-qMS; the correlation coefficient of each FAME was 0.999 and the lowest concentration quantified was 0.01 μg/mL. The range of recovery of the FAMEs was 82.1%–98.7% (relative standard deviation 2.2%–6.8%). The limits of quantification (LOQ) were 1.25–5.95 μg/L. The number of carbon atoms in the FAs identified ranged from 12 to 20; hexadecanoic and octadecanoic acids were the most abundant. Eighteen samples of SO purchased from Qinghai, Anhui and Jiangsu provinces of China were categorized into three groups by principal component analysis (PCA) according to the contents of the most abundant FAs. The results showed SOs samples were rich in FAs with significantly different profiles from different origins. The method described here can be used for quality control and SO differentiation on the basis of the FA profile.     

Identification and Characterization of Phospholipids with Very Long Chain Fatty Acids in Brewer’s Yeast

Yeast lipids and fatty acids (FA) were analyzed in Saccharomyces pastorianus from seven breweries and in the dietary yeast supplement Pangamin. GC–MS identified more than 30 FA, half of which were very‐long chain fatty acids (VLCFA) with hydrocarbon chain lengths of ≥22 C atoms. Positional isomers ω‐9 and ω‐7 were identified in FA with C18–C28 even‐numbered alkyl chains. The most abundant ω‐7 isomer was cis‐vaccenic acid. The structure of monounsaturated FA was proved by dimethyl disulfide adducts (position of double bonds and cis geometric configuration) and by GC–MS of pyridyl carbinol esters. Ultra‐high performance liquid chromatography‐tandem mass spectrometry with negative electrospray ionization identified the phospholipids phosphatidylethanolamine, phosphatidylinositol and phosphatidylcholine, with more than 150 molecular species. Wild‐type unmutated brewer's yeast strains conventionally used for the manufacture of food supplements were found to contain VLCFA.     

Analysis of Fatty Acids in 12 Mediterranean Fish Species: Advantages and Limitations of a New GC-FID/GC–MS Based Technique

When fatty acids in fish are analyzed, results in percentage form (profile analysis) are mostly reported. However, the much more useful results expressed as mg/100 g (absolute analysis) is the main information required. Absolute methods based on calibration curves are of good accuracy but with a high degree of complexity if applied to a great number of analytes. Procedures based on the sequence profile analysis–total FA determination–absolute analysis may be suitable for routine use, but suffer from a number of uncertainties that have never been really resolved. These uncertainties are mainly related to the profile analysis. In fact, most profile analyses reported in the literature disagree about the number and type of fatty acids monitored as well as about the total percentage to assign to their sum so leading to possible inaccuracies; in addition the instrumental response factor for all FAME (fatty acid methyl esters) is often considered as a constant, but this is not exactly true. In this work, a set of 24 fatty acids was selected and studied on 12 fish species in the Mediterranean area (variable in lipid content and month of sampling): in our results, and in these species, this set constitutes, on average, 90 ± 3 % of the total fatty acid content. Moreover the error derived from the assumption of a unique response factor was investigated. Two different detection techniques (GC-FID and GC–MS) together with two capillary columns (different in length and polarity) were used in order to acquire complementary data on the same sample. With the protocol here proposed absolute analyses on the 12 cited species are easily achievable by the total FA determination procedure. The accuracy of this approach is good in general, but in some cases (DHA for example) is lower than the accuracy of calibration-based methods. The differences were evaluated on a case by case basis.     

β-Ketoacyl-acyl Carrier Protein Synthase I (KASI) Plays Crucial Roles in the Plant Growth and Fatty Acids Synthesis in Tobacco

Fatty acids serve many functions in plants, but the effects of some key genes involved in fatty acids biosynthesis on plants growth and development are not well understood yet. To understand the functions of 3-ketoacyl-acyl-carrier protein synthase I (KASI) in tobacco, we isolated two KASI homologs, which we have designated NtKASI-1 and NtKASI-2. Expression analysis showed that these two KASI genes were transcribed constitutively in all tissues examined. Over-expression of NtKASI-1 in tobacco changed the fatty acid content in leaves, whereas over-expressed lines of NtKASI-2 exhibited distinct phenotypic features such as slightly variegated leaves and reduction of the fatty acid content in leaves, similar to the silencing plants of NtKASI-1 gene. Interestingly, the silencing of NtKASI-2 gene had no discernibly altered phenotypes compared to wild type. The double silencing plants of these two genes enhanced the phenotypic changes during vegetative and reproductive growth compared to wild type. These results uncovered that these two KASI genes had the partially functional redundancy, and that the KASI genes played a key role in regulating fatty acids synthesis and in mediating plant growth and development in tobacco.     

Nonwood Fibers—A Potential Source of Raw Material in Papermaking

World demand for paper has increased at an average annual rate of 4.7% over the past 40 years. Although future growth will reduce to 2–3% the existing wood resources may be inadequate to meet this growing demand for paper especially in the Asia-Pacific region and Eastern Europe. In addition, logging is coming under increasing pressure from environmentalists concerned about habitat destruction and other longer-term impacts of forest harvesting. It is, therefore, necessary to consider alternative fiber sources to meet the possible shortfall of wood fiber for papermaking. Suitable nonwood fibers are abundantly available in many countries and are the major source of fiber for papermaking in some developing nations.     

A Comprehensive GC–MS Sub-Microscale Assay for Fatty Acids and its Applications

Fatty acid analysis is essential to a broad range of applications including those associated with the nascent algal biofuel and algal bioproduct industries. Current fatty acid profiling methods require lengthy, sequential extraction and transesterification steps necessitating significant quantities of analyte. We report the development of a rapid, microscale, single-step, in situ protocol for GC–MS lipid analysis that requires only 250 μg dry mass per sample. We furthermore demonstrate the broad applications of this technique by profiling the fatty acids of several algal species, small aquatic organisms, insects and terrestrial plant material. When combined with fluorescent techniques utilizing the BODIPY dye family and flow cytometry, this micro-assay serves as a powerful tool for analyzing fatty acids in laboratory and field collected samples, for high-throughput screening, and for crop assessment. Additionally, the high sensitivity of the technique allows for population analyses across a wide variety of taxa.     

Direct Determination of Glycidyl Esters of Fatty Acids in Vegetable Oils by LC–MS

An LC–MS method using a single quadrupole mass spectrometer was developed for direct analysis of glycidyl esters of fatty acids in vegetable oils. Without any sample clean-up, this method provided acceptable recovery of seven glycidyl esters, comparable results to a previously-published method utilizing two solid-phase extraction steps, and consistent detection parameters after greater than 200 injections without any cleaning operations performed. This method could readily be implemented as a screening assay for glycidyl esters in most oil laboratories.     

Effect of Saturated/Unsaturated Fatty Acid Ratio on Physicochemical Properties of Palm Olein–Olive Oil Blend

Although blending polyunsaturated oil with more saturated or monounsaturated oils has been studied extensively, there is no similar information regarding the partial replacement of palm olein with olive oil (OO). Therefore the main objective of this study was to investigate the effects of OO partial replacement (0, 25, 50, 75, 90 and 100% w/w) on the chemical stability of palm olein oil (POO). The physicochemical properties of oil samples namely iodine value, peroxide value (PV), anisidine value, TOTOX value (total oxidation value, TV), free fatty acid (FFA), cloud point, color and viscosity were considered as response variables. Significant differences among the oil blend properties were determined at the significance level of P < 0.05. Apart from FFA, all the response variables were significantly influenced by type and concentration of oils. The oil blend containing 10% POO and 90% OO showed the highest TV (6.10); whereas the blend containing 90% POO and 10% OO exhibited the least TV (2.41). This study indicated that the chemical stability of oil blend significantly (P < 0.05) increased with increasing the proportion of polyunsaturated/monounsaturated fatty acid.