For Free Fatty Acids Research Articles

Esterification of free fatty acids in non- edible oils using partially sulfonated polystyrene for biodiesel feedstock

Partially sulfonated polystyrene (PSS), synthesized from expanded polystyrene waste (EPS), was used as a catalyst for free fatty acid (FFA) conversion in non-edible oils. Acidic and water absorbing properties of the PSS facilitated the catalytic action for the FFA conversion by esterification reaction. The reaction was done on simulated acid oil (WCO) containing oleic acid and sunflower oil, and rubber seed oil (RSO). Effects of temperature, catalyst amount and alcohol to acid molar ratio were studied. FFA conversion increased with each of these factors. The advantage of this heterogeneous catalyst is that it is efficient as commercial ion exchange resin and easily removable from the reaction mixture. PSS is found to substantially reduce the acid value of WCO and RSO from 17 to 3.2mg KOH/g and from 28.8 to 4.8mg KOH/g respectively at 75°C. The WCO and RSO with reduced acid value may be used as a feedstock for biodiesel production.

Promising Olive Cultivars for Oil Production in Hawaii

To determine promising olive (Olea europaea) cultivars for oil production in Hawaii, seven trees each of 10 cultivars (Arbequina, Arbosana, Coratina, Frantoio, Koroneiki, Leccino, Mission, Moraiolo, Pendolino, and Taggiasca) were planted in Feb. and July 2011 at the Lalamilo Experiment station on Hawaii Island (lat. 20.0176°N, long. 155.6827°W, elevation 2700 ft). In addition, two trees each of these 10 cultivars were planted in June 2011, with the exception of Arbequina, which was planted in July 2012, at the Maui Agricultural Research Center in Kula, Maui (lat. 20.7564°N, long. 156.3289°W, elevation 3100 ft). At Lalamilo, after ≈2 years of growth in the field (2013), three cultivars of olives (Arbequina, Arbosana, and Koroneiki) flowered, fruited, and produced oil yields of greater than 20%. These same cultivars flowered and fruited in 2014 and 2015. There was no significant difference among cultivars in fresh weight fruit yield averaged over 2 years (2013 and 2014), ranging from 2.14 to 2.45 kg/tree. During December to March, calculation of chilling hours below 12.5 °C was 141 hours during 2012–13 and 161 hours during 2013–14. The other seven cultivars did not flower and fruit during these 2 years of growth at Lalamilo, perhaps due to a greater requirement for chilling hours. At Kula, after 3 years of growth (2015), nine cultivars of olives with the exception of Moraiolo flowered and fruited. Mean fresh weight fruit yield in 2015 ranged from 0.25 to 22.06 kg/tree for various cultivars grown in Kula, Maui. In 2013, the oil from three cultivars grown at Lalamilo was analyzed for free fatty acids (FFA), peroxide value (PV), ultraviolet absorption for conjugated double bonds, 1,2-diacylglycerol (DAG), and pyropheophytins (PPP). Oil quality was within the range of extra-virgin olive oil. There is a need to investigate further the effects of temperature and management on flowering and fruiting of olive cultivars grown in Hawaii at various elevations. In particular, ‘Arbequina’, ‘Arbosana’, and ‘Koroneiki’ appear to have a lower requirement for chilling hours than other cultivars tested.

Lipase Catalyzed Incorporation of Conjugated Linoleic Acid by Transesterification into Sunflower Oil Applying Immobilized Lipase (Lipozyme Thermomyces lanuginosus and Rhizomucor mehei)

Conjugated Linoleic Acid (CLA), Glycerol (G) and sunflower oil blends with varying concentration were subjected to enzymatic esterification using a 1, 3- specific immobilized lipase. CLA was used as acyl due to its purported health benefits. The transesterified lipids were evaluated for free fatty acids (FFA) and composition of fatty acids by gas chromatography. Lipozyme RM IM is preferred for enzymatic esterification because of more CLA incorporation and less free fatty acids at the end of reaction than Lipozyme TL IM. Response surface methodology (RSM) was used to determine the effects of three variables Glycerol concentration, reaction temperature, and amount of enzyme on the lipase catalyzed incorporation of CLA into Sunflower oil. The optimum condition for maximum CLA incorporation (96.7% Incorporation Yield), maximum reaction rate (0.0271 M/h) and minimum free fatty acids production (Acid No.2.73) was obtained, at 75 C by using 3 % enzyme and 0.27 M glycerol.

DETERMINATION OF Cu, Fe, Mn, Zn AND FREE FATTY ACIDS IN PEQUI OIL

Pequi (Caryocar brasiliense Camb.), a typical fruit of the Brazilian Cerrado, is an important source of micronutrients and fatty acids. In this work, a new approach for the acid digestion (using H2SO4, HNO3 and H2O2) of pequi oil samples and the determination of Cu, Fe, Zn and Mn by flame atomic absorption spectrometry (F AAS) was employed. Capillary zone electrophoresis (CZE) was used for free fatty acid (FFA) determination after simple and fast extraction with heated ethanol. Good results regarding precision (RSD < 10%, in most cases), sensitivity and adequate LOD and LOQ values were obtained. The accuracy was evaluated using spike tests and the recoveries were from 97 to 107%. The analytes were investigated in four different pequi oil samples. Fe was the trace element with the highest concentration (from 1.99 to 10.3 mg/100 g), followed by Zn, Mn and Cu (1.15 to 3.19, 0.42 to 0.91 and 0.31 to 0.56 mg/100 g, respectively). The main FFA found were oleic acid and palmitic acid (1.61 to 10.7 and 0.82 to 2.69 g/100 g, respectively), while linoleic acid (0.50 g/100 g) was detected in only one sample. The pequi oil chemical composition showed good characteristics to be used as a food additive, in cosmetic formulations and for traditional medicine.

English

The study was carried out between March, 2012 and January, 2013 at the University for Development Studies, Nyankpala campus, Ghana. The objectives of the study were to characterize farmer shea nut varieties to determine the physical and chemical characteristics of the shea nut butter and determine appropriate shea nut quality for the domestic market. The shea nuts were characterized as T1 (small nuts), T2 (medium nuts), T3 (big nuts) and T4 (very big nuts). These nuts were arranged in a completely randomized design and replicated three times for shea nut butter extraction and assessed for free fatty acid (FFA), moisture content and sensory quality. The study reveals strong correlation between shea nut size and butter quality with bigger shea nuts recording lower FFA and moisture content hence higher butter quality. There were differences in colour and texture of the shea butter extracted from the different shea nut sizes. It is recommended that, further research be carried out at different geographical locations to assess the effect of shea nut size on butter quality. Key words: Shea kernel, free fatty acid, moisture content, sensory quality.

The Level of Circulating Octanoate Does Not Predict GhrelinO-Acyl Transferase (GOAT)-Mediated Acylation of Ghrelin During Fasting

Acyl-ghrelin is a 28-amino acid peptide released from the stomach. Ghrelin O-acyl transferase (GOAT) attaches an 8-carbon medium-chain fatty acid (MCFA) (octanoate) to serine 3 of ghrelin. This acylation is necessary for the activity of ghrelin. Animal data suggest that MCFAs provide substrate for GOAT and an increase in nutritional octanoate increases acyl-ghrelin. To address the question of the source of substrate for acylation, we studied whether the decline in ghrelin acylation during fasting is associated with a decline in circulating MCFAs. Eight healthy young men (aged 18-28 years, body mass index range, 20.6-26.2 kg/m(2)) had blood drawn every 10 minutes for acyl- and desacyl-ghrelin and every hour for free fatty acids (FFAs) during the last 24 hours of a 61.5-hour fast and during a fed day. FFAs were measured by a highly sensitive liquid chromatography-mass spectroscopy method. Acyl- and desacyl-ghrelin were measured in an in-house assay; the results were published previously. Ghrelin acylation was assessed by the ratio of acyl-ghrelin to total ghrelin. With the exception of MCFAs C8 and C10, all other FFAs, the MCFAs (C6 and C12), and the long-chain fatty acids (C14-C18) significantly increased with fasting (P < .05). There was no significant association between the fold change in ghrelin acylation and circulating FFAs. These results suggest that changes in circulating MCFAs are not linked to the decline in ghrelin acylation during fasting and support the hypothesis that acylation of ghrelin depends at least partially on the availability of gastroluminal MCFAs or the regulation of GOAT activity.

Constraint‐based modeling of heterologous pathways: Application and experimental demonstration for overproduction of fatty acids in Escherichia coli

Constraint-based modeling has been shown, in many instances, to be useful for metabolic engineering by allowing the prediction of the metabolic phenotype resulting from genetic manipulations. But the basic premise of constraint-based modeling-that of applying constraints to preclude certain behaviors-only makes sense for certain genetic manipulations (such as knockouts and knockdowns). In particular, when genes (such as those associated with a heterologous pathway) are introduced under artificial control, it is unclear how to predict the correct behavior. In this paper, we introduce a modeling method that we call proportional flux forcing (PFF) to model artificially induced enzymatic genes. The model modifications introduced by PFF can be transformed into a set of simple mass balance constraints, which allows computational methods for strain optimization based on flux balance analysis (FBA) to be utilized. We applied PFF to the metabolic engineering of Escherichia coli (E. coli) for free fatty acid (FFA) production-a metabolic engineering problem that has attracted significant attention because FFAs are a precursor to liquid transportation fuels such as biodiesel and biogasoline. We show that PFF used in conjunction with FBA-based computational strain optimization methods can yield non-obvious genetic manipulation strategies that significantly increase FFA production in E. coli. The two mutant strains constructed and successfully tested in this work had peak fatty acid (FA) yields of 0.050 g FA/g carbon source (17.4% theoretical yield) and 0.035 g FA/g carbon source (12.3% theoretical yield) when they were grown using a mixed carbon source of glucose and casamino acids in a ratio of 2-to-1. These yields represent increases of 5.4- and 3.8-fold, respectively, over the baseline strain.

Esterification pretreatment of free fatty acid in biodiesel production, from laboratory to industry

In the US, biodiesel producers usually follow the 19.8:1 methanol-to-FFA molar ratio for free fatty acid (FFA) esterification, as suggested by the National Renewable Energy Laboratory (NREL) without optimization studies. In this paper, both laboratory studies and industrial practices of the esterification process were compared, and an optimization study of a used vegetable oil with 5% FFA was conducted. The optimal conditions of this oil, i.e., methanol-to-FFA molar ratio of 40:1, and sulfuric acid usage of 10%, fell out of the suggested range of 19.8:1. The activation energy of the esterification reaction is 20.7kJ/mol at the optimized condition and 45.9kJ/mol at the 19.8:1 methanol to FFA ratio. It was found that the 19.8:1 methanol-to-FFA molar ratio worked well only within the FFA range of 15–25% while the suggested 5% sulfuric acid worked well only within the FFA range of 15–35%. Outside these ranges, especially at FFA levels less than 15%, optimization study is necessary. Regression models of methanol and acid dosing have been utilized in two industrial scale biodiesel producing facilities and have successfully reduced the FFA level to less than 0.5%.

Desorption Electrospray Ionization Mass Spectrometry Reveals Lipid Metabolism of Individual Oocytes and Embryos

Alteration of maternal lipid metabolism early in development has been shown to trigger obesity, insulin resistance, type 2 diabetes and cardiovascular diseases later in life in humans and animal models. Here, we set out to determine (i) lipid composition dynamics in single oocytes and preimplantation embryos by high mass resolution desorption electrospray ionization mass spectrometry (DESI-MS), using the bovine species as biological model, (ii) the metabolically most relevant lipid compounds by multivariate data analysis and (iii) lipid upstream metabolism by quantitative real-time PCR (qRT-PCR) analysis of several target genes (ACAT1, CPT 1b, FASN, SREBP1 and SCAP). Bovine oocytes and blastocysts were individually analyzed by DESI-MS in both positive and negative ion modes, without lipid extraction and under ambient conditions, and were profiled for free fatty acids (FFA), phospholipids (PL), cholesterol-related molecules, and triacylglycerols (TAG). Principal component analysis (PCA) and linear discriminant analysis (LDA), performed for the first time on DESI-MS fused data, allowed unequivocal discrimination between oocytes and blastocysts based on specific lipid profiles. This analytical approach resulted in broad and detailed lipid annotation of single oocytes and blastocysts. Results of DESI-MS and transcript regulation analysis demonstrate that blastocysts produced in vitro and their in vivo counterparts differed significantly in the homeostasis of cholesterol and FFA metabolism. These results should assist in the production of viable and healthy embryos by elucidating in vivo embryonic lipid metabolism.

Editorial: Molecular Endocrinology Special Feature Minireviews for August 2013: GPCRs in Endocrine Physiology and Pathophysiology

G protein-coupled receptors (GPCRs) are a large superfamily of plasma membrane receptors, which mediate a number of physiological regulatory mechanisms. They are also major drug targets used for a variety of diseases. Of particular importance to the field of endocrinology, GPCRs have recognized roles in the physiology, pathophysiology, and therapy of diabetes mellitus and metabolic syndrome. A GPCR forum held at ENDO2013 chaired by Agnes Schonbrunn, Deborah L. Segaloff, and Robert Peter Millar addressed this topic under the title, New light on GPCRs in the pathophysiology of diabetes and metabolic disorders. In this special issue, the editors of Molecular Endocrinology invited the speakers of this event to present their views and most recent results in this field. Human pancreatic islets of Langerhans express a large number of GPCRs. Most of these receptors are present in insulin-producing -cells, which is the main population of human islet cells. In these cells, as well as in other islet cells, GPCRs mediate sympathetic and parasympathetic actions and the effects of incretins, such as glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide. However, the physiological and therapeutic relevance of the many GPCRs expressed in pancreatic islets are still unknown. GLP-1 mimetics are used successfully in the treatment of type 2 diabetes mellitus (T2DM), demonstrating the relevance of targeting specific GPCRs to the treatment of this disease. Therefore, GPCRs become a major target of drug development for antidiabetic therapy because they can increase insulin secretion, potentiate the effects of glucose, and increase -cell mass by stimulating cell proliferation or decreasing apoptosis. GLP-1 receptors are important targets for the therapy of T2DM. Patrick Sexton and colleagues describe the divergent effects of the responses of endogenous and exogenous GLP-1 receptor ligands on the function of the receptor. Their work suggests that naturally occurring polymorphisms of this receptor have the potential to produce differential responses to distinct GLP-1 receptor ligands. Furthermore, these findings reveal the importance of individualized therapy, which takes these variations into consideration. Receptors for free fatty acids (FFAs) are either validated (FFA1) or potential (FFA2-FFA4) therapeutic targets for the treatment of T2DM and other diseases. Consistently, FFA1 receptors are highly expressed in pancreatic islets, whereas other FFA receptors are also expressed in this tissue. Graeme Milligan and his coworkers present data about the limitations of rodent models to study the human relevance of FFA receptors and demonstrate that their polymorphisms can have functional consequences. M3 muscarinic receptors are the major mediators of the effects of acetylcholine released during parasympathetic receptor on insulin and glucagon secretions. Jurgen Wess and colleagues present their findings, which demonstrated physiological roles of M3 receptors in rodent models and present an innovative approach using an M3 receptor-based designer receptor, which enabled them to chronically activate in -cells a Gq-coupled receptor by an otherwise pharmacologically inert drug. Their data show that chronic stimulation of this pathway protected mice against experimentally induced diabetes and glucose intolerance.

A highly active nanocomposite silica-carbon supported palladium catalyst for decarboxylation of free fatty acids for green diesel production: Correlation of activity and catalyst properties

A class of Pd catalyst supported on a silica-activated carbon nanocomposite for free fatty acid (FFA) decarboxylation was developed, and displayed excellent activity and operation stability selectively for the green diesel hydrocarbons formation in the absence of H2 under mild reaction conditions. Six catalysts containing 5wt% Pd were prepared by systematically varying the silica content in the support. In addition to the effect of particle size, the impact of catalyst preparation method on the activity and selectivity was elucidated. A 5wt% Pd/Si-C-4 catalyst maintained stable activity for 16 days under reaction conditions of 1.5MPa and 300°C. Although a continuous supply of H2 was not necessary, H2 treatment was essential to restore the catalytic activity and the desired product selectivity. Characterization of the catalyst revealed that the highly active Pd/Si-C-4 catalyst has easily accessible and well-distributed metallic Pd nanoparticles inside the hybrid mesopores.

In vitro study of intestinal lipolysis using pH-stat and gas chromatography

Developing healthy products requires in-depth knowledge of digestion. This study focuses on lipid digestion in relation to emulsion properties typically followed by pH-stat. Although this is a fast and easy method to follow the overall digestion, it provides no details on lipid digestion products. Thus, the aims of the present study were to use gas chromatography (GC) to determine all products present during lipolysis, i.e. monoglycerides (MG), diglycerides (DG) and triglycerides (TG), and to compare this method with the pH-stat method for free fatty acids (FFA). Fine, medium and coarse emulsions stabilized with two different emulsifiers (whey protein isolate (WPI) or gum arabic) were digested under in vitro intestinal conditions. Although the amount of FFA increased for both methods for WPI stabilized emulsions, the amount of FFA was 2–3 times higher when determined by GC compared with pH-stat. GC analysis showed decreasing amounts of MG and DG with increasing droplet size for both emulsions. Molar ratios of FFA/DG and MG/DG were twofold higher for WPI than for gum arabic stabilized emulsions. This indicates that the total production of lipolytic products (i.e. FFA+MG+DG) depends on the droplet size and the emulsifier but their proportions only depend on the emulsifier. Although pH-stat provides a fast measure of FFA release, it is influenced by the emulsifier type at the oil–water interface and therefore care should be taken when interpreting pH-stat results. We suggest combining this method with GC for accurate FFA determination and further evaluation of all lipolytic products.

SHELF STABILITY AND SENSORY ATTRIBUTES OF A DEEP-FRIED PRODUCT INCORPORATED WITH PRE-TREATED SOY FLOUR

ABSTRACT The effect of incorporating soy on quality of a deep-fried product was investigated. Defatted soy flour was pretreated with microwave heat, pressure heat and enzyme and incorporated at 20% level to the product prepared with Bengal gram. Product without soy served as control. They were stored in different packaging materials at room and low temperatures for 4 weeks and analyzed every week for free fatty acids (FFA), peroxide value (PV) and acceptability. Fat absorption reduced by 31% and 11–19% in products prepared with untreated and pretreated soy, respectively, in comparison with control. The FFA content of low temperature-stored product at week IV was 0.92% compared with products stored at room temperature (1.56–1.81%). Incorporation of soy flour did not alter the FFA content in stored products but decreased PV. Low temperature-stored products were acceptable at week IV, whereas others developed an off flavor at the end of week III. PRACTICAL APPLICATIONS Deep-fried foods have a large share of the snack foods industry and are specially liked for their crunchy texture and unique flavor. Generally, however, they have high fat content and need to be consumed in restricted amounts for health reasons. A reduction in fat content of fried foods without compromising the sensory quality is a major challenge for the food manufacturer. The study explores the possibility of utilizing pretreated defatted soy flour in a deep-fried snack product with the purpose of reducing fat absorption and improving sensory and keeping quality. Incorporation of soy flour resulted in reduced fat absorption in fried products and pretreatment of soy flour improved the sensory quality of products. The study has potential application in snack food industry for reducing fat content in fried products and for improving sensory attributes by incorporating defatted soy flour, which in turn has many health benefits.

Short-term oral exposure to white wine transiently lowers serum free fatty acids

In humans little is known as to whether oral sensory stimulation with alcohol elicits cephalic phase responses. This study sought to determine whether oral alcohol exposure, in the form of white wine, provokes cephalic phase responses in normal-weight and overweight women. In a semi-randomized, crossover trial, eleven normal-weight and eleven overweight women sham-fed, after an overnight fast under three separate conditions 4 weeks apart, cake (750kJ), 25cL white wine (750kJ; ∼26g alcohol) and 25cL water. Blood was drawn prior to and for 30min after two 3-min episodes of modified sham-feeding (MSF). Blood samples were analyzed for free fatty acid (FFA), triglyceride, glucose, pancreatic polypeptide (PP), insulin and alcohol concentrations. Incremental area under the curves (IAUC) of FFA concentrations differed significantly between the three treatments but not between BMI categories. After MSF with wine, FFA concentrations dropped to a minimum of 77±3% of baseline concentrations at t=12±2min after baseline and returned to baseline after ∼30min, whereas after MSF with cake and water, FFA concentrations gradually increased. In conclusion, short-term oral white wine exposure substantially and temporarily decreases FFA concentrations suggesting a cephalic phase response of alcohol. This effect occurred regardless of BMI.

The effect of modifying dietary protein and carbohydrate in weight loss on arterial compliance and postprandial lipidemia in overweight women with polycystic ovary syndrome

In overweight women with polycystic ovary syndrome, weight loss improves arterial compliance and postprandial lipidemia. Modifying dietary carbohydrate or protein in weight loss provided similar improvements in arterial compliance and postprandial lipidemia.

Monitoring of Fat Content, Free Fatty Acid and Fatty Acid Profile Including trans Fat in Pakistani Biscuits

AbstractThe fat contents of 12 brands of biscuits were extracted and evaluated for free fatty acids (FFA) and their fatty acid composition (FAC). The oil content and FFA varied from 13.7 to 27.6% and 0.2 to 1.0%, respectively. The FAC was analyzed by gas chromatography–mass spectroscopy with particular emphasis on trans fatty acids (TFA). Total saturated, unsaturated, cis‐monounsaturated and polyunsaturated fatty acids were determined in the range of 37.9–46.9, 53.0–62.0, 12.3–43.7 and 0.1–9.2%, respectively. The high amount of TFA was observed in all biscuit samples and varied from 9.3 to 34.9%. The quantity and quality of the lipid fraction of the biscuits indicated that the all analyzed biscuits are a rich source of fat, saturated fatty acids and trans fatty acids, consequently not suitable for the health of consumers. The high content of trans fatty acids and palmitic acid also indicated that blends of RBD palm oil and partially hydrogenated oil had been used in the biscuit manufacturing.

Oral Detection of Short-, Medium-, and Long-Chain Free Fatty Acids in Humans

There is increasing evidence supporting an oral chemosensory detection system for free fatty acids (FFA). The presumptive transduction mechanisms have different ligand specificities. Psychophysical studies with FFA varying in chain length and saturation may aid in identifying the presence and functionality of these mechanisms in humans. Oral detection thresholds were measured for linoleic, stearic, lauric, and caproic acids in 32 healthy adults by an ascending, 3-alternative, forced-choice, sip and spit procedure. Thresholds were obtained for all fatty acids from all participants, but the distributions were wide and nonnormal. Thresholds were not correlated between fatty acids nor with thresholds for sucrose (taste), butanol (olfactory), mineral oil, or gum acacia (both somatosensory). These data demonstrate human oral sensitivity to short-, medium-, and long-chain FFA and suggest the presence of multiple transduction mechanisms. The findings are consistent with, but do not definitively demonstrate, a role for taste that may have a genetic basis.

Determination of Free Fatty Acids in Edible Oils with the Use of a Variable Filter Array IR Spectrometer

AbstractThe feasibility of employing a portable variable filter array (VFA) IR spectrometer equipped with a transmission flow cell to quantitatively analyze edible oils or biodiesel feedstocks for free fatty acids (FFA) was evaluated. The approach to FFA determination employed was based on a previously reported FTIR method that involves the extraction of FFAs into methanol containing the base NaHNCN, which converts the FFAs to their salts, followed by measurement of the carboxylate absorbance at ~1,573 cm−1 in the spectrum of the methanol phase. When this methodology was implemented on the low‐resolution VFA‐IR spectrometer, the analytical performance was comparable to that of conventional FTIR instrumentation at FFA concentrations of &lt;1%. However, at higher FFA levels, the relatively weak pulsed IR source of the VFA‐IR spectrometer was found to provide insufficient energy for accurate measurement of the carboxylate absorption superimposed on the strong methanol absorption at ~1,450 cm−1. By changing the extraction solvent to ethanol (EtOH), good spectra and calibrations could be obtained over an FFA range of 0–5%, having an overall SD of ±0.07% FFA. Based on this assessment, a VFA‐IR spectrometer provides an economical instrumental means for at‐line monitoring of FFA levels in crude and refined edible oils and biodiesel feedstocks, capable of analyzing ~20–30 prepared samples per hour.

Effects of pomegranate sauce on quality of marinated anchovy during refrigerated storage

Effects of pomegranate sauce on the quality of marinated anchovy during storage at 4 °C were investigated. Anchovy were marinated with 30 g/L acetic acid and 150 g/L salt, put into glass jars, filled with either sunflower oil or pomegranate sauce and stored at 4 °C. Total volatile basic nitrogen (TVB-N) and trimethylamine (TMA-N) values increased during the storage. Higher values for free fatty acid (FFA), conjugated diens (k-dien) and para-anisidine ( p-Av) were found in samples with sunflower oil than those with pomegranate sauce. Samples in pomegranate sauce showed better oxidative stability. Higher taste and flavour and lower appearance scores were found for samples in pomegranate sauce than those samples in sunflower oil. It was found that pomegranate sauce was as least effective as the traditional sunflower oil to keep quality. Pomegranate sauce also produced desirable taste and flavour but the coloration should be studied in further studies.

Endothelial Nitric Oxide Synthase (eNOS) Knockout Mice Have Defective Mitochondrial β-Oxidation

Recent observations indicate that the delivery of nitric oxide by endothelial nitric oxide synthase (eNOS) is not only critical for metabolic homeostasis, but could also be important for mitochondrial biogenesis, a key organelle for free fatty acid (FFA) oxidation and energy production. Because mice deficient for the gene of eNOS (eNOS(-/-)) have increased triglycerides and FFA levels, in addition to hypertension and insulin resistance, we hypothesized that these knockout mice may have decreased energy expenditure and defective beta-oxidation. Several markers of mitochondrial activity were assessed in C57BL/6J wild-type or eNOS(-/-) mice including the energy expenditure and oxygen consumption by indirect calorimetry, in vitro beta-oxidation in isolated mitochondria from skeletal muscle, and expression of genes involved in fatty acid oxidation. eNOS(-/-) mice had markedly lower energy expenditure (-10%, P < 0.05) and oxygen consumption (-15%, P < 0.05) than control mice. This was associated with a roughly 30% decrease of the mitochondria content (P < 0.05) and, most importantly, with mitochondrial dysfunction, as evidenced by a markedly lower beta-oxidation of subsarcolemmal mitochondria in skeletal muscle (-30%, P < 0.05). Finally, impaired mitochondrial beta-oxidation was associated with a significant increase of the intramyocellular lipid content (30%, P < 0.05) in gastrocnemius muscle. These data indicate that elevated FFA and triglyceride in eNOS(-/-) mice result in defective mitochondrial beta-oxidation in muscle cells.