Formation of trans-epoxy fatty acids correlates with formation of isoprostanes and could serve as biomarker of oxidative stress.

In mammals, epoxy-polyunsaturated fatty acids (epoxy-PUFA) are enzymatically formed from naturally occurring all-cis PUFA by cytochrome P450 monooxygenases leading to the generation of cis-epoxy-PUFA (mixture of R,S- and S,R-enantiomers). In addition, also non-enzymatic chemical peroxidation gives rise to epoxy-PUFA leading to both, cis- and trans-epoxy-PUFA (mixture of R,R- and S,S-enantiomers). Here, we investigated for the first time trans-epoxy-PUFA and the trans/cis-epoxy-PUFA ratio as potential new biomarker of lipid peroxidation. Their formation was analyzed in correlation with the formation of isoprostanes (IsoP), which are commonly used as biomarkers of oxidative stress. Five oxidative stress models were investigated including incubations of three human cell lines as well as the in vivo model Caenorhabditis elegans with tert-butyl hydroperoxide (t-BOOH) and analysis of murine kidney tissue after renal ischemia reperfusion injury (IRI). A comprehensive set of IsoP and epoxy-PUFA derived from biologically relevant PUFA (ARA, EPA and DHA) was simultaneously quantified by LC-ESI(-)-MS/MS. Following renal IRI only a moderate increase in the kidney levels of IsoP and no relevant change in the trans/cis-epoxy-PUFA ratio was observed. In all investigated cell lines (HCT-116, HepG2 and Caki-2) as well as C. elegans a dose dependent increase of both, IsoP and the trans/cis-epoxy-PUFA ratio in response to the applied t-BOOH was observed. The different cell lines showed a distinct time dependent pattern consistent for both classes of autoxidatively formed oxylipins. Clear and highly significant correlations of the trans/cis-epoxy-PUFA ratios with the IsoP levels were found in all investigated cell lines and C. elegans. Based on this, we suggest the trans/cis-epoxy-PUFA ratio as potential new biomarker of oxidative stress, which warrants further investigation.

Ácidos grasos trans y ácido linoleico conjugado en alimentos: origen y propiedades biológicas / Trans fatty acids and conjugated linoleic acid in food: origin and biological properties

Los ácidos grasos trans (AGT) son componentes lipídicos minoritarios que se encuentran en distintos alimentos, entre ellos, aquellos derivados de animales rumiantes, que han merecido atención por su relación con el riesgo de incidir en enfermedades cardiovasculares. El origen de los AGT en los alimentos se encuentra mayoritariamente en los procesos de hidrogenación industrial de aceites vegetales insaturados y en las reacciones enzimáticas de biohidrogenación que tienen lugar, de forma natural, en el tracto digestivo de los rumiantes. Aunque las moléculas que se generan por ambos mecanismos son similares, la distribución isomérica de los AGT es muy diferente, lo que puede generar diferencias a la hora de evaluar los efectos biológicos derivados de su consumo. Las grasas vegetales hidrogenadas son abundantes en ácido elaídico (trans-9 18:1) y trans-10 18:1 entre otros. En contraste, el ácido vacénico (trans-11 18:1) es el principal AGT presente en la leche y otros productos derivados de rumiantes, siendo además precursor fisiológico del ácido linoleico conjugado, un componente al que se atribuyen numerosos efectos beneficiosos para la salud. En este artículo se actualizan los efectos biológicos y las potenciales propiedades bioactivas de estos ácidos grasos

Trans fatty acids (TFA) are minor lipid components present in different foods, including ruminant derived products, which have received great attention due to their relationship with cardiovascular disease risk. The origin of TFA in food is mainly related to the industrial hydrogenation processes of unsaturated vegetable oils, but they can also occur naturally in the digestive tract of ruminants by enzymatic biohydrogenation reactions. Both mechanisms generate similar TFA compounds. However, TFA consumption may exert different biological effects depending on the isomeric distribution, which is strongly influenced by the dietary source (i.e., industrial or natural). Industrial partially hydrogenated vegetable fats are rich in elaidic (trans-9 18:1) and trans-10 18:1 fatty acids, among others. In contrast, vaccenic acid (trans-11 18:1) is the major TFA isomer detected in milk and other ruminant derived products. Vaccenic acid is the physiological precursor of conjugated linoleic acid, a bioactive lipid with beneficial effects on human health. This article provides updated information on the biological effects and potential bioactive properties of TFA considering both, their chemical structure and provenance

[Trans fatty acids and conjugated linoleic acid in food: origin and biological properties]. / Ácidos grasos trans y ácido linoleico conjugado en alimentos: origen y propiedades biológicas.

INTRODUCTION: Trans fatty acids (TFA) are minor lipid components present in different foods, including ruminant derived products, which have received great attention due to their relationship with cardiovascular disease risk. The origin of TFA in food is mainly related to the industrial hydrogenation processes of unsaturated vegetable oils, but they can also occur naturally in the digestive tract of ruminants by enzymatic biohydrogenation reactions. Both mechanisms generate similar TFA compounds. However, TFA consumption may exert different biological effects depending on the isomeric distribution, which is strongly influenced by the dietary source (i.e., industrial or natural). Industrial hydrogenated vegetable fats are rich in elaidic (trans-9 18:1) and trans-10 18:1 fatty acids, among others. In contrast, vaccenic acid (trans-11 18:1) is the major TFA isomer detected in milk and other ruminant derived products. Vaccenic acid is the physiological precursor of conjugated linoleic acid, a bioactive lipid with beneficial effects on human health. This article provides updated information on the biological effects and potential bioactive properties of TFA considering both, their chemical structure and provenance.

INTRODUCCIÓN: Los ácidos grasos trans (AGT) son componentes lipídicos minoritarios que se encuentran en distintos alimentos, entre ellos, aquellos derivados de animales rumiantes, que han merecido atención por su relación con el riesgo de incidir en enfermedades cardiovasculares. El origen de los AGT en los alimentos se encuentra mayoritariamente en los procesos de hidrogenación industrial de aceites vegetales insaturados y en las reacciones enzimáticas de biohidrogenación que tienen lugar, de forma natural, en el tracto digestivo de los rumiantes. Aunque las moléculas que se generan por ambos mecanismos son similares, la distribución isomérica de los AGT es muy diferente, lo que puede generar diferencias a la hora de evaluar los efectos biológicos derivados de su consumo. Las grasas vegetales hidrogenadas son abundantes en ácido elaídico (trans-9 18:1) y trans-10 18:1 entre otros. En contraste, el ácido vacénico (trans-11 18:1) es el principal AGT presente en la leche y otros productos derivados de rumiantes, siendo además precursor fisiológico del ácido linoleico conjugado, un componente al que se atribuyen numerosos efectos beneficiosos para la salud. En este artículo se actualizan los efectos biológicos y las potenciales propiedades bioactivas de estos ácidos grasos.

Characterization of two Agrostis-Festuca alpine pastures and their influence on cheese composition.

Recently, there has been a renewed interest in mountain farming, and several studies have been carried out on milk and cheese obtained in the unique environmental conditions of the Alps, a 1300 km mountain chain, located in the north of Italy. In this paper, the influence, on some cheese constituents, of two very similar mountain grasslands, both dominated by Festuca - Agrostis , was investigated. The two pastures were located in the same area in the southeastern Italian alpine region and differed in sunshine orientation and exposure. Milk obtained from cows grazing on these pastures was used to produce a semi-hard traditional cheese. The differences observed between the cheeses of the two areas for both some hydrocarbons (1-phytene and 2-phytene) and trans-fatty acids can be explained by a different rumen environment created by the botanical composition of the two pastures. The multidisciplinary approach can be considered a successful strategy, suitable for studying markers of authenticity.

The impact of substituting SFA in dairy products with MUFA or PUFA on CVD risk: evidence from human intervention studies.

With the substantial economic and social burden of CVD, the need to modify diet and lifestyle factors to reduce risk has become increasingly important. Milk and dairy products, being one of the main contributors to SFA intake in the UK, are a potential target for dietary SFA reduction. Supplementation of the dairy cow's diet with a source of MUFA or PUFA may have beneficial effects on consumers' CVD risk by partially replacing milk SFA, thus reducing entry of SFA into the food chain. A total of nine chronic human intervention studies have used dairy products, modified through bovine feeding, to establish their effect on CVD risk markers. Of these studies, the majority utilised modified butter as their primary test product and used changes in blood cholesterol concentrations as their main risk marker. Of the eight studies that measured blood cholesterol, four reported a significant reduction in total and LDL-cholesterol (LDL-C) following chronic consumption of modified milk and dairy products. Data from one study suggested that a significant reduction in LDL-C could be achieved in both the healthy and hypercholesterolaemic population. Thus, evidence from these studies suggests that consumption of milk and dairy products with modified fatty acid composition, compared with milk and dairy products of typical milk fat composition, may be beneficial to CVD risk in healthy and hypercholesterolaemic individuals. However, current evidence is insufficient and further work is needed to investigate the complex role of milk and cheese in CVD risk and explore the use of novel markers of CVD risk.

Isolation and optimisation of the oleaginous yeast Sporobolomyces roseus for biosynthesis of 13C isotopically labelled 18-carbon unsaturated fatty acids and trans 18:1 and 18:2 derivatives through synthesis.

An oleaginous and psychrotrophic strain (F38-3) of Sporobolomyces roseus Kluyver & van Niel was isolated from a salt marsh environment in Nova Scotia, Canada following a screening program to select for high producers of 18-carbon unsaturated fatty acids. Fatty acid production was characterised as a function of temperature at 20 g glucose L(-1), and optimal yields were obtained at 14°C, achieving 5.7 g dw biomass and 39.2% total fatty acids by dry weight, with 18:1, 18:2 and 18:3 all-cis fatty acids accounting for 49.4%, 14.3% and 6.7% of total fatty acids (TFA), respectively--the highest reported for this species. Production of 18:3 was inversely correlated to growth temperature, rising from 2% of TFA at 30°C to 8.9% at 6°C. Cultivation of isolate F38-3 on universally (13)C (U-(13)C) labelled glucose and subsequent transesterification and isolation of the fatty acid methyl esters (FAMEs) by preparative chromatography yielded pure, highly (13)C-enriched (>90%) 18:1, 18:2 and 18:3 all-cis FAMEs. The U-(13)C 18:1 FAME was catalytically converted to U-(13)C 18:1 trans-9 and purified to >99.5% purity. The U-(13)C 18:2 was converted by alkaline isomerisation into a 50/50 mixture of 18:2 cis-9, trans-11 and 18:2 trans-10, cis-12 isomers and purified to >95.0% purity. Overall, 10%, by weight, of labelled glucose fed to isolate F38-3 was recovered as fatty acid methyl esters and 7.5% as 18-carbon unsaturated fats, and the final isomerisation reactions resulted in yields of 80% or greater. The ultimate goal of the work is to develop methodologies to produce (13)C-labelled metabolic tracers as tools to study the metabolism of trans fats.

Effect of sunflower, linseed and fish oils on the production of trans fatty acids in vitro.

In vitro anaerobic incubations were used to determine the effect of different oils (LO-linseed, SO-sunflower, FO-fish oil) on trans fatty acid production in rumen fluid and to test if combining of monensin (MON) with the oils affects the interactions on trans fatty acid concentrations in mixed cultures of ruminal microorganisms. Two different sources of rumen fluid were used; the inoculum from the sheep fed hay and barley (80:20%)--the inoculum A and the inoculum from the sheep fed alfalfa and barley (80:20 %)--the inoculum B. The analyses showed that inoculum B contained more short chain fatty acids (SCFA), medium chain fatty acids (MCFA) and saturated fatty acids (SFA) than inoculum A. In contrast, inoculum A contained more unsaturated fatty acids (UFA) than inoculum B. The results show, that the oils affected the biohydrogenation of fatty acids (FA) by increasing the concentration of C18:0 (3-7 times) and trans C18:1 isomers (2-9 times). The concentration of two main intermediates of FA biohydrogenation-- cis 9, trans 11 C18:2 (CLA) and trans 11C18:1 (TVA) were increased with the oils, but FO was more efficient than other plant oils on CLA and TVA production. The monensin treatment had similar effect on FA metabolism as the oil treatment in comparison to unincubated control. The interactions of monensin treatment with the oils were characterized with decrease (LO+MON, SO+MON) or increase (FO+MON) of the proportions of C18:0 and trans C18:1 isomers in comparison to oil treatment. The highest concentrations of two main isomers--CLA,TVA were found in the samples containing fish oil and monensin. In conclusion, fish oil treatment and monensin with fish oil treatment was more efficient than other plant oils in the effect on trans fatty acid production (mainly CLA and TVA) in fermentation fluid in vitro.

Formation of trans fatty acids is not involved in growth-linked membrane adaptation of Pseudomonas putida.

Fatty acid compositions in growing and resting cells of several strains of Pseudomonas putida (P8, NCTC 10936, and KT 2440) were studied, with a focus on alterations of the saturation degree, cis-trans isomerization, and cyclopropane formation. The fatty acid compositions of the strains were very similar under comparable growth conditions, but surprisingly, and contrary to earlier reports, trans fatty acids were not found in either exponentially growing cells or stationary-phase cells. During the transition from growth to the starvation state, cyclopropane fatty acids were preferentially formed, an increase in the saturation degree of fatty acids was observed, and larger amounts of hydroxy fatty acids were detected. A lowered saturation degree and concomitant higher membrane fluidity seemed to be optimal for substrate uptake and growth. The incubation of cells under nongrowth conditions rapidly led to the formation of trans fatty acids. We show that harvesting and sample preparation for analysis could provoke the enzyme-catalyzed formation of trans fatty acids. Freeze-thawing of resting cells and increased temperatures accelerated the formation of trans fatty acids. We demonstrate that cis-trans isomerization only occurred in cells that were subjected to an abrupt disturbance without having the possibility of adapting to the changed conditions by the de novo synthesis of fatty acids. The cis-trans isomerization reaction was in competition with the cis-to-cyclopropane fatty acid conversion. The potential for the formation of trans fatty acids depended on the cyclopropane content that was already present.

Short communication: Diurnal profiles of conjugated linoleic acids and trans fatty acids in ruminal fluid from cows fed a high concentrate diet supplemented with fish oil, linseed oil, or sunflower oil.

Trans-18:1 and 18:2 isomer composition in ruminal fluid during the daily feeding cycle was examined in 3 cows fed a high concentrate diet (35:65) with 5% (DM basis) sunflower oil (SO), 5% linseed oil (LO), or 2.5% fish oil (FO) in a 3 x 3 Latin square with 3 4-wk periods. Grass hay and concentrate mixtures were fed at 0900, 1300, and 1700 h daily. Ruminal fluid was collected at 0900, 1100, 1300, 1500, 1700, 2000, and 0000 h. Feeding SO resulted in the greatest mean concentrations (% of total fatty acids) of trans10,cis12-18:2 and cis9,trans11-18:2. In particular, trans10,cis12-18:2 with SO was greater at 1500 (0.29%), 2000 (0.34%), and 0000 h (0.25%) relative to 0900 h (0.07%). Cis9,trans11-18:2 concentration increased from 0.47% at 0900 h to a peak of 2.06% at 1100 h; it remained greater than the percentage determined at 0900 h at 1300 (1.4%) through 0000 h (1.1%). Concentration of trans11,cis15-18:2 was greatest with LO, ranging from 3.3% (0900 h) to a peak of 11.4% at 2000 h. Mean trans10-18:1 concentration ranked by diet was SO > FO > LO. Peak trans10-18:1 with SO was observed at 1700 h (14.9%) compared with 0900 h (5.1%). Trans11-18:1 did not differ with diet or time. Stearic acid decreased over time with all diets reaching minimum concentrations at 1700 to 2000 h relative to 0900 h. Feeding FO, however, decreased mean 18:0 concentration 4-fold compared with LO or SO. The moderate effect on concentration of trans-18:1 coupled with accumulation of 18:2 intermediates and the decrease of 18:0 over time suggest that oils reduced the biohydrogenation of 18:2 isomers to trans-18:1.