Conjugated Linoleic Acid Isomers Research Articles

Production of Conjugated Linoleic Acid (CLA) by Lactiplantibacillus plantarum: A Review with Emphasis on Fermented Foods.

The term Conjugated Linoleic Acid (CLA) refers generically to a class of positional and geometric conjugated dienoic isomers of linoleic acid. Among the isomers of linoleic acid cis9, trans11-CLA (c9, t11-CLA) and trans10, cis12-CLA (t10, c12-CLA) are found to be biologically active isomers, and they occur naturally in milk, dairy products and meat from ruminants. In addition, some vegetables and some seafoods have also been reported to contain CLA. Although the CLA levels in these natural sources are insufficient to confer the essential health benefits, anti-carcinogenic or anti-cancer effects are of current interest. In the rumen, CLA is an intermediate of isomerization and the biohydrogenation process of linoleic acid to stearic acid conducted by ruminal microorganisms. In addition to rumen bacteria, some other bacteria, such as Propionibacterium, Bifidobacterium and some lactic acid bacteria (LAB) are also capable of producing CLA. In this regard, Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) has demonstrated the ability to produce CLA isomers from linoleic acid by multiple enzymatic activities, including hydration, dehydration, and isomerization. L. plantarum is one of the most versatile species of LAB and the bacterium is widely used in the food industry as a microbial food culture. Thus, in this review we critically analyzed the literature produced in the last ten years with the aim to highlight the potentiality as well as the optimal conditions for CLA production by L. plantarum. Evidence was provided suggesting that the use of appropriate strains of L. plantarum, as a starter or additional culture in the production of some fermented foods, can be considered a critical factor in the design of new CLA-enriched functional foods.

Dietary supplementation of phytoncide and soybean oil increases milk conjugated linoleic acid and depresses methane emissions in Holstein dairy cows

The objective of this study was to determine whether adding phytoncide oil (PO) and soybean oil (SBO) to the dairy cow diet could increase milk conjugated linoleic acid (CLA) and depress methane (CH4) emissions in Holstein dairy cows. Rumen fermentation was conducted at four levels of SBO (0, 1, 2, and 4%, on DM basis) and two levels of PO (0 and 0.1%, on DM basis) with in vitro experiment. To evaluate blood parameters, fecal microbe population, milk yield and fatty acid compositions, and CH4 production, in vivo experiment was conducted using 38 Holstein dairy cows divided into two groups of control (fed TMR) and treatment (fed TMR with 0.1% PO and 2% SBO as DM basis). In the in vitro study (Experiment 1), PO or SBO did not affect rumen pH. However, SBO tended to decrease ruminal ammonia-N (p = 0.099). Additionally, PO or SBO significantly decreased total gas production (p = 0.041 and p = 0.034, respectively). Both PO and SBO significantly decreased CH4 production (p < 0.05). In addition, PO significantly increased both CLA isomers (c9, t11 and t10, c12 CLA) (p < 0.001). Collectively, 0.1% PO and 2% SBO were selected resulting in most effectively improved CLA and decreased CH4 production. In the in vivo study (Experiment 2), 0.1% PO with 2% SBO (PSO) did not affect complete blood count. However, it decreased blood urea nitrogen and magnesium levels in blood (p = 0.021 and p = 0.01, respectively). PSO treatment decreased pathogenic microbes (p < 0.05). It increased milk yield (p = 0.017) but decreased percentage of milk fat (p = 0.013) and MUN level (p < 0.01). In addition, PSO treatment increased both the concentration of CLA and PUFA in milk fat (p < 0.01). Finally, it decreased CH4 emissions from dairy cows. These results provide compelling evidence that a diet supplemented with PSO can simultaneously increase CLA concentration and decrease CH4 production with no influence on the amount of milk fat (kg/day) in Holstein dairy cows.

CONJUGATED LINOLEIC ACID AND FATTY ACID ISOMERS IN SELECTED COLD PRESSED OILS: ANALYSIS BY GC/FID TECHNIQUE

The conjugated linoleic acid (CLA) isomers in cold pressed oils [pomegranate seed oil (PGSO), linseed oil (LSO), black cumin seed oil (BCSO), nettle seed oil (NSO),grape seed oil (GSO), sesame seed oil (SSO),safflower oil (SFO), pumpkin seed oil (PSO), wheat germ oil (WGO), fig seed oil (FSO), coriander oil (CO), walnut oil (WO) and coconut oil (CNO)] extracted with lab–scale screw press machine were further subjected to gas chromatography/flame ionization detection (GC/FID) analysis. The composition of polyunsaturated fatty acids (PUFAs), monounsaturated fatty acids (MUFA) and saturated fatty acids (SFAs) of the samples was also determined. The five different positional and geometric isomers of CLA [cis–9, cis–11 CLA, cis–9, trans–11 CLA, trans–9, cis–11 CLA, trans–9, trans–11 CLA and trans–10, cis–12 CLA] were also well separated by a highly polar column (100m×0.2μm×0.25mm i.d; HP–88 cyanopropyl) and an applied GC temperature program. It was concluded that the samples were all rich in total CLA (∑CLA) and they were found between 0.14% for PSO and 2.11% for SSO. The most abundant CLA isomer was in general to be cis–9, trans–11 CLA form, which represented the content of isomer between 3.15% and 72.08% of ∑CLA. Besides, the ∑SFA values were detected between 2.43% and 93.14%, ∑MUFA values were between 4.60% and 71.11% and ∑PUFA values were between 1.79% and 87.59%. Therefore, this study might offer valuable information for the introduction of new food sources, as well as incorporation into medicinal purposes and food formulations which have the potential to be commercially valuable.

Computational Analysis and Heterologous Expression of BBI-like Proteins from Food-Grade Bifidobacterium Species Reveal Possibly a Key Factor in Conjugated Linoleic Acid Bioconversion.

Bifidobacteria are well-known probiotics that can produce multiple health-promoting bioactivities, including the bioconversion of conjugated linoleic acid (CLA). However, there is a lack of insights into the genetic diversity of functional proteins in Bifidobacterium at the species level, particularly owing to the vastly different CLA conversion capabilities of these strains. Here, we performed a comprehensive bioinformatics analysis and the in vitro expression of bbi-like sequences that are widely distributed among CLA-producing Bifidobacterium strains. The BBI-like protein sequences from all four species of bifidobacterial CLA-producing strains were predicted to be stable integral membrane proteins with a transmembrane topology number of seven or nine. All BBI-like proteins were found to be expressed in the Escherichia coli BL21(DE3) hosts with a pure c9, t11-CLA-producing activity. Furthermore, their activities significantly differed in the same genetic background, and their sequence differences were indicated to be important potential factors contributing to the high activity levels in CLA-producing Bifidobacterium breve strains. The use of microorganisms, such as food-grade or industrial-grade strains, to obtain single CLA isomers will accelerate CLA-related food and nutrition research and further enrich the scientific theory of bifidobacteria as probiotics.

Bioproduction of conjugated linolenic acid by L. plantarum using linolenic acid as a substrate

Whole cell biocatalysis is an environment-friendly method for selective synthesis of bioactive isomers of conjugated linolenic acid (CLnA), the potential anticarcinogenic, antiatherogenic and a number of other therapeutic qualities bearing substance. Present study was design to use Lactobacillus plantarum as a whole cell biocatalyst for production of CLnA using linolenic acid (α-LNA) as a substrate. After optimization of biotransformation conditions such as pH, time and α-LNA concentration, the maximum CLnA conversion 70 μg ml−1 was achieved at 48 h with 0.71 mg ml−1 α-LNA under aerobic conditions. HPLC analysis with the diode array detector showed distinct absorption maxima at 270 nm and 267 nm for two separated CLnA isomers. GC-MS analysis of DMOX derivatives of obtained product reveals cis-9, trans-11, cis-15 as one of the major isomer. These results highlight the ability of L. plantarum to produce specific CLnA isomers of different biological and physiological activities at optimized conditions. Hence the microbially produced CLnAs can be good sources for nutritional and health benefits, as functional foods or fermented dairy products, for human and animal intake.

Conjugated linoleic acid trans-10, cis-12 increases the expression of genes from cell cycle progression and cis-9, trans-11 stimulates apoptotic genes in different mammary tumor explants of female dogs.

The conjugated linoleic acid (CLA) isomer cis-9, trans-11 is an anticarcinogen that inhibits cell proliferation and/or induces apoptosis of tumor cells. The objective of this study was to evaluate the expression of genes responsible for cell cycle regulation and apoptosis in tumor explants of mammary anaplastic carcinoma (AC) and mammary tubulopapillary carcinoma (TC) cultured in vitro with the CLA isomers cis-9, trans-11 and trans-10, cis-12. In this study we used mammary explants from two adult female dogs that revealed two types of malignant tumors: (a) anaplastic mammary carcinoma (AC) and (b) mammary tubulopapillary carcinoma (TC). The explants (n = 6 per treatment) had an average weight of 80.0 ± 2.0mg and were cultured for 24h in 35mm culture plates under the following treatments: (a) Control: Culture medium + fatty acid free bovine serum albumin (BSA); (b) Culture medium + cis-9, trans-11 CLA (75 µM) diluted with fatty acid free bovine serum albumin (BSA), and; (c) Culture medium + trans-10, cis-12 CLA (75 µM) diluted with fatty acid free bovine serum albumin (BSA). After that, total RNA was extracted, complementary DNA was synthesized (cDNA), and quantitative analysis by real-time polymerase chain reaction (RT-qPCR) was conducted. Data were analyzed using the MIXED procedure of SAS. Compared with the Control, the CLA trans-10, cis-12 treatment decreased expression of the gene encoding the p53 by 20% (P = 0.02), Caspase-3 by 25% (P = 0.06) and Bax by 51% (P = 0.001) in AC. The CLA cis-9, trans-11 increased the gene expression of proapoptotic protein Bax in TC by 68% (P = 0.01), but increased the expression of the antiapoptotic Bcl2 gene in AC by 72% (P = 0.006). The CLA cis-9, trans-11 stimulates apoptotic genes in mammary tubulopapillary carcinoma, but has a contrary effect on the anaplastic carcinoma, and the CLA trans-10, cis-12 stimulates cell cycle progression genes and may have an antiapoptotic effect, mainly in mammary anaplastic carcinoma.

Probiotics from the bovine raw milk of Lahaul valley showed cis-9, trans-11 conjugated linoleic acid isomer and antioxidant activity with food formulation ability

Dairy foods, especially of bovine origin, are the most common carriers of probiotics and conjugated linoleic acid (CLA)-producing microorganisms. The present study isolated probiotic strains from raw cow milk of the Lahaul valley capable of producing cis-9, trans-11 CLA isomer, besides exhibiting antioxidant properties with food formulation abilities. After preliminary probiotic screening, 34 strains were obtained, with 21 qualifying all the probiotic attributes and passing safety analysis. The seven most statistically promising probiotics were further explored for CLA production (16.87–28.01 μg/mL), and showed cis-9, trans-11 CLA isomer production as confirmed by GC-MS. To establish genomic evidence for safety analysis, whole genome sequencing was conducted for Enterococcus faecalis LJM:05, resulting in the deduction of antibiotic-resistance and virulence genes (lower identity scores). Based on the superior probiotic attributes, antioxidant, and CLA-producing properties, the strain Latilactobacillus curvatus LGM:16 was used to develop the curdled milk, with a legal, viable count (7.55 log10 CFU/mL) under pH ∼4.15 for 28 days of storage periods (4 °C). Scanning electron microscopic examination revealed microstructural changes after fermentation in the curdled milk compared to the control. The findings highlighted the role of indigenous probiotic strains with superior probiotic features, antioxidant activity, and CLA-producing ability in functional food formulation.

Pomegranate Seed Oil as a Source of Conjugated Linolenic Acid (CLnA) Has No Effect on Atherosclerosis Development but Improves Lipid Profile and Affects the Expression of Lipid Metabolism Genes in apoE/LDLR-/- Mice.

The aim of this study was to evaluate the anti-atherosclerotic effect of pomegranate seed oil as a source of conjugated linolenic acid (CLnA) (cis-9,trans-11,cis-13; punicic acid) compared to linolenic acid (LnA) and conjugated linoleic acid (CLA) (cis-9,trans-11) in apoE/LDLR-/- mice. In the LONG experiment, 10-week old mice were fed for the 18 weeks. In the SHORT experiment, 18-week old mice were fed for the 10 weeks. Diets were supplied with seed oils equivalent to an amount of 0.5% of studied fatty acids. In the SHORT experiment, plasma TCh and LDL+VLDL cholesterol levels were significantly decreased in animals fed CLnA and CLA compared to the Control. The expression of PPARα in liver was four-fold increased in CLnA group in the SHORT experiment, and as a consequence the expression of its target gene ACO was three-fold increased, whereas the liver's expression of SREBP-1 and FAS were decreased in CLnA mice only in the LONG experiment. Punicic acid and CLA isomers were determined in the adipose tissue and liver in animals receiving pomegranate seed oil. In both experiments, there were no effects on the area of atherosclerotic plaque in aortic roots. However, in the SHORT experiment, the area of atherosclerosis in the entire aorta in the CLA group compared to CLnA and LnA was significantly decreased. In conclusion, CLnA improved the lipid profile and affected the lipid metabolism gene expression, but did not have the impact on the development of atherosclerotic plaque in apoE/LDLR-/- mice.

RAPID METHOD FOR SIMULTANEOUS DETERMINATION OF CIS-9, TRANS-11 AND TRANS-10, CIS-12 CONJUGATED LINOLEIC ACID ISOMERS IN MILK BY GC-FID

Ruminant milk is the lead source of conjugated linoleic acid (CLA) in the human diet, with cis-9, trans-11 CLA being the major among all. Small amounts of trans-10, cis-12 CLA are also found in synthetic supplements. Since both isomers are biologically active with potential health benefits, there is great interest in quantifying them for quality control routines. An alternative method for the analysis of the aforementioned CLAs by fast gas chromatography (GC) is discussed in the present study. The fatty acid methyl ester mixture obtained by alkaline catalysis was injected into a GC equipped with a flame ionization detector (FID) and fitted with an ionic liquid SLB-IL111 chromatographic column (15 m × 0.10 mm × 0.08 µm). Separation was achieved in less than 5 min using a 168 °C isotherm run. Both CLA isomers were quantified by using of single point standard addition statistical approach. Results were contested to those obtained using the recommended 100 m long CP-Sil88 capillary column and none evidence of significant differences was found within 95% confidence interval. Therefore, the proposed method could be valuable to focused regulatory routines of large numbers of samples with greater analytical frequency.

Supplementation with Sunflower/Fish Oil-Containing Concentrates in a Grass-Based Beef Production System: Influence on Fatty Acid Composition, Gene Expression, Lipid and Colour Stability and Sensory Characteristics of Longissimus Muscle.

Beef contains an array of conjugated linoleic acid (CLA) isomers for which positive effects have been reported in animal models of human disease. The objectives were to develop a CLA-enriched beef production system and to assess its quality. Sixty Spring-born heifers were housed in Autumn and offered unwilted grass silage and a barley/soyabean concentrate or wilted grass silage and a concentrate containing sunflower oil and fish oil. In May, both groups were offered either pasture for 22 weeks, restricted pasture and sunflower oil and fish oil for 22 weeks, or pasture for 11 weeks and restricted pasture and sunflower oil and fish oil for the final 11 weeks. The predominant CLA isomer in beef was cis9, trans11 representing on average, 80% total CLA. The modified winter diet followed by supplementation for 22 weeks resulted in beef that had a CLA concentration that was higher, at a comparable intramuscular fatty acid concentration, than previously reported. The lipid and colour stability (over 10 days in modified atmosphere packaging) and sensory characteristics were generally not negatively affected. There were minor effects on the expression of candidate genes involved in lipid metabolism. Consumption of this beef would make a substantial contribution to the quantity of CLA suggested to have a positive effect on consumer health.

Rapid synthesis of conjugated linoleic acid from fruit processing residues seed oil by alkali‐dimethyl sulfoxide isomerization

AbstractRapid synthesis of conjugated linoleic acid (CLA) by the alkali‐dimethyl sulfoxide (DMSO) isomerization method is presented in this study. Seeds from fruit processing residues in Thailand were screened as an alternative potential source of linoleic acid (LA)‐rich oil. Passion fruit with 72.10% LA and 26.18% oil in the seeds was selected as starting oil. The Alkali‐DMSO isomerization was carried out by mixing DMSO with alkali hydroxides and polyhydric alcohols. Under a mass ratio of 1:0.5:1:1 oil/NaOH/propylene glycol/DMSO, at 180°C, production of the major isomers of CLA (c9,t11, and t10,c12) was complete within 3 min, resulting in a 20‐fold increase in productivity compared to the conventional method. The reaction time decreased because of an increment in the basic strength of the alkali‐DMSO catalyst. CLA from alkali‐DMSO isomerization and commercial supplements showed a similar isomer distribution and yielded similar CC50 values when evaluated against the Vero cell line. This method might be an alternative in industrial production as it is fast and simply applied to the conventional system. In addition, seed oils from fruit processing residues can be used as an alternative to traditional LA‐rich oils.

Characterization of polar and non-polar lipids of Hermetia illucens and Tenebrio molitor meals as animal feed ingredients

Although increasingly used in animal feeding, the black soldier fly (Hermetia illucens - HI) and mealworm (Tenebrio molitor - TM) larvae and pupae require the characterization and modulation of their fat profile for use in the feed industry. In this study, we provide a comprehensive fatty acid profile of the total lipid triglycerides and phospholipids, and sterol composition. For a more detailed characterization of the nutritional properties, we defined the profile of esterified fatty acids (FAs) in the sn-2 position of the triglycerides, which represents the most adsorbed fraction by animals during digestion. Twenty-four batches (12 batches per species divided into 6 batches of larvae and 6 batches of pupae) of 70 subjects were used for the analysis. For each of the developmental stages, individuals in the intermediate stage were considered (2 months and 8 days for TM larvae and pupae respectively; 9 days and 7 days for HI larvae and pupae respectively). Some fatty acids (decanoic, lauric, or myristic acid) were more abundant in HI than in TM. These FAs were produced de novo, indicating that HI produce them from carbohydrates as a source of acetyl-CoA. Hermetia illucens showed a higher level of conjugated linoleic acid (CLA) isomers, a class of fatty acids with positive health effects. In contrast, TM showed a similar fatty acid profile to the plant lipid source (such as soy and sunflower) due to the high level of linoleic acid. Analysis of the triglyceride fraction showed a different affinity for the three glyceride positions. In particular, lauric acid, polyunsaturated fatty acids n-3 and CLA isomers esterified preferentially in sn-2, and therefore were the most adsorbed by the animal during digestion. As for the phospholipids, HI showed a higher level, above all in terms of the phosphatidylethanolamine fraction. Interestingly there was no sphingomyelin in the pupae of both species. The high level of phospholipids is a useful feature for the emulsion of fats in animal feed diets. Lastly, we observed a considerable amount of phytosterol in the sterol profile of both species, which makes these insects a valid alternative to other food sources of plant origin for animal nutrition. We believe that our findings provide new nutritional information for the use of TM and HI in the animal diet.

Differential Inflammatory Responses in Cultured Endothelial Cells Exposed to Two Conjugated Linoleic Acids (CLAs) under a Pro-Inflammatory Condition.

Conjugated linoleic acid (CLA) isomers have been shown to possess anti-atherosclerotic properties, which may be related to the downregulation of inflammatory pathways in different cell types, including endothelial cells (ECs). However, whether different CLA isomers have different actions is not entirely clear, with inconsistent reports to date. Furthermore, in cell culture studies, CLAs have often been used at fairly high concentrations. Whether lower concentrations of CLAs are able to affect EC responses is not clear. The aim of this study was to evaluate the effects of two CLAs (cis-9, trans-11 (CLA9,11) and trans-10, cis-12 (CLA10,12)) on the inflammatory responses of ECs. ECs (EA.hy926 cells) were cultured under standard conditions and exposed to CLAs (1 to 50 μM) for 48 h. Then, the cells were cultured for a further 6 or 24 h with tumour necrosis factor alpha (TNF-α, 1 ng/mL) as an inflammatory stimulant. ECs remained viable after treatments with 1 and 10 μM of each CLA, but not after treatment with 50 μM of CLA10,12. CLAs were incorporated into ECs in a concentration-dependent manner. CLA10,12 increased the levels of ICAM-1, IL-6, and RANTES in the culture medium, while CLA9,11 had null effects. Both CLAs (1 μM) decreased the appearance of NFκB1 mRNA, but only CLA9,11 maintained this downregulation at 10 μM. CLA10,12 had no effect on THP-1 cell adhesion to ECs while significantly decreasing the percentage of ECs expressing ICAM-1 and also levels of ICAM-1 expression per cell when used at 10 µM. Although CLA9,11 did not have any effect on ICAM-1 cell surface expression, it reduced THP-1 cell adhesion to the EA.hy926 cell monolayer at both concentrations. In summary, CLA10,12 showed some pro-inflammatory effects, while CLA9,11 exhibited null or anti-inflammatory effects. The results suggest that each CLA has different effects in ECs under a pro-inflammatory condition, highlighting the need to evaluate the effects of CLA isomers independently.

Valorization of waste chicken skin for conjugated linoleic acid (CLA) production: optimization using response surface methodology.

The valorization of waste chicken skin fat (WCSF) for conjugated linoleic acid (CLA) production was performed by photoisomerization and optimized the process conditions for high CLA production by response surface methodology. The fat extraction yield from waste chicken skin was approximately 52%. The linoleic acid content of the fat obtained from waste chicken skin was increased by the fractionation process approximately 2 times, up to 52%. Optimum iodine amount and processing time for predicted maximum total CLA amount were determined as 0.87% and 116.36h, respectively. The maximum total CLA amount was predicted and produced experimentally as 32.14% and 29.01%, respectively. Additionally, iodine amount, processing time, and their interaction significantly affected the amount and variety of produced CLA isomers. The results indicated that dominant isomers produced by photoisomerization of WCSF were trans, trans isomers. However, the amount of cis 9, trans 11 and trans 10, cis 12 CLA isomers could be more increased by optimizing the production parameters. The present study indicated that waste chicken skin could be valorized in CLA production by photoisomerization and obtained high value-added product, and also a more economical and faster CLA production could be realized.

Heterologous expression of a novel linoleic acid isomerase BBI, and effect of fusion tags on its performance

Functional proteins with the ability to isomerise free linoleic acid (LA) to conjugated linoleic acid (CLA) are termed linoleic acid isomerases (LAI). BBI is a novel LAI from Bifidobacterium breve with unique advantages in the production of a single CLA isomer; however, its complex membrane-bound form hampers over-expression of the protein in its natural host. To overcome this challenge, heterologous expression of BBI in Pichia pastoris was studied. Further, to investigate the influence of His-tags on the heterologous expression of BBI, three P. pastoris recombinant strains carrying either a C-terminal His-tag, an N-terminal His-tag, or none were constructed. The expression of recombinant proteins was analysed by dot and western blotting, and the enzyme activity was determined by GC-MS. The results show that the three P. pastoris recombinant strains successfully expressed the recombinant protein and had LAI activity. Compared with those BBIs without a His-tag or carrying a His-tag on the C-terminus, the BBI carrying an N-terminal His-tag had reduced expression and enzyme activity and that was also explained by the protein modelling analysis. Moreover, this study highlights the advantages of using P. pastoris for BBI expression to achieve efficient production of c9, t11-CLA monomers; the highest conversion rate of the substrate LA was over 80%, resulting in the production of 0.81 mg of c9, t11-CLA per mg of crude enzyme.

Trans 10, cis 12, but Not cis 9, trans 11 Conjugated Linoleic Acid Isomer Enhances Exercise Endurance by Increasing Oxidative Skeletal Muscle Fiber Type via Toll-like Receptor 4 Signaling in Mice.

Conjugated linoleic acid (CLA) has been implicated in regulating muscle fiber. However, which isomer elicits this effect and the underlying mechanisms remain unclear. Here, male C57BL6/J mice and C2C12 cells were treated with two CLA isomers, and the exercise endurance, skeletal muscle fiber type, and involvement of Toll-like receptor 4 (TLR4) signaling were assessed. The results demonstrated that dietary t10, c12, but not c9, t11-CLA isomer enhanced exercise endurance of mice (from 115.88 ± 11.21 to 130.00 ± 15.84 min, P < 0.05) and promoted the formation of oxidative muscle fiber type of gastrocnemius muscle (from 0.15 ± 0.04 to 0.24 ± 0.05, P < 0.05). Consistently, t10, c12-CLA isomer increased the mRNA expression of oxidative muscle fiber type in C2C12 myotubes (from 1.00 ± 0.08 to 2.65 ± 1.77, P < 0.05). In addition, t10, c12-CLA isomer increased TLR4 signaling expression in skeletal muscle and C2C12 myotubes. More importantly, knockdown of TLR4 eliminated the t10, c12-CLA isomer-induced enhancement of exercise endurance in mice and elevation of oxidative muscle fiber type in C2C12 myotubes and gastrocnemius muscle. Together, these findings showed that t10, c12, but not c9, t11-CLA isomer enhances exercise endurance by increasing oxidative skeletal muscle fiber type via TLR4 signaling.

Cancer Influences the Elemental Composition of the Myocardium More Strongly than Conjugated Linoleic Acids-Chemometric Approach to Cardio-Oncological Studies.

The aim of the study was to verify in a cardio-oncological model experiment if conjugated linoleic acids (CLA) fed to rats with mammary tumors affect the content of selected macro- and microelements in their myocardium. The diet of Sprague–Dawley females was supplemented either with CLA isomers or with safflower oil. In hearts of rats suffering from breast cancer, selected elements were analyzed with a quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). In order to better understand the data trends, cluster analysis, principal component analysis and linear discriminant analysis were applied. Mammary tumors influenced macro- and microelements content in the myocardium to a greater extent than applied diet supplementation. Significant influences of diet (p = 0.0192), mammary tumors (p = 0.0200) and interactions of both factors (p = 0.0151) were documented in terms of Fe content. CLA significantly decreased the contents of Cu and Mn (p = 0.0158 and p = 0.0265, respectively). The level of Ni was significantly higher (p = 0.0073), which was more pronounced in groups supplemented with CLA. The obtained results confirmed antioxidant properties of CLA and the relationship with Se deposition. Chemometric techniques distinctly showed that the coexisting pathological process induced differences to the greater extent than diet supplementation in the elemental content in the myocardium, which may impinge on cardiac tissue’s susceptibility to injuries.

Mechanisms of Anticancer Activity of a Fatty Acid Mixture Extracted from Hen Egg Yolks Enriched in Conjugated Linoleic Acid Diene (CLA) against WM793 Melanoma Cells.

The conjugated linoleic acid (CLA) diene is a biologically active compound with proven health-promoting effects. In terms of anticancer properties, it has been shown that CLA reduces the proliferation of cancer cells. In this study, it has been demonstrated that a mixture of fatty acids, isolated from chicken egg yolk enriched in CLA isomers by biofortification, reduces (by 30.5%) the proliferation of human melanoma cancer cells line WM793 to a greater extent than a mixture of fatty acids not containing these isomers. At the same time, the tested fatty acid mixtures show no effect on human normal BJ fibroblast cells. For the first time, the genes with increased expression have been identified and the proteins have been activated by the fatty acid mixture of CLA-enriched egg yolk, mainly responsible for mitochondrial pathway-dependent apoptosis.

Evaluation of the influence of diet supplementation with conjugated linoleic acid isomers on elemental composition in the cardio-oncological nutritional programming rat’ model

BackgroundThe ‘developmental origin of health and disease’ (DOHaD) hypothesis assumes that due to the action of some stimuli during fetal life the long-term physiological changes occurs and may affects the risk of various diseases. The main aim of this study was to assess impact of supplementation of maternal’ and early postnatal diet with conjugated linoleic acids (CLA) isomers on selected elements content in hearts of their female offspring with chemically induced breast cancer. MethodsElemental composition was determined by quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). ResultsThe effect of maternal’ diet on the elements content was more pronounced than the progeny diet modifications. Significant correlations among determined elements, especially macroelements, and lipidomic parameters, in the experimental factors dependent manner were observed. It can be concluded that supplementation of maternal and progeny diets with CLA isomers may significantly influence the contents of both macro- and microelements in cardiac tissue of newborns. ConclusionOur results also indicate, that dynamic and intricate balance among various elements in body may be affected by the lipid dietary supplements also in the pathological state. Utility of cardio-oncological approach in developmental programming study was confirmed.

Milk from cows fed clover-rich silage compared with cows fed grass silage is higher in n-3 fatty acids

The present study investigated the effect of a high proportion of different forage species in the diet, parity, milking time, and days in milk (DIM) on milk fatty acid (FA) profile, and transfer efficiency of C18:2n-6, C18:3n-3, n-6, and n-3 in dairy cows. Swards with perennial ryegrass [early maturity stage (EPR) and late maturity stage (LPR)], festulolium, tall fescue (TF), red clover (RC), and white clover (WC) were cut in the primary growth, wilted, and ensiled without additives. Thirty-six Danish Holstein cows in an incomplete Latin square design were fed ad libitum with total mixed rations containing a high forage proportion (70% on dry matter basis). The total mixed rations differed only in forage source, which was either 1 of the 6 pure silages or a mixture of LPR silage with either RC or WC silage (50:50 on dry matter basis). Proportion of C18:2n-6 in milk FA was affected by diet, and RC and WC diets resulted in the highest proportion of C18:2n-6 in milk FA (21.6 and 21.8 g/kg of FA, respectively). The highest and lowest milk C18:3n-3 proportion was observed in WC and LPR, respectively. In addition, WC diet resulted in highest transfer efficiency of C18:3n-3 from feed to milk (12.2%) followed by RC diet (10.7%), whereas EPR diet resulted in the lowest transfer efficiency of C18:3n-3 (3.45%). The highest milk proportion of cis-9,trans-11 conjugated linoleic acid (CLA) was observed in cows fed TF (3.20 g/kg of FA), which was 23 to 64% higher than the proportion observed in the cows fed the other diets. The highest α-tocopherol concentration (µg/mL) in milk was observed in EPR (1.15), LPR (1.10), and festulolium (1.06). Primiparous cows showed higher proportion of cis-9,trans-11 CLA (2.63 g/kg of FA) than multiparous cows (2.21 g/kg of FA). Cows early in lactation had a higher proportion of long-chain FA in milk than cows later in lactation, as long-chain FA decreased with 0.184 g/kg of FA per DIM, whereas medium-chain FA increased with 0.181 g/kg of FA per DIM. Proportion of C18:2n-6 in milk from evening milking was higher than in milk from morning milking (16.7 vs. 15.8 g/kg of FA). In conclusion, the results showed that milk FA profile of cows was affected by forage source in the diet, and RC and WC increased the health-promoting FA components, particularly n-3, whereas the TF diet increased proportion of CLA isomers in milk. Proportion of CLA isomers in milk FA from primiparous cows was higher than in milk from multiparous cows. In addition, evening milk contained more FA originating from diets compared with morning milk.