Iso- but Not Anteiso-Branched Chain Fatty Acids Exert Growth-Inhibiting and Apoptosis-Inducing Effects in MCF-7 Cells.

The present study compared the growth-inhibitory effects of four common branched chain fatty acids (BCFAs) found in beef and dairy fats including iso 15:0, anteiso 15:0, iso 17:0, and anteiso 17:0. MCF-7 human breast cancer cells were exposed for 72 h to media containing increasing doses (50 to -400 µM) of the four BCFA. Cell viability was not affected by any of the BCFA treatments at doses less than 200 µM. Culturing cells with 200 µM of iso-15:0 or iso-17:0 reduced cell viability by 27 ± 2.8 and 43 ± 8.3% at 24 h, 35 ± 4.6 and 49 ± 9.1% at 48 h, and 44 ± 6.8 and 57 ± 8.8% at 72 h posttreatment. In contrast, culturing cells with 200 µM of anteiso-15:0 or anteiso-17:0 did not affect cell viability for any durations tested. The incorporation of iso 15:0 and iso 17:0 into cells (19.1 ± 1.3 and 21.2 ± 1.4 µmol/mg protein, respectively) was greater (P < 0.01) than that of anteiso 15:0 and anteiso 17:0 (11.8 ± 0.7 and 13.8 ± 0.8 µmol/mg protein, respectively). Iso-15:0 and iso-17:0 downregulated (P < 0.01) the expression of antiapoptotic Bcl-2 (0.71 ± 0.6-fold and 0.64 ± 0.09-fold, respectively) and upregulated (P < 0.01) the expression of proapoptotic Bax (1.72 ± 0.14-fold and 2.15 ± 0.24-fold, respectively) compared to the control, whereas their corresponding anteiso isomers did not affect the expression of any apoptosis-related genes. Our findings suggest that the branching structure influences anticarcinogenic effects of BCFAs, with iso being more potent than anteiso.

The Growth-Inhibiting Effects of Beef Fatty Acids on MCF-7 Cells Are Influenced Mostly by the Depot Location and Inconsistently by the Biohydrogenation Intermediate Content.

Biohydrogenation intermediates (BHI) including conjugated linoleic acid (CLA) isomers are formed during ruminal biohydrogenation of polyunsaturated fatty acids (PUFA) in ruminants. Although many studies have examined the anticarcinogenic effects of CLA, few studies have reported the anticarcinogenic properties of BHI in their natural form found in dairy and beef fats. The present study compared the growth-inhibitory effects of fatty acids from beef perirenal fat (PRF) or subcutaneous fat (SCF) with low or high levels of BHI in MCF-7 human breast cancer cells. Cells were exposed for 72 h to media containing increasing doses (50 to 400 µM) of different beef fat treatments. Fatty-acid analysis showed that BHI were readily incorporated into cell phospholipids (PL) in a treatment-dependent manner, but higher BHI in PL did not consistently inhibit growth. Culturing with low-BHI PRF or high-BHI PRF did not lead to growth inhibition, but low-BHI SCF inhibited growth, and inhibition was further increased by high-BHI SCF. Other classes of fatty acids may, therefore, be interacting with BHI resulting in differential effects on growth inhibition in human breast cancer cells.

Anti-mutagenic Properties of Mono- and Dienoic Acid Biohydrogenation Products from Beef Fat.

Unsaturated fatty acid biohydrogenation products from beef fat and pure fatty acids were subjected to the Ames Salmonella mutagenicity testing, including monounsaturated fatty acids [MUFA: oleic acid, vaccenic acid, elaidic acid; beef fatty acid fractions rich in trans (t)11/t13-t14-18:1 (t11,13,14-Frac), t10-18:1 (t10-Frac)] and dienoic fatty acids [linoleic acid, conjugated linoleic isomers cis (c)9,t11-18:2 and t10,c12-18:2, and a mixed beef dienoic fatty acid fraction high in c9,t13-/t8,c12/t11c15-18:2 (MD)]. Significantly higher anti-mutagenic effects of oleic acid, vaccenic acid, t11, 13, 14-Frac, and t10-Frac against daunomycin were observed at 2.5 mg. All dienoic acids except MD significantly reduced daunomycin mutagenicity at ≥0.25 mg. Anti-mutagenicity of oleic and vaccenic acids against 2-aminoanthracene was found at 2.5 and 0.25 mg, respectively. All dienoic acids significantly reduced 2-aminoanthracene mutagenicity at ≥0.25 mg. Findings of this study show that unsaturated fatty acids, including trans-fatty acids commonly found in beef, can act as strong anti-mutagens.

Trans10,cis15 18:2 Isolated from Beef Fat Does Not Have the Same Anti-Adipogenic Properties as Trans10,cis12-18:2 in 3T3-L1 Adipocytes.

During ruminal biohydrogenation of α-linolenic acid, a non-conjugated non-methylene interrupted dienoic acid is formed containing a t10 double bond, namely t10,c15-18:2. The present study was designed to examine whether t10,c15-18:2 would exert similar anti-adipogenic effects compared to t10,c12-18:2 in 3T3-L1 adipocytes. Differentiated 3T3-L1 adipocytes were treated with 35 or 70 µM of LNA, t10,c12-18:2, t10,c15-18:2, or bovine serum albumin (BSA) vehicle control for 120 h. Cellular triacylglycerol and protein were quantified using commercial colorimetric kits. Cells were analyzed for fatty acid composition and gene expression using gas chromatography and quantitative PCR, respectively. Trans10,cis12-18:2 decreased (P < 0.05) the adipocyte triacylglycerol (TAG) content, which was mainly related to a reduction in saturated fatty acids (SFA; e.g., 16:0 and 15:0) and cis monounsaturated fatty acids (c-MUFA; e.g., c9-16:1 and c9-18:1). Trans10,cis12 also decreased (P < 0.05) the expression of genes related to fatty acid synthesis (ACACA, FASN), delta-9 desaturation (SCD1), fatty acid elongation (ELOVL5), and fatty acid uptake (LPL) and upregulated (P < 0.05) the expression of the rate-liming enzyme involved in fatty acid ß-oxidation (CPT1). In contrast, LNA and t10,c15-18:2 did not affect the gene expression and cellular content of the TAG, SFA, c-MUFA, or SCD1 indices in adipocytes. Our findings suggest that t10,c15-18:2, despite having structural similarity to t10,c12-18:2 (presence of a trans-10 double bond), does not exert anti-adipogenic effects in 3T3-L1 adipocytes.

A trans10-18:1 enriched fraction from beef fed a barley grain-based diet induces lipogenic gene expression and reduces viability of HepG2 cells.

Beef fat is a natural source of trans (t) fatty acids, and is typically enriched with either t10-18:1 or t11-18:1. Little is known about the bioactivity of individual t-18:1 isomers, and the present study compared the effects of t9-18:1, cis (c)9-18:1 and trans (t)-18:1 fractions isolated from beef fat enriched with either t10-18:1 (HT10) or t11-18:1 (HT11). All 18:1 isomers resulted in reduced human liver (HepG2) cell viability relative to control. Both c9-18:1 and HT11were the least toxic, t9-18:1had dose response increased toxicity, and HT10 had the greatest toxicity (P<0.05). Incorporation of t18:1 isomers was 1.8-2.5 fold greater in triacylglycerol (TG) than phospholipids (PL), whereas Δ9 desaturation products were selectively incorporated into PL. Culturing HepG2 cells with t9-18:1 and HT10 increased (P<0.05) the Δ9 desaturation index (c9-16:1/16:0) compared to other fatty acid treatments. HT10 and t9-18:1 also increased expression of lipogenic genes (FAS, SCD1, HMGCR and SREBP2) compared to control (P<0.05), whereas c9-18:1 and HT11 did not affect the expression of these genes. Our results suggest effects of HT11 and c9-18:1 were similar to BSA control, whereas HT10 and t-9 18:1 (i.e. the predominant trans fatty acid isomer found in partially hydrogenated vegetable oils) were more cytotoxic and led to greater expression of lipogenic genes.

The scope for manipulating the polyunsaturated fatty acid content of beef: a review.

Since 1950, links between intake of saturated fatty acids and heart disease have led to recommendations to limit consumption of saturated fatty acid-rich foods, including beef. Over this time, changes in food consumption patterns in several countries including Canada and the USA have not led to improvements in health. Instead, the incidence of obesity, type II diabetes and associated diseases have reached epidemic proportions owing in part to replacement of dietary fat with refined carbohydrates. Despite the content of saturated fatty acids in beef, it is also rich in heart healthy cis-monounsaturated fatty acids, and can be an important source of long-chain omega-3 (n-3) fatty acids in populations where little or no oily fish is consumed. Beef also contains polyunsaturated fatty acid biohydrogenation products, including vaccenic and rumenic acids, which have been shown to have anticarcinogenic and hypolipidemic properties in cell culture and animal models. Beef can be enriched with these beneficial fatty acids through manipulation of beef cattle diets, which is now more important than ever because of increasing public understanding of the relationships between diet and health. The present review examines recommendations for beef in human diets, the need to recognize the complex nature of beef fat, how cattle diets and management can alter the fatty acid composition of beef, and to what extent content claims are currently possible for beef fatty acids.

Improving beef hamburger quality and fatty acid profiles through dietary manipulation and exploitation of fat depot heterogeneity.

BACKGROUND: Hamburger is the most consumed beef product in North America, but lacks in nutritional appeal due to its high fat content and high proportion of saturated fatty acids (SFA). Objectives of the present study were to improve the FA profiles of hamburgers made with perirenal fat (PRF) and subcutaneous fat (SCF) when feeding steers different diets along with examining differences in sensory attributes and oxidative stability. Diets included a control diet containing 70:30 red clover silage: barley based concentrate, a diet containing sunflower-seed (SS) substituted for barley, and diets containing SS with 15% wheat dried distillers' grain with solubles (DDGS-15) or 30% DDGS (DDGS-30). Hamburgers were made from triceps brachii and either PRF or SCF (80:20 w/w). RESULTS: Perirenal fat versus SCF hamburgers FA had 14.3% more (P <0.05) 18:0, 11.8% less cis (c)9-18:1 (P <0.05), and 1.82% more total trans (t)-18:1 mainly in the form of t11-18:1. During sensory evaluation, PRF versus SCF hamburgers had greater (P <0.05) mouth coating, but the difference was less than one panel unit. Examining effects of steer diet within PRF hamburgers, feeding the SS compared to the control diet increased (P <0.05) t-18:1 by 2.89% mainly in the form of t11-18:1, feeding DGGS-15 diet led to no further changes (P >0.05), but feeding DDGS-30 diet reduced the proportions of (P <0.05) of t-18:1 chiefly t11-18:1. Feeding SS and DDGS diets had small but significant (P <0.05) effects on hamburger sensory attributes and oxidative stability. CONCLUSIONS: Feeding high-forage diets including SS and 15% DDGS, and taking advantage of the FA heterogeneity between fat depots offers an opportunity to differentially enhance beef hamburgers with 18:2n-6 biohydrogenation products (i.e., t11-18:1) with potential human health benefits without compromising their sensory attributes and oxidative stability during retail display.

Red blood cell trans-18:1 isomeric profile correlates with subcutaneous fat and muscle profiles in beef cattle.

Due to significant variation in polyunsaturated fatty acid biohydrogenation products in beef it would be useful to determine if levels of trans-18:1 isomers in samples collected ante-mortem are correlated with those collected post-mortem. Beef blood (RBC), subcutaneous fat (SC) and muscle (intramuscular fat; IM) samples were collected from an experiment with dietary vitamin E with/without flaxseed (n=80) and fatty acids analyzed. Across treatments, correlation analysis of total and individual trans-18:1 isomers were performed between tissues. Correlations between SC and IM were highly significant for all individual and total trans-18:1. RBC trans-18:1 were also well correlated with other tissues except for vaccenic acid. Levels of 10t-, 12t- and 13t/14t- were amongst the best correlated between RBC and SC and IM profiles. Levels of 6t/7t/8t-, 9t-, and 15t-18:1 showed significant but lower correlation factors particularly between RBC and SC. These results confirm the possibility of utilizing blood as a non-destructive sample to predict the total and isomeric profile of trans-18:1 in beef.

Comparing subcutaneous adipose tissue in beef and muskox with emphasis on trans 18:1 and conjugated linoleic acids.

Muskox (Ovibos moschatus) are ruminant animals native to the far north and little is known about their fatty acid composition. Subcutaneous adipose tissue (backfat) from 16 wild muskox was analyzed and compared to backfat from 16 barley fed beef cattle. Muskox backfat composition differed substantially from beef and the most striking difference was a high content of 18:0 (26.8 vs. 9.77%). This was accompanied by higher levels of most other saturated fatty acids except beef had more 16:0. Muskox backfat also had a lower level of cis-18:1 and this was related to a lower expression of steroyl-CoA desaturase mRNA. Beef backfat had a higher level of total trans-18:1 (4.25 vs. 2.67%). The most prominent trans-18:1 isomers in beef backfat were 10t-18:1 (2.13%) and 11t-18:1 (0.77%) whereas the most prominent isomers in muskox backfat were 11t-18:1 (1.41%), 13t/14t- (0.27%) and 16t-18:1 (0.23%). The total conjugated linoleic acid (CLA) content was higher in beef backfat than muskox (0.67 vs. 0.50%) with 9c,11t-18:2 as the most abundant CLA isomer. The second most abundant CLA isomer in beef backfat was 7t,9c-18:2 (0.10%) whereas in muskox it was 11t13c-18:2 (0.04%). Muskox backfat had a higher content of 18:3n-3 and its elongation and desaturation products 20:5n-3, 22:5n-3 and 22:6n-3 and a lower n-6/n-3 ratio. Overall, the high forage diet of muskox seemed to produce a healthier fatty acid profile and highlighted the need to develop feeding strategies for intensively raising beef that will not negatively impacting fatty acid composition.