Plasma phospholipid and dietary fatty acids as predictors of type 2 diabetes: interpreting the role of linoleic acid 1-3

Background: Dietary fatty acids may be associated with diabetes but are difficult to measure accurately.

Objective: We aimed to investigate the associations of fatty acids in plasma and diet with diabetes incidence.

Design: This was a prospective case-cohort study of 3737 adults aged 36-72 y. Fatty acid intake (/kJ) and plasma phospholipid fatty acids (%) were measured at baseline, and diabetes incidence was assessed by self-report 4 y later. Logistic regression excluding (model 1) and including (model 2) body mass index and waist-hip ratio was used to calculate odds ratios (ORs) for plasma phospholipid and dietary fatty acids.

Results: In plasma phospholipid, positive associations with diabetes were seen for stearic acid [OR model 1, highest versus lowest quintile: 4.14 (95% CI: 2.65, 6.49), P for trend < 0.0001] and total saturated fatty acids [OR model 1: 3.76 (2.43, 5.81), P for trend < 0.0001], whereas an inverse association was seen for linoleic acid [OR model 1: 0.22 (0.14, 0.36), P for trend < 0.0001]. Dietary linoleic [OR model 1: 1.77 (1.19, 2.64), P for trend = 0.002], palmitic [OR model 1: 1.65 (1.12, 2.43), P for trend = 0.012], and stearic [OR model 1: 1.46 (1.00, 2.14), P for trend = 0.030] acids were positively associated with diabetes incidence before adjustment for body size. Within each quintile of linoleic acid intake, cases had lower baseline plasma phospholipid linoleic acid proportions than did controls.

Conclusions: Dietary saturated fat intake is inversely associated with diabetes risk. More research is required to determine whether linoleic acid is an appropriate dietary substitute.

Failure of conjugated linoleic acid supplementation to enhance biosynthesis of docosahexaenoic acid from α-linolenic acid in healthy human volunteers

A rate-limiting step in docosahexaenoic acid (DHA) formation from α-linolenic acid (ALA) involves peroxisomal oxidation of 24:6n-3 to DHA. The aim of the study was to determine whether conjugated linoleic acid (CLA) would enhance conversion of ALA to DHA in humans on an ALA-supplemented diet. The subjects (n=8 per group) received daily supplementation of ALA (11g) and either CLA (3.2g) or placebo for 8 weeks. At baseline, 4 and 8 weeks, blood was collected for plasma fatty acid analysis and a number of physiological measures were examined. The ALA-supplemented diet increased plasma levels of ALA and eicosapentaenoic acid (EPA). The addition of CLA to the ALA diet resulted in increased plasma levels of CLA, as well as ALA and EPA. Plasma level of DHA was not increased with either the ALA alone or ALA plus CLA supplementation. The results demonstrated that CLA was not effective in enhancing DHA levels in plasma in healthy volunteers.

Effects of dietary fat and conjugated linoleic acid on plasma metabolite concentrations and metabolic responses to homeostatic signals in pigs

Sixteen female cross-bred (Large White × Landrace) pigs (initial weight 65 kg) with venous catheters were randomly allocated to four treatment groups in a 2×2 factorial design. The respective factors were dietary fat (25 or 100 g/kg) and dietary conjugated linoleic acid (CLA; 0 or 10 g CLA-55/kg). Pigs were fed every 3 h (close to ad libitum digestible energy intake) for 8 d and were bled frequently. Plasma glucose and non-esterified fatty acid (NEFA) responses to insulin and adrenaline challenges were determined on day 8. Plasma concentrations of NEFA were significantly increased (10·5 and 5·4 % for low- and high-fat diets respectively, P=0·015) throughout the experiment, suggesting that there was a possible increase in fat mobilisation. The increase in lipolysis, an indicator of ß-adrenergic stimulated lipolysis, was also evident in the NEFA response to adrenaline. However, the increase in plasma triacylglycerol (11·0 and 7·1 % for low- and high-fat diets respectively, P=0·008) indicated that CLA could have reduced fat accretion via decreased adipose tissue triacylglycerol synthesis from preformed fatty acids, possibly through reduced lipoprotein lipase activity. Plasma glucose, the primary substrate for de novo lipid synthesis, and plasma insulin levels were unaffected by dietary CLA suggesting that de novo lipid synthesis was largely unaffected (P=0·24 and P=0·30 respectively). In addition, the dietary CLA had no effect upon the ability of insulin to stimulate glucose removal.

Dietary conjugated linoleic acid differentially alters fatty acid composition and increases conjugated linoleic acid content in porcine adipose tissue

Conjugated linoleic acids (CLA) have been shown to decrease body fat content in pigs. It is possible that feeding pigs diets rich in CLA may increase carcass lipid CLA to levels that could provide health benefits when included as a part of a healthy diet. Therefore, the aim of the present study was to determine whether dietary CLA supplementation has any effect on the fatty acid composition of subcutaneous and intramuscular adipose tissue in pigs. Thirty-five female cross bred (Large White X Landrace) pigs (initial weight 57·2 kg and initial P2 back fat 11·5 mm) were used in the present study. Pigs were housed individually and randomly allocated to one of six dietary treatments (0·00, 1·25, 2·50, 5·00, 7·50 and 10·00 g CLA55 (55 g CLA isomers/100 g total fatty acids; Natural Lipids Ltd, Hovdebygda, Norway)/kg)
and fed their respective diets for 8 weeks. Twelve CLA isomers in the diet and in pig tissue lipids were separated by Agþ-HPLC. CLA was incorporated at fivefold higher levels in subcutaneous fat as compared with intramuscular fat and in a dose-dependant manner. Overall, the transfer efficiency of CLA was maximized at 5·00 g CLA55/kg. However, there was clear selectivity in the uptake or incorporation of cis,trans-9,11 isomer over the trans,cis-10,12 isomer. In general, CLA supplementation produced significant changes in skeletal muscle and adipose tissue fatty acid composition, indicating that dietary CLA had a potent affect on lipid transport and metabolism in vivo. Significant increases in myristic, palmitic and palmitoleic acids and a reduction in arachidonic acid were observed, suggesting an alteration in
activity of Δ5-, Δ6- and Δ9-desaturases in pig adipose tissue. In conclusion, feeding pigs diets supplemented with CLA increases carcass lipid CLA, but also results in changes in the fatty acid profile in pig fat that could potentially outweigh the benefits of CLA.

Conjugated linoleic acid decreases fat accretion in pigs evaluation by dual-energy X-ray absorptiometry

Thirty female Large White × Landrace pigs (average weight 57·2 (SD 1·9) kg) were allocated to one of six dietary treatments containing 0, 1·25, 2·5, 5·0, 7·5 or 10·0 g 55 % conjugated linoleic acids (CLA) isomers (CLA-55)/kg diet and fed for 8 weeks. Each pig was scanned at 0, 28 and 56 d and again at post slaughter using dual-energy X-ray absorptiometry (DXA) to determine the temporal pattern of body composition responses. Values determined by DXA were adjusted using regression equations generated from validation experiments between chemically and DXA-predicted values. Overall, there was a significant linear reduction in fat content with the increasing levels of CLA in the diet (P=0·007, P=0·011, P=0·008 at week 4, week 8 and for the carcass, respectively). The greatest improvement was recorded at the early stages of CLA supplementation and for the highest dose of CLA (week 4, -19·2 % compared with week 8, -13·7 %). In the first 4 weeks of feeding CLA, pigs receiving 10 g CLA-55/kg diet deposited 93 g less fat/d than pigs fed basal diets (P=0·002) compared with only 6 g less fat than control animals in the final 4 weeks. Lean content and lean deposition rate were maximised at 5 and 2·5 g CLA-55/kg diet for the first 4 weeks (P=0·016) and the final 4 weeks of treatment (P=0·17), respectively. DXA estimates of bone mineral content and bone mineral density were not affected by CLA supplementation throughout the experiment. These data demonstrate that dietary CLA decreases body fat in a dose-dependent manner and that the response is greatest over the initial 4 weeks of treatment.

Effects of dietary conjugated linoleic acid on haematological and humoral responses in the grower pig

The effects of conjugated linoleic acid (CLA) on the levels of total serum leucocytes, granulocytes including neutrophils, basophils, and eosinophils, as well as on monocytes and leucocytes were measured in pigs selected from a clean (minimal disease) herd. Thirty pigs were fed different rates of dietary CLA (0, 1.25, 2.5, 5.0, 7.5, and 10.0 g CLA-55/kg diet) for 8 weeks. Blood samples were collected at the end of the study for assessment of haematological and humoral responses to CLA supplementation. No difference in total white blood cells including the neutrophil, monocyte, and lymphocyte counts was observed among different dietary groups. A dose-dependent reduction (P = 0.02) in eosinophil concentrations suggests that CLA exerts anti-inflammatory activities. A 2-fold increase in the level of basophils was recorded in pigs fed lower levels of CLA (1.25 and 2.5 g CLA/kg diet) but the levels decreased gradually (P = 0.05) and were below the detection limit at the highest rate (10 g/kg) of CLA supplementation. The level of IgG was reduced by over 50% in CLA-fed pigs (P < 0.001), although the response was quadratic in nature (P < 0.001). T-cell population analysis showed that CD4+ cells tended (P = 0.06) to be reduced linearly with increasing inclusion of CLA in the diet. Our results suggest that dietary CLA modulates haematological and humoral responses in a dose-dependent manner.

Dietary conjugated linoleic acid improves carcass leanness without altering meat quality in the growing pig

One constraint facing the pig industry is that ad libitum feeding can often result in high levels of body fat and technologies which can reduce the ratio of lean to fat deposition in the pig are continually being explored. Conjugated linoleic acids have been shown to decrease body fat content in pigs. Therefore, the aim of this study was to determine whether dietary conjugated linoleic acids supplementation has any effect on meat quality and carcass characteristics in finisher pigs. Sixty female crossbred (Large White × Landrace) pigs (average initial weight 56.6 ± 1.9 kg and average initial P2 backfat 11.4 ± 1.3 mm) were used in the present study. Pigs were individually housed and randomly allocated to 1 of 6 dietary treatments: 0, 0.125, 0.25, 0.50, 0.75 or 1.0% (w/w) of conjugated linoleic acids-55. The wheat-based diets were formulated to contain 14.3 MJ DE and 9.3 g available lysine per kg and were fed ad libitum for 8 weeks. Pigs were slaughtered and meat quality was determined on the longissimus thoracis using standard techniques. Dietary conjugated linoleic acids reduced subcutaneous back fat in a linear manner with effects being most pronounced in the middle back fat layer. There was also a linear (P<0.001) decrease in intramuscular fat with increasing dietary conjugated linoleic acids supplementation. However, there was no effect of conjugated linoleic acids on subjective measures of marbling of the loin. Also, loin muscle ultimate pH (P = 0.94), lightness values (P = 0.46) subjective colour scores (P = 0.79), cooking loss (P = 0.71), drip loss (P = 0.40), shear force (P = 0.61) and subjective measures of wetness/firmness (P = 0.19) were unaffected. Dietary conjugated linoleic acids did not alter oxidation, as measured by the level of TBARs at day 1 post-slaughter (P = 0.38) or after 9 days of simulated retail display (P = 0.35). These data confirm that dietary conjugated linoleic acids can improve carcass quality by decreasing back fat depths without having any detrimental effects on meat quality.

1-14C-Linoleic Acid distrubution in various tissue lipids of guinea pigs following an oral dose

A recent study on the metabolism of 1-¹⁴C-α-linolenic acid in the guinea pig revealed that the fur had the highest specific activity of all tissues examined, 48 h after dosing. The present study investigated the pattern of tissue lipid labeling following an oral dose of 1-¹⁴C-linoleic acid after the animals had been dosed for the same time as above. Guinea pigs were fed one of two diets with a constant linoleic acid content (18% total fatty acids) and a different content of α-linolenic acid (0.3 or 17.3%) from weaning for 3 wk and 1-¹⁴C-linoleic acid was given orally to each animal for 48 h prior to sacrifice. The most highly labeled tissues (dpm/mg of linoleic acid) were liver, followed by brain, lung and spleen, heart, kidney and adrenal and intestines, in both diet groups. The liver had almost a three-fold higher specific activity than skin and fur which was more extensively labeled than the adipose and carcass. Approximately two-thirds of the label in skin plus fur was found in the fur which, because of a low lipid mass, would indicate that the fur was highly labeled. All tissues derived from animals on the diet with the low α-linolenic acid level were significantly more labeled than the tissues from the animals on the high α-linolenic acid diet, by a factor of 1.5 to 3. The phospholipid fraction was the most highly labeled fraction in the liver, free fatty acids were the most labeled fraction in skin & fur, while triacyglycerols were the most labeled in the carcass and adipose tissue. In these tissues, more than 90% of the radioactivity was found in fatty acids with 2-double bonds in the tissue lipids. These data indicate that the majority of label found in guinea pig tissues 48 h after dosing was still associated with a fatty acid fraction with 2-double bonds, which suggests there was little metabolism of linoleic acid to more highly unsaturated fatty acids in this time frame. In this study, the labeling of guinea pig tissues with linoleic acid, 48 h after dosing, was quite different from the labeling with α-linolenic acid reported previously. The retention of the administered radioactivity from ¹⁴C-linoleic acid in the whole body lipids was 1.6 times higher in the group fed the low α-linolenic acid diet (diet contained a total of 1.8 g PUFA/100 g diet)compared with the group fed the high α-linolenic acid diet (diet contained 3.6 g PUFA/100 g diet). The lack of retention of ¹⁴C-labeled lipids in the whole body would be consistent with an increased rate of β-oxidation of the labeled fatty acid on the diet rich in PUFA, a result supported by other studies using direct measurement of labeled carbon dioxide.

Effect of feeding systems on omega-3 fatty acids, conjugated linoleic acid and trans fatty acids in Australian beef cuts: potential impact on humnan health

The influence of feeding systems on the levels of functional lipids and other fatty acid concentrations in Australian beef was examined. Rump, strip loin and blade cuts obtained from grass feeding, short-term grain feeding (80 days; STGF) and long-term grain feedlot rations (150-200 days; LTFL) were used in the present study. The typical Australian feedlot ration contains more than 50% barley and/or sorghum and balanced with whole cottonseed and protein meals were used as feed for STGF and LTFL regimens. Meat cuts from 18 cattle for each feeding regimen were trimmed of visible fat and  connective tissue and then minced (300 g lean beef); replicate samples of 7g were used for fatty acid (FA) analysis. There was a significantly higher level of total omega-3 (n-3) and long chain n-3 FA in grass-fed beef (P <0.0001) than the grain-fed groups regardless of cut types. Cuts from STGF beef had significantly reduced levels of n-3 FA and conjugated linoleic acid (CLA) and similar levels of saturated, monounsaturated and n-6 FA compared with grass feeding (P <0.001). Cuts from LTFL beef had higher levels of saturated, monounsaturated, n-6 FA and trans 18:1 than similar  cuts from the other two groups (P <0.01), indicating that increased length of grain feeding was associated with more fat deposited in the carcass. There was a step-wise increase in trans 18:1 content from grass to STGF to LTGF, suggesting grain feeding elevates trans FA in beef, probably because of increased intake of 18:2n-6. Only grass-fed beef reached the target of more than 30mg of long chain n-3 FA/100 g muscle as recommended by Food Standard Australia and New Zealand for a food to be considered a source of omega- 3 fatty acids. The proportions of trans 18:1 and n-6 FA were higher (P<0.001) for both grain-fed beef groups than grass-fed beef. Data from the present study show that grain feeding decreases functional lipid  components (long chain n-3 FA and CLA) in Australian beef regardless of meat cuts, while increasing total trans 18:1 and saturated FA levels.

Conjugated linoleic acid suppresses myogenic gene expression in a model of human muscle cell inflammation

Proinflammatory cytokines, such as tumor necrosis factor (TNF)-{alpha}, contribute to muscle wasting in inflammatory disorders, where TNF{alpha} acts to regulate myogenic genes. Conjugated linoleic acid (CLA) has shown promise as an antiproliferative and antiinflammatory agent, leading to its potential as a therapeutic agent in muscle-wasting disorders. To evaluate the effect of CLA on myogenesis during inflammation, human primary muscle cells were grown in culture and exposed to varying concentrations of TNF{alpha} and the cis-9, trans-11 and trans-10, cis-12 CLA isomers. Expression of myogenic genes (Myf5, MyoD, myogenin, and myostatin) and the functional genes creatine kinase (CK) and myosin heavy chain (MHC IIx) were measured by real-time PCR. TNF{alpha} significantly downregulated MyoD and myogenin expression, whereas it increased Myf5 expression. These changes corresponded with a decrease in both CK and MHC IIx expression. Both isomers of CLA mimicked the inhibitory effect of TNF{alpha} treatment on MyoD and myogenin expression, whereas myostatin expression was diminished in the presence of both isomers of CLA either alone or in combination with TNF{alpha}. Both isomers of CLA decreased CK and MHC IIx expression. These findings demonstrate that TNF{alpha} can have specific regulatory effects on myogenic genes in primary human muscle cells. A postulated antiinflammatory role of CLA in myogenesis appears more complex, with an indication that CLA may have a negative effect on this process.

Fatty acid composition in tissues of mice fed diets containing conjugated linolenic acid and conjugated linoleic acid

The influence of 1% alpha-eleostearic acid (α-ESA, cis9,trans11,trans 13-18:3) and 1% punicic acid (PA, cis9,trans11,cis13-18:3) on fatty acid composition in mouse tissues was compared with conjugated linoleic acid (CLA, mixture of primarily cis9,trans11- and trans10,cis12-18:2) in the present study. The content (% total fatty acids) of 18:2n-6 was significantly reduced in the heart and adipose tissues, and total polyunsaturated fatty acids (PUFAs) and n-6 PUFA were significantly reduced in adipose tissue by α-ESA, PA and CLA feeding. The content of 22:6n-3 and total n-3 PUFA were significantly increased in the liver, kidney and heart by PA feeding, but not by α-ESA. In contrast to PA, supplementation with CLA significantly decreased 22:6n-3 in the liver, kidney and heart. The content of 20:4n-6 was significantly decreased in the liver and kidney by CLA feeding, but not by α-ESA and PA. The present results indicate that α-ESA, PA and CLA have differential effects on 22:6n-3 and 20:4n-6 content in mouse tissues.

Effects of conjugated linoleic acid on myogenic and inflammatory responses in a human primary muscle and tumor coculture model

The antiproliferative and anti-inflammatory properties of conjugated linoleic acid (CLA) make it a potentially novel treatment in chronic inflammatory muscle wasting disease, particularly cancer cachexia. Human primary muscle cells were grown in coculture with MIA PaCa-2 pancreatic tumor cells and exposed to varying concentrations of c9,t11 and t10,c12 CLA. Expression of myogenic (Myf5, MyoD, myogenin, and myostatin) and inflammatory genes (CCL-2, COX-2, IL-8, and TNF-) were measured by real-time PCR. The t10,c12 CLA isomer, but not the c9,t11 isomer, significantly decreased MIA PaCa-2 proliferation by between 15% and 19%. There was a marked decrease in muscle MyoD and myogenin expression (78% and 62%, respectively), but no change in either Myf5 or myostatin, in myotubes grown in coculture with MIA PaCa-2 cells. CLA had limited influence on these responses. A similar pattern of myogenic gene expression changes was observed in myotubes treated with TNF- alone. Several-fold significant increases in CCL-2, COX-2, IL-8, and TNF- expression in myotubes were observed with MIA PaCa-2 coculture. The c9,t11 CLA isomer significantly decreased basal expression of TNF- in myotubes and could ameliorate its tumor-induced rise. The study provides insight into the anti-inflammatory and antiproliferative actions of CLA and its application as a therapeutic agent in inflammatory disease states.

Compositions of cis-9, trans-11 conjugated linoleic acid and vaccenic acid and uses thereof

The invention relates to use of the cis-9, trans-11 isomer of conjugated linoleic acid or a salt or ester thereof (cis-9, trans-11 CLA) and vaccenic acid or a salt or ester thereof (VA) to treat or prevent conditions associated with one or more of leukocyte infiltration, eosinophilia, airway remodelling, bronchoconstriction, mucus hypersecretion, and lung and skin inflammation. The present invention also relates to a composition comprising cis-9, trans-11 CLA and VA and use of the composition to treat or pre-vent conditions associated with one or more of leukocyte infiltration, eosinophilia, airway remodelling, bronchoconstriction, mucus hypersecretion, and lung and skin inflammation. In particular, the medicinal uses, compositions and methods of the invention may be used to treat or prevent conditions such as asthma and dermatitis, and related disorders.