Alpha-linolenic acid but not conjugated linolenic acid is hypocholesterolaemic in hamsters

Conjugated linolenic acid (CLN) refers to a group of octadecatrienoic acid isomers that have three double bonds in conjugation. Both pomegranate and tung seed oils are rich in CLN but the major isomer in the former is cis9,trans11,cis13 while in the latter it is cis9,trans11,trans13. The present study examined the effects of CLN, isolated from either pomegranate seed oil or tung seed oil, and alpha-linolenic acid (LN), isolated from flaxseed oil, on serum cholesterol levels in male hamsters (body weight 105 g; age 10 weeks) fed a 0.1% cholesterol and 10% lard diet, for a period of 6 weeks. All hamsters were allowed free access to food and fluid. The blood samples were taken by bleeding from the retro-orbital sinus into a heparinized capillary tube under light ether anaesthesia after overnight fasting at weeks 0, 2, 4 and 6. It was found that supplementation of CLN at levels of 12.2-12.7 g/kg diet exhibited no significant effect on serum cholesterol level while LN at a similar level of supplementation had serum cholesterol reduced by 17-21% compared with the control diet containing no LN and CLN. Supplementation of CLN and LN significantly decreased hepatic cholesterol but no effect was observed on heart and kidney cholesterol levels. It was concluded that LN possessed hypocholesterolaemic activity while CLN had no effect on blood cholesterol, at least in hamsters.     

Effects of conjugated linoleic acid on linoleic and linolenic acid metabolism in man

Evidence from animal studies suggests that conjugated linoleic acid (CLA) modulates plasma and tissue appearance of newly synthesized PUFA. The effects of a 1.2g (0.5 % energy) daily intake of the cis-9,trans-11 (c9,t11) isomer of CLA, trans-10,cis-12 (t10,c12) isomer of CLA or olive oil (placebo) on linoleic acid (LA) and linolenic acid (LNA) metabolism in healthy human volunteers was investigated. Fifteen subjects were fed an experimental diet and supplemented with c9,t11-CLA, t10,c12-CLA or placebo for 7 d before consuming a tracer dose of U-[(13)C]LA (50 mg) and U-[(13)C]LNA (50 mg). Blood samples were taken at 0, 2, 4, 6, 8, 24, 48, 72 and 168 h and analysed using high-precision MS. No differences between the groups in peak plasma [(13)C]LA (10.3-11.6 % of dose), [(13)C]LNA (2.5-2.9 % of dose), [(13)C]arachidonic acid (0.09-0.12 % of dose), [(13)C]EPA (0.04-0.06 % of dose) or [(13)C]DHA (0.06-0.10 % of dose) were detected. Concentration v. time curves (area under the curve) also showed no significant differences between groups. This suggests that, in healthy human subjects consuming a diet with adequate intake of essential fatty acids, CLA does not affect metabolism of LA or LNA.     

Ferulic acid is quickly absorbed from rat stomach as the free form and then conjugated mainly in liver

Ferulic acid (FA) is one of the most abundant phenolic antioxidants in the human diet. Many studies have documented its beneficial properties. It is therefore essential to understand the absorption and metabolism of FA in detail. The purpose of this study was to confirm the hypothesis that FA is absorbed in rat stomach and metabolized mainly in the liver. We determined the recovery of FA and its metabolites (FA sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after 2.25 micromol FA was administered in 0.5 mL physiological saline and incubated for 25 min in situ in the stomach of rats. Within 25 min, 74 +/- 11% of the administered FA disappeared from the stomach; later, FA was recovered in both free and conjugated forms in plasma, bile, and urine. On the other hand, only free FA was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as 49 +/- 5% of the total FA (all forms of FA). However, the proportion of free FA in the celiac arterial plasma, bile, and urine decreased to 5-8%. These results indicate that FA can be quickly absorbed from the rat stomach, and then is likely metabolized mainly in the liver. Such novel information would be helpful in the use of FA as a nutrient supplement. For example, oral administration of FA in capsule form or in a form bonded with sugar esters may provide a more appropriate concentration of FA in the circulation, which may improve its proposed efficacy in preventing chronic disease.     

Trans-vaccenic acid is desaturated to conjugated linoleic acid in mice

Mice were fed pure trans11 octadecenoic acid (trans-vaccenic acid; TVA) to determine whether it is desaturated to cis9, trans11 octadecadienoic acid, a predominant isomer of conjugated linoleic acid (CLA). In a preliminary trial, 12% of the TVA consumed during a 2-wk feeding period was recovered in the carcass as CLA. As a proportion of TVA in the tissues available for bioconversion, 48.8% was desaturated. We tested whether desaturation could be modified by supplementing no modifier, 0.5% clofibric acid to stimulate desaturation, or increasing the polyunsaturated fatty acids (PUFA) (10% corn oil vs. 4% corn oil) to inhibit desaturation in diets with or without 1% TVA. These diets were fed to six groups of mice in a 3x2 factorial arrangement of treatments. Feeding 1% TVA with 10% corn oil decreased feed intake (2.70 vs. 3.73 g/d, SEM 0.23; P<0.05). Bioconversion of dietary TVA was 12.0, 7.5 and 5.1% for mice fed no modifier of desaturation, clofibrate and increased PUFA, respectively. Conversion based on TVA available for desaturation was 52.6, 55.5 and 37.0%, respectively. Thus, clofibrate did not increase bioconversion, but increasing PUFA decreased conversion by 30%. To test whether TVA decreases food intake directly or after conversion to CLA, four groups of mice were fed diets containing 1% stearic, TVA, elaidic or conjugated linoleic acid. Dietary CLA decreased food intake and body fat, but did not change body protein. CLA was found in the carcass only when TVA or CLA was fed. CLA was found in both triacylglycerol and phospholipids when CLA was fed, but only in triacylglycerol when TVA was fed, suggesting that bioconversion occurred in the adipose tissue. In three trials, conversion of dietary TVA to CLA was 11.4+/-1.25%; conversion of stored TVA was 50.8+/-1.91%. Similar bioconversion of TVA in humans would increase current estimates of CLA available for the general population by 6- to 10-fold.     

Alpha-eleostearic acid (9Z11E13E-18:3) is quickly converted to conjugated linoleic acid (9Z11E-18:2) in rats

We previously showed that alpha-eleostearic acid (alpha-ESA; 9Z11E13E-18:3) is converted to conjugated linoleic acid (CLA; 9,11-18:2) in the liver and plasma of rats that were given diets including 1% alpha-ESA for 4 wk. In this study, we investigated this phenomenon in detail. First, the chemical structure of CLA produced by alpha-ESA administration was determined. After alpha-ESA was orally administered to rats, CLA in rat liver was isolated by HPLC. The positional and geometric isomerism was determined using GC-EI/MS and (13)C-NMR, respectively, and the CLA generated in rats after alpha-ESA feeding was confirmed to be 9Z11E-CLA. Next, the concentrations of alpha-ESA and CLA were determined 0, 3, 6, and 24 h after oral administration of alpha-ESA to rats. Moreover, we also investigated whether enteric bacteria are involved in the conversion of alpha-ESA to CLA using germ-free rats. alpha-ESA was orally administered to germ-free and normal rats and alpha-ESA and CLA were detected in the organs of both groups. In addition, to confirm that this reaction was enzyme-mediated, alpha-ESA was reacted with tissue homogenates (liver, kidney, and small intestine mucous) and coenzymes (NADH, NAD(+), NADPH, and NADP(+)), and the enzyme activities were estimated from the amount of CLA produced. CLA was detected when alpha-ESA was reacted with liver, kidney, and small intestine mucous homogenates and a coenzyme (NADPH). These results indicated that alpha-ESA is converted to 9Z11E-CLA in rats by a Delta13-saturation reaction carried out by an NADPH-dependent enzyme.     

Conjugated linoleic acid is related to bone mineral density but does not affect parathyroid hormone in men

The relationships between conjugated linoleic acid (CLA) status, bone, body composition, and the effect of CLA on calciotropic hormones are unclear. A cross-sectional study was designed to examine the association between c9, t11 CLA status in erythrocyte membranes (RBC) and body composition. This preceded a dose-response trial investigating if c9, t11 CLA affected parathyroid hormone (PTH). It was hypothesized that (1) higher c9, t11 CLA status in RBC will be associated with a lower fat and higher bone mass and that (2) PTH will be reduced by 30% after supplementation of c9, t11 CLA. Fifty-four men (age, 19-53 years) were included in the cross-sectional analysis, of which 31 were studied in the dose-response trial and randomized to 1 of 3 groups: placebo (n = 10), 1.5 g/d (n = 11), or 3.0 g/d (n = 10) of c9, t11 CLA for 16 weeks. Men with RBC c9, t11 CLA status above the median had higher whole body bone mineral density (BMD) (1.359 ± 0.024 vs 1.287 ± 0.023 g/cm²; P = .04) and whole body lean mass (WBL) percentage (78.8% ± 0.9% vs 75.3% ± 1.0%; P = .01), whereas body mass index (24.8 ± 0.5 kg/m² vs 27.3 ± 0.9 kg/m²; P = .01) and whole body fat mass percentage (17.3% ± 0.9% vs 21.3% ± 1.1%; P = .007) were lower. In regression analysis, RBC c9, t11 CLA status accounted for a significant proportion (r² = 0.10) of the variation in whole body BMD (P = .03). There were no time or treatment differences among any bone or biomarkers of bone metabolism including PTH. These findings indicate that RBC c9, t11 CLA status, a reflection of long-term (~4 months) dietary CLA intake, positively relates to BMD. However, c9, t11 CLA supplementation does not appear to affect PTH in healthy men.     

Transfer of absorbed cis-9, trans-11 conjugated linoleic acid into milk is biologically more efficient than endogenous synthesis from absorbed vaccenic acid in lactating cows

Cis-9, trans-11, the major isomer of conjugated linoleic acid (CLA) in bovine milk fat, is derived from ruminal biohydrogenation of 18:2 (n-6) and endogenous conversion of trans-11 18:1 (vaccenic acid; VA) in the mammary gland. Most evidence to date suggests that endogenous synthesis is the major source of cis-9, trans-11 CLA, but the extent of VA desaturation is less well defined. Four lactating cows were used in consecutive 4 x 4 Latin squares to examine changes in milk fatty acid composition and secretion in response to abomasal infusions of lipid supplements enriched with cis-9, trans-11 CLA (88.8%) or VA (29.4%). Treatments were infused over 4-d, followed by a 3-d washout, during 7 d experimental periods and administered to deliver 0, 3, 6, and 12 g cis-9, trans-11 CLA/d (Expt. 1) or 0, 7.5, 15 and 30 g VA/d (Expt. 2). Infusions of cis-9, trans-11 CLA increased linearly milk cis-9, trans-11 CLA concentrations from 0.68 to 1.46 g/100 g fatty acids. Abomasal infusions of VA increased linearly milk VA and cis-9, trans-11 CLA content from 1.22 to 2.72 and 0.61 to 1.24 g/100 g fatty acids, respectively. Changes in milk fatty acid secretion indicated that 28.9% of VA was converted to cis-9, trans-11 CLA. Results provide evidence that conversion by Delta9-desaturase to cis-9, trans-11 CLA in the lactating cow is independent of postruminal VA supply. In conclusion, endogenous synthesis via VA was equivalent to approximately 21% of the response to increases in cis-9, trans-11 CLA available for absorption.     

Bioconversion of vaccenic acid to conjugated linoleic acid in humans

BACKGROUND: Vaccenic acid (11-trans octadecenoic acid; VA), a major trans fatty acid in the fat of ruminants, is produced in the rumen and converted in tissues to rumenic acid (9-cis, 11-trans octadecenoic acid; RA), an isomer of conjugated linoleic acid, by Delta(9)-desaturase. There are indications that this conversion also occurs in humans. OBJECTIVE: The aim of this controlled intervention was to study the conversion of VA to RA in humans after consumption of diets with increasing amounts of VA. DESIGN: Thirty healthy subjects consumed a baseline diet rich in oleic acid for 2 wk. The subjects were then divided into 3 groups (n = 10 per group) and provided a diet containing 1.5, 3.0, or 4.5 g VA/d for 9 d. All diets contained equal amounts of macronutrients and differed only in their fatty acid compositions. The fats were mixed into conventional foods, and nearly all food was provided during the study. RESULTS: The proportion of VA in serum total fatty acids increased 94%, 307%, and 620% above baseline with the 1.5-, 3.0-, and 4.5-g diets, respectively. This was associated with a linear increase in the proportion of RA. The conversion rate was 19% on average, with significant interindividual differences with all 3 intakes of VA. The urinary excretion of 8-iso-prostaglandin F(2alpha) increased in all groups (P < 0.001). CONCLUSIONS: The results quantify the desaturation of VA to RA in humans. Conversion is likely to contribute significantly to the amount of RA available to the body, and dietary intakes of VA should thus be taken into account when predicting RA status.     

The change in conjugated linoleic acid concentration in blood of Japanese fed a conjugated linoleic acid diet

Conjugated linoleic acid (CLA) is a collective term used for fatty acids with a conjugated double bond that are geometrical and positional isomers of linoleic acid. Anti-obesity and anti-cancer properties, an immunopotentiation effect, and promotion of bone formation by CLA have been shown in cell culture and animal studies. A mixture of 9c11t- and 10t12c-CLA is now used as a health food supplement after testing in clinical trials. These trials focused on improvement of lipid metabolism by CLA, whereas few studies have examined absorption and metabolism of CLA in humans. In addition, there is no report concerning absorption and metabolism of CLA in Japanese. This study was designed to examine CLA concentration in blood, the elimination rate of CLA, and metabolic differences between 9c11t-CLA and 10t12c-CLA in blood in Japanese who ingested CLA (about 2 g/d, equal weights of 9c11t-CLA and 10t12c-CLA) for 3 wk. Blood samples were collected 1 wk before the 3-wk period, on the first and last days of the period, and 1 wk after the end of the period, and the CLA concentration and distribution in blood were investigated. The CLA concentration in blood was significantly increased by CLA ingestion and reached 36 μmol/L. The CLA concentration in blood one week after the intake period was significantly lower than that at the end of CLA intake. The 10t12c-CLA level in plasma decreased faster than that of 9c11t-CLA. This suggests faster metabolism (fatty acid β oxidation) of 10t12c-CLA compared with 9c11t-CLA.     

Dietary conjugated linoleic acid reduces adiposity in lean but not obese Zucker rats

Recent studies have demonstrated a reduction in body fat in growing animals fed conjugated linoleic acid (CLA). Two experiments were conducted to extend these observations to obese rats so that the mechanism of the actions of CLA might be more easily elucidated. In experiment 1, male lean and obese Zucker rats were fed diets containing either 0 or 0.5% CLA for 5 wk. There was no effect of diet on growth rate or food intake. Dietary CLA reduced retroperitoneal and inguinal fat pad weights in the lean rats but increased fat pad weights in the obese genotype (diet x genotype interaction; P < 0.05). Determination of fat pad cellularity indicated that these changes in fat pad weight were due to a reduction or increase in average fat cell size for the lean and obese Zucker rats, respectively. In experiment 2, we sought to reproduce these effects on fat pad size, as well as to determine the effect of dietary CLA on the catabolic response to bacterial endotoxin injection in obese Zucker rats. Growing female lean and obese Zucker rats were fed diets containing 0 or 0.5% CLA for 8 wk. On d 28, each rat was injected intraperitoneally with lipopolysaccharide from Escherichia coli serotype 055:B5 (1 mg/kg body weight) and body weight was determined over the next 96 h. There was a diet x genotype interaction (P < 0.05) for the body weight response to lipopolysaccharide 24 h postinjection. Lean rats fed CLA lost less weight than did lean controls, but obese rats fed CLA lost more weight than did obese controls. As in the first experiment, there was a diet x genotype (P < 0.05) for the effect of treatment on retroperitoneal fat pad weights determined at the end of the experiment. Lean rats fed CLA had smaller RP fat pads than did lean controls, but obese rats fed CLA once again had heavier RP fat pads than did obese controls. These results indicate that CLA reduces body fat and catabolic response to endotoxin injection in lean Zucker rats but not in the obese genotype. The observed interaction between diet and genotype warrants additional investigation into the specific mechanism(s) of the biological activities of CLA.     

Cis-9, trans-11 conjugated linoleic acid is synthesized from vaccenic acid in lactating women

We studied the incorporation of the trans-11 vaccenic-1-(13)C acid ((13)C-VA) into milk and endogenous synthesis of cis-9, trans-11 conjugated linoleic acid (CLA) in lactating women. Subjects (n = 4) were 247 +/- 30 d postpartum, weighed 70.8 +/- 3.7 kg, breast-fed at least 6 times/d and consumed self-selected diets. After an overnight fast, they consumed the (13)C-VA (2.5 mg/kg body wt). Milk samples were obtained by complete breast expression at 0, 2, 4, 8, 12, 18, 24, and 48 h post-(13)C-VA ingestion. Lipid was extracted using chloroform:methanol. Fatty acids were methylated and converted to dimethyl disulfide and Diels-Alder derivatives before analysis by gas chromatography mass spectrometry. The mean (13)C-enrichment of milk VA was 3.1% at 8 h and reached maximal enrichment of 7.6% at 18 h. The (13)C enrichment of milk cis-9, trans-11 CLA reached a maximum of 0.4% at 18 h, confirming its conversion of VA to the Delta9-desaturase enzyme product. In the subjects examined, a portion (<10%) of the cis-9, trans-11 CLA present in milk was endogenously synthesized from VA.     

Carp oil or oleic acid,but not linoleic acid or linolenic acid,inhibits tumor growth and metastasis in Lewis lung carcinoma-bearing mice

I examined the effects of carp oil, oleic acid, linoleic acid and linolenic acid on tumor growth and metastasis to the liver in mice implanted intrasplenically with highly metastatic Lewis lung carcinoma (LLC) tumors. Carp oil (0.1 or 0.2 mL per mouse) significantly reduced tumor growth and metastasis to the liver. Carp oil at 100 or 1000 mg/L inhibited the DNA synthesis in LLC cells, the capillary-like tube formation of human dermal microvascular endothelial cells (HMVEC) at 1000 mg/L and the adherence of LLC cells to HMVEC at 10 to 1000 mg/L (in vitro). Carp oil (0.2 mL per mouse) inhibited the angiogenesis induced by Matrigel supplemented with vascular endothelial growth factor (VEGF) and heparin (in vivo). Antitumor and antimetastatic actions of carp oil might be partly attributable to the inhibition of DNA synthesis in LLC cells and angiogenesis through the inhibition of the adherence of LLC cells to the microvascular endothelium. Oleic acid (0.1 or 0.2 mL per mouse) significantly inhibited the metastasis to the liver, but it had no effect on the primary solid-tumor growth. Oleic acid inhibited the angiogenesis in both in vitro and in vivo models. Oleic acid at 1000 micromol/L inhibited the DNA synthesis in LLC cells but did not affect the DNA synthesis in HMVEC. These inhibitory actions of oleic acid may be attributable to the inhibition of angiogenesis induced by the tumor. Linoleic acid and linolenic acid had no effect on tumor growth or metastasis to the liver.     

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 Ag+-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 delta5-, delta6- and delta9-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.     

Control of rat mammary epithelium proliferation by conjugated linoleic acid.

Past research showed that mammary gland morphogenesis in the pubescent rat was retarded by the feeding of conjugated linoleic acid (CLA). A major objective of the present study was to examine the proliferative activity and the expression of cell cycle regulatory proteins in the developing mammary epithelium of rats fed a mixture of CLA isomers (primarily as free fatty acid c9, t11-CLA and t10,c12-CLA) or a highly enriched natural source of c9,t11-CLA (as triacylglycerol in butterfat). In both experiments, the diets, with or without CLA, were started at weaning and continued for four weeks. The two CLA preparations were equally effective in suppressing bromodeoxyuridine labeling and the expression of cyclin D1 and cyclin A (determined by immunohistochemistry) in the terminal end buds and alveolar clusters of the mammary epithelium while it undergoes extensive ductal branching during pubescence. There was a trend of an increase, although not statistically significant, in the proportion of cells expressing the p16 and p27 cdk inhibitors. A separate experiment was designed to evaluate the effect of c9,t11-CLA (as a free fatty acid of > 90% purity) treatment on the rate of proliferation of the mammary epithelium as the animal matured from weanling to adult. The bromodeoxyuridine labeling data indicated that the mammary epithelium appeared to lose its sensitivity to CLA control of proliferation as it completely filled the fat pad and became quiescent. These observations suggest that the responsiveness of mammary epithelial cells to CLA intervention may be dependent on their proliferative status.     

Recent advances in conjugated linoleic acid research

New results on the physiological properties of conjugated linoleic acid have been published by several working groups, especially showing the effects of single conjugated linoleic acid isomers on carcinogenesis and body composition. Recently, other studies have shown that conjugated linoleic acid has an influence on diabetes mellitus, platelet aggregation and the immune system. Conjugated linoleic acid was found to modify prostaglandin metabolism and delta9-desaturase activity and influence apoptosis. Furthermore, improved analytical methods using 13C nuclear magnetic resonance and silver ion high performance liquid chromatography are available to investigate the composition of conjugated linoleic acid mixtures and the exact structure of separated isomers. Also, the synthesis of isolated isomers is described, as published by different authors, in order to determine further the effects of each single conjugated linoleic acid isomer. In addition, new data on the contents of conjugated linoleic acid in foods, human adipose tissue and fluids are given in this review. More data need to be obtained using isolated isomers, with particular emphasis on studies in humans.     

Conjugated linoleic acid reduces early aortic atherosclerosis greater than linoleic acid in hypercholesterolemic hamsters

Thirty-six male F₁B hamsters, 10 weeks of age, were divided into 3 groups of 12 based on similar body weights. The experimental diets comprised of a chow-based hypercholesterolemic diet supplemented with 20% coconut oil, 2% safflower oil, and 0.12% cholesterol (HCD); the HCD plus either 1% CLA as the free fatty acid (CLA), or 1% LA as the free fatty acid (LA) and were fed for 12 weeks. Plasma total cholesterol (TC) and nonHDL-C (very low- and low-density lipoprotein cholesterol) were significantly lower in the CLA and LA relative to the HCD (P < 0.05). The CLA had significantly less maximum number of dienes formed relative to the LA and HCD (P < 0.05). The CLA developed significantly less early aortic atherosclerosis relative to both the HCD and LA (P < 0.05). Thus it appears CLA reduces the development of early aortic atherosclerosis to a greater degree than LA possibly through changes in LDL oxidative susceptibility in hypercholesterolemic hamsters.     

cis-9, trans-11 conjugated linoleic acid is synthesized directly from vaccenic acid in lactating dairy cattle

The utilization of (13)C-labeled vaccenic acid (VA) by lactating dairy cows to synthesize cis-9, trans-11 conjugated linoleic acid (CLA) was investigated. Primiparous ruminally cannulated Holstein cows (n = 3) were abomasally infused with 1.5 g of VA-1-(13)C. Blood and milk samples were taken frequently before and after VA infusion. Milk and plasma lipid were extracted using chloroform:methanol. Plasma lipid was separated into triacylglycerol (TG), cholesterol ester (CE), phospholipid (PL), nonesterified fatty acid (NEFA), and mono- and diacylglycerol (MDG) fractions. Lipid was methylated, converted to dimethyl disulfide and Diels-Alder adducts, and analyzed by GC-MS. Increased enrichment of (13)C was determined using a 2-sample t test for each sample time compared with -24 h, with significance declared at P < 0.05. Enrichment in milk fat VA was detected at 4 (3.0%), 8 (8.3%), 12 (4.1%), 16 (2.2%), and 20 h (0.8%). Enrichment in VA was also detected in plasma TG, NEFA, PL, and MDG. Enrichment in milk fat cis-9, trans-11 CLA, the Delta9-desaturase product of VA, was detected at 4 (2.6%), 8 (6.6%), 12 (3.4%), 16 (1.7%), and 24 h (0.7%). Enrichment was not detected in cis-9, trans-11 CLA for any plasma lipid fraction. Modeling of the data showed the exponential decay in (13)C enrichment over time for both VA and cis-9, trans-11 CLA in milk fat. Conversion of dietary VA to cis-9, trans-11 CLA endogenously was confirmed with the mammary gland being the primary site of Delta9-desaturase activity; approximately 80% of milk fat cis-9, trans-11 CLA originated from VA.     

Changes in body composition with conjugated linoleic acid

Conjugated linoleic acid has been shown to reduce body fat accumulation in several animal models. We have conducted several studies in AKR/J mice showing that CLA reduces body fat accumulation whether animals are fed a high-fat or low-fat diet, with no effect on food intake. One mechanism by which CLA reduces body fat is by increased energy expenditure, which is observed within one week of CLA feeding and is sustained for at least six weeks. The increased energy expenditure is sufficient to account for the decreased fat accumulation. Increased uncoupling protein gene expression does not appear to be involved in the increased energy expenditure. We have observed increased fat oxidation but no decrease in de novo fat biosynthesis with CLA feeding. We have also observed increased liver weights and plasma insulin levels with higher doses of CLA. In all of the studies we have conducted to date we have used a CLA preparation that contains several isomers, primarily c9,t11 and t10,c12. It was assumed that the active form was c9,t11, as CLA was identified as an anticarcinogenic compound from cooked beef, of which the c9,t11 form accounts for 60% to 80% of the CLA. Most of the studies conducted so far must be repeated using the purified isomers in order to determine which isomers are responsible for each of the identified actions of CLA.     

Conjugated linoleic acid increased C-reactive protein in human subjects

We previously showed that conjugated linoleic acid (CLA) increases 15-keto-dihydro-prostaglandin F2alpha, a marker for cyclooxygenase-mediated lipid peroxidation and thus an indicator of cyclooxygenase-mediated inflammation. The aim of the present study was to investigate the effects of CLA on other indicators of inflammation in human subjects, including C-reactive protein, TNF-alpha, TNF-alpha receptors 1 and 2, and vascular cell adhesion molecule-1. In a double-blind, placebo-controlled study, fifty-three human subjects were supplemented with a mixture (4.2 g/d) of the isomers cis-9,trans-11 CLA and trans-10,cis-12 CLA or control oil for 3 months. CLA supplementation increased levels of C-reactive protein (P=0.003) compared with the control group. However, no changes in TNF-alpha, TNF-alpha receptors 1 and 2, and vascular cell adhesion molecule-1 were detected.