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The effect of dietary linoleic acid on rat platelet sialic acid.

The total sialic acid content of blood platelets from rats raised for 8 weeks or 12 months on a diet containing 1% linoleic acid (1LA) was significantly lower (by over 30%) than that from those raised on an isocalorific diet containing 6% linoleic acid (6LA). The transfer of sialic acid to endogenous glycoprotein acceptor was also significantly lower (up to almost 4-fold) in 1LA platelet and megakaryocyte-rich preparations but the transfer to exogenous glycoprotein acceptor was similar in both 1LA and 6LA platelets. The megakaryocyte-rich fraction of 1LA animals showed a reduced phosphodolichol-sensitive N-acetylglucosaminyl (but not mannosyl) transfer to endogenous glycoprotein compared with 6LA animals. No significant difference was found between the megakaryocytes of 1LA and 6LA animals in the incorporation of radioactive mannose and glucosamine into the glycoprotein of the whole cells. It was concluded that the decreased transfer of sialic acid to glycoproteins of platelets and megakaryocyte of animals on the 1LA diet was due to the decreased availability of sialyl acceptor. The formation of N-linked oligosaccharide was the same in both 1LA and 6LA megakaryocytes, and thus any differences in phosphodolichol-mediated N-glycosylation did not account for this decreased availability of sialyl acceptor.

Acetylglucosamine↗

Reduction of murine mammary tumor metastasis by conjugated linoleic acid.

Recent studies have shown that conjugated linoleic acid (CLA) can inhibit the initiation and thus, incidence of mammary tumors in rodents. The concentration of CLA required for these effects was as low as 0.1% of the diet, with no increased effects above 1%. To date, there is little evidence that CLA has any effect on growth or metastasis of mammary tumors. In this report, we demonstrate that CLA, at the concentrations used in previous studies, had a significant effect on the latency, metastasis, and pulmonary tumor burden of transplantable murine mammary tumors grown in mice fed 20% fat diets. The latency of tumors from mice fed CLA was significantly increased when compared with the 0% CLA control diet. The volume of pulmonary tumor burden, as a result of spontaneous metastasis, decreased proportionately with increasing concentrations of dietary CLA. With 0.5 and 1% CLA, pulmonary tumor burden was significantly decreased compared to mice treated with the eicosanoid inhibitor, indomethacin and fed diets containing no CLA. Tumors of mice fed as little as 0.1% CLA and as much as 1% had significantly decreased numbers of pulmonary nodules when compared with diets containing no CLA. The decrease in the number of pulmonary nodules by CLA was nearly as effective as indomethacin, a known suppressor of tumor growth and metastasis in this malignant model. These data suggest that effects of CLA on mammary tumorigenesis may go beyond the reported alterations in tumor incidence and effect later stages, especially metastasis.

Animals↗

The characterization of the linoleic acid desaturation and elongation system in ovine placental tissue.

The desaturation and elongation of linoleic acid has been studied in homogenates and in subfractions of ovine placental tissue. The reaction was characterized in terms of pH and temperature optima, time course and protein concentration. Activity was found to be confined to the 11,000 g supernatant fraction of the tissue and the results suggest that the enzymes are membrane bound. The cytosolic fraction and ATP were required for full activity and the reaction was inhibited by cyanide. The properties of the reaction are compared with those of other desaturation systems and their implications with regard to possible reaction mechanisms are discussed.

Animals↗

Effect of dietary linolenic and linoleic acids upon growth and lipid metabolism of rainbow trout (Salmo gairdneri).

Nine diets, each containing different levels of linoleic acid (18:2 omega 6) and linolenic (18:3 omega 3) were fed to duplicate groups of rainbow trout for 14 weeks. The growth rate, feed efficiency, accumulated mortality, and fatty acid composition of neutral fat and phospholipids of these groups of fish were determined. The growth was slow in the groups of fish receiving diets containing (A) low concentration of 18:3 omega 3and (B) high concentration (5%) of 18:2 omega 6. The accumulated mortality was high in these groups of fish. The diet containing 1% of 18:3 omega 3 alone supported rapid fish growth with low mortality . The feed efficiency of this diet was also high. The metabolism of 18:2 omega 6 and 18:3 omega 3 in fish and their conversion to more unsaturated fatty acids typical of fish lipids was investigated.

Animals↗

Effects of conjugated linoleic acid supplementation on blood lipids and adiposity of rats fed diets rich in saturated versus unsaturated fat.

Conjugated linoleic acid (CLA) may decrease adiposity and improve blood lipid profiles under some conditions. The goal of this study was to determine the effects of CLA supplementation on blood lipid profiles and adiposity of rats fed a diet containing a primarily saturated fat versus a diet containing a primarily unsaturated fat. Twenty-eight male Sprague-Dawley rats were randomly assigned to one of four diets containing coconut oil, coconut oil with CLA, corn oil or corn oil with CLA. After 28 days, blood was collected and serum concentrations of total cholesterol (TC), HDL-cholesterol (HDL-C), and triacylglycerols (TG) were assessed. Food intake, body weights, and epididymal fat pads were measured. No significant differences (p>0.05) were noted among groups for amount of food consumed, weight gained, food efficiency ratio or serum TG concentrations. TC concentrations were lower (p<0.05) in the CLA-supplemented rats that were fed coconut oil but not those consuming corn oil. Serum HDL-C was lower (p<0.05) in rats consuming corn oil but was not significantly different (p>0.05) for CLA supplemented groups. Epididymal fat pads weighed significantly more (p<0.05) in the coconut oil fed group compared to the corn oil fed group, but there was no significant difference (p>0.05) between the corn oil and coconut oil + CLA group. Overall, this study suggests that CLA is more beneficial for control of blood lipids and adiposity when supplemented to a diet rich in saturated versus unsaturated fat.

Adipose Tissue↗

Cholesterol synthesis in the liver, kidneys and brain after injection of uniformly 14C-labeled oleic or linoleic acid to developing rats.

6-day-old suckling rats, born to females kept on a fat-free diet, were used to determine cholesterol and fatty acid specific radioactivity (SRA) in the liver, kidneys and brain, after injection of 3 microCi uniformly 14C-labeled linoleic acid (ULI) or oleic acid (UOL). 1 h after injection, cholesterol SRA was highest in the liver and kidneys, and then decreased when UOL was injected. Cholesterol SRA peaked 3 h after injection of ULI in liver and kidneys. The delay in appearance of the ULI (over UOL) peak of cholesterol SRA may be due to differences in the rates of oxidation of these two labeled fatty acids into acetyl-CoA ester, according to the structural role of ULI. In the period between 3 and 56 h, cholesterol was more radioactive in the three tissues after injection of ULI than after injection of UOL. The radioactivity of saturated fatty acids was low in the ULI and UOL groups of these very young animals. Therefore, cholesterol synthesis seemed to happen at a faster rate than the other lipid syntheses in liver and kidneys, where the rate of linoleic acid elongation into arachidonic acid was also slower than cholesterol synthesis. Different results were obtained in the brain, where arachidonic acid SRA increased rapidly after ULI injection.

Animals↗

[Free radical oxidation of biological membrane lipids. VIII. Effect of linoleic acid and its oxidation products on the functional parameters of mitochondria].

Effects of linoleic acid and its oxidation products (with 75--80% content of hydroperoxide) on rat liver mitochondria were studied by polarographic and spectrophotometric methods. Both agents according to their concentration are able to uncouple oxidation and phosphorylation and to inhibit the electron transport in the initial region of the respiration chain as well. Unlike the unoxidized linoleic acid, its oxidation products increase the activity of some enzymatic systems of mitochondria, and at high concentrations they produce a more pronounced effect on the electron transport.

Animals↗

The influence of a maternal diet rich in linoleic acid on brain and retinal docosahexaenoic acid in the rat.

Female rats were given throughout pregnancy and lactation a semi-synthetic diet, in which the fat was provided entirely by a soft margarine rich in linoleic acid (18:2 omega 6) or by mixture of butter and lard, and the influence on the fatty acid composition of fetal and pup brain and retinal phosphoglycerides was studied. The percentage of docosahexaenoic acid (22:6 omega 3) was much lower but that of docosapentaenoic acid (22:5 omega 6) was correspondingly higher in the brains of the day-22 fetuses and the 21-d-old pups from the margarine group compared with those from the group given the animal fats. Similar changes were noted in the synaptosomal and retinal phosphoglycerides, being most marked in the ethanolamine phosphoglycerides. The remaining pups from two groups were weaned on day 21 post partum on to the same stock diet that contained preformed 22:6 omega 3. After 9 weeks of this diet, the differences between the two groups in the fatty acid composition of the brain phosphoglycerides were barely discernible. The percentage of 22:5 omega 6 had decreased and had been replaced by 22:6 omega 3. It is concluded that the consequences of consuming a diet rich in linoleic acid and almost devoid of 22:6 omega 3 on brain fatty acid composition deserve consideration in man.

Animals↗

Effect of linoleic acid hydroperoxide on production of matrix metalloproteinases by human skin fibroblasts.

The effect of linoleic acid hydroperoxide on in vitro production of matrix metalloproteinases (MMPs) by human skin fibroblasts was studied. The addition of linoleic acid hydroperoxide significantly increased the production of MMP-1 (tissue collagenase) and MMP-3 (stromelysin), while it rather decreased that of MMP-2 (gelatinase of 72 kDa; so-called "type IV collagenase"). The effect of lipid peroxides to alter collagen metabolism was discussed from pathogenic points of view.

Cells, Cultured↗

Effects of dietary linoleic acid and gamma linolenic acid on platelets of patients with multiple sclerosis.

The effects of dietary evening primrose oil (rich in linoleic acid and gamma-linolenic acid) were studied on platelets of multiple sclerosis (MS) patients and controls. It was found that platelet aggregation (ADP, thrombin and collagen), platelet fibrinogen binding and platelet glycoprotein (sialic acid and N-acetyl glucosamine) content were not significantly modified by evening primrose oil in MS patients and controls. Moreover, platelet fibrinogen binding and platelet glycoprotein (sialic acid and N-acetyl glucosamine) content were determined for the first time in MS patients and found similar to controls. Platelets of MS patients aggregated more to thrombin and collagen compared to controls, but the difference was only significant with thrombin aggregation after the oil treatment. This study does not show a significant effect of evening primrose oil on platelets of MS patients.

8,11,14-Eicosatrienoic Acid↗

Effect of a high linoleic acid diet on delta 9-desaturase activity, lipogenesis and lipid composition of pig subcutaneous adipose tissue.

The effects of three diets were compared: a high linoleic acid diet (diet M containing 4% maize oil), diet T containing 4% beef tallow, and C, a conventional control diet, on delta 9-desaturase activity and lipogenesis in pig subcutaneous adipose tissue. Diet M increased lipogenesis (estimated from the activities of acetyl-CoA-carboxylase, malic enzyme and glucose-6-phosphate dehydrogenase), and decreased delta 9-desaturase activity, in comparison to the other diets. Linoleic acid content was higher in the pigs fed diet M than in the other pigs (amounting to 26% of total tissue fatty acids versus 15%, respectively). The lower monounsaturated fatty acid content in adipose tissue of pigs fed diet M compared to pigs fed other diets could be associated with the lower delta 9-desaturase activity and the lower oleic acid content of diet M. The present study suggests that delta 9-desaturase could be involved in the regulation of monounsaturated fatty acid content and hence in the quality of pig adipose tissue.

Acetyl-CoA Carboxylase↗

Conjugated linoleic acid effects on circulating hormones, metabolites and lipoproteins, and its proportion in fasting serum and erythrocyte membranes of swine.

BACKGROUND: Conjugated linoleic acids (CLA)# refer to a group of linoleic acid (18:2)-derived isomers with conjugated double bonds mostly at carbon atoms 9 and 11 or 10 and 12, and with all possible cis and trans combinations. CLA is a newly recognized nutrient that functions to regulate energy retention and metabolism and that causes a serum lipoprotein profile considered to be less atherogenic. However, rodent models that have been frequently used for these studies are only of limited use because of distinct differences in physiology, compared with man. Additionally, possible differences in food intake between the experimental groups remained often unconsidered in those studies. Thus, it can not be excluded that the beneficial effects of CLA reported in a series of studies may be due, at least partially, to differences in nutrient and energy ingested. AIMS OF THE STUDY: This prompted us to undertake an investigation on the action of CLA by using a pig model and a feeding regimen with controlled amounts of food and antioxidants. The parameters used to assess CLA-specific action were selected hormones and metabolites involved in energy metabolism, individual lipoproteins and the appearance of CLA in fasting serum and erythrocyte membranes. Blood as an easily available biological sample was used for investigation. METHODS: For that purpose 16 adult female pigs were divided into two groups of 8 each, and were isoenergetically fed diets containing 0 (control diet) or 1% level of CLA (by weight) for 6 weeks. Plasma concentrations of total and free thyroxine (T4), total and free triiodothyronine (T3), and insulin were measured by radioimmunoassays. Plasma non-esterified fatty acids (NEFA), beta-hydroxybutyrate, circulating blood ATP and other clinical chemical variables were determined using enzymatic assays. The concentration of alpha-tocopherol was determined by high performance liquid chromatography. The lipoproteins VLDL (density <1.019 kg/L), LDL (density 1.019 - 1.063 kg/L), and HDL (density >1.063 kg/L) were isolated by step-wise ultracentrifugation. Fatty acids of the dietary oils, serum and blood cell membranes were separated and quantified by gas chromatography. RESULTS: At week 6, body weights of the pigs fed the CLA-supplemented diet were not different from that of the controls. CLA-treated pigs exhibited a 37% higher concentration of fasting serum insulin than their controls receiving no CLA (P = 0.11). Circulating free and total T4 and T3 as well as serum levels of beta-hydroxybutyrate, alpha-tocopherol, protein, glucose, urea, creatinine and circulating blood ATP remained unaffected by CLA supplementation. Serum concentrations of non-esterified fatty acids were reduced by 38% in CLA-treated pigs relative to the controls, although this difference was not significant. CLA-treated pigs tended to have lower leukocyte counts in blood than their controls (P <0.1). Erythrocyte and platelet counts, the hematocrit and hemoglobin concentration were similar between the groups. Serum of CLA-treated pigs showed a trend toward increased levels of triacylglycerols, cholesterol and phosphatidylcholine in the very low density and low density lipoproteins (LDL), without distinct changes in the high density lipoprotein fraction (HDL). The LDL cholesterol to HDL cholesterol ratio was significantly increased by CLA. When pigs were fed CLA at a dietary level of 1%, limited proportions of CLA appeared in fasting serum (1.6%) and erythrocyte membranes (1.1%). CONCLUSIONS: Under the present experimental conditions there appeared to be parallels between the effects of CLA and the reported effects of trans fatty acids in the mode of action on lipoproteins and insulin. The failure to demonstrate significant beneficial effects of CLA on the lipoprotein profile which have been observed in other studies requires further research.

3-Hydroxybutyric Acid↗

Effect of catecholamines and beta-blockers on linoleic acid desaturation activity.

The effect of catecholamines and adrenergic blocking agents on the oxidative desaturation of linoleic acid in rat liver microsomes was studied. Epinephrine (1 mg/kg/body weight) produced a significant decrease on the conversion of [1-14C]linoleic acid to gamma-linolenic acid. The effect of epinephrine was blocked by single injections of the beta blockers propranolol (10 mg/kg body weight) or dichloroisoproterenol 30 min before the hormone treatment. Isoproterenol (100 microgram/kg body weight) produced a significant decrease on the activity of the linoleyl-CoA desaturase. The effect of the catecholamines was postulated to be mediated through beta receptors by an enhancement of the intracellular levels of cyclic AMP.

Adrenergic beta-Antagonists↗

Isolation of linoleic acid as an estrogenic compound from the fruits of Vitex agnus-castus L. (chaste-berry).

A methanol extract of chaste-tree berry (Vitex agnus-castus L.) was tested for its ability to displace radiolabeled estradiol from the binding site of estrogen receptors alpha (ERalpha) and beta (ERbeta). The extract at 46 +/- 3 microg/ml displaced 50% of estradiol from ERalpha and 64 +/- 4 microg/ml from ERbeta. Treatment of the ER+ hormone-dependent T47D:A18 breast cancer cell line with the extract induced up-regulation of ERbeta mRNA. Progesterone receptor (PR) mRNA was upregulated in the Ishikawa endometrial cancer cell line. However, chaste-tree berry extract did not induce estrogen-dependent alkaline phosphatase (AP) activity in Ishikawa cells. Bioassay-guided isolation, utilizing ER binding as a monitor, resulted in the isolation of linoleic acid as one possible estrogenic component of the extract. The use of pulsed ultrafiltration liquid chromatography-mass spectrometry, which is an affinity-based screening technique, also identified linoleic acid as an ER ligand based on its selective affinity, molecular weight, and retention time. Linoleic acid also stimulated mRNA ERbeta expression in T47D:A18 cells, PR expression in Ishikawa cells, but not AP activity in Ishikawa cells. These data suggest that linoleic acid from the fruits of Vitex agnus-castus can bind to estrogen receptors and induce certain estrogen inducible genes.

Breast Neoplasms↗

Assimilation of 14C-linoleic acid by the rat fetus.

Following ingestion, by the 20-day pregnant mother, of (1-14C)linoleic acid, the retention and distribution of 14C are determined in th fetus, placenta and mother, after 1, 3 and 6 h. Comparison of the specific activities of linoleic and arachidonic acids in the maternal plasma, liver and placenta, and the fetal plasma, liver and carcass leads to the conclusion of a selective materno-fetal transfer of arachidonic acid, relative to linoleic acid, through the placenta.

Animals↗

Analysis of the contribution of the beta-oxidation auxiliary enzymes in the degradation of the dietary conjugated linoleic acid 9-cis-11-trans-octadecadienoic acid in the peroxisomes of Saccharomyces cerevisiae.

Beta-oxidation of the conjugated linoleic acid 9-cis,11-trans-octadecadienoic acid (rumenic acid) was analyzed in vivo in Saccharomyces cerevisiae by monitoring polyhydroxyalkanoate production in the peroxisome. Polyhydroxyalkanoate is synthesized by the polymerization of the beta-oxidation intermediates 3-hydroxyacyl-CoAs via a bacterial polyhydroxyalkanoate synthase targeted to the peroxisome. The amount of polyhydroxyalkanaote synthesized from the degradation of rumenic acid was found to be similar to the amount synthesized from the degradation of 10-trans,12-cis-octadecadienoic acid, oleic acid or 10-cis-heptadecenoic acid. Furthermore, the degradation of 10-cis-heptadecenoic acid was found to be unaffected by the presence of rumenic acid in the media. Efficient degradation of rumenic acid was found to be independent of the Delta(3,5),Delta(2,4)-dienoyl-CoA isomerase but instead relied on the presence of Delta(3),Delta(2)-enoyl-CoA isomerase activity. The presence of the unsaturated monomer 3-hydroxydodecenoic acid in polyhydroxyalkanoate derived from rumenic acid degradation was found to be dependent on the presence of a Delta(3),Delta(2)-enoyl-CoA isomerase activity. Together, these data indicate that rumenic acid is mainly degraded in vivo in S. cerevisiae through a pathway requiring only the participation of the auxiliary enzymes Delta(3),Delta(2)-enoyl-CoA isomerase, along with the enzyme of the core beta-oxidation cycle.

Linoleic Acids↗

Conjugated linoleic acid supplementation in humans--metabolic effects.

Supplementation with conjugated linoleic acid (CLA) induces a number of physiological effects in experimental animals, including reduced body fat content, decreased aortic lipid deposition, and improved serum lipid profile. Controlled trials on the effects of CLA in humans have hitherto been scarce. The aim of this study was to evaluate the effects of supplementation with CLA in healthy humans on anthropometric and metabolic variables and on the fatty acid composition of serum lipids and thrombocytes. Fifty-three healthy men and women, aged 23-63 yr, were randomly assigned to supplementation with CLA (4.2 g/d) or the same amount of olive oil during 12 wk in a double-blind fashion. The proportion of body fat decreased (-3.8%, P< 0.001) in the CLA-treated group, with a significant difference from the control group (P = 0.050). Body weight, body mass index, and sagittal abdominal diameter were unchanged. There were no major differences between the groups in serum lipoproteins, nonesterified fatty acids, plasma insulin, blood glucose, or plasminogen activator inhibitor 1 (PAI-1). In the CLA group the proportions of stearic, docosatetraenoic, and docosapentaenoic acids increased in serum lipids and thrombocytes, while proportions of palmitic, oleic, and dihomo-gamma-linolenic acids decreased, causing a decrease of the estimated delta-6 and delta-9 and an increase in the delta-5 desaturase activities. These results suggest that supplementation with CLA may reduce the proportion of body fat in humans and that CLA affects fatty acid metabolism. No effects on body weight, serum lipids, glucose metabolism, or PAI-1 were seen.

Adult↗