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Enzymatic oxydation of linoleic acid: formation of bittertasting fatty acids.

Linoleic acid was oxidized with a protein fraction from soya beans (25 degrees C; 2h), in which lipoxygenase and peroxydase activities occurred. The fatty acids formed were isolated and, after emulsification with a sugar ester, were evaluated for bitter taste. The main components of the bitter-tasting fractions was a mixture of 9.12.13-trihydroxyoctadec-10- and 9.10.13-trihydroxyoctadec-11-enoic acids. The taste threshold lies in the range 0.6-0.9 mumol/ml. Two further trihydroxy-acids and two oxodihydroxy-acids were also identified in the bitter-tasting fraction.

Fatty Acids

Differential stimulatory and inhibitory responses of human MCF-7 breast cancer cells to linoleic acid and conjugated linoleic acid in culture.

Consumption of dietary fat has been linked to the high incidence of certain cancers. However, recent research has stimulated interest in conjugated linoleic acid (CLA), a newly recognized anticarcinogenic fatty acid. Human MCF-7 breast cancer cells were incubated for 12 d in culture medium supplemented with various concentrations (1.78-7.14 x 10(-5) M) of linoleic acid (LA) or CLA. Linoleic acid initially stimulated MCF-7 cell growth with an optimal effect at concentrations of 3.57-7.14 x 10(-5) M, but was inhibitory at similar concentrations after 8 and 12 d of incubation. In contrast, CLA was inhibitory to cancer cell growth at all concentrations and times tested. Cell growth inhibition by CLA was dose- and time-dependent. Growth retardation at the prescribed LA and CLA concentrations ranged, respectively, from 4 to 33% and 54 to 100% following 8 to 12 d of treatment. At similar LA and CLA concentrations, cytostatic and cytotoxic effects of CLA were more pronounced (8-81%) than LA. These in vitro results suggest that CLA is cytotoxic to MCF-7 cells.

Breast Neoplasms

The incorporation of orally fed radioactive gamma-linolenic acid and linoleic acid into the liver and brain lipids of suckling rats.

The incorporation of radioactivity from orally administered gamma-linolenic acid-1--14C and linoleic acid--3H into the liver, plasma, and brain lipids of suckling rats was studied. Significantly more radioactivity from the former compound was incorporated into the liver and brain lipids 22 hr after dosing. The distribution of the radioactivity in the fatty acids of the liver and brain lipids was different for each isotope. Most of the -3H was still associated with linoeic acid, whereas most of the -14C was in the 20:3 and 20:4omega6 fractions. These results suggest that the desaturation of linoleic to gamma-linolenic acid in vivo is a rate-limiting step in the conversion of linoleic to arachidonic acid.

Administration, Oral

Changes in the fluorescence and absorbance of lipoxygenase-1 induced by 13-Ls-hydroperoxylinoleic acid and linoleic acid.

1. The addition of 13-Ls-hydroperoxylinoleic acid to lipoxygenase-1 (linoleate: oxygen oxidoreductase EC 1.13.11.12) from soybeans at pH 9 and 25 degrees C causes a quenching of the fluorescence of the enzyme at 328 nm when exciited at 280 nm and gives rise to an increase of the absorbance of the enzyme in the 300 nm to 450 nm region. 2. In the absence of 02, addition of linoleic acid to enzyme treated with 13-Ls-hydroperoxylinoleic acid, causes an increase of the fluorescence at 328 nm and a decrease of the absorbance in the 300 nm to 450 nm region. 3. The fluorescence changes are suggested to be directly coupled to the absorbance changes via a non-radioactive energy transfer process. 4. It is proposed that the observed fluorescence and absorbance changes are related to changes in the formal change of iron in the protein.

Hydrogen Peroxide

Importance of the spleen for the immuno-inhibitory aciton of linoleic acid in mice.

Certain immuno-inhibitory effects of a polyunsaturated fatty acid, linoleic acid (C18:2), no longer occur after splenectomy of young adult CBA mice. This observation suggest that the spleen is a major intermediary in the action of C18:2 on the lymphoreticular system. Pathways of this action remain to be elucidated. Possibilities include C18:2-stimulated suppressor cell generation in the spleen, or excess biosynthesis of immuno-inhibitory prostaglandins by splenic macrophages.

Animals

A simple method for the preparation of pure 9-D-hydroperoxide of linoleic acid and methyl linoleate based on the positional specificity of lipoxygenase in tomato fruit.

Incubation of linoleic acid with crude homogenate of tomato fruit gave a high yield (69%) of linoleic acid hydroperoxides with a ratio of 9- to 13-hydroperoxide isomers of 96:4. After chromatography of the products, as free acids or methyl esters, hydroperoxides with 9- to 13-isomeric ratios of greater than 99:1 were obtained. The major product was characterized as 9-D-hydroperoxy-octadeca-trans-10,cis-12-dienoic acid. The results demonstrate the positional specificity of lipoxygenase from tomato fruit.

Linoleic Acids

Reaction of linoleic acid hydroperoxide with thiobarbituric acid.

The linoleic acid hydroperoxide obtained by enzymatic peroxidation of linoleic acid was found to react with thiobarbituric acid to yield a red pigment. The optimum pH for the reaction was found to be 4.0. In the early stages of peroxidation of linoleic acid, thiobarbituric acid value, the amount of conjugated diene, oxygen consumption, and peroxide value were in parallel with one another. The data were compared with those on peroxidation of linolenic acid and arachidonic acid.

Chemical Phenomena

Micelle and acid-soap formation of linoleic acid and 13-L-hydroperoxylinoleic acid being substrates of lipoxygenase-1.

Surface tension measurements of linoleic acid solutions in 0.1 M sodiumborate buffer pH 10 at 23 degrees C showed that at increasing the linoleic acid concentration a sharp transition from monomers to micelles occurs at 167 micrometer. At pH 9 and 8 formation of acid-soap dimers from monomers starts at 60 micrometer and 21 micrometer respectively. The concentration range at which only monomers exist is therefore markedly reduced. For 13-L-hydroperoxylinoleic acid at pH 10 acid-soap formation still takes place, starting at approx. 220 micrometer. The total lipid concentration at which acid-soap or micelle formation starts in mixtures of linoleic acid and 13-L-hydroperoxylinoleic acid has been determined in relation to the molar ratio of both acids.

Kinetics

[Qualitative and quantitative studies on the metabolism of fatty acids, in particular on that of linoleic acid in calves].

24 calves of the breed "Deutsche Schwarzbunte Rasse" were used for qualitative and quantitative investigations on the metabolism of fatty acids, particularly that of linoleic acid. The trial comprised 6 calves in 4 groups. The experimental rations consisted of powdered skim milk, oat meal, milk sugar, coco fat and varying proportions of linoleic acid (1.0%, 1.5%, 2.5%). The control group received the basal ration without supplements of linoleic acid. After a short period of adjustment the metabolic trial was started with 9 periods each lasting 7 days. The composition of fatty acids in the faecal lipids was analysed by gas chromatography. The studies led to the conclusion that the fatty acids contained in the faecal fat were not immediately derived from the dietary fat. The results of the trial also showed that the major proportion of all the fatty acids present in the faecal lipids consisted of only 4 fatty acids, viz. oleic acid, palmitic acid, lauric acid and myristic acid. Changes observed for the content of linoleic acid in faecal lipids were negligible so that the assumption may be made that all of the linoleic acid added was completely digested.

Animal Feed

Enterotoxigenic intestinal bacteria in tropical sprue. IV. Effect of linoleic acid on growth interrelationships of Lactobacillus acidophilus and Klebsiella pneumoniae.

The factors responsible for colonization of the small intestine by enterotoxigenic coliform bacteria in Puerto Ricans with tropical sprue are unknown, but epidemiological observations have suggested that they may be related to an increased dietary intake of long-chain unsaturated fatty acids, particularly linoleic acid, which is known to exert an inhibitory effect on the growth of gram-positive organisms that normally comprise the flora of the small intestine. We have examined, by using a glucose-limited continuous-culture system, what effect this fatty acid exerts on the growth relationships of enteric gram-positive and coliform bacteria. In this system, colonization by an invading strain of Klebsiella pneumoniae was prevented by the presence of an established culture of Lactobacillus acidophilus, principally by virtue of a lowered pH of the medium that was incompatible with Klebsiella growth. However, when the population density of L. acidophilus was reduced by the presence of a sufficient concentration of linoleic acid, the invading K. pneumoniae successfully colonized the system and, once established, suppressed the growth of L. acidophilus. These observations indicate that, under the conditions of our chemostat, gram-positive enteric bacteria suppress coliform growth and that this effect is reversible by the presence of linoleic acid. It remains to be established, however, what pertinence these in vitro observations have to conditions within the intestinal tract of persons living in the tropics.

Enterotoxins

Substitution reactions of linoleic acid hydroperoxide isomerase.

Linoleic acid hydroperoxide isomerase was extracted from corn germ and partially purified by differential centrifugation. This enzyme catalyzed the isomerization of linoleic acid hydroperoxide.(see article) Isomerase also catalyzed the substitution of various reagents at the carbon bearing the hydroperoxide group. These fatty acid products had the following functional groupings: (see article) where X is either oleoyloxy, ethylthio, or methoxy resulting from the presence of oleic acid, ethanethiol, or methanol, respectively. A crude wheat germ extract containing both lipoxygenase and isomerase enzymes reacted with linoleic acid to yield alpha-ketols, gamma-ketols, and a substitution product, the linoleoyloxy ester of alpha-ketol. Characterization of these products from wheat germ enzymes showed that the substitution reaction was not unique to corn germ. Because anions of the reagents tested are typical nucleophiles, the substitution reactions may proceed by a nucleophilic mechanism as mediated by the isomerase enzyme.

Isomerases

Comparative effect of glucagon, dibutyryl cyclic AMP, and epinephrine on the desaturation and elongation of linoleic acid by rat liver microsomes.

The effect of glucagon, dibutyryl cyclic adenosine 3',5'-monophosphate, and epinephrine on the biosynthesis of polyunsaturated fatty acids of the linoleic acid family was studied. The incubations were performed with rat liver microsomes and labeled linoleic acid under desaturating and elongating conditions. Under desaturating conditions linoleic acid was converted to gamma-linolenic acid, whereas under elongating conditions it was converted to 20:2omega6. Glucagon, dibutyryl cyclic AMP, and epinephrine decreased the oxidative desaturation of linoleic acid to gamma-linolenic acid while the elongating reaction was not modified in the experimental conditions tested. Consequently, the results support the hypothesis that the oxidative desaturation of linoleic acid to gamma-linolenic acid is the main controllable step in the biosynthesis of polyunsaturated fatty acids of the linoleic acid family in the microsomes.

Animals

Biohydrogenation and availability of linoleic acid in lactating cows.

Linoleic acid biohydrogenation, absorption and availability for maintenance and milk production in dairy cows fed high grain (60--85% of dry matter) diets were quantitatively estimated by isotope dilution, using two methods of dosing. [1-14C]Linoleic acid-labeled chylomicra and very low density lipoproteins (VLDL) were obtained from lymph of a calf fed [1-14C]linoleic acid and fitted with a thoracic duct-venous shunt. Labeled chylomicra were injected intravenously into two cows: a Jersey (trial 1), and a Holstein (trial 2). Labeled VLDL was injected intravenously into a Holstein cow (trial 3). In trials 4 and 5 the [1-14C]linoleic acid was placed into the omasal canal of two rumen-fistulated Holstein cows. Linoleic acid biohydrogenation (%), absorption (g/day), and availability above requirements for milk production (mg/kg body wt 3/4) were: 68.1 +/- 2.28, 52.1 +/- 2.92, and 244 +/- 19.4 (mean +/- SE), respectively. The biohydrogenation data indicate that both methods of dosing the cows were equally dependable. The estimates of linoleic acid biohydrogenation are consistent with limited data previously reported, indicating that the isotope dilution technique used is a reliable method to estimate linoleic acid absorption in lactating cows. Linoleic acid available to the lactating cow above milk production requirements was more than double the requirement of weanling female rats, when compared on the basis of metabolis body size.

Animal Feed

Tumor promoting phorbol diesters stimulate release of radioactivity from [3H]-arachidonic acid labeled- but not [14C]linoleic acid labeled-cells. Indomethacin inhibits the stimulated release from [3H] arachidonate labeled cells.

The tumor promoting phorbol diester, 12-O-tetradecanoyl-phorbol-13-acetate, stimulates MDCK cells to deacylate cellular phospholipids and to produce prostaglandins when measured as the release of arachidonic acid and its metabolites into the culture fluid. Indomethacin, at levels of 2.8 x 10(-8) to 2.8 x 10(-6) M, inhibits the release of radioactivity from [3H]arachidonate labeled cells stimulated by 12-O-tetradecanoyl-phorbol-13-acetate treatment in a concentration dependent manner. At these concentrations, the conversion of released [3H]arachidonic acid into prostaglandins E2 and F2alpha and the production of PGE2 measured serologically also is suppressed in a concentration dependent manner. Indomethacin, at these levels, has no effect on the acylation of [3H]arachidonic acid into cellular lipids. The tumor promoting phorbol diester does not stimulate the release of radioactive materials from MDCK cells labeled with [14C]linoleic acid, although prostaglandin production by these cells is stimulated.

Animals