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Effect of extracellular fatty acids on lipid metabolism in cultured rabbit articular chondrocytes.

Rabbit articular chondrocytes were cultured for 8 h in the presence of various concentrations (5-500 microM) of [14C] oleic, [14C] linoleic, and [3H] arachidonic acids. The radioactive unsaturated fatty acids were incorporated into triacylglycerol (TG) and phosphatidylcholine (PC) in a concentration-dependent manner; more fatty acids were incorporated into TG than into PC, at higher concentrations of extracellular fatty acids. Among these fatty acids, arachidonic acid was incorporated into TG much more than into PC, in spite of a very low concentration of arachidonic acid in TG. After transfer of the labeled cells to maintenance medium, the radioactivity in TG declined rapidly and [3H] arachidonic acid radioactivity in PC increased continuously during the chase time periods. Palmitoyl-unsaturated species were mainly formed in PC when cultured at a concentration of 5 microM of each fatty acid. However, when cultured at 500 microM, unsaturated-unsaturated species, specific for each unsaturated fatty acid were actively formed. These findings indicate that (1) fatty acid composition of TG and PC in articular chondrocytes is influenced by the degree of fatty acid supply, (2) formation and turnover of TG plays a role in fatty acid metabolism of cells, and (3) fatty acid pairing in PC is modulated by extracellular fatty acid concentrations.

Animals↗

Acyl specificity of phospholipases A2 and C in thrombin-stimulated human platelets.

Platelets were labelled separately with six different, radioactive unsaturated fatty acids. The cells were isolated from the radioactive precursors and treated with and without 2 U/ml of thrombin. The formation of radioactive free fatty acid+oxygenated fatty acids and of radioactive radioactive phosphatidic acid+diacylglycerol was taken as a measure of the PLA(2) and PLC reactions, respectively. We found that that in intact platelets PLA(2) prefers phospholipid molecular species containing unsaturated acyls, most likely in the sn-2 position, in the priority order: 20:4>20:5>18:2 = 18:3 = 22:6>>18:1, while PLC prefers its substrates in the priority order 20:5>20:4>18:2>18:3 = 22:6>18:1.

Blood Platelets↗

Lipid biosynthesis in developing mustard seed: formation of triacylglycerols from endogenous and exogenous Fatty acids.

Cotyledons of developing mustard (Sinapis alba L.) seed have been found to synthesize lipids containing the common plant fatty acids and very long-chain monounsaturated (icosenoic, erucic, and tetracosenic) and saturated (icosanoic, docosanoic, and tetracosanoic) fatty acids from various radioactive precursors. The in vivo pattern of labeling of acyl lipids, either from fatty acids synthesized ;endogenously' from radioactive acetate or malonate, or from radioactive fatty acids added ;exogenously', indicates the involvement of the following pathways in the biosynthesis of triacylglycerols. Palmitic, stearic, and oleic acid, synthesized in the acyl carrier protein-track, are channeled to the Coenzyme A (CoA)-track and converted to triacylglycerols via the glycerol-3-phosphate pathway. Pools of stearoyl-CoA and oleoyl-CoA are elongated to very long-chain saturated and monounsaturated acyl-CoA, respectively. Most of the very long-chain saturated acyl-CoAs acylate preformed diacylglycerols. Very long-chain monounsaturated acyl-CoAs are converted to triacylglycerols, partly via phosphatidic acids and diacylglycerols, and partly by acylation of preformed diacylglycerols.

Journal Article↗

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↗

Myocardial uptake of radioactively labelled free fatty acids.

Structural variations in the carbon chain of free fatty acids influence the uptake of free fatty acids in the myocardium. To enable the use of free fatty acids in nuclear cardiology, various methods of introducing gamma-emitting isotopes have been evaluated. The uptake of various free fatty acids is described and structure-activity relationships deduced.

Animals↗

Elongation of (omega-14C)oleic acid and (omega-14C)nervonic acid.

During feeding experiments with [omega-14C]oleic acid and [omega-14c]nervonic acid to adult rats, 14C-labelled C26, C28 and C30 fatty acids were recovered from the intestinal mucosa, liver, plasma, kidney and stools. The structures of these fatty acids were determined by g.l.c., radio-g.l.c. and mass spectrometry. The Schmidt and Ginger degradation methods indicated that most of the 14C found in these extra-long fatty acids remained in the omega position. These radioactive extra-long fatty acids were found mainly in the polar lipids of rats killed 3 or 15 h after being fed on labelled oleic acid or nervonic acid. Rats killed 63 h later yielded only traces of these extra-long fatty acids. When the rats were given antibiotics or received the same radioactive fatty acids by intravenous injection, the labelled extra-long fatty acids could not be detected in any of the tissues. We conclude that they were probably synthesized by elongation of oleic acid and nervonic acid by intestinal micro-organisms (probably yeasts) and then absorbed by the intestinal mucosa.

Animals↗