Phosphoenolpyruvate carboxykinase and the synthesis of glyceride-glycerol from pyruvate in adipose tissue.
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The nature of quantity of the free lipids of Pseudomonas maltophilia have been examined. Lipid components identified as phosphatidyl ethanolamine, phosphatidyl glycerol, diphosphatidyl glycerol, phosphatidylamino acid derivatives and phosphatidic acid accounted for about one-half of the total lipids and are generally typical of gram-negative bacteria. The remaining lipids were identified as a complex family of aliphatic hydrocarbon isomers (C22--C32) and a mixture of glycolipoproteins. These components are generally atypical lipids of bacteria. The glycolipoproteins were composed of a broad range of amino acids, ketodeoxyotulonic acid, mannose, glucose, 4-aminopentose, glucosamine, phosphates, and a relatively large quantity of fatty acids, principally OH-13:0. The total esterified fatty acids consist of cyclopropane, iso- and anteiso-branched and normal saturated and unsaturated chains in the range from C14 to C20. Amide bound fatty acids consist of OH 11:0, OH-12:0, OH-13:0, and methyl-branched OH-13 in addition to methyl-branched and normal saturated and unsaturated chains.
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Weanling male rats were fed a balanced diet containing 15 p. 100 peanut oil by weight. The digestibility and fecal excretion of oil fatty acids were studied for 8 days at the beginning of the diet, which was then continued up to 4 months. At the end of the experiment, perirenal adipose tissues were removed and their triacylglycerols isolated and studied. They were fractionated into 8 fractions according to unsaturation and then analyzed by gas-liquid chromatography (GLC) for fatty acids and triacylglycerols. Each fraction was submitted to pancreatic lipolysis; the mono- and diacylglycerols formed were isolated and their fatty acids analyzed by GLC. The diacylglycerols were further fractionated according to unsaturation and their fatty acid composition analyzed. The triacylglycerol structure of the adipose tissue was determined using mathematical methods previously described. The long-chain saturated fatty acids (from stearic to lignoceric) were poorly absorbed and not incorporated into the fat tissue, whereas oleic acid, the major fatty acid of the oil (54 p. 100), was highly incorporated (57 p. 100) into triacylglycerols. As in the diet oil, unsaturated fatty acids were mainly esterified at the 2-position. Trioleoylglycerol was the major (21 p. 100) triacylglycerol, as in the oil (25 p. 100). Only 14 isomers accounted for more than 1 p. 100 and together represented more than 80 p. 100 of the total. Of the 14 isomers, the first four (10 p. 100 or more) accounted for more than one-half of oil triacylglycerols, namely 56 p. 100. The triacylglycerol structure of studied adipose tissue was very similar to that of the diet peanut oil.
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The digestibility and fecal excretion patterns of erucic (cis n-9 docosenoic) and brassidic (trans n-9 docosenoic) acids, esterified in triglycerides of different structure, were compared in the rat. Synthetic triglycerides were prepared either by mixing or interesterifying trierucin or tribrassidin with an equal amount of corn oil. Rapeseed and peanut oils were used as references. The apparent digestibility of trierucin mixed or interesterified with corn oil was similar to that of peanut oil but higher than that of rapeseed oil. The high amount of 2-monoerucin resulting from diets containing trierucin explained the improved digestibility of erucic acid as compared to that of rapeseed oil which had a very small erucic acid content on the 2-position. It also seemed that 2-monoerucin enhanced the digestibility of saturated fatty acids. Tribrassidin mixed with corn oil had a poor digestibility because pancreatic lipase hydrolysis was delayed in the intestinal lumen. The high melting point of the homogeneous triglyceride was responsible for that phenomenon. The interesterification of tribrassidin with corn oil, which drastically lowered the melting point of the mixture from 52 to 37 degrees C, improved both the hydrolysis of the triglycerides and the digestibility of the C22: 1 trans fatty acid as well as that of the other fatty acids in the diets. Nevertheless, the unabsorbed brassidic acid in every case was principally excreted in the form of a compound insoluble in organic neutral solvent. In that respect, brassidic acid behaved like shorter chain saturated fatty acids such as palmitic and stearic acids.
From L-serine 2-(linoleoylamino)-1,3-propanediol, a monoglyceride analogue, was synthesized and characterized by thin-layer and gas chromatography, infra red spectroscopy and mass spectrometry. After oral application to rats there was an accumulation, almost exclusively in the liver, of triglyceride, diglyceride, phosphatidylcholine and phosphatidylethanolamine analogues with amide-bound linoleic acid. Small amounts of triglyceride analogues could also be detected in the adipose tissue. The analogues were chromatographically isolated using in addition enzymatic and non-enzymatic hydrolysis procedures. Results disclosed that the amide-bound fatty acid of 2-(linoleoylamino)-1,3-propanediol can be absorbed and metabolized intact. Phosphoglyceride analogues are probably formed by the CDP-choline and CDP-ethanolamine pathway fromdiglyceride analogues. Phosphatidylcholine and its analogues disclosed a similar composition of the 1-O-acyl fatty acids. Apparently analogues have in vivo similar substrate properties to hydrolases and acyltransferases as their natural counterparts.
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Lifibrol (U-83860), K12.148) is a lipid-lowering drug that has the potential to accumulate in the liver and induce hepatic peroxisome proliferation. To investigate the identity and potential human relevance of persistent lifibrol-related residues in rat liver, rat and human hepatocyte primary cultures were treated with 30 microM of [14C]lifibrol. After a steady uptake for 24 hr, cellular levels of radioactivity became stable for the next 24-48 hr. A nonradioactive lifibrol chase caused an efflux of intracellular radioactivity. Cellular autoradiography revealed the association of radioactivity with small lipid drops at 6 hr exposure and with large lipid drops at 24 hr exposure. HPLC analysis of media revealed that lifibrol acyl glucuronide and a t-butylcarboxylic acid metabolite (U-94613) were the major metabolites of rat and human hepatocytes, respectively. Using an HPLC method suitable for nonpolar metabolites, the analysis of rat and human cell extracts revealed a broad band of multiple, radioactive peaks that had a similar retention and UV spectrum to a synthetic standard of lifibrol cholesterol ester. Folch extracts of liver from rats treated with [14C]lifibrol or unlabeled lifibrol and [14C]acetate had a unique radioactive TLC band that had similar HPLC retention to hepatocyte residues. The group of nonpolar peaks from the hepatocytes was purified by HPLC. Conversion of the lifibrol sec-hydroxy group to a nicotinate ester afforded particle beam-electron impact mass spectra of the cholesterol ester standard and hepatocyte residues. The derivatized rat hepatocyte residue did not contain detectable lifibrol cholesterol ester (M+.816), but did contain molecular ion clusters corresponding to a mixed triglyceride of lifibrol and two fatty acids. Lifibrol-specific product ions of molecular ion clusters centered at M+.1021, 1047, and 1073 were observed at m/z 448, 430, and 310. The major lifibrol-containing triglycerides had a fatty acid composition of C16-C20 with 0-6 unsaturations.
Cells of Streptococcus faecalis incorporate radioactive glycerol exclusively into the glycerol moieties of monoglucosyl diglyceride, diglucosyl diglyceride, phosphatidyl diglucosyl diglyceride, phosphatidylglycerol, diphosphatidylglycerol, phosphatidic acid, diglyceride, and a phospholipid tentatively identified as an amino acyl phosphatidylglycerol. Phosphatidylglycerol is one of the major radioactive lipids synthesized and contains an equal amount of radioactivity in each of its two glycerol moieties.