Absorption and metabolism of 1-14C-methyl linoleate hydroperoxide

The absorption and metabolism of 1-¹⁴C-methyl linoleate hydroperoxide by rats was investigated. After intubation with 2 mg of peroxide, peak¹⁴CO₂ production occurred at 90 min and 25% of the dose was expired in 24 hr. Fortyfive per cent stil remained in the gastrointestinal tract after 24 hr, most of it bound to the stomach epithelium in the form of intact peroxide. Lymph was collected from the thoracic duct 2 hr after intubation and examined for labeled metabolites. Seven per cent of the radioactivity in the lymph was present in a free 1-¹⁴C-hydroxy fatty acid and 31% in its methyl ester. Fifty-seven per cent occurred in lymph triglycerides where it was equally distributed between a 1-¹⁴C-trienoic fatty acid and an unidentified 1-¹⁴C-oxy acid. The radioactivity in liver lipids was associated mainly with randomly labeled normal fatty acids. No¹⁴C-hydroxy acids were detected in liver lipids and no evidence was obtained for the absorption of unchanged peroxide. The hydroxy and trienoic acids appear to be formed during absorption by a reduction-dehydration reaction sequence.     

Absorption and metabolism of 1-14C-hydroxy octadecadienoate in the rat

The metabolism of 1-¹⁴C-9(13)-hydroxy octadecadienoic acid methyl ester (1-¹⁴C-HAME) by the rat was investigated in vivo and in liver slices. A 1.5 mg dose of 1-¹⁴C-HAME administered by stomach tube was efficiently hydrolyzed and absorbed from the intestinal tract. In comparison with 1-¹⁴C-methyl linoleate (1-¹⁴C-ML), 1-¹⁴C-HMAE was more extensively oxidized to¹⁴CO₂ in vivo and in vitro. After 1-¹⁴C-HAME administration as much as 50% of the radioactivity in the adipose tissue triglycerides was associated with¹⁴C-hydroxy fatty acids. The remaining activity was present in randomly labeled normal fatty acids. No evidence was obtained for the incorporation of¹⁴C-hydroxy acids into liver lipids; most of the radioactivity from 1-¹⁴C-HAME in this organ was recovered in saturated and monoenoic fatty acids. About 10% of the radioactivity 24 hr after 1-¹⁴C-HAME administration was associated with triglyceride trienoic acids, indicating that at least a portion of this acid was dehydrated in the liver. An unidentified polar acid was detected in the urine of the 1-¹⁴C-HAME-treated animals.     

Tissue culture of cocoa beans (Theobroma cacao L.): Incorporation of acetate and laurate into lipids of cultured cells

Suspension cultures of cocoa bean tissue readily incorporated exogenous acetate into lipids. The distribution of radioactivity from acetate in individual lipid classes after 48 hr was 20, 5, 1, 15, 25, and 35% in triglycerides, diglycerides, free fatty acids, sterol esters, sterols and polar lipids, respectively. The labeled acetate was rapidly incorporated into various fatty acids within 2 hr. The [1-¹⁴C] saturated fatty acids declined slightly after 4 hr, whereas [1-¹⁴C] oleate declined significantly after 2 hr. There was a concomitant increase in [1-¹⁴C] linoleate. The radioactivity associated with linolenate was relatively high up to 4 hr, declined by 24 hr, and then increased again. The kinetics of fatty acid labeling suggested that biosynthesis of linolenic acid in cocoa bean suspension culture may occur via the desaturation of linoleic acid and the chain elongation of dodecatrienoic acid. The patterns of fatty acid radiolabeling following incubation of cells with [1-¹⁴C] laurate was consistent with this mechanism.     

Dietary fatty acids: Their metabolic fate and influence on fatty acid biosynthesis

Adult rats fed a low-fat diet or diets containing 15% of either tripalmitin, triolein or trilinolein were injected intraperitoneally with H³-labeled acetate. Those which received fat were also given by mouth, simultaneously with acetate, the 1-C¹⁴-labeled sodium salt of the respective dietary fatty acid. The fate of the tagged material was followed by time-spaced biopsies of subcutaneous adipose tissue and by collection of the expired C¹⁴O₂. After 72 hr, 51, 64, and 52% of the dietary palmitate, oleate, and linoleate, respectively, were catabolized, as indicated by the corresponding percentages of the label having been excreted as C¹⁴O₂. Dietary linoleate was relatively less incorporated into body triglycerides than palmitate and oleate. Animals ingesting diets of 15% triolein had only about one-half the amount of phospholipids in their tissues as had the other groups. The distribution of both the C¹⁴ and H³ labels in the tissue triglycerides showed that all diets containing fats decreased fatty acid synthesis but did not inhibit conversion of palmitate to oleate. Conversions of oleate or linoleate appeared to be through acetate. As a result of these factors, the fatty acid composition of the tissue triglycerides after 3 months’ ingestion of tripalmitin was essentially the same as that of the low-fat group, whereas the ingestion of triolein produced triglycerides with a very high content of oleic acid. Trilinolein ingestion produced effects similar to triolein but to a less pronounced degree. Both the respiratory C¹⁴O₂ and the C¹⁴- and H³-labeled fatty acids in subcutaneous adipose tissue exhibited a second rise in specific activity 12 to 24 hours after the administration of the label. Presented at the AOCS meeting, Chicago, 1964.     

The metabolism of 1-14C-palmitic acid in the testis of the rat

The metabolism of 1-¹⁴C-palmitic acid was studied in rat testis at various intervals after intratesticular injection and after a 2 hr incubation period. Significant catabolism occurred as evidenced by production of¹⁴CO₂ in both in vivo and in vitro experiments. Of the¹⁴C activity retained in the injected testis, less than 15% remained as free fatty acid after 2 hr and less than 5% by the end of 2 weeks. Activity of¹⁴C appeared in phosphatides faster than in the triglyceride fraction, and with time (1–2 weeks) the activity in phosphatides decreased relative to that in triglycerides. In phosphatides of these experiments ca. 80% of the¹⁴C was in the palmitic acid fraction and the balance predominantly (98%) in palmitate with the balance in stearate and oleate. By the first and second weeks almost half of the¹⁴C-palmitate present in the testes had been newly synthesized from¹⁴C-acetyl CoA resulting from oxidation of the administered 1-¹⁴C-palmitate. In the incubated samples the only fatty acid with¹⁴C activity in any lipid fraction was palmitate. In these experiments ca 90% of the¹⁴C was in the free fatty acid fraction, 7% in the phosphatides and 2% in the triglycerides.     

Lipid metabolism in plasma,liver, and adipose tissue of rats with experimental chronic nephrotic syndrome

Plasma, liver, and adipose tissue lipid composition and synthesis from [1-¹⁴C] acetate were studied three months following induction of nephrotic syndrome in rats by injection of antiglomerular basement membrane protein. Plasma triglyceride concentrations and specific radioactivities were elevated, and the triglycerides contained increased proportions of oleic acid. Plasma cholesterol and phospholipid concentrations were also increased, but free fatty acid levels were not. Liver triglyceride concentrations were decreased and incorporation of [1-¹⁴] acetate into liver triglycerides was also depressed below that of normal controls. Nephrotic rat liver triglycerides contained a higher proportion of oleic acid and lower arachidonic acid than did controls. Incorporation of [1-¹⁴C] acetate into adipose tissue lipids of the nephrotic rats was increased, and the proportion of palmitic acid was decreased. In the chronic nephrotic rat, the major source of the increased plasma triglycerides may be fatty acids mobilized from adipose tissue stores.     

Tissue culture of cocoa bean (Theobroma cacao L.): Incorporation of fatty acids into lipids of cultured cells

Suspension cell cultures of cocoa bean rapidly incorporated palmitic, stearic, oleic and linoleic acids into cellular lipids. Thus, 75 and 20% of [1-¹⁴C] palmitic acid was incorporated into polar lipids and triglycerides, respectively, after 48 hr. When [1-¹⁴C] oleic and [1-¹⁴C] linoleic acid were added separately, polar lipids consistently contained most of the radioactive fatty acids. Ca. 60% of the stearic acid accumulated as unesterified fatty acid in the cells. Palmitic and stearic acid were not desaturated, but oleic acid and linoleic acid were further desaturated. The kinetics of conversion of oleic acid and linoleic acid suggested a sequential desaturation pathway of 18∶1→18∶2→18∶3 in cocoa bean cell suspensions.     

Regulation of the metabolism of linoleic acid to arachidonic acid in rat hepatocytes

When 5×10⁶ hepatocytes were incubated for 40 min with from 0.15 to 0.60 mM [1-¹⁴C]linoleic acid, [1-¹⁴C]6,9,12-octadecatrienoic acid, or [1-¹⁴C]8,11,14-eicosatrienoic acid, there was a concentration-dependent acylation of radioactive metabolites into both triglycerides and phospholipids. When the concentration of either [1-¹⁴C]linoleic acid or [1-¹⁴C]8,11,14-eicosatrienoic acid exceeded 0.3 mM, there was no further increase in the metabolism of either fatty acid to other (n−6) metabolites. When the concentration of [1-¹⁴C]6,9,12-octadecatrienoic acid exceeded 0.15 mM, there was an apparent substrate-induced inhibition in its metabolism to 8,11,14-eicosatrienoic acid. With all three substrates (0.3 mM), there was time-dependent metabolism to other (n−6) acids. Cells then were incubated simultaneously with 0.3 mM [1-¹⁴C]linoleic acid along with 0.15 to 0.45 mM 6,9,12-octadecatrienoic acid or 8,11,14-eicosatrienoic acid. These exogenous nonradioactive (n−6) acids suppressed but did not abolish the conversion of [1-¹⁴C]linoleate to radioactive arachidonate. These findings suggest that some linoleate is converted to arachidonate without intracellular mixing of 6,8,12-octadecatrienoic or 8,11,14-eicosatrienoic acids. This hypothesis is supported by the finding that exogenous linoleate did not markedly affect the metabolism of [1-¹⁴C]6,9,12-octadecatrienoic or [1-¹⁴C]8,11,14-eicosatrienoic acid by microsomal chain elongating or desaturating enzymes.     

Metabolism of14C-labelled oleic acid,erucic acid and nervonic acid in rats

1-¹⁴C-Oleic acid, 2-¹⁴C-erucic acid and 2-¹⁴C-nervonic acid were administered to rats by tail-vein and the distribution of radioactivity in liver lipids was determined at intervals from 15 min to 6 hr after injection. High levels of activity were found after short time intervals which were mainly associated with triglycerides in the case of oleic acid and with free fatty acids in the case of erucic acid and nervonic acid. The activity in these lipids decreased with time and was later exceeded by that in more polar lipids. In rats given erucic acid or nervonic acid, sphingolipids were more highly labelled than glycerophosphatides. Nervonic acid showed little tendency to form a complex with serum albumin and erucic acid complexed less readily than palmitic acid. Presented at the AOCS Meeting in Houston, April 1965.     

The metabolism of linoleic and arachidonic acids in rat testis

Linoleic and arachidonic acids, labeled with¹⁴C and injected intratesticularly, were used to study with time the interconversion of polyunsaturated fatty acids in rat testis and their incorporation into the major lipid classes. With both substrates¹⁴C activity was readily incorporated into longer chain, more highly unsaturated fatty acids. After the injection of 1-¹⁴C-linoleic acid the major portion of the¹⁴C was found in palmitic, linoleic, 8,11,14-eicosatrienoic, 5,8,11,14-eicosatetraenoic, 7,10,13,16-docosatetraenoic and 4,7,10,13,16-docosapentaenoic acids. Hydrogenation of the total fatty acids isolated from rat testes after intratesticular injection of 1-¹⁴C-linoleate revealed that the polyenoic acids hydrogenating to lignoceric acid (previously characterized as 9,12,15,18-tetracosatetraenoate and 6,9,12,15,18-tetracosapentaenoate) had a relatively high specific activity. After the injection of 1-¹⁴C-arachidonate significant¹⁴C activity was found in palmitate, 7,10,13,16-docosatetraenoate, 4,7,10,13,16-docosapentaenoate, 9,12,15,18-tetracosatetraenoate and 6,9,12,15,18-tetracosapentaenoate. The biosynthesis of the ω⁶ polyunsaturated fatty acids in rat testis is discussed in relation to these data. Investigation of the distribution of label in the complex lipid fractions demonstrated the majority of the¹⁴C activity to be present in phosphatides and triglycerides after injection of either of these¹⁴C substrates with only small quantities being present as nonesterified acids. At the time periods studied the polyenoic acids of triglycerides had a higher specific activity than the corresponding acids of phosphatides with the exception of linoleate. Presented in part at the Meeting of the American Institute of Nutrition, Atlantic City, April 1968 and at the AOCS Meeting in New York, April 1969. These data were taken from a thesis submitted by R. B. Bridges in partial fulfillment of the requirements for the Ph.D. degree, Vanderbilt University.     

Placental transport oftrans fatty acids in the rat

Placental transport of 9-trans [1-¹⁴C] octadecenoic (elaidic) and 9-trans,12-trans [1-¹⁴C] octadecadienoic (linoelaidic) acids was demonstrated in rats. On the 18th day of gestation, a¹⁴C-labeled albumin complex of elaidic or linoelaidic acid was injected into the jugular vein of pregnant rats. For comparison, 9-cis [1-¹⁴C] octadecenoic (oleic) or 9-cis,12-cis [1-¹⁴C] octadecadienoic (linoleic) acid also was injected into the maternal circulation of rats. All animals were sacrificed 1 hr following injection. Lipid composition and distribution of label were determined in maternal plasma, placental and fetal tissues. Differences in specific activities of plasma, placental and fetal total lipids indicated a decreasing concentration gradient for bothcis andtrans isomers of octadecenoic and octadecadienoic acids. Distribution of radioactivity in various lipid components was determined by thin layer chromatography. Irrespective of the label, the highest percentage of total radioactivity was carried by triglycerides (TG) in maternal plasma (∼60–80%), and was incorporated mainly in phospholipids (PL) of fetal tissues (∼50–60%). A nearly equal distribution of the label was found between PL and TG of placental lipids (∼40%). Radioactivity of fatty acid methyl esters (FAME) determined by radiogas liquid chromatography indicated that after injection of linoelaidate, radioactivity of maternal plasma, placental and fetal tissue FAME was associated only witht,t-18∶2. Following injection of elaidate, all the radioactivity in placental FAME was associated witht-18∶1; however, in fetal tissues, the label was distributed between 16∶0 andt-18∶1. These findings suggest that, in contrast to linoelaidic acid, rat fetal tissues can metabolize elaidic acid via β oxidation to form acetyl CoA and palmitic acid.     

Relative incorporation of acetate and glucose into glycerides ofGlomerella cingulata

Cultures of the fungusGlomerella cingulata were pulse-labeled for 30 min with 1-¹⁴C-acetate at ages from 2 to 12 days old. The greatest incorporation into triglycerides and phosphoglycerides occurred in the youngest cultures, but more of the radioactive acetate was diverted to the phosphoglycerides than to the triglycerides in these cultures. In another series of experiments the incorporation of 1-¹⁴C-acetate or 2-¹⁴C-glucose into the triglycerides and phosphoglycerides of the fungus was investigated at 15-minute intervals for 1 hr. Hydrolysis of the two classes of glycerides revealed some labeling of glycerol from acetate; but, not surprisingly, acetate was incorporated into the fatty acid moieties to a greater extent than into the glycerol moieties, and there was relatively greater incorporation of 2-¹⁴C-glucose into glycerol than into fatty acids. Some relationships of these results to the growth and development ofG. cingulata are suggested, and implications relative to control mechanisms are pointed out.     

Selectivity in incorporation,utilization and retention of oleic and linoleic acids by human skin fibroblasts

Fetal human fibroblasts were grown in culture medium containing 10% fetal bovine serum supplemented with [1-¹⁴C] linoleate or [1-¹⁴C] oleate. At all concentrations of exogenous fatty acids, the incorporation of oleate was greater than that of linoleate. With increased medium fatty acid concentrations, linoleate in triacylglycerol (TAG) could be increased from 13 to 75% of the total incorporated; at each concentration, relatively more linoleate than oleate was in TAG. When the cells were exposed to exogenous oleate/linoleate mixtures, the composition of the mixture determined the extent of incorporation of both fatty acids. When the mixture was primarily linoleate, scarce oleate was used preferentially for phospholipids (PL); no such specificity for scarce linoleate was observed. Addition of exogenous fatty acids resulted in a shift of previously incorporated¹⁴C fatty acids from phospholipid into TAG; retention of oleate in PL was greater than that of linoleate. Incorporation of oleate into phospholipids was also higher than that of linoleate from exogenous fatty acid mixtures which were 80% saturated. It is suggested that normal human fibroblasts have adapted to the low levels of exogenous polyunsaturated fatty acids in culture media by increased use of oleate in phospholipid. Even when the cells aresupplemented with linoleate, the preferential use of oleate in phospholipid groups is retained. Presented in part at the ASBC Meeting, Dallas, april 1979.     

Incorporation of radioactive polyunsaturated fatty acids into liver and brain of developing rat

The incorporation of radioactivity from orally administered linoleic acid-1-¹⁴C, linolenic acid-1-¹⁴C, arachidonic acid-³Hg, and docosahexaenoic acid-¹⁴C into the liver and brain lipids of suckling rats was studied. In both tissues, 22 hr after dosing, 2 distinct levels of incorporation were observed: a low uptake (from 18∶2-1-¹⁴C and 18∶3-1-¹⁴C) and a high uptake (from 20∶4-³H₈ and 22∶6-¹⁴C). In adult rats, the incorporation of radioactivity into brain lipids from 18∶2-1-¹⁴C and 20∶4-³H was considerably lower than the incorporation into the brains of the young rats. In the livers of the suckling rats, the activity from the 18 carbon acids was associated mostly with the triglyceride fraction, whereas the activity from the 20∶4-³H₈ and 22∶6-¹⁴C was concentrated in the phospholipid fraction. In the brain lipids, the activity from the different fatty acids was associated predominantly with the phospholipids. In the liver and brain phospholipid fatty acids, some of the activity in the 18∶2-1-¹⁴C and 18∶3-1-¹⁴C experiments was associated with 20 and 22 carbon polyunsaturated fatty acids; however, radioactivity from orally administered 20∶4-³H₈ and 22∶6-¹⁴C was incorporated intact into the tissue phospholipid to a much greater extent compared with the incorporation of radioactivity into 20∶4 and 22∶6 in the experiments where 18∶2-1-¹⁴C and 18∶3-1-¹⁴C, respectively, were administered. Possible reasons for these differences are discussed. Rat milk contains a wide spectrum of polyunsaturated fatty acids, including linoleate, linolenate, arachidonate, and docosahexaenoate. During the suckling period in the rat, there is a rapid deposition of 20∶4 and 22∶6 in the brain. The results of the present experiments suggested that dietary 20∶4 and 22∶6 were important sources of brain 20∶4 and 22∶6 in the developing rat.     

Transfer of arachidonate from phosphatidylcholine to phosphatidylethanolamine and triacylglycerol in guinea pig alveolar macrophages

Guinea pig alveolar macrophages were labeled by incubation with either arachidonate or linoleate. Arachidonate labeled phosphatidylcholine (PC), phosphatidylethanolamine (PE) and triglycerides (TG) equally well, with each lipid containing about 30% of total cellular radioactivity. In comparison to arachidonate, linoleate was recovered significantly less in PE (7%) and more in TG (47%). To investigate whether redistributions of acyl chains among lipid classes took place, the macrophages were incubated with 1-acyl-2-[1-¹⁴C]arachidonoyl PC or 1-acyl-2-[1-¹⁴C]linoleoyl PC. After harvesting, the cells incubated with 1-acyl-2-[1-¹⁴C]linoleoyl PC contained 86% of the recovered cellular radioactivity in PC, with only small amounts of label being transferred to PE and TG (3 and 6%, respectively). More extensive redistributions were observed with arachidonate-labeled PC. In this case, only 60% of cellular radioactivity was still associated with PC, while 22 and 12%, respectively, had been transferred to PE and TG. Arachidonate transfer from PC to PE was unaffected by an excess of free arachidonate which inhibited this transfer to TG for over 90%, indicating that different mechanisms or arachidonoyl CoA pools were involved in the transfer of arachidonate from PC to PE and TG. Cells prelabeled with 1-acyl-2-[1-¹⁴C]arachidonoyl PC released¹⁴C-label into the medium upon further incubation. This release was slightly stimulated by zymosan and threefold higher in the presence of the Ca²⁺-ionophore A₂₃₁₈₇. Labeling of macrophages with intact phospholipid molecules appears to be a suitable method for studying acyl chain redistribution and release reactions.     

Metabolism of n−3 polyunsaturated fatty acids by the isolated perfused rat liver

The hepatic metabolism of oleic acid and n−3 fatty acids (eicosapentaenoic acid, EPA and docosahexaenoic acid, DHA), and secretion of very low density lipoprotein (VLDL) were studied in isolated perfused rat livers from normal chow fed male rats. The basal perfusion medium contained 30% bovine erythrocytes, 6% bovine serum albumin (BSA), and 100 mg/dL glucose, in Krebs-Henseleit bicarbonate buffer (pH 7.4) which was recycled through the liver for 2 hr. Individual fatty acids (EPA, DHA or oleic acid), as complexes with 6% BSA, or albumin alone, were infused at a rate of 70 μmol/hr. When any of these fatty acids was infused at this rate, the ambient concentration in the medium was maintained at 0.3–0.4 μmol/mL, indicative of similar hepatic rates of uptake for each fatty acid (i.e., approximately 6 μmol/g liver/hr). When fatty acid was not infused, the ambient free fatty acid level was 0.16 μmol/mL. The concentrations of infused free fatty acids increased appropriately in the perfusion medium; however, with infusion of EPA, DHA, or oleate, the concentrations of perfusate palmitate and linoleate were the same as when fatty acid was not infused. Additionally, the perfusate concentration of oleate in the free fatty acid fraction was not affected by infusion of EPA and DHA. These data indicate a constant outflow of endogenous fatty acid unaffected by the presence of the exogenously supplied fatty acid. The net secretion rate of VLDL lipids and protein was stimulated by infusion of oleate, whereas when EPA was infused, secretion rates were lower and similar [except for VLDL cholesterol (C), which was greater] to those occuring when fatty acid was not provided. DHA stimulated the secretion of VLDL triacylglycerol (TG), phospholipid (PL) and C to a similar rate, as did oleate, but secretion of VLDL cholesteryl ester (CE) and protein was lower and similar to that with EPA. VLDL and hepatic TG and CE were enriched with the infused fatty acids, compared to experiments without fatty acids, as determined by gas chromatography. Enrichment of PL, however, was significant only in liver upon infusion of EPA. The formation of¹⁴CO₂ and perchloric acid soluble products from [1-¹⁴C]EPA, considered separately, did not differ statistically from that obtained with [1-¹⁴C]oleate, although the mean values were higher with [1-¹⁴C]EPA. However, the sum of oxidation products derived from EPA was significantly greater than that from oleate. Incorporation of [1-¹⁴C]EPA into TG and CE, but not into PL, was lower as compared to that from [1-¹⁴C]oleate. These lower rates of incorporation of [1-¹⁴C]EPA into VLDL lipids therefore paralleled the mass fatty acid enrichment-patterns. It may be concluded that EPA is used to a similar extent as oleate for synthesis of PL, but is a poorer substrate for synthesis of TG. The reduced output of newly synthesized (radioactive) PL reflected the lower hepatic output of VLDL. Since hepatic uptake of EPA, DHA or oleate was identical, utilization of EPA for TG synthesis was less than that of oleate or DHA. Further-more, utilization of endogenous fatty acids for TG synthesis and secretion of the VLDL was reduced in the presence of EPA. The decreased TG synthesis resulted in reduced formation of VLDL for transport of TG from the liver. These effects taken together with an apparently increased oxidation of EPA provide substantial evidence for a decrease in formation of VLDL and transport of TG, PL, C and CE into the circulation in response to EPA. DHA, however, appears to be an adequate substrate for TG synthesis and stimulates VLDL secretion. The reduced transport of CE may reflect lower selectivity of DHA by acyl-CoA; cholesterol acyltransferase for CE formation.     

Distribution of14C after oral administration of (U-14C)labeled methyl linoleate hydroperoxides and their secondary oxidation products in rats

To study the toxicity of low molecular weight (LMW) compounds formed during the autoxidation of oils,¹⁴C-labeled primary monomeric compounds (methyl linoleate hydroperoxides) and secondary oxidation products, i.e., polymer and LMW compounds prepared from autoxidized methyl [U-¹⁴C]linoleate hydroperoxides (MLHPO) were orally administered to rats, and their radioactive distributions in tissues and organs were compared. The polymeric fraction consisted mainly of dimers of MLHPO. For the LMW fraction, 4-hydroxy-2-nonenal, 8-hydroxy methyl octanoate and 10-formyl methyl-9-decenoate were identified as major constituents by gas chromatography-mass spectrometry (GC-MS) after chemical reduction and derivatization. When LMW compounds were administered to rats,¹⁴CO₂ expiration and the excreted radioactivity in urine in 12 hr were significantly higher than those from polymer or MLHPO administration. Maximum¹⁴CO₂ expiration appeared 2–4 hr after the dose of LMW compounds. Radioactivity of the upper part of small intestines six hr after the dose of LMW compounds was higher than the values from administered polymer or MLHPO. The remaining radioactivity in the digestive contents and feces 12 hr after administration of LMW compounds was much lower than the values observed from administered polymer or MLHPO. Among internal organs, the liver contained the highest concentration of radioactivities from polymer, MLHPO and LMW fractions, and an especially higher level of radioactivity was found in liver six hr after the administration of LMW compounds. Six hours after the dose of LMW compounds, a relatively higher level of radioactivity also was detected in kidney, brain, heart and lung. These results show that the LMW compounds from MLHPO autoxidation are more easily absorbed in rat tissues than polymer and MLHPO.     

Positional specificity oftrans fatty acids in fetal lecithin

Differences in the positional incorporation of 9-trans[1-¹⁴C] octadecenoic (elaidic) and 9-trans,12-trans[1-¹⁴C] octadecadienoic (linoelaidic) acids in fetal lecithin of rats were demonstrated. On the 20th day of gestation, a¹⁴C-labeled albumin complex of elaidic or linoelaidic acid was injected into the jugular vein of pregnant rats. For comparative purposes, 9-cis[1-¹⁴C] octadecenoic (oleic) or 9-cis,12-cis[1-¹⁴C] octadecadienoic (linoleic acid) was injected into the maternal circulation of rats. Animals were killed 6 hr later. Distribution of label in total lipids and phospholipids (PL) of fetal tissue was measured by TLC. Irrespective of the label, the highest percentage of total radioactivity was associated with PL-59 to 67%. Within PL, the major portion of radioactivity was found in choline phosphoglycerides (CPG)-53 to 67%, and in ethanolamine phosphoglycerides (EPG)-18 to 33%. While linoelaidic acid was predominantly esterified in the 2-position of CPG, elaidic acid was nearly equally distributed between positions 1 and 2 of lecithin. Distribution of radioactivity within fatty acid methyl esters (FAME) of CPG measured by radio-GLC suggested that oleic and possibly linoleic acids may be converted to nervonic and arachidonic acid, respectively, in the rat by the 20th day of gestation. Following injection of elaidate, radioactivity of FAME was distributed between palmitate and elaidic acid indicating that rat fetal tissue may metabolize elaidic acid via β-oxidation. In contrast, following injection of linoelaidate, radioactivity of FAME was primarily associated withtt-18∶2, suggesting little biotransformation to other fatty acids by fetal tissues.     

Stimulation of hepatic lipogenesis by eicosa-5,8,11,14-tetraynoic acid in mice fed a high linoleate diet

Liver slices, from mice fasted for one day and then refed for three days either a 15% corn oil diet or a 15% corn oil diet containing eicosa-5,8,11,14-tetraynoic acid (TYA), were incubated with [1-¹⁴C] acetate or [³H]H₂O to determine lipogenic capacity. Dietary TYA produced a twofold stimulation in fatty acid and cholesterol synthesis. TYA also caused an increase in the relative proportion of linoleate (C_18∶2) and a decrease in that of arachidonate (C_20∶4) in liver. Thus, (a) despite high levels of C_18∶2, hepatic lipogenesis can be increased, and (b) even short term feeding of TYA can alter the hepatic fatty acid composition presumably by inhibition of arachidonate synthesis from linoleate.     

Formation of diglycerides of long turnover time from labeled acetate and glucose in rat tissues

Rat adipose tissue pieces were incubated with acetate-2-¹⁴C and glucose-¹⁴C(U), respectively, and liver slices with acetate-2-¹⁴C. The labeled tissues were then reincubated in inactive medium, and the changes of radioactivity in the different lipid classes were determined. In all three experiments a significant amount of radioactivity was incorporated in the diglycerides. During 1 hr of reincubation in inactive medium the radioactivity of diglycerides decreased from 35 to 26% of the total lipid activity in the adipose tissue labeled with acetate. In the adipose tissue labeled with glucose radioactivity fell from 25 to 19%. In liver slices 11% of the labeled acetate was incorporated in the diglycerides, and during the 2 hr of reincubation this value fell to its half. The radioactivity of the uniformly labeled glucose was distributed equally in the fatty acids and the glycerol. The distribution of radioactive glycerol between diglycerides and triglycerides was similar to that of the labeled fatty acids. Triglyceride synthesis seems to always be accompanied by the formation of diglycerides with a lastint turnover time.