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PubMed1980

Plasma free fatty acid umbilical venous-arterial concentration differences and placental transfer of [14C]palmitic acid in pigs.

The passage of plasma free fatty acids across the pig placenta was investigated in four anaesthetized sows with a total of 41 fetal piglets. The plasma free fatty acid concentrations in fetal piglets were very low and appeared independent of maternal concentrations. In each piglet there was a small positive umbilical venous-arterial concentration difference. There were large differences between the composition of maternal and fetal plasma free fatty acid. The fetal free fatty acid contained relatively large amounts of arachidonic and docosahexaenoic acids. [14C]palmitic acid given intravenously to the sows appeared rapidly in fetal plasma free fatty acids but the amount of radioactivity was low. Analysis of the specific activity of palmitic acid in fetal and maternal blood indicated that little more than 10% of fatty acids coming from the placenta were derived directly from maternal plasma free fatty acids. It is concluded that, in the anaesthetized pig at Caesarean section, fatty acids enter the fetal circulation from the placenta and are taken up by the fetus at rates exceeding the requirements for structure and storage, but the major part of these free fatty acids are not derived directly from maternal free fatty acids.

Animals↗

PubMed2000

Intestinal fatty acid binding protein may favor differential apical fatty acid binding in the intestine.

The intestinal mucosa metabolizes fatty acids differently when presented to the lumenal or basolateral membrane. Expression of both liver and intestinal fatty acid binding proteins (L- and I-FABPs) uniquely in the enterocyte offers a possible explanation of this phenomenon. An organ explant system was used to analyze the relative binding of fatty acids to each protein. More fatty acid was bound to L-FABP than to I-FABPs (28% vs. 6% of cytosolic radioactivity), no matter on which side the fatty acid was added. However, a 2-3-fold increase in fatty acid binding to the intestinal paralog was noted after apical addition of palmitic or oleic acid in mucosa from chow fed rats. When oleic acid was added apically, a 1.4-fold increase in binding to I-FABP was observed in mucosa derived from chronically fat fed rats, consistent with the previously observed 50% increase in the content of that protein. Immunocytochemical localization of both FABPs in vivo demonstrated an apical cytoplasmic localization in the fasting state, and redistribution to the entire cytoplasm after fat feeding. These data are consistent with the hypothesis that I-FABP may contribute to the metabolic compartmentalization of apically presented fatty acids in the intestine.

Animals↗

PubMed1987

In vitro clearance of chylomicron triglycerides containing (omega-3) eicosapentaenoate.

Rat mesenteric lymph chylomicrons, containing triglycerides enriched with either [14C]oleic acid or [14C]eicosapentaenoic acid, were prepared by ultracentrifugation of lymph samples collected for 6 h after a single duodenal infusion of an emulsion containing 0.3 mmol of either fatty acid. After determination of protein and of total fatty acid content and composition, enriched chylomicrons were suspended in Krebs-bicarbonate buffer. Non-working hearts were perfused in a recirculating system for 45 min using the enriched chylomicron preparations. At 15 min intervals during perfusion, the media were assayed for total radioactivity, 14CO2 and 14C-labeled fatty acids associated with triglycerides, unesterified fatty acids, phospholipids, mono- and diglycerides. After perfusion, the hearts were extracted and assayed for total lipid radioactivity and isotope distribution among heart lipid fractions. With this membrane-supported lipoprotein lipase system, clearances of chylomicron triglycerides containing either fatty acid were identical, as were the myocardial uptakes of the fatty acids and oxidations to 14CO2. Furthermore, except for a significantly greater incorporation of eicosapentaenoate into myocardial phospholipids, tissue isotope distributions of the two labeled fatty acids were also the same. These studies suggest that at least the initial phases of peripheral clearance of chylomicrons enriched in omega-3 fatty acids is as efficient as with those containing oleate.

Animals↗

PubMed1992

Direct effects of temperature on phospholipid and polyunsaturated fatty acid metabolism in isolated brain cells from rainbow trout, Oncorhynchus mykiss.

1. The direct effects of temperature on the metabolism of [1-14C]18:2(n-6), [1-14C]18:3(n-3), [1-14C]20:4(n-6) and [1-14C]20:5(n-3) were studied in isolated brain cells from rainbow trout, Oncorhynchus mykiss. 2. Recovery of radioactivity from all the polyunsaturated fatty acids (PUFA) in total lipid was significantly greater at 5 and 15 degrees C than at 25 degrees C. 3. The lower incubation temperatures decreased the relative net incorporation of all the 14C-labelled PUFA into phosphatidylcholine (PC) and increased the relative incorporation of the PUFA into the other phosphoglycerides, especially phosphatidylethanolamine (PE). 4. The effects on PC were generally more significant between 25 and 15 degrees C, whereas the effects on PE were generally significant both between 25 and 15 degrees C and between 15 and 5 degrees C. 5. This suggests that the lysophospholipid acyltransferases responsible for the incorporation of PUFA into different phosphoglycerides may have differential sensitivities to temperature. 6. In contrast, the acyltransferase activities showed fatty acyl preferences that were independent of temperature. 7. Although a trend towards decreased activity at 5 degrees C was apparent, temperature generally had little significant effect on the relative percentages of the PUFA metabolized via the desaturase pathways.

Acclimatization↗

PubMed1989

Quantitative analysis of myocardial kinetics of 15-p-[iodine-125] iodophenylpentadecanoic acid.

Myocardial extraction and the characteristic tissue clearance of radioactivity following bolus injections of a radioiodinated (125I) long chain fatty acid (LCFA) analog 15-p-iodophenylpentadecanoic acid (IPPA) were examined in the isolated perfused working rat heart. Radioactivity remaining in the heart was monitored with external scintillation probes. A compartmental model which included nonesterified tracer, catabolite, and complex lipid compartments successfully fitted tissue time-radioactivity residue curves, and gave a value for the rate of IPPA oxidation 1.8 times that obtained from steady-state release of tritiated water from labeled palmitic acid. The technique was sensitive to the impairment of LCFA oxidation in hearts of animals treated with the carnitine palmitoyltransferase I inhibitor, 2[5(4-chlorophenyl)pentyl]oxirane-2-carboxylate (POCA). IPPA or similar modified fatty acids may be better than 11C-labeled physiological fatty acids such as palmitate in this type of study, because efflux of unoxidized tracer and catabolite(s) from the heart are kinetically more distinct, and their contributions to the early data can be reliably separated. This technique may be suitable for extension to in vivo measurements with position tomography and appropriate modified fatty acids.

Animals↗

PubMedDate not supplied

Fatty acid biosynthesis in Wallerian degeneration of rat sciatic nerve.

In-vitro fatty acid biosynthesis was studied in normal rat sciatic nerve and during wallerian degeneration. Normal nerve incorporated 1,3-C14-malonyl-CoA and 1-C14-acetyl-CoA into fatty acids by a de-novo biosynthetic pathway. The reaction product with highest radioactivity was palmitic acid, and the free fatty acids of nerve contained 90% of the total fatty acid label. During wallerian degeneration, there was a rapid increase of fatty acid biosynthesis that reached a peak between 16 and 24 days after nerve section. Values declined to normal levels at approximately 50 days.

Acetyl Coenzyme A↗

PubMed1977

Incorporation of a fatty acid containing a photosensitive group into lipids of cultured cardiac cells from chick embryo.

Radioactive 12-(4-azido-2-nitrophenoxy)-stearic acid (NAP-stearate) was synthetized; it behaves as a competitive inhibitor of long-chain fatty acids for the entry into cultured cardiac cells from chick embryo. After uptake, [3H] NAP-stearate was incorporated by an energy-dependent process into neutral and polar lipids. Photoactivation as a function of time leads to a covalent labelling of the cells: up to 31 per cent of the radioactivity was recovered in the 105 000 g cell pellet, mainly in proteins. These experiments show that fatty acids containing photosensitive groups would potentially allow to localize the proteins involved in the binding and/or in the transport of fatty acids.

Animals↗

PubMed1978

Fatty acid biosynthesis in the leaves of barley, wheat and pea.

1. The incorporation of radioactivity from [1-14C]acetate into the leaf lipids of barley, pea and wheat has been studied in pulse-labelling experiments. 2. There was little increase in the total labelling of lipids after the leaves were transferred to non-radioactive medium. However, there was an increase in the relative labelling of unsaturated fatty acids. In addition, there was an increase in the relative labelling of diacylgalactosylglycerol. 3. The principal radioactively labelled acyl lipids were diacylgalactosylglycerol and phosphatidylcholine. Phosphatidylcholine showed a decreasing proportion of [14C]oleate and an increasing amount of [14C]linoleate with time. Diacylgalactosylglycerol also had decreasing amounts of [14C]oleate but, in addition, had an increasing proportion of [14C]linolenate with time. 4. The absence of significant amounts of [14C]linolenate in phosphatidylcholine appeared to exclude a role for this phospholipid in linoleate desaturation. 5. The specific radioactivities of oleate and linoleate in phosphatidylcholine, diacylgalactosylglycerol and diacylgalabiosylglycerol were very similar in any single experiment. It was concluded that these fatty acids can rapidly exchange between the three intact lipids.

Diglycerides↗

PubMedDate not supplied

Exposure to free fatty acid increases the transfer of albumin across cultured endothelial monolayers.

An initial exposure to high concentrations of free fatty acid increased the transfer of albumin across cultured endothelial monolayers. The rate and amount of albumin transfer was dependent on the oleic acid concentration to which the cultures were initially exposed, with 300 microM producing the maximum transfer. The albumin transfer also increased with the increasing time of exposure to oleic acid, the maximum effect occurring during the first 24 hours. An exposure to 300 microM linoleic acid produced an even greater increase in albumin transfer than did 300 microM oleic acid. The increased albumin transfer observed when cells were exposed to high concentrations of free fatty acid was largely reversible after reincubation of the cell monolayers in free fatty acid-poor media. In parallel experiments, radioactive oleic acid incorporation into cell triglycerides increased linearly as the fatty acid concentration was raised, with cell triglyceride content increasing up to sevenfold after incubation in a medium containing 300 microM oleic acid. A significant amount of oleic acid was incorporated into phospholipids, and the fatty acid composition of the endothelial triglycerides and phospholipids was modified. All these effects of oleic occurred without altering the incorporation of leucine into the cell protein. These results indicate that exposure to high concentrations of free fatty acid can alter endothelial cell lipid composition, and that this increases the albumin transfer across endothelium. This process might permit more macromolecules to enter the arterial wall.

Albumins↗

PubMed1985

Biosynthesis of fatty acids by lactating human breast epithelial cells: an evaluation of the contribution to the overall composition of human milk fat.

The objective of this study was to characterize the fatty acid biosynthetic pathway of the lactating human breast. Mixed cell populations, obtained by centrifugation of human milk, were enriched in breast epithelial cells by a selective adsorption procedure. Confirmation of the identity of the breast epithelial cells was obtained immunohistochemically. These viable breast epithelial cells incorporated radioactively labeled acetate predominantly into fatty acids with less than 16C atoms. The presence of the two key enzymes characteristic of the medium-chain fatty acid biosynthetic pathway of nonruminants, fatty acid synthetase, and thioesterase II, was demonstrated both qualitatively, by immunohistochemistry, and quantitatively, by enzyme assay. The results indicate that the lipogenic system of the human breast is qualitatively very similar to that of rats, mice, and rabbits, which also secrete milk fats containing medium-chain fatty acids. Quantitatively, however, the mammary fatty acid biosynthetic pathway appears to be less active in humans than in these other species.

Adult↗

PubMed1991

Absorption and intestinal catabolism of fatty acids in the rat: effect of chain length and unsaturation.

Simultaneous portal blood absorption and intestinal mucosal catabolism of labelled fatty acids were investigated. Anaesthetized adult Wistar rats were infused intraduodenally either with 90 mumol of capric (C10:0), oleic (C18:1), linoleic (C18:2) or arachidonic (C20:4) 1-14C acids or with 30 mumol of each labelled fatty acid in addition to 30 mumol of oleic acid and 30 mumol of monopalmitin. For mixed infusates, experiments were carried out with two additional long-chain fatty acids: palmitic (C16:0) and erucic (C22:1) 1-14C acids. Radioactivity was quantified in the lipids and in the catabolic products in portal blood recovered at 5 min intervals for 1 h after infusion. At the end of the experiment, the disappearance of radioactivity from the mucosa was quantified. When labelled fatty acids were infused alone, 49% of the radiolabelled lipid disappearing from the mucosa was recovered in the blood for C10:0, but only 7.8% for C18:1, 6.4% for C18:2 and 10.6% for C20:4. With mixed infusates, 41% of the radiolabelled lipid disappearing from the mucosa was recovered in the blood for C10:0 compared with 12% for C18:1, 10.2% for C18:2, 10.5% for C20:4 and 2.7% for C16:0 and 2% for C22:1. Labelled catabolites appear with the same profiles as those of the respective fatty acids in blood. These studies confirm a minor absorption into blood of long-chain fatty acids compared to the medium-chain fatty acids and highlight differences in the catabolism of the fatty acids according to their chain length and their degree of unsaturation. The differences might be related to the differences in the fatty acid hydrosolubility and to their different affinities for the I- and L-cytosolic fatty acid binding proteins. These phenomena may be important in nutrition in relation to the availability of essential fatty acids.

Absorption↗

PubMed1967

The incorporation of acetate, stearate and D(-)-beta-hydroxybutyrate into milk fat by the isolated perfused mammary gland of the goat.

1. Mammary glands of lactating goats were perfused for 12.5-15hr. with heparinized whole blood and infused with a substrate mixture of glucose, acetate and amino acids (and sometimes chylomicra) containing either [1-(14)C]acetate, d(-)-beta-hydroxy[1-(14)C]butyrate or [U-(14)C]stearate. 2. There was a substantial net uptake of acetate by the glands and transfer of radioactivity into milk fat. Acetate was extensively utilized for the synthesis of milk fatty acids of chain length up to C(14) and to a smaller extent for the synthesis of palmitate. 3. There was a small and variable net uptake of stearate and beta-hydroxybutyrate and negligible oxidation of these substrates. However, tissue uptake was demonstrated by a substantial fall in specific radioactivity across the glands and an extensive transfer of radioactivity into milk fatty acids. 4. With beta-hydroxybutyrate the labelling of milk fat was very similar to that with acetate, but the distribution of radioactivity suggested a cleavage into C(2) fragments of about 40%. 5. Labelled stearate gave rise to highly labelled stearate and oleate in the milk fat. Small amounts of radioactivity were detected in stearate of plasma triglycerides and oleate of plasma free fatty acids. 6. In experiments where there was a decline in milk-fat secretion late in perfusion, the milk fatty acids showed a marked decline in the proportion of stearate and oleate and a rise in the proportion of myristate and palmitate. This did not occur in experiments where milk-fat secretion was maintained at a higher level. 7. The present results confirm that there is a large pool of long-chain fatty acids in mammary tissue that can act as an endogenous source of these substrates.

Acetates↗

PubMed1985

The effect of penicillin on fatty acid synthesis and excretion in Streptococcus mutans BHT.

Treatment of exponentially growing cultures of Streptococcus mutans BHT with growth-inhibitory concentrations (0.2 microgram/ml) of benzylpenicillin stimulates the incorporation of [2-14C] acetate into lipids excreted by the cells by as much as 69-fold, but does not change the amount of 14C incorporated into intracellular lipids. At this concentration of penicillin cellular lysis does not occur. The radioactive label is incorporated exclusively into the fatty acid moieties of the glycerolipids. The increase in the radioactive content of the extracellular lipids reflects an actual net increase in the total fatty acid content as determined by a chemical assay. During a 4-hr incubation in the presence of penicillin, the extracellular fatty acid ester concentration (per mg cell dry weight) increases 1.5 fold, even though there is no growth or cellular lysis. No change is observed in the intracellular fatty acid ester content. An indication of the relative rate of fatty acid synthesis was most readily obtained by placing S. mutans BHT in a buffer containing 14C-acetate. Under these nongrowing conditions free fatty acids are the only lipids labeled, a factor which simplifies the assay. The addition of glycerol to the buffer causes all of the nonesterified fatty acids to be incorporated into glycerolipid. The cells excrete much of the lipid whether glycerol is present or not. Addition of penicillin to the nongrowth supporting buffer system does not stimulate the incorporation of [14C]-acetate into fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗

PubMedDate not supplied

Correlation of heterogeneous blood flow and fatty acid uptake in the normal dog heart.

Blood flow heterogeneity in normal myocardium may be caused by heterogeneous metabolic demand. We studied, from 80 tissue samples of the left ventricle (LV) of eight anesthetised, open-chest dogs (with prior beta-blockade (metoprolol) in four dogs), the radioactivity of 201Thallium-chloride (201Tl), an indicator of blood flow, and of the fatty acid 131-Iodine-heptadecanoic acid (131I-HDA), an indicator of metabolic demand, 3 min after intravenous injection. Global LV uptake (in percent of injected dose x 10(-2), per g tissue; mean +/- SD) was 4.94 +/- 0.71 for 201Tl and 4.48 +/- 0.58 for 131I-HDA in the dogs without beta-blockade, and 2.08 +/- 0.26 and 1.69 +/- 0.20, respectively, in dogs with beta-blockade (p < 0.05). Beta-blockade thus decreased the fraction of cardiac output delivered to the LV, concurrently with a decreased heart rate and arterial blood pressure (p < 0.05) and, thus, global metabolic demand and fatty acid uptake. Regional radioactivities per gram were normalized for mean LV radioactivities and heterogeneity was expressed as the coefficient of variation (CV). For pooled data (n = 320) in dogs without beta-blockade, regional 201Tl and 131I-HDA radioactivities varied from a factor of 0.1 to 1.6 and 0.3 to 1.8 of mean radioactivities, with a CV of 22.9 and 19.4%, respectively, and correlated (r = 0.77, p < 0.005). For pooled data (n = 320) in dogs with beta-blockade, regional 201Tl and 131I-HDA radioactivities varied from a factor of 0.2 to 1.5 and 0.2 to 1.6 of mean radioactivity and CV was 23.6% and 24.8%, respectively; r = 0.92 (p < 0.005). The endo/epi ratio for both radioactivities exceeded unity in each dog. In normal myocardium, blood flow and fatty acid uptake are thus heterogeneous, both transmurally and circumferentially, and matched, concomitantly with coupling of global blood flow to global metabolic demand and fatty acid uptake. This supports the idea that heterogeneous myocardial O2 supply reflects heterogeneous metabolic demand.

Animals↗

PubMed1975

Nonequilibration of acetyl units from fatty acids and glucose in mammary glands of lactating mice.

Lipid synthesis by lactating mouse mammary gland slices from [1-carbon14] propionate, [1-carbon14] valerate, and [1-carbon-14] heptanoate, in the presence of glucose and from [uniformly labeled carbon-14] glucose in the presence of propionate, valerate, and heptanoate was studied. Regardless of the labeled substrate, almost all of the radioactivity incorporated into the total lipid fraction was in triglycerides. In experiments with labeled glucose and either the propionic. the valeric, or the heptylic acid, the carbon-14 fatty acids in triglycerides were almost exclusively composed of even-chain fatty acids. When the substrate was either [1-carbon-14] propionate, [1-carbon-14] valerate, or [1-carbon-14] heptanoate, in the presence of glucose, the pattern changed to one in which a larger proportion of the radioactivity was in the odd-chain fatty acids. These findings suggest that in mouse mammary gland, acetyl units produced via beta-oxidation of fatty acids may not equilibrate with those formed from oxidation of pyruvate which is derived from glucose.

Acetates↗

PubMed1996

Determination of DNA adducts of malonaldehyde in humans: effects of dietary fatty acid composition.

The effects of dietary fatty acid composition on the endogenous formation of DNA adducts of malonaldehyde (MA), the major product of lipid peroxidation, were investigated in humans. A group of 59 healthy individuals of both sexes and different ages was initially fed a milk fat-based diet rich in saturated fatty acids for 14 days. Following this initial period, after which the group was considered homogeneous with respect to diet, 30 randomly chosen subjects were given a sunflower oil-based (rich in polyunsaturated fatty acids) (SO) diet and the remaining 29 individuals a low erucic acid rapeseed oil-based (rich in monounsaturated fatty acids) (RO) diet for 25 days. The fatty acid composition of plasma lipid fractions and the level of DNA adducts of MA in total white blood cells were then determined at the end of the SO and RO dietary periods. DNA adduct levels were measured by 32p-postlabelling using reversed-phase HPLC with on-line detection of radioactivity. Higher concentrations of polyunsaturated fatty acids in plasma triglycerides and higher levels of DNA adducts of MA were found in the subjects on the SO diet when compared with those in the RO dietary group. A large inter-individual variation in adduct levels was observed. The average adduct level in the SO diet group was 7.4 +/- 8.7 adducts/10(7) nucleotides (n = 23). This level was 3.6-fold higher than that found in individuals in the RO diet group (P < 0.001). Our results, in conjunction with the mutagenic and carcinogenic properties of MA, thus suggest the interaction of lipid peroxidation products such as MA with DNA as one plausible mechanism explaining the involvement of dietary fat in carcinogenesis.

Adolescent↗

PubMed1967

The oxidation and utilization of palmitate, stearate, oleate and acetate by the mammary gland of the fed goat in relation to their overall metabolism, and the role of plasma phospholipids and neutral lipids in milk-fat synthesis.

1. Measurements were made of milk yield, mammary blood flow and arteriovenous differences of each plasma lipid fraction, and their specific radioactivities, during the infusion of [U-(14)C]stearate, [U-(14)C]oleate, [U-(14)C]palmitate and [1-(14)C]acetate into fed lactating goats. 2. Entry rates of fatty acids into the circulation were 4.2mg./min./kg. body wt. for acetate, and 0.18, 0.28 and 0.42mg./min./kg. for stearate, oleate and palmitate respectively. Acetate accounted for 23% of the total carbon dioxide produced by the whole animal, and contributed to the oxidative metabolism of the mammary gland to about the same extent. Corresponding values for each of the long-chain acids were less than 1%. 3. There were no significant arteriovenous differences of phospholipids, sterols or sterol esters, and their fatty acid composition showed no net changes during passage through the mammary gland. 4. There were large arteriovenous differences of plasma triglycerides, and their fatty acid composition showed marked changes across the gland. The proportions of palmitate and stearate fell, and that of oleate increased. 5. Arteriovenous differences of plasma free fatty acids (FFA) were small and variable, but a large fall in the specific radioactivity of each of the long-chain acids examined indicated substantial uptake of plasma FFA, accompanied by roughly equivalent FFA release from mammary tissue. The uptake of FFA was confirmed by the extensive transfer of radioactivity into milk. The FFA of milk were similar in composition and radioactivity to the milk triglyceride fatty acids, and quite unlike plasma FFA. 6. The formation of large amounts of oleic acid (18-21 mg./min.) from stearic acid was demonstrated. 7. During the terminal stages of the [(14)C]acetate infusion, milk triglyceride fatty acids of chain length C(4)-C(14) showed specific radioactivities that were 75-90% of that of blood acetate, and that of palmitate was roughly one-quarter of this value. Oleate and stearate were unlabelled. 8. The results confirmed that milk fatty acids of chain length C(4)-C(14) arise largely from blood acetate, and palmitate is derived partly from acetate and partly from plasma triglyceride, the latter fraction being almost the sole precursor of oleate and stearate.

Journal Article↗

PubMed1987

Metabolism of saturated fatty acids by Paramecium tetraurelia.

Paramecium requires oleate for growth. The phospholipids of the ciliate contain high concentrations of palmitate and 18- and 20-carbon unsaturated fatty acids. We previously showed that radiolabeled oleate is desaturated and elongated to provide these 18- and 20-carbon unsaturated acids. We now report on saturated fatty acid (SFA) metabolism in Paramecium. Radiolabeled palmitate and stearate were incorporated directly into cellular phospholipids with little or no desaturation and/or elongation. Radiolabeled acetate, malonate, pyruvate, citrate, or glucose added to cultures were not incorporated into cellular phospholipid fatty acids indicating that these exogenously supplied putative precursors were not utilized for fatty acid synthesis by Paramecium. Radiolabel from octanoate or hexanoate appeared in fatty acyl groups of phospholipids, possibly by partial beta-oxidation and reincorporation of the label. Under oleate-free conditions in which cultures do not grow, radiolabel from these shorter chain SFA were beta-oxidized and preferentially used for the formation of arachidonate, the major end-product of fatty acid synthesis in Paramecium. Cerulenin inhibited culture growth apparently by inhibiting de novo fatty acid synthesis. Cerulenin-treated cells did not incorporate radioactivity from [1-14C]octanoate into esterified palmitate. However, total saponifiable phospholipid fatty acids, including SFA, per cell increased under these conditions.

Acetates↗