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PubMed1975

Biosynthesis of phosphoglycerides in skeletal muscle in control rats and in rats deficient in essential fatty acids.

1. The specific radioactivities of individual molecular species of phosphoglycerides in the skeletal muscles of control rats and of rats deficient in essential fatty acids have been determined 3 h after intraperitoneal injection of ortho[32P] phosphate. 2. It has been demonstrated that the high average specific radioactivity of phosphoglycerides in muscles of rats deficient in essential fatty acids is due to both increased amounts and increased turnover of 1-palmitoyl-2-oleoyl phosphatidylcholine and phosphatidylethanolamine. 3. The 1-stearoyl-2-arachidonoyl phosphatidylcholine was found to turn over faster than the 1-palmitoyl-2-arachidonoyl species. In rats deficient in essential fatty acids, the 1-stearoyl-2-(5,6,11-eicosatrienoyl) phosphatidylcholine turned over more rapidly than the 1-palmitoyl-2-(5,8,11-eicosatrienoyl) species. Both findings are in constant with similar findings for liver.

Animals↗
PubMed1981

Products of fatty acid synthesis by a particulate fraction from germinating pea (Pisum sativum L.).

The synthesis of lipids and acyl thioesters was studied in microsomal preparations from germinating pea (Pisum sativum cv. Feltham First) seeds. Under conditions of maximal synthesis (in the presence of exogenous acyl-carrier protein) acyl-acyl-carrier proteins accounted for about half the total incorporation from [14C]malonyl-CoA. Decreasing the concentrations of exogenous acyl-carrier protein lowered the overall synthesis of fatty acids by decreasing, almost exclusively, the radioactivity associated with acyl-acyl-carrier proteins. A time-course experiment showed that acyl-acyl-carrier proteins accumulated most of the radioactive label at the beginning of the incubation but, eventually, the amount of radioactivity in that fraction decreased, while a simultaneous increase in the acyl-CoA and lipid fractions was noticed. Addition of exogenous CoA (1 mM) produced a decrease of total incorporation, but an increase in the radioactivity incorporated into acyl-CoA. The microsomal preparations synthesized saturated fatty acids up to C20, including significant proportions of pentadecanoic acid and heptadecanoic acid. Synthesis of these 'odd-chain' fatty acids only took place in the microsomal fraction. In contrast, when the 18,000g supernatant (containing the microsomal and soluble fractions) was incubated with [14C]malonyl-CoA, the radioactive fatty acid and acyl classes closely resembled the patterns produced by germinating in the presence of [14C]acetate in vivo. The results are discussed in relation to the role of acyl thioesters in the biosynthesis of plant lipids.

Fabaceae↗
PubMed1988

Effects of dietary casein and soybean protein on triglyceride turnover in rat liver.

We previously found that hepatic lipogenic enzyme induction, fatty acid synthesis, and triglyceride level were markedly lower in rats fed soybean protein than in those fed casein (Iritani et al. (1986): J. Nutr., 116: 190). After labeling triglycerides with tritiated water, the effects of dietary protein on the triglyceride degradation have been investigated. After the injection of tritiated water into rats, the radioactivities of fatty acids and triglycerides reached a plateau in 1-2 days and were markedly lower in the soybean protein group than in the casein group. The decreasing rates of triglyceride radioactivities were similar between the casein and soybean protein groups. The enzyme activities in glycerolipid synthesis were similar between the groups. Therefore, the lowering effects of soybean protein on triglyceride levels appear to be ascribed to triglyceride synthesis (due to fatty acid synthesis) rather than to the degradation.

Animals↗
PubMed1993

[Metabolism of nuclear neutral lipids and rat thymocyte chromatin under normal conditions and after gamma-irradiation].

The levels and specific radioactivities of cholesterol and free fatty acids in nuclei and chromatin of thymocytes from normal and gamma-irradiated (10 Gy) rats have been studied. The radioactivity of the total lipid fraction of gamma-irradiated cells was decreased significantly in the absence of inhibition of [2-14C]acetate incorporation into the total protein and lipid reactions and the [3H]uracil incorporation into the acid-insoluble RNA. The concentration of free fatty acids in the nuclei increased significantly after irradiation. The specific radioactivity of cholesterol in chromatin was higher than in the nuclei. The differences in specific radioactivities of free fatty acids were less pronounced. After irradiation the ratio of specific radioactivities of free fatty acids in chromatin to that in the nuclei showed a tendency to increase.

Animals↗
PubMed1978

Lack of enterohepatic circulation of intact biliary phosphatidylcholine in the rat.

The bile phosphatidylcholine was radioactively labelled by intraperitoneal or intravenous injection of [3H]- or [14C]-palmitic acid into bile fistula rats. The radioactive bile was infused intraduodenally into other bile fistula rats. On an average, only 1.0% of the infused radioactive phosphatidylcholine fatty acid reappeared in bile phosphatidylcholine, whereas about 68% of simultaneously infused [3H]-taurocholate was recovered over the same period of time (6 hours). There is thus no significant enterohepatic circulation of intact biliary phosphatidylcholine.

Animals↗
PubMed1975

[The lipid metabolism of the small intestine and its correlation to the lipid and lipoprotein metabolism of the total organism].

It is attempted to discuss the intestinal lipid metabolism particularly as it relates to systemic lipid and lipoprotein metabolism. In an introductory review of recent concepts of intestinal fat absorption special reference is given to the significance of the mixed lipid micelle during the intraluminal phase of fat absorption, to special morphological and functional features of the lumen-absorptive cell interface and to the proposed role of a fatty acid binding protein in the intracellular transport of long chain fatty acids. Furthermore the metabolic fate of absorbed fatty acids within the absorptive epithelial cell is reviewed and pointed out, that certain proteins (f.i. apoprotein B) are of crucial significance for the export of intestinal lipids into lymph. Absorption of dietary fat, however, is not the only function of the intestine in lipid metabolism. Intestinal mucosa is also capable of de novo synthesis of lipids, a function which is elaborated upon in the discussion of intestinal biosynthesis of cholesterol. Since in addition biosynthesis of cholesterol in the liver has been shown to be under sensitive control of cholesterol in intestinal lymph lipoproteins and the intestine represents the only organ, in which serum cholesterol can be excreted (after conversion to bile salts in the liver), the intestine occupies the central role in cholesterol metabolism. The intensive interrelation of intestinal functions and systemic lipoprotein metabolism is underscored by the more recent finding, that the intestine also synthesizes endogenous lipoproteins, which are secreted constantly into intestinal lymph even in the fasting state, thus contributing to plasma very low density lipoproteins. The intestine is the only organ, in which extrahepatic production of endogenous lipoproteins has been demonstrated. In the special section of this paper own original work is presented, elaborating on intestinal metabolism of plasma free fatty acids (FFA) and dealing with the important question, whether plasma FFA are incorporated into intestinal lymph lipoproteins. Studies were conducted in rats initially, which were given a rapid i.v. injection of labelled fatty acids. Mucosal radioactivity of the small intestine was greatest 2 minutes after i.v. 14C-palmitate, and accounted for 1% of administered isotope. Of mucosal 14C, 42% were present in water soluble metabolites, including CO2 and ketoacids, 28% in phospholipids and only 16% in triglycerides. The specific activity of mucosal triglyceride fatty acids (TGFA) was 11 times that of serum TGFA, confirming triglyceride synthesis by intestinal mucosa. The unexpectedly low percent incorporation of 14C-palmitate into triglycerides and its preferential conversion to water soluble metabolites, reflecting oxidation, were confirmed in double label experiments (3H-palmitate intraluminally, 14C-palmitate i.v.), which showed marked differences in the metabolism of fatty acids entering mucosa simultaneously from the two sources. While i.v...

Adult↗
PubMed1986

Metabolism of tween-Fatty Acid esters by cultured soybean cells : kinetics of incorporation into lipids, subsequent turnover, and associated changes in endogenous Fatty Acid synthesis.

Uptake of Tween-fatty acid esters and incorporation of the fatty acids into lipids by soybean (Glycine max [L.] Merr.) suspension cultures was investigated, together with subsequent turnover of the incorporated fatty acids and associated changes in endogenous fatty acid synthesis. Tween uptake was saturable, and fatty acids were rapidly transferred from Tweens to all acylated lipids. Patterns of incorporation into glycerolipids were similar in cells treated with Tweens carrying [1-(14)C]-fatty acids and in cells treated with [1-(14)C]acetate, indicating that exogenous fatty acids were used for glycerolipid synthesis essentially as if they had been made by the cell. In Tween-treated cells neutral lipids (which include Tweens) initially accounted for the majority of lipid radioactivity. Radioactivity was then rapidly transferred to glycerolipids. A transient pool of free fatty acids accounting for up to 10% of lipid radioactivity was observed. This was consistent with the hypothesis that fatty acids are transferred from Tweens to lipids by deacylation of the Tweens, creating a pool of free fatty acids which are then used for lipid synthesis. Sterols were only slightly labeled in cells treated with Tweens, but accounted for nearly 50% of lipid radioactivity in cells treated with acetate. This suggested very little degradation and reutilization of the radioactive fatty acids in cells treated with Tweens. In cells treated with either [1-(14)C]acetate or Tween-[1-(14)C]-18:1, 70% of the initial fatty acid radioactivity remained in fatty acids after a 100 hour chase. By contrast, fatty acids not normally present disappeared more rapidly, suggesting differential treatment of such fatty acids compared with those normally present. Cells which had incorporated large amounts of exogenous fatty acids altered fatty acid synthesis in three distinct ways: (a) amounts of [1-(14)C]acetate incorporated into fatty acids were reduced; (b) cells incorporating exogenous unsaturated fatty acids increased the proportion of [1-(14)C]acetate partitioned into saturated fatty acids, while the converse was true of cells which had incorporated exogenous saturated fatty acids; (c) desaturation of 18:1 to 18:2 and 18:3 was reduced in cells which had incorporated unsaturated fatty acids. These results suggest that Tween-fatty acid esters will be useful for supplying fatty acids to cells for a variety of studies related to fatty acid or membrane metabolism.

Journal Article↗
PubMed1984

Effect of propionic acid on kinetics of acetate and oleate and on plasma and milk fatty acid composition of goats.

Entry rates of acetate and oleate and their incorporation into lipids of blood plasma and fatty acids of milk were studied in lactating goats fed a concentrate-roughage ration with propionic acid infused intraruminally at 0, 5.52, and 13.74 g/h by primed constant intravenous infusion of [1-carbon-14] acetate and [9, 10-hydrogen-3] oleate. Means for infusion rates were acetate, 60, 52, and 39 micrograms/ml blood plasma; propionate 9, 12, and 22 micrograms/ml; oleate, 19, 14, and 12 micrograms/ml; acetate entry rate, 3.9, 2.7, and 1.8 mmol/h per kg bodyweight; oleate entry rate, 47, 29, and 19 mumol/h per kg bodyweight; acetate oxidation rate, 2.0, 1.7, and 1.4 mmol/h per kg, and its contribution to the total carbon dioxide production, 16, 14, and 11%. Propionic acid increased incorporation of carbon-14 and hydrogen-3 into plasma lipids, elevated proportions of 7:0, 9:0, 11:0, 13:0, 15:0, and 17:0 fatty acids in milk, and tended to lower others. Specific radioactivities of milk fatty acids during infusion of propionic acid were elevated by 1.8 to 2.8 times, and total fatty acids in milk and plasma were lowered by 22 and 38%. Data support the glucogenic theory that propionic acid either directly or through gluconeogenesis stimulates insulin secretion, which in turn inhibits release of fatty acids from adipose tissue, resulting in milk fat depression.

Acetates↗
PubMed1976

In vitro and in vivo synthesis of long-chain fatty acids from (1-14C) acetate in the renal papillae of rats.

1. The relationship between the rate of [1-14C] acetate incorporation into the fatty acids of renal papillary lipids and the acetate concentration in the medium has been measured. 2. [1-14C] acetate was incorporated mainly into fatty acids of phospholipids and triacylglycerols. Only a few per cent of the radioactivity was found in the free fatty acid fraction. 3. The major part of the [1-14C] acetate was found to be incorporated by a chain elongation of prevalent fatty acids. The major component of the poly-unsaturated fatty acids in triacylglycerols and the major product of fatty acid synthesis from [1-14C] acetate in vitro was demonstrated by mass spectrometry to be docosa-7,10,13,16-tetraenoic acid. 4. The radioactivity of docosa-7,10,13,16-tetraenoic acid accounted for 40% of total radioactivity in triacylglycerol fatty acids (lipid droplet fraction) and 20% of total radioactivity in membrane phospholipid fatty acids.

Animals↗
PubMed1983

High-energy diets produce different effects on fatty acid synthesis in brown adipose tissue, white adipose tissue and liver in the rat.

The influence of feeding rats a high-energy diet for 7 days on fatty acid synthesis in brown adipose tissue, white adipose tissue and liver of the rat was investigated. The incorporation of 3H2O and [U-14C]glucose into fatty acid was measured in vivo. The rats fed the high-energy diets had higher rates of fatty acid synthesis in white adipose tissue than the controls fed on chow, while fatty acid synthesis in brown adipose tissue and liver was either decreased or unchanged relative to that of controls fed on chow. After an oral load of [U-14C]glucose the incorporation of radioactivity into tissue fatty acid was several-fold higher in brown adipose tissue than in white adipose tissue in rats fed on chow. In rats fed the high-energy diets, incorporation of radioactivity into fatty acid in brown adipose tissue was decreased while that into white adipose tissue was either increased (Wistar rats) or unchanged (Lister rats).

Adipose Tissue↗
PubMed1988

Differential effects of chloroquine on the phospholipid metabolism of Plasmodium-infected erythrocytes.

The effect of the antimalarial drug chloroquine (CQ) on the phospholipid metabolism in Plasmodium knowlesi-infected simian erythrocytes has been studied by incubating cells with different labeled precursors and various concentrations of CQ. The drug induced considerable modifications of this metabolism but at the same time decreased nucleic acid and protein synthesis as well as the output of 14CO2 from radioactive glucose. Phosphatidylcholine biosynthesis was severely reduced. However, under these conditions, CQ had the early effect of markedly increasing phosphatidylinositol labeling from radioactive inositol, fatty acids, 1-(14C)palmitoyl-lysophosphatidylcholine, but not from glycerol. Synthesis of phosphatidylserine from (14C)serine and of phosphatidylethanolamine from labeled glycerol, ethanolamine, and serine was increased, especially at high CQ concentrations when the whole metabolism of the parasite was severely reduced. These effects reflect a deep differential effect of CQ on the intense phospholipid metabolism of the Plasmodium-infected erythrocytes, which might involve a redirecting of phospholipid metabolism similar to that induced by other cationic amphiphilic drugs, and a compensatory synthesis resulting from the severe blockage of phosphatidylcholine synthesis.

Animals↗
PubMed1992

Solubility of long-chain fatty acids in phosphate buffer at pH 7.4.

The solubility of the saturated fatty acids lauric, myristic, palmitic, and stearic acid and the unsaturated oleic acid at 37 degrees C in phosphate buffer (pH 7.4) was estimated by using two independent methods. The one was a conventional solubility technique measuring the concentration of dissolved fatty acid in buffer by using radioactive compounds. The other was a dialysis exchange technique monitoring possible aggregation of solvated fatty acid anions by measuring the rate of diffusion of labelled compound across a dialysis membrane under conditions of chemical equilibrium. It was found that the results were strongly dependent on the radiochemical purity of the fatty acids. Using highly purified samples of radioactively labelled fatty acids, the solubility of monomeric laurate was shown to be greater than 500 microM, whereas the solubility of monomeric myristate was found to be 20-30 microM. Palmitate, stearate, and oleate solutions, on the other hand, showed a tendency to aggregation even at concentrations below 1 microM. Special attention was given to palmitate, as a reference compound for long-chain fatty acids, and the solubility of monomeric palmitate was estimated to be lower than 10(-10) M.

Autoradiography↗
PubMed1973

Abnormal fatty acid metabolism in peripheral nerves of patients with pernicious anemia.

Fatty acid synthesis from radiopropionate was evaluated in sural nerve biopsy slices from five normal controls and nine patients with pernicious anemia. The nerves were incubated in [(14)C]propionate, the lipids were extracted, and the fatty acid methyl esters were chromatographed by gas-liquid chromatography. In the normal nerves the radiolabel was found primarily in short chain (C12 and C14) fatty acids. The nerves from pernicious anemia patients showed two fatty acids peaks that were not discernible in the normal nerves, and these fatty acids had retention times intermediate to those of myristic (C14.0) and palmitic (C16.0) acids and palmitoleic (C16.1) and stearic (C18.0) acids, respectively. These two peaks (a C15 and C17 fatty acid) contained the bulk of the radioactivity recovered in the fatty acid fraction after incubation with [(14)C]propionate. Catalytic reduction and rechromatography failed to alter the retention time of these compounds suggesting that they are not unsaturated fatty acids. The nerves from the pernicious anemia patients had a decrease in the mean content of normal fatty acids when compared with the nerves from control patients as well as a decrease in the mean synthesis of normal fatty acids as estimated by isotope incorporation after incubation with [(14)C]propionate or (3)H(2)O. Analysis of myelin isolated from the nerves indicated that the changes at least in part were in that fraction.

Acetates↗
PubMed2000

Effect of triiodothyronine on phospholipid metabolism in skeletal muscles of the rat.

The aim of the present study was to examine the effect of treatment with triiodothyronine (T3) on certain aspects of phospholipid metabolism in skeletal muscles. Rats were injected with triiodothyronine (T3) daily (10 microg x 100 g(-1) b.w., s.c.) for six days. Saline-treated rats served as controls. 24 h after the last dose of T3, 14C palmitic acid suspended in the serum of a donor rat, was administered intravenously. Thirty min later, samples of the soleus, white and red section of the gastrocnemius and blood from the abdominal aorta were taken. The muscle phospholipids were extracted and separated into different fractions by means of thin layer chromatography. The following fractions were obtained: shingomeylin, phosphatidylcholine phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and cardiolipin. The phospholipids were quantified and their radioactivity was measured. The plasma free fatty acid concentration and radioactivity was also determined. Treatment with T3 reduced the content of phosphatidylinositol and phosphatidylserine in each muscle type, whereas the concentration of other phospholipids remained stable. T3 increased markedly incorporation of the blood-borne fatty acids into each phospholipid fraction in the muscles. It is concluded that an excess of T3 influences the metabolism of phospholipids in skeletal muscles.

Animals↗
PubMed1979

Effects of hypo- and hyper-thyroidism on in vivo lipogenesis in fed and fasted rats.

Thyroidectomized rats (T) daily injected with either 0, 0.1, 1.8 or 25 microgram of L-thyroxine/100 g body wt. were compared with intact controls (C). The appearance of radioactivity in fatty acids 30 min after the i.p. injection of (3-14C)pyruvate was reduced in adipose tissue and enhanced in liver of T+25, being no differences between the other groups and C. (14C)-Fatty acids are reduced with 3 h of fasting only in the adipose tissue of T+1.8 and C, while 24 h produces a reduction in liver in the T+1.8, T+25 and C, and in adipose tissue in the T+1.8 and C animals. The highest percentage of radioactivity was observed in the liver glyceride glycerol fraction, being greater in T+25 than in the other groups. Fasting produces an increment in the (14C)-glyceride glycerol fraction. Being significant only in thehypothyroid animals in both liver and adipose tissue. The most sensitive parameter to fasting was the formation of (14C)-non-saponifiable lipid in both the C and T+1.8 animals, while it does not change in T+0 or T+0.1, but is enhanced within 24 h in the adipose tissue of T+25. It is proposed that most of the observed changes are due to the other endocrine disfunction s which appear in hypo- and hyperthyroidism, as the in vivo results do not comply with in vitro effects of thyroxine onlipogenesis of others.

Animals↗
PubMedDate not supplied

[Study of the role of nicotinamide coenzymes in the regulation of glyceroneogenesis from pyruvate in rat epididymis fat tissue].

The paper deals with a regulatory effect of the redox state of nicotinamide coenzymes on glyceroneogenesis in the epididymal fatty tissues involving incorporation of [2-14C] pyruvate into synthetized de novo blood glucose, glycerol and fatty acids of triacyglycerines. Large values of the NAD+/NADH and NADP+/NADPH ratios in cytoplasm and mitochondria promote a high rate of lipogenesis and glucose oxidation processes, which is pronounced in a more intense 14C incorporation into fatty acids than in triacylglycerol glycerols. A decrease in the NAD+/NADH ratio and an increase in the reducing ability of NAD-pairs under fasting intensify glyceroneogenesis in the fatty tissue. The incorporation of [14C] pyruvate into blood glucose in 3.6 times as high, the radioactivity of fatty acids lowers. Nicotinamide administered to animals after fastening inhibits glyceroneogenesis in the fatty tissue, lowering considerably the incorporation of [14C] pyruvate into triacylglycerol glycerol and blood glucose.

Adipose Tissue↗
PubMed1980

Involvement of phospholipids in polyunsaturated Fatty Acid synthesis in developing soybean cotyledons.

Developing soybean (cv. Dare) cotyledons harvested at 30 days after flowering were pulse-labeled with [1-(14)C]oleoyl-CoA. The metabolic interrelation of radiolabeled unsaturated fatty acids between the major glycerolipid classes was determined at various time intervals. At chase time zero, [(14)C]oleic acid accounted for 99.2% of the total glycerolipid radioactivity, and phospholipids contained 92% of the total incorporated radioactivity. With time, phospholipids were metabolized in triacylglycerol biosynthesis and radioactivity was detected in polyunsaturated fatty acids. The hypothesis that phospholipids were metabolic intermediates in polyunsaturated fatty acid biosynthesis was tested by comparing the theoretical and the actual amount of radiolabeled oleic acid that was associated with triacylglycerol as a function of time. The radioactive oleic acid found in triacylglycerol at various intervals was derived from phospholipids via a diacylglycerol intermediate. Assuming no phospholipid desaturation, the potential or theoretical amounts of [(14)C]oleic acid that could be transferred to triacylglycerol from phospholipids was defined by a system of differential equations. The results demonstrated that the decline in [(14)C]oleic acid from phospholipid after long chase intervals was equal to the total amount of radioactive unsaturated fatty acids found in neutral lipids. The difference between the theoretical and actual amounts of [(14)C]oleic acid present in triacylglycerol after long time intervals was equal to the amount of radioactivity present in polyunsaturated fatty acids. Based upon those findings in soybeans, the desaturation of oleic acid associated with phospholipids was highly probable.

Journal Article↗