Fatty-acid desaturation and microsomal lipid fatty-acid composition in experimental hyperthyroidism.

We have studied the influence of experimental hyperthyroidism in the rat on the synthesis of unsaturated fatty acids and on liver microsomal lipid fatty-acid composition. Tri-iodothyronine treatment (25 micrograms/100 g body weight) daily for 3 weeks caused no significant changes in delta 9 (stearate) desaturation but a 24% decrease in delta 6 (linoleate) desaturation. Much larger doses of tri-iodothyronine increased delta 9 desaturation. Liver microsomal fatty-acid composition in hyperthyroidism is altered with significantly increased proportions of stearate and arachidonate and decreased proportions of palmitate, palmitoleate, linoleate (C18:2) and eicosa-8,11,14-trienoate (C20:3). These changes, other than the decreases proportion of C20:3 fatty acid, which may be due to the diminished delta 6 desaturase activity, cannot be attributed to changes in fatty-acid desaturation. Most of these changes were also found to be due not simply to the decreased weight gain or the increased food intake of the hyperthyroid animals. Only the decreased C18:2 fatty-acid proportions could be mimicked by restricting food intake of control animals and none of the changes were prevented by restricting food intake of hyperthyroid animals. Thus most of the changes in microsomal lipid fatty-acid composition are likely to be due to a thyroid hormone effect on peripheral lipid mobilization or lipid degradation.     

Fatty acid composition and lipid synthesis in developing safflower seeds

Linoleic acid predominated in every lipid class during the whole period of seed development of safflower, while linolenic acid decreased with increasing maturation and it was not detected in mature seeds. Just before the initiation of triacylglycerol accumulation, the fatty acid composition of triacylglycerols changed more rapidly than those of phospholipids and glycolipids. Saturated fatty acids tended to accumulate at the 1- and 3-positions of the glycerol molecule and the more highly unsaturated acids at the 2-position. The fatty acid compositions at the 1- and 3-positions were similar in all cases investigated, but in none of the triacylglycerols was the distribution completely symmetrical. The positional distribution of linolenic acid in triacylglycerols prepared from the immature seeds 2 days after flowering and from the leaves was unusual; in spite of its highest degree of unsaturation, it was preferentially esterified at the 1- and 3-positions. When triacylglycerol was most rapidly accumulated (14–18 days after flowering), the incorporation of acetate-[U- ¹⁴C] into total lipids was also maximum and dienoic fatty acids were the principal acids labelled. Diacylglycerols and compound lipids reached the highest rate of synthesis 15 days after flowering, and then a maximum incorporation into triacylglycerol occurred 18 days after flowering. Incubation temperature affected the synthesis of individual lipid classes. Triacylglycerol was more rapidly synthesized at 32° than at 10°, while diacylglycerols and compound lipids were accumulated under the low-temperature condition. A rise of incubation temperature caused a depression in dienoic acid synthesis.     

Fatty acid composition of lipid A in Pseudomonas fluorescens

The fatty acid composition of lipid A was studied using gas-liquid chromatography (GLC) and GLC-mass spectrometry in Pseudomonas fluorescens strains of biovars A, B, C, i, F and G, the type strain ATCC 13525 (biovar A) inclusive. The following fatty acids were identified as predominant in the composition of lipid A in the strains representing biovars A, B, C, i, F and G: 3-hydroxydecanoic (3-OH C10:0), 2-hydroxydodecanoic (2-OH C12:0), 3-hydroxydodecanoic (3-OH C12:0), dodecanoic (C12:0), hexadecanoic (C16:0), octadecanoic (C18:0), hexadecenoic (C16:1) and octadecenoic (C18:1) acids. Lipid A of a biovar G strain differed noticeably from other strains in its fatty acid composition. Its main components were as follows: 3-hydroxytetradecanoic (3-OH C14:0), 3-hydroxypentadecanoic (3-OH C15:0) and dodecanoic (C12:0) fatty acids. The coefficients of similarity were determined for lipid A specimens isolated from the studied strains of P. fluorescens by calculating their fatty acid composition with a computer.     

Studies on Tetrahymena microsomal electron transport systems: solubilization of microsomal electron transport enzymes involved in fatty acid desaturation

Tetrahymena microsomes were solubilized with five different detergents and the effect on electron transport enzymes involved in fatty acid desaturation was studied. Cytochrome b560ms and NADPH-cytochrome c reductase were solubilized with a low concentration detergent (0.25%), in the order of sodium deoxycholate greater than Renex 690 greater than Triton X-100 greater than octylglucoside greater than sodium cholate, whereas all of these detergents at the high concentration (1%) could solubilize preferentially both enzymes (70-100%). Increasing the concentration of various detergents from 0.5 to 1.0% did not produce an incremental change in NADH-ferricyanide reductase solubilization. NADH-cytochrome c reductase system, which would be catalyzed by the cooperation action of NADH-ferricyanide and cytochrome b560ms, was relatively inactivated by all detergents. Compared to the other four detergents, octylglucoside has a much higher recovery of stearoyl-CoA desaturase activities in the supernatant. Our study suggests that octylglucoside may be more useful for the isolation in active form of cyanide-sensitive factor (CSF) from Tetrahymena microsomes.     

Fatty acid desaturation in the intestinal mucosa

Information as to the ability of the enterocyte to desaturate fatty acids is lacking. This is important in understanding whether the source of intestinal arachidonic (20:4(n-6) acid is biliary or from de novo synthesis. Delta 9- and delta 6-desaturase enzymes were assayed in homogenates of rat jejunum, ileum and liver. Rat small intestine possesses desaturase activity to convert palmitic (16:0) to palmitoleic (16:1) and linoleic (18:2(n-6) to linolenic (18:3(n-6) acid. Enzyme activities were highest in liver relative to activity in jejunal and ileal homogenates. It is concluded that delta 9- and delta 6-desaturase activities may have an important role in determining physico-chemical properties and thus transport properties of enterocyte membranes.     

An indirect thyroxine stimulation of microsomal fatt acid desaturation in vitro.

Metabolically active intestinal epithelial cells were isolated using collagenase plus hyaluronidase. Oxygen consumption was measured and was found to be inhibited by KCN, antimycin A, and rotenone. Cells from 3-methylcholanthrene(MC)-treated rats metabolized benzo(α)pyrene (BP) at a rate that was 30-fold greater than control cells. The addition of salicylamide to the incubation medium inhibited conjugation of BP metabolites and facilitated the accumulation of fluorescent and ethylacetate extractable metabolites. Metyrapone, SKF 525-A, α-naphthoflavone (α-NF), and rotenone inhibited BP-metabolism in intestinal cells from MC-treated rats, with α-NF being the most inhibitory. In intestinal cells from control animals, metyrapone and SKF 525-A both inhibited BP metabolism, while α-NF and rotenone both produced an increase in the formation of fluorescent BP products. The distribution of metabolites from MC-treated rats was determined by high-pressure liquid chromatography and compared with authentic BP derivatives. Incubations were conducted for 5 and 30 min in the presence and absence of salicylamide, and 30-min samples incubated in the absence of salicylamide were hydrolyzed with β-glucuronidase or aryl sulfatase. In the absence of salicylamide, large amounts of conjugates were formed, the formation of which were inhibited by salicylamide addition. A product corresponding to the 4,5-oxide constituted the major metabolite after a 5-min incubation, while little dihydrodiol formation occurred. Large amounts of phenolic BP derivatives were also formed. After 30-min incubations, the percentage of products corresponding to the 4,5-oxide decreased, and an increase in dihydrodiol formation was observed. The slow metabolism of the 4,5-oxide and the slow accumulation of dihydrodiols is due to the presence of low epoxide hydrase activity in the intestine. Intestinal cells are capable of xenobiotic metabolism, and offer a convenient method of studying intestinal drug metabolizing processes which may significantly contribute to the overall xenobiotic metabolis in the body.     

Seipin deficiency alters fatty acid Δ9 desaturation and lipid droplet formation in Berardinelli-Seip congenital lipodystrophy

Berardinelli-Seip congenital lipodystrophy (BSCL) is a rare recessive disease characterized by near absence of adipose tissue and severe insulin resistance. In most cases, BSCL is due to loss-of-function mutations in the genes encoding either seipin of unknown function or 1-acyl-glycerol-3-phosphate O-acyltransferase 2 (AGPAT2) which catalyses the formation of phosphatidic acid from lysophosphatidic acid. We studied the lipid profile of lymphoblastoid cell-lines from 20 BSCL patients with null mutations in the genes encoding either seipin (n = 12) or AGPAT2 (n = 8) in comparison to nine control cell-lines. In seipin deficient cells, we observed alterations in the pattern of lipid droplets which were decreased in size and increased in number as compared to control cells. We also observed alterations in the triglycerides content as well as in the fatty acid composition from triglycerides and phosphatidylethanolamine, with an increased proportion of saturated fatty acids at the expense of the corresponding monounsaturated fatty acids, reflecting a defect in Δ9-desaturase activity. In AGPAT2 deficient cells, no specific alterations in lipid droplet pattern nor in fatty acid composition was observed but the cellular level of lysophosphatidic acid was increased as compared to that of control and seipin deficient cells. These results indicate that seipin like AGPAT2 is involved in lipid metabolism but exerts a different function. Seipin intervenes at a proximal step in triglycerides and phospholipids biosynthesis being involved in the pathway that links fatty acid Δ9 desaturation to lipid droplet formation. These findings provide new insights into how seipin deficiency causes severe lipodystrophy.     

Fatty acid,polar lipid and wall amino acid composition of Gardnerella vaginalis

Representative strains of Gardnerella vaginalis were degraded using both an alkaline and an acid methanolysis and the fatty acid methyl esters released examined by thin-layer and gas chromatography. The profiles obtained were both qualitatively and quantitatively similar and were comprised of straight chain saturated and unsaturated non-hydroxylated fatty acids with hexadecanoic acid (16:0) and octadecenoic acid (18:1) the major components. All of the strains contained very characteristic polar lipid patterns consisting of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, five partially identified glycolipids and an uncharacterised phospholipid. Analyses of wall amino acid preparations using gas chromatography showed that Gardnerella vaginalis strains contain major amounts of alanine, glycine, glutamic acid and lysine. The chemical data support the integrity of the genus Gardnerella.     

Effect of glutamic acid on the fatty acid and lipid composition of Choanephora cucurbitarum.

The fatty acid composition of the total, neutral, sterol, free fatty acid, and polar-lipid fractions in the mycelium of Choanephora curcurbitarum was determined. The major fatty acids in all lipid fractions were palmitic, oleic, linoleic, and gamma-linolenic acid. Different lipid fractions did not show any particular preference for any individual fatty acid; however, the degree of unsaturation was different in different lipid fractions. Free fatty acid and polar lipid fractions contained a higher proportion of gamma-linolenic acid than did triglyceride and sterol fractions. Addition of glutamic acid to the malt-yeast extract and medium resulted in the biosynthesis of a number of long-chain fatty acids beyond the gamma-linolenic acid. These fatty acids, e.g., C22:1, C24:0, and C26:0, were never observed to be present in the fungus when grown on a malt-yeast extract medium without glutamic acid. Furthermore, thin-layer chromatographic analysis showed a larger and denser spot of diphosphatidyl glycerol from the mycelium grown on glutamic acid medium than from the control mycelium. The possible significance of this finding is discussed.     

Fatty acid,menaquinone and polar lipid composition of Rothia dentocariosa

The lipid compositions of Rothia dentocariosa was investigated. All of the strains tested possessed closely related lipid profiles consisting of predominantly straight-chain saturated and methyl branched long-chain fatty acids, unsaturated menaquinones with seven isoprene units and a polar lipid composition comprising diphosphatidylglycerol, phosphatidylglycerol and a diglycosyldiacylglycerol. The results of the present study indicate Rothia dentocariosa is a good and distinct taxon. The lipid data however does not support the classification of Rothia dentocariosa in the family Actinomycetaceae.     

Liver microsomal membrane lipid composition in marasmic-kwashiorkor

The microsomal membrane cholesterol and phospholipid content and phospholipid composition of marasmic kwashiorkor rats have been compared with those of normal rats. A Significant increase in the cholesterol/phospholipid ratio, as well as in the sphingomyelin/phosphatidyl-choline ratio was observed in the marasmic-kwashiorkor rat. These effects would tend to decrease the fluidity of the phospholipid bilayer of the endoplasmic reticulum membrane and may thus affect drug metabolism.It is well known that a change in the quality or quantity of dietary protein causes an alteration in the rates of metabolism of many xenobiotics by the mammalian liver (1–3). These metabolic alteration have been attributed mainly to changes in the levels of microsomal membrane proteins themselves, especially that of cytochrome P-450 (4–6). However, a recent report by Suzuki et al. (7) indicates that the more subtle features of drug metabolism such as interactions between NADPH-cytochrome P-450 reductase, cytochrome P-450, cytochrome b and other specific drug metabolzing enzymes in the membrane of the endoplasmic reticulum might well be affected by the fluidity of the phospholipid bilayer.There is still a high incidence of protein-energy malnutrition (PEM) diseases such as kwashiorkor in many part of the world (8). The membrane lipid composition from microsomes of marasmic-kwashiorkor rats have therefore been investigated with a view to finding out if there are any changes in these components due to protein deficiency which could contribute to the decreased metabolism of xenobiotics in this condition.     

Phospholipid composition and fatty acid desaturation in the roots of rye during acclimatization of low temperature

When the roots of rye plants grown at 20°C were cooled to 8°C the concentration of phospholipid in them more than doubled over a 7 d period in comparison with that in roots remaining at 20°C. The relative abundance of lecithin (PC) declined while that of phosphatidyl ethanolamine (PE) increased; this change was completed after 2 d cooling. Labelling with ³²P suggested that turnover of phospholipids may be inhibited by low temperature. Acyl lipids contained an increased proportion of linolenic acid (18:3) and reduced proportion of linoleic acid (18:2) when roots were cooled at 8°C for 7 d. The ratio of these acids is a relatively more sensitive indicator of desaturation than is the double bond index. Cooling brought about no change in the abundance of the principal saturated acid, palmitic (16:0). In the first 3 days of cooling PC and PE desaturated markedly while there was no change in galactosyl and neutral lipids. Desaturation did not appear to be greatly sensitive to the concentration of dissolved O₂ and was only partly inhibited in 8°C solutions where the oxygen concentration was lowered to 0.5–2.0%. Positional analysis of acyl chains in PC and PE showed that more than 90% of all 16:0 is associated with position I while 65% of the 18:2+18:3 is associated with position II. When roots are cooled the abundance of 18:3 increases in both chains but the relative distribution of saturated and unsaturated fatty acids remains constant in positions I and II. At both 20°C and 8°C there is a high probability that a saturated chain in position I will be paired with the polyunsaturated one in position II.Abbreviations PC Lecithin - PE phosphatidyl ethanolamine - TLC thin layerchromatography - BHT butylatedhydroxytoluene     

Ethanol sensitivity and fatty acid composition of chick liver mitochondrial and microsomal membranes

Fatty acid composition of hepatic mitochondrial and microsomal membranes was studied in 2-day-old chicks exposed to ethanol for 60 h (short treatment) or 18 days (chronic treatment). Short ethanol treatment induced in mitochondria an increase in the 18:1/18:0 ratio as a consequence of both an increase in the percentage of oleic and a decrease in that of stearic acid. Likewise, a clear decrease in the polyunsaturated fatty acids and in the 20:4/18:2 ratio was found in mitochondria after short ethanol administration. Microsomal membranes were practically unaffected by this treatment. However, chronic ethanol exposure produced a significant increase in the percentages of polyunsaturated fatty acids in both mitochondria and microsomes as well as a decrease in the 18:1/18:0 ratio. These results suggest that delta 9 desaturase modifies its activity in response to ethanol treatment with a different pattern to those showed by delta 6 and delta 5 desaturase activities.     

Fatty acid transport into the brain: of fatty acid fables and lipid tails

The blood-brain barrier formed by the brain capillary endothelial cells provides a protective barrier between the systemic blood and the extracellular environment of the central nervous system. Brain capillaries are a continuous layer of endothelial cells with highly developed tight junctional complexes and a lack of fenestrations. The presence of these tight junctions in the cerebral microvessel endothelial cells aids in the restriction of movement of molecules and solutes into the brain. Fatty acids are important components of biological membranes, are precursors for the biosynthesis of phospholipids and sphingolipids and are utilized for mitochondrial β-oxidation. The brain is capable of synthesizing only a few fatty acids. Hence, most fatty acids must enter into the brain from the blood. Here we review current mechanisms of transport of free fatty acids into cells and describe how free fatty acids move from the blood into the brain. We discuss both diffusional as well as protein-mediated movement of fatty acids across biological membranes.