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At least 19 recordsLinked to original sources

Effects of dietary lipids on behaviour, lipid biosynthesis and lipid composition, in rat platelets.

Rats of either sex were fed for 18 and 34 weeks respectively diets containing 40% (by weight) lipids with polyunsaturated fatty acids representing 1.34% or 13.2% of total calories. Platelet reactivity to thrombin, platelet fatty acid composition and incorporation of [14C]acetate into platelet lipids were investigated. Diets rich in saturated fatty acids markedly increased platelet sensitivity to thrombin. The concentration of 20:3 and 22:3 of the (n - 9) series and of 20:3 and 22:5 of the (n - 6) series were increased at the expense of 18:2 and 22:4 of the (n - 6) family in platelet lipids. 20:4 (n - 6) was unchanged. The fatty acid changes were more pronounced in male rats and after 34 weeks. [14C]Acetate incorporation into total platelet lipids and particularly into choline phosphoglycerides and ceramides was lower in animals fed saturated fats. This diet reduced the synthesis of 16:0 and of 22:4(n - 6) in platelet total fatty acids, while that of 22:3(n - 9) was markedly enhanced. This study showed that long-term feeding of high-saturated-low-polyunsaturated fat diets in rats induced marked changes in platelet lipid synthesis and composition, in both sexes. The lipid synthesis modification appears to be more pronounced in males than in females. The changes in the fatty acids 20:3(n - 9), 22:3(n - 9) and 22:4(n - 6) appeared to be closely related to platelet behaviour. The balance between the content and synthesis of these last fatty acids might be of significance for the effect of diet on thrombogenesis.

Acetates↗

[Influence of carbon source on lipid biosynthesis by Candida gluilliermondii].

The biosynthesis of lipids by Candida guilliermondii was studied during the growth of the yeast on carbon substrates which resulted in different pathways of lipid biosynthesis. The highest content of lipids was found during the growth on octadecane. The quantitative ratio between fractions of neutral lipids and phospholipids depended on the carbon substrate whereas the quanlitative composition of the fractions remained the same. The fatty acid composition of lipid fractions was studied, and possible pathways of their biosynthesis are discussed. Apparently, the assimilated hydrocarbon is oxidized to acetyl residues in certain structures of the yeast cells while, in other structures, the hydrocarbon substrate undergoes only monoterminal oxidation yielding an aliphatic alcohol and an acid which are directly used in large amounts for the synthesis of wax and triglycerides.

Acetates↗

Influence of sex and dietary fats on platelet lipid biosynthesis in rat.

The platelet biosynthesis of total lipids, lipid fractions and fatty acids was determined by incorporation of [14C]acetate in normal and castrated rats of both sexes. Comparison was made between animals fed laboratory chow alone, and animals receiving, in addition, for 4 days by stomach tube a saturated (cream) or polyunsaturated (sunflower seed oil) fat. In male rats, the polyunsaturated fat increased slightly the total platelet lipid biosynthesis. The saturated fat drastically reduced it by 43% in comparison to the polyunsaturated fat-fed animals. Normal female rats did not exhibit a similar difference in the platelet lipid synthesis. However, the inhibitory effect of saturated fat on lipid synthesis could be observed in castrated females, although it was less pronounced (27% reduction) than in castrated or normal males (43%). Administration to castrated males of estradiol for 1 month almost completely inhibited the difference induced by the feeding of the different fats in the lipid platelet synthesis of male rats. This difference in the platelet lipid biosynthesis between male and female rats, normal and castrated, was observed mostly in the phospholipid (especially phosphatidylcholine), monoacylglycerol and triacylglycerol fractions and affected primarily the synthesis of the three main saturated fatty acids, 14:0, 16:0 and 18:0. Thus, it seems that, in rat, the short-term administration of a saturated fat induces drastic changes in the platelet lipid biosynthesis, but only in males. The protection observed in females appears to be essentially dependent upon estrogens.

Acetates↗

Epidermal lipid biosynthesis in acne.

Lipid biosynthesis from glucose was increased in severe acne both in dermis and in epidermis which included the top portions of the pilosebaceous follicles. Both tissues retained their characteristic lipid labelling pattern. The magnitude of the increase in dermis and epidermis was similar, suggesting that epidermis may be subject to the same sebotrophic-lipogenic stimulus which affects sebaceous glands.

Acne Vulgaris↗

Differential lipid biosynthesis underlies a tradeoff between reproduction and flight capability in a wing-polymorphic cricket.

The biochemical basis of life-history tradeoffs is a poorly studied aspect of life-history evolution. We used radiotracer and endocrine approaches to investigate the extent to which morphs of a wing-polymorphic insect differ in the biosynthesis of lipid classes important for dispersal capability vs. reproduction (ovarian growth). The flight-capable genotype of Gryllus firmus biosynthesized a greater amount of total lipid and triglyceride (main flight fuel), which was preferentially allocated to somatic tissue during early adulthood. By contrast, the flightless genotype biosynthesized a significantly greater amount of phospholipid (important in egg development), which was preferentially allocated to ovaries. Topical application of a juvenile-hormone mimic to the flight-capable morph caused it to express all aspects of lipid metabolism seen in the flightless morph. Differences in biosynthesis between morphs (i) occur coincident with 100-400% greater ovarian growth in the flightless morph, (ii) result from alterations of both de novo biosynthesis of fatty acid and downstream partitioning of fatty acids into triglyceride vs. phospholipid, and (iii) possibly result from genetically polymorphic hormonal regulators with negative pleiotropic effects on lipid biosynthesis and ovarian growth. The present study provides direct documentation of genetically based alterations of in vivo flux through pathways of intermediary metabolism leading to the differential production of end products central to the specialization of phenotypes for alternate life histories.

Animals↗

Glycogen content and lipid biosynthesis in the lungs of fetuses of diabetic rats.

The fetal pulmonary glycogen content and the pulmonary biosynthesis of neutral lipids and phospholipids were estimated at gestational days 18, 20, and 22 in the offspring of normal, manifest diabetic, and insulin-treated diabetic rats. In all groups the fetal pulmonary glycogen concentration was highest on gestational day 20 and decreased during the subsequent 2 days. At the same time the biosynthesis of neutral lipids and phospholipids increased in the fetal lungs. The fetuses of the manifest diabetic rats showed an increased glycogen concentration and decreased total lipid biosynthesis compared with the other two groups. Insulin treatment of the rat mothers largely normalized the pulmonary lipid biosynthesis and glycogen accumulation in their fetuses. These data suggest that the diabetic maternal environment induces a transient block in the fetal utilization of pulmonary glycogen for the biosynthesis of lipids, in particular for the production of surfactant phospholipids.

Animals↗

Localization of sites of lipid biosynthesis in mammalian epidermis.

The end-product of epidermal differentiation is a stratified layer of corneocytes whose extracellular lipid bilayers provide a permeability barrier. It is generally accepted that the epidermis synthesizes most if not all of the lipids found in this tissue and that extra-epidermal tissues contribute very little to this lipid content. Moreover, the individual epidermal strata in which epidermal lipid biosynthesis occurs are not known. To address this question, we examined [3H]H2O incorporation into nonsaponifiable and saponifiable lipids in individual epidermal cell layers 3 hr after intraperitoneal injection into neonatal mice, and compared this to protein and DNA synthesis using intraperitoneal [3H]leucine and [3H]thymidine incorporation, respectively. Lipid biosynthesis was also assessed by [14C]acetate incorporation into lipid fractions in organ cultured skin and in epidermal subpopulations. The in vivo studies demonstrated that the biosynthetic activity of both saponifiable and nonsaponifiable lipids was comparable to, if not greater, in the stratum granulosum (SG) than in basal/spinous (SB + SS) layer, despite significantly lower levels of both protein and DNA synthesis in the SG. On a mass basis, the SG accounts for about four times the biosynthetic activity of the combined SB + SS layers. The lipid biosynthetic activity in vitro also was two- to fivefold higher in the SG, regardless of whether the epidermis was separated into individual cell layers before or after incubations with radiolabel. Moreover, this difference could not be ascribed to increased acetate pools or to elevated metabolism in the SG versus the SB + SS since the rates of CO2 production were much lower in the SG fraction. The increase in lipid biosynthesis in SG over SB + SS was greatest for phospholipids, followed by glycosphingolipids, and free sterols but was observed in almost all lipid classes. These studies show not only that mammalian epidermis is an active site of de novo lipid biosynthesis, but also that this activity remains high in the stratum granulosum, while other forms of metabolic activity are diminishing. These observations are consistent with the knowledge that lipids extruded from the stratum granulosum layer provide the hydrophobic permeability barrier, and further suggest that elevated synthetic activity in the stratum granulosum would allow rapid replenishment in the event that the barrier is damaged.

Animals↗

Characterization of the precursor of tetraether lipid biosynthesis in the thermoacidophilic archaeon Thermoplasma acidophilum.

Polar lipid biosynthesis in the thermoacidophilic archaeon Thermoplasma acidophilum was analyzed using terbinafine, an inhibitor of tetraether lipid biosynthesis. Cells of T. acidophilum were labeled with [(14)C]mevalonic acid, and their lipids were extracted and analyzed by two-dimensional thin-layer chromatography. Lipids labeled with [(14)C]mevalonic acid, [(14)C]glycerol, and [(32)P]orthophosphoric acid were extracted and hydrolyzed under different conditions to determine the structure of polar lipids. The polar lipids were estimated to be archaetidylglycerol, glycerophosphatidylcaldarchaetidylglycerol, caldarchaetidylglycerol, and beta- l-gulopyranosylcaldarchaetidylglycerol, the main polar lipid of T. acidophilum. Pulse and chase experiments with terbinafine revealed that one tetraether lipid molecule is synthesized by head-to-head condensation of two molecules of archaetidylglycerol and that a sugar group of tetraether phosphoglycolipid is expected to attach to the tetraether lipid core after head-to-head condensation in T. acidophilum. A precursor accumulated in the presence of terbinafine with a fast-atom-bombardment mass spectrometry peak m/z 806 was compatible with archaetidylglycerol. The relative height of the peak m/z 806 decreased after removal of the inhibitor. The results suggest that most of the precursor, archaetidylglycerol, is in fully saturated form.

Chromatography, Thin Layer↗

Early action of prolactin on ornithine decarboxylase activity is not essential for the subsequent actions of prolactin on casein and lipid biosynthesis.

The actions of prolactin (PRL) on casein and lipid biosynthesis in cultured mouse mammary gland explants require the ongoing synthesis of the polyamines. This is supported by the fact that (MGBG) methylglyoxal bis(guanylhydrazone), a drug that inhibits the conversion of putrescine to spermidine, abolishes the effects of PRL on casein and lipid biosynthesis; the inhibitory effects of MGBG are reversed by the addition of spermidine to the culture medium. alpha-Difluoro methyl ornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase activity, reduces the PRL-stimulated ornithine decarboxylase (ODC) activity by more than 95%, and yet does not suppress the effects of PRL on RNA, casein or lipid synthesis. These observations suggest that PRL's early action on ODC activity is not essential for the subsequent actions of PRL on the synthesis of certain of the components of milk.

Acetates↗

Temperature-sensitive Saccharomyces cerevisiae mutant defective in lipid biosynthesis.

A temperature-sensitive mutant of Saccharomyces cerevisiae (DAM303) is described that exhibits an early defect in lipid biosynthesis at the restrictive growth temperature, 37 degrees C. This strain rapidly lost viability after 1 h of incubation at 37 degrees C, and this was accompanied by a significantly reduced incorporation of 32Pi into cellular lipid and an accumulation of [1-14C]acetate into the free fatty acid fraction. The temperature-sensitive DAM303 mutation failed to complement the sec13 mutation described by Novick et al. (Cell 21:205-215, 1980), and from analysis of invertase secretion in the temperature-sensitive DAM303 strain, it is clear that the loss of invertase secretion in the mutant occurs after the loss of phospholipid synthesis. Although the precise nature of the temperature-sensitive lesion in the DAM303 strain has still to be identified, the results from the study of this mutant indicate that a defect in lipid biosynthesis can be correlated with subsequent alterations in extracellular protein secretion and loss of other macromolecular functions including DNA, RNA, and protein syntheses. From studies of this mutant, two procedures of enriching for other temperature-sensitive mutants with defects in lipid biosynthesis have emerged: inositol overproduction and screening for increased buoyant densities.

Centrifugation, Density Gradient↗

Control analysis of lipid biosynthesis in tissue cultures from oil crops shows that flux control is shared between fatty acid synthesis and lipid assembly.

Top-Down (Metabolic) Control Analysis (TDCA) was used to examine, quantitatively, lipid biosynthesis in tissue cultures from two commercially important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). A conceptually simplified system was defined comprising two blocks of reactions: fatty acid synthesis (Block A) and lipid assembly (Block B), which produced and consumed, respectively, a common and unique system intermediate, cytosolic acyl-CoA. We manipulated the steady-state levels of the system intermediate by adding exogenous oleic acid and, using two independent assays, measured the effect of the addition on the system fluxes (J(A) and J(B)). These were the rate of incorporation of radioactivity: (i) through Block A from [1-(14)C]acetate into fatty acids and (ii) via Block B from [U-(14)C]glycerol into complex lipids respectively. The data showed that fatty acid formation (Block A) exerted higher control than lipid assembly (Block B) in both tissues with the following group flux control coefficients (C):(i) Oil palm: *C(J(TL))(BlkA)=0.64+/-0.05 and *C(J(TL))(BlkB)=0.36+/-0.05(ii) Olive: *C(J(TL))(BlkA)=0.57+/-0.10 and *C(J(TL))(BlkB)=0.43+/-0.10where *C indicates the group flux control coefficient over the lipid biosynthesis flux (J(TL)) and the subscripts BlkA and BlkB refer to defined blocks of the system, Block A and Block B. Nevertheless, because both parts of the lipid biosynthetic pathway exert significant flux control, we suggest strongly that manipulation of single enzyme steps will not affect product yield appreciably. The present study represents the first use of TDCA to examine the overall lipid biosynthetic pathway in any tissue, and its findings are of immediate academic and economic relevance to the yield and nutritional quality of oil crops.

Culture Techniques↗

Impaired membrane traffic in defective ether lipid biosynthesis.

The first steps of ether lipid biosynthesis are exclusively localized to peroxisomes and hence some peroxisomal disorders are characterized by a severe deficiency of plasmalogens, the main ether lipids in humans. Here we report on gene defects of plasmalogen biosynthesis, chromosomal localization of the corresponding genes and, as a consequence of plasmalogen deficiency, on structural alterations of caveolae, clathrin-coated pits, endoplasmic reticulum and Golgi cisternae, as well as on the reduced rate of transferrin receptor cycling. The data suggest that plasmalogens, analogous to cholesterol, are essential for correct membrane functioning and their deficiency results in impaired membrane trafficking.

Acyltransferases↗

Oral contraceptive and Platelet lipid biosynthesis in female rats: dose-response relationship.

Female rats were treated with different doses of an oral contraceptive (ethinyl estradiol + lynestrenol) and lipid biosynthesis was studied in blood platelets by acetate incorporation into different fractions separated by thin layer chromatography. A marked increase in lipid biosynthesis was observed, especially in the sterol fractions (cholesterol and lanosterol-dihydrolanosterol). It was dose-dependent, observed after a lag-phase, maximal in 3 days and normalized in 8 days. Thus, the oral contraceptive studied here appears to modify platelet lipid biosynthesis for the entire life of the platelets.

Acetates↗

Cell adhesion to fibronectin regulates membrane lipid biosynthesis through 5'-AMP-activated protein kinase.

We have shown that attachment to a fibronectin substrate stimulates two pathways of lipid biosynthesis in cultured human fibroblasts. Detachment of these cells (mechanically, with trypsin, or by RGDS peptides) caused a significant decrease in their 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity and in their incorporation of [3H]acetate into fatty acids. This inhibition was substantially reversed by the reattachment of cells to fibronectin substrates, but not to poly-L-lysine substrates or to fibronectin in solution. Inhibiting phosphoprotein phosphatase activity with okadaic acid blocked the recovery of both biosynthetic activities. Both 3-hydroxy-3-methylglutaryl-coenzyme A reductase and fatty acid biosynthesis are known to be inhibited by the action of 5'-AMP-activated protein kinase, which is activated by an increase in the level of AMP relative to ATP. For example, in our system, sodium azide and 2-deoxy-D-glucose increased the ratio of cellular AMP to ATP and caused a decrease in lipid biosynthesis. We then verified the prediction that detachment of cells from substrates also caused an increase in the AMP/ATP ratio. We therefore conclude that the attachment of cells to fibronectin promotes lipid biosynthesis, presumably in coordination with the cellular growth response evoked by attachment to the extracellular matrix.

AMP-Activated Protein Kinases↗

Lipid biosynthesis, oxidative enzyme activities and cellular changes in growing olive fruit.

Fruit of Olea europea L. was examined by light and electron microscopy to determine whether commencement of lipid accumulation depended upon the fruit achieving structural maturity. Maturation of fruit develops progressively from the smallest changes towards the largest in cellular structures. Important metabolic and structural changes have been observed: oil body formation, changes in the structural and reserve lipid biosynthesis and in the fatty acid of total lipid content, as well as in G6PDH and LOX activities. The labelling of fruit lipids by previously incubating the leaves with (1-14C)-acetate and (1,5-14C)-citrate or by putting the labelled substrates directly on the fruit surface, shows a 14C assimilate derived from acetate greater than that from citrate; the incorporation of the latter is higher in the methanol-water fractions. At the beginning of fruit development the lipid biosynthesis with both substrates is greater in polar lipids; on the contrary, the incorporation of 14C into neutral lipids increases during fruit maturation. Additionally, a maximum of substrate export from leaves to fruit coincides with an increase in the lipoxygenase and, above all, in the glucose-6-phosphate dehydrogenase activities. The transported 14C from leaves begins its activity before the small oil bodies close to the tonoplast can be observed in the fruit, and well before the beginning of maturation. The results suggest that structural development and some other rate controlling metabolic steps can govern the initiation of lipid accumulation in olive fruit.

Glucosephosphate Dehydrogenase↗

The significance of adipost tissue and liver as sites of lipid biosynthesis in the turkey.

A study was conducted to determine the significance of adipose tissue and liver as sites of lipid biosynthesis in the turkey. Seven-week-old turkey males were injected intravenously with either acetate-U-14C or glucose-U-14C. Ten, 30, 60, and 120 minutes after injection, adipose tissue, liver and plasma samples were taken. Total lipids were extracted from these samples and 14C incorporation into the lipids was determined. Acetate-U-14C incorporation into liver lipid was 516.2, 493.2, 382.7 and 364.7 x 103 d.p.m./gm. lipid as compared to 2.7, 2.9, 3.1 and 1.3 x 103 d.p.m./gm. lipid for adipose tissue after 10, 30, 60 and 120 minutes, respectively. The incorporation of glucose-U-14C into liver lipid was 27.4, 63.4, 157.8 and 243.6 x 103 d.p.m./gm. lipid after 10, 30, 60 and 120 minutes, respectively; whereas glucose-U14C incorporation into adipose tissur was 1.1, 1.3, 3.1 and 5.9 x 103 d.p.m./gm. lipid after the same time intervals. The results of this study showed the liver is the primary site of lipid biosynthesis and acetate was converted into lipid more rapidly than glucose.

Acetates↗