Purification and Properties of a Mitochondrial Lipoprotein Inhibitor of Sterol Synthesis

A lipoprotein inhibitor of hydroxymethylglutaryl CoA reductase (EC 1.1.1.34) and of cholesterol synthesis by rat liver homogenates, was isolated from the mitochondria of starved rats’ livers. The isolated lipoprotein complex contained a low molecular weight protein and fatty acids. The fatty acids identified were arachidonic, linoleic, oleic, stearic and palmitic. The saturated fatty acids and oleic acid did not inhibit. Inhibition of the enzyme was to a large extent related to the degree of fatty acid unsaturation.     

Selective retention of n-3 and n-6 fatty acids in human milk lipids in the face of increasing proportions of medium chain-length (C10-14) fatty acids

This paper reports the results of our analysis of the impact high levels of de novo fatty acids have on the proportions of essential and non-essential fatty acids in human milk lipids. The data for seven fatty acids (linoleic, alpha-linolenic, arachidonic (AA), docosahexaenoic (DHA), palmitic, stearic and oleic) were derived from several studies conducted in Nigeria. The proportion by weight of each of these fatty acids was plotted versus the proportion of C10-14 fatty acids. As the proportion of C10-14 fatty acids increased from 15 to 65%, there was not a proportional decrease in the percentages of all seven fatty acids, but, instead, preferential incorporation of the essential fatty acids, AA and DHA into the triacylglycerol component of the milk. At the same time, the proportions of stearic and oleic acid declined by 69% and 86%, respectively. However, the proportions of linoleic acid, palmitic acid, DHA, AA and alpha-linolenic acid, in milk lipids decreased by only 44%, 40%, 39%, 28% and 2.3%, respectively. These observations indicate that as the contribution of C10-14 fatty acids increases, essential fatty acids are preferentially incorporated into milk triacylglycerols at the expense of oleic acid and stearic acid.     

Neutral lipids, phospholipids and higher fatty acids in bull's testes

The lipid fractions were studied in the testicular tissue of mature bulls, of the lowland black-and-white breed. It was found that the main component of neutral lipids was cholesterol (48%) followed by triglycerides (24%), cholesterol esters (16%) and free fatty acids (12%). In cholesterol esters the main component was palmitic acid (41%) followed by oleic acid (22%), stearic acid (14%) and linoleic acid (14%). In phospholipids the main fraction was composed of lecithins (48%) followed by phosphatidylethanolamine (20%) and phosphatidic acids and phosphatidylglycerol (13%). Palmitic acid was found mainly in the fractions of lecithins and sphingomyelins, stearic acid in fractions of phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol. Linoleic acid was found in the fractions of phosphatidylethanolamine, phosphatidylcholine and sphingomyelin. Arachidonic, docosatetraenoic and docosapentaenoic acids in the fractions of phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and phosphatidylcholine.     

Relationship between polyreactive properties of immunoglobulins and level of lipids in them

Presented paper deals with the relationship between immunoglobulin polyreactive properties and its lipid composition. Serum blood immunoglobulin fraction of an intact rabbit as an experimental model was used. Immunoglobulins (Ig) obtained by this way were transformed into polyreactive immunoglobulins (PRIg) by treatment with chaotropic agent KSCN or reactive oxygen species (ROS) with usage of Fe2+, EDTA and ascorbic acid. It was demonstrated that native Ig were able to bind with immobilized antigen (ovalbumin) and this ability dramatically increased after transformation of Ig into PRIg. The high immunoreactivity of PRIg was associated with marked fall (by 2-3 fold) of total phospholipids as well as individual ones--sphingomyelin and phosphatidylcholine. The main fatty acids of the Ig and PRIg phospholipid fractions in the sequence to decreasing decrease were stearic, palmitic, oleic and linoleic acids. The treatment of Ig by chaotropic agent and ROS led to decrease of stearic acid and enhancement of oleic, linoleic and arachidonic acids. The level of free cholesterol of Ig did not differ from that of PRIg. At the same time the content of cholesterol esters of PRIg was substantially diminished if compare with Ig. The main fatty acids of the Ig and PRIg cholesterol ester fraction in the sequence to decreasing were arachidonic, stearic, oleic, linoleic and palmitoleic acids. Transformation of Ig into PRIg was accompanied by enhancement of stearic acid level and loss of docosapentaenoic, arachidonic and palmitoleic acids. The results presented here support the idea about non peroxidative manner of the phospholipid and cholesterol ester extrusion from Ig molecule under its transformation into PRIg. Rather the last process could be explained by the term of concurrent physico-chemical interaction of Ig molecule with chaotropic agent or ROS leading to fall of lipid content. The presented data for the first time provide us an opportunity to conclude that transformation of Ig into PRIG is associated with the marked loss of essential phospholipids and cholesterol esters by the Ig molecule. The probable implication of this process in development of immune imbalance under certain diseases associated with oxidative stress have been discussed.     

The long-chain fatty acid composition of yeasts used in the brewing industry

Summary The cellular long-chain fatty acids of 69 strains of yeasts, representing 29 species associated with the brewing industry, were extracted by saponification and analyzed asmethyl esters by gas chromatography. The strains were characterized by the presence of palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acid as the major fatty acids. The strains were divided into six groups on the basis of their fatty acid content. With this method it was possible to differentiate between the yeasts on species and, in some instances, on strain level.     

Sulfoquinovosyldiglyceride and Triglycosyldiglyceride in Rice Grains

Sulfoquinovosyldiglyceride and triglycosyldiglyceride were isolated from rice grains, mainly by column chromatography and thin-layer chromatography. The major component fatty acids were palmitic, oleic, stearic and linoleic acids in sulfoquinovosyldiglyceride whereas they were linoleic, palmitic and oleic acids in triglycosyldiglyceride, in decreasing order of amount. The component sugar in sulfoquinovosyldiglyceride was sulfoquinovose, while those in triglycosyldiglyceride were galactose and glucose, the latter being predominant.     

Composition and synthesis of fatty acids in atherosclerotic aortas of the pigeon

The composition, synthesis, and esterification of fatty acids were studied in aortas of White Carneau and Show Racer pigeons after perfusion of the aortas with a medium containing acetate-1-(14)C. For both breeds of pigeons the principal change in aortic fatty acids, in response to an atherogenic diet, was a marked increase in the percentage of oleic acid in the cholesteryl ester fraction. In atherosclerotic aortas incorporation of acetate-1-(14)C into the phospholipid and glyceride fractions increased 2-fold, while a much greater increase (up to 10-fold) was seen in incorporation into cholesteryl esters. In those birds receiving the atherogenic diet, palmitic acid accounted for approximately 50% of the fatty acid radioactivity, compared with approximately 25% from control aortas. Calculation of fatty acid synthesis showed the major newly synthesized fatty acids to be stearic acid in the phospholipid fraction; stearic, palmitic, and oleic acids in the glycerides; and oleic acid in the cholesteryl esters. The pattern of fatty acid synthesis was closely similar to the actual fatty acid composition of the aorta. In atherosclerotic aortas an increased synthesis of all fatty acids was seen, but the greatest increase was seen in the synthesis of oleic acid and its esterification to cholesterol.     

Steroid 5alpha-reductase inhibitory activity and hair regrowth effects of an extract from Boehmeria nipononivea

The acetone extract of Boehmeria nipononivea showed both potent 5alpha-reductase inhibitory activity and hair regrowth promotion effects on mice. 5alpha-Reductase inhibitory activity-guided fractionation led to six active fatty acids: alpha-linolenic, linoleic, palmitic, elaidic, oleic and stearic acids. The extract of B. nipononivea, and alphalinolenic, elaidic and stearic acids exhibited a hair regrowth effect.     

The characterization and distribution of hexahydropolyprenyl esters in cultures of Aspergillus fumigatus Fresenius

The total mycelial lipid of Aspergillus fumigatus was analysed and over half of its hexahydropolyprenol was shown to be esterified with fatty acids. Comparison of the fatty acid content of the prenyl esters with the sterol ester and the total lipid indicated a marked predominance of saturated fatty acids in the polyprenyl esters. The predominant acids esterified to the prenols were palmitic acid, linoleic acid, oleic acid, lignoceric acid, stearic acid and palmitoleic acid. Most of the unesterified polyprenol was found in the mitochondrial fraction, but the esterified prenol was equally distributed throughout the cell fractions. This distribution was unlike that found for ergosteryl ester in the same tissue.     

Differential absorption and esterification of dietary long‐chain fatty acids by larvae of the dragonfly,Aeshna cyanea

In order to evaluate whether dietary long‐chain fatty acids were differentially absorbed, Aeshna cyanea larvae received 5 μl oral doses containing combinations of two radiolabeled fatty acids at nearly equal radioactive and nmolar concentrations: (1) ³H‐oleic and ¹⁴C‐palmitic acids; (2) ³H‐oleic and ¹⁴C‐stearic acids; and (3) ³H‐palmitic and ¹⁴C‐stearic acids. After 3 h or 1 day, hemolymph samples, midgut tissue, midgut contents and fat body tissue were collected and assayed for labeled fatty acids. The ³H/¹⁴C ratios indicated that there was a preference for absorption of the monounsaturated oleic acid over both saturated palmitic and stearic acids and that the shorter palmitic acid was absorbed at a higher rate than the longer stearic acid. There were also differences in the ³H/¹⁴C ratios of the various lipid classes of the midgut wall, hemolymph, and fat body that reflected differential esterifications and transport of these fatty acids. Arch. Insect Biochem. Physiol. 40:183–193, 1999. © 1999 Wiley‐Liss, Inc.     

Kinetic profile of the cellular lipid composition in an oleaginous Yarrowia lipolytica capable of producing a cocoa-butter substitute from industrial fats

Cell growth, lipid accumulation and cellular lipid composition of Yarrowia lipolytica growing on mixtures of industrial fats containing stearic, oleic, linoleic and palmitic acid have been studied. During growth, the strain incorporated oleic and linoleic acids more rapidly than the saturated fatty acids. Relatively high lipid accumulation (up to 0.44 g of lipids per g of dry matter) was observed when stearic acid was included in the culture medium. In contrast, substrates rich in oleic acid did not favor cellular lipid accumulation. The accumulated lipids, mainly composed of triacylglycerols (45-55% w/w), demonstrated a different total fatty acid composition compared with that of the substrate; in all cases, the microorganism showed the unusual capacity to increase its cellular stearic acid level, even if this fatty acid was not found in high concentrations in the substrate. This permitted the synthesis of interesting lipid profiles with high percentages of stearic acid and non-negligible percentages of palmitic and oleic acid, with a composition resembling that of cocoa-butter.     

Effects of fatty acids on lysis of Streptococcus faecalis.

Palmitic, stearic, oleic, and linoleic acids at concentrations of 200 nmol/ml all inhibited autolysin activity 80% or more in whole cells or cell-free extracts. This concentration of the saturated fatty acids palmitic acid and stearic acid had little or no effect on the growth of whole cells or protoplasts. However, the unsaturated fatty acids oleic acid and linoleic acid induced lysis in both situations. This lytic effect is apparently not related to any uncoupling activity or inhibition of energy catabolism by unsaturated fatty acids. It is concluded that unsaturated fatty acids induce cell and protoplast lysis by acting as more potent membrane destabilizers than saturated fatty acids.     

Fatty Acid Transport and Utilization for the Developing Brain

Abstract: To determine the transport and utilization of dietary saturated, monounsaturated, and n-6 and n-3 polyunsaturated fatty acids for the developing brain and other organs, artificially reared rat pups were fed a rat milk substitute containing the perdeuterated (each 97 atom% deuterium) fatty acids, i.e., palmitic, stearic, oleic, linoleic, and linolenic, from day 7 after birth to day 14 as previously described. Fatty acids in lipid extracts of the liver, lung, kidney, and brain were analyzed by gas chromatography-mass spectrometry to determine their content of each of the deuterated fatty acids. The uptake and metabolism of perdeuterated fatty acid lead to the appearance of three distinct groups of isotopomers: the intact perdeuterated, the newly synthesized (with recycled deuterium), and the natural unlabeled fatty acid. The quantification of these isotopomers permits the estimation of uptake and de novo synthesis of these fatty acids. Intact perdeuterated palmitic, stearic, and oleic acids from the diet were found in liver, lung, and kidney, but not in brain. By contrast, perdeuterated linoleic acid was found in all these organs. Isotopomers of fatty acid from de novo synthesis were observed in palmitic, oleic, and stearic acids in all tissues. The highest enrichment of isotopomers with recycled deuterium was found in the brain. The data indicate that, during the brain growth spurt and the prelude to myelination, the major saturated and monounsaturated fatty acids in brain lipids are exclusively produced locally by de novo biosynthesis. Consequently, the n-6 and n-3 polyunsaturated fatty acids must be transported and delivered to the brain by highly specific mechanisms.     

Lipid Metabolism of Rumen Ciliates and Bacteria: II. Uptake of Fatty Acids and Lipid Analysis of Isotrichia intestinalis and Rumen Bacteria with Further Information on Entodinium simplex

The total lipid and free fatty acid contents of Isotricha intestinalis, Entodinium simplex, and the rumen bacterial flora of the respective protozoa were determined. Warburg manometric data showed that the sodium salts of tributyrin, oleic, and acetic acids stimulated gas production in I. intestinalis, whereas tributyrin was stimulatory with E. simplex and less active with oleic and acetic acids. Rumen bacteria provided fatty acids produced lower manometric gaseous increases when compared with the protozoa. Volatile fatty acids were produced by I. intestinalis and rumen bacteria with tributyrin, but not with tripalmitin. Sodium oleate gave little volatile fatty acid response with I. intestinalis or rumen bacteria. Washed suspensions of I. intestinalis and rumen bacteria concentrated C¹⁴-labeled oleic, palmitic, stearic, and linoleic acids within the cells during short incubation periods. Autoradiographs demonstrated the conversion of C¹⁴-labeled oleic, palmitic, stearic, linoleic, and acetic acids in the rumen protozoa and bacterial cells.     

Platelet membrane fatty acids in thrombocytosis due to myeloproliferative disorders

The fatty acid composition of platelet membranes has been analysed in patients with thrombocytosis due to myeloproliferative disorders, who had not taken any drugs. A significant increase in palmitic and oleic acid, together with a decrease in stearic, linoleic and arachidonic acids was observed. The fatty acid pattern of platelet membranes was also analysed in patients during treatment with ASA (acetylsalicylic acid). ASA ingestion completely normalizes the platelet content of palmitic acid and partially that of stearic and arachidonic acid, whereas it has no effect on the level of linoleic acid and raises that of oleic acid. The altered pattern of fatty acids observed in patients may interfere with platelet function by decreasing membrane fluidity. Treatment of patients with ASA seems to act on platelet membranes by partially normalizing the fatty acid composition.     

Spectrum of total fatty acids in cerebrospinal fluid determined by gas chromatography

A simple gas—liquid chromatographic method for the determination of the spectrum of fatty acids in a small volume of cerebrospinal fluid (CSF) is presented. In a group of 49 neurological patients it has been found that in the CSF of the controls (n = 12) there are the following main fatty acids: oleic (27.28%), palmitic (23.23%), stearic (12.21%), linoleic (7.66%), myristic (5.02%), and palmitoleic (4.51%). Altogether 28 fatty acid methyl esters (FAMEs) from 12:0 to 22:2 have been tentatively identified. The majority of them appeared irregularly, sometimes in less than in 10% of cases. The composition of FAMEs in the CSF of patients with lumbar discopathy and with acute ischaemic cerebrovascular accidents does not differ from the control group. A significant difference (P<0.01) has been found in the group of hypophyseal adenomas in which the amounts of practically all saturated FAMEs with an odd carbon number are elevated. The same applies to the 18:0 and 20:0 compounds. In the group of atrophic and degenerative CNS processes the palmitic and stearic acids predominated to the detriment of oleic and linoleic acids.     

Cholesterol esters in developing rat brain: concentration and fatty acid composition

Abstract— The contents and the fatty acid composition of cholesterol esters were analysed in developing rat brain. The total content did not exceed 20 μg/brain throughout development. Elimination of serum by adequate perfusion was essential for accurate results. Two separate events appeared to affect the levels of cholesterol esters in developing rat brain, one probably reflecting general developmental changes and the other apparently related to myelination. On either a unit weight or a whole brain basis, the curves appeared to be a superimposition of the two events. There was an underlying developmental change, which was characterized on a unit weight basis by the highest level of cholesterol esters immediately after birth and a steady decline to the adult level by 30 days of age or which on the basis of whole brain was characterized by a steady increase throughout the development. A period of transient increase was superimposed on this underlying developmental change between the ages of 7 and 27 days and corresponded to the period of active myelination. The major fatty acids of rat brain cholesterol esters were palmitic, palmitoleic, oleic and arachidonic acids. Palmitic and palmitoleic acids decreased in proportion while oleic acid increased, as the animal matured. The fatty acid composition of serum cholesterol esters was distinctly different from that of brain cholesterol esters; those from serum contained much higher proportions of linoleic and arachidonic acids and much less palmitoleic and oleic acids.     

Effect of fatty acids on the synthesis and secretion of apolipoprotein B by rat hepatocytes

The modulation of apolipoprotein B synthesis and secretion by fatty acids in rat hepatocytes was studied. Maximum apolipoprotein B production was obtained in the case of oleic acid followed by linoleic, stearic and palmitic/linolenic acid when compared to control which was not supplemented with any fatty acids. Oleic acid was found to exert a concentration dependent increase in the secretion of [³H] apolipoprotein B into the medium while that associated with the cell layer was not affected. Pulse chase experiments in the presence of oleic acid showed that it caused an increase in the secretion of apolipoprotein B into the medium.¹⁴C-acetate incorporation into cholesterol and cholesteryl ester associated with the cell layer and secreted very low density lipoproteins also showed an increase in the presence of oleic acid indicating an increase in cholesterogenesis. The effect of oleic acid on [³H] apolipoprotein B and very low density lipoproteins secretion appeared to be mediated through cholesterol as (i) ketoconazole, an inhibitor of cholesterol synthesis caused significant reduction in the stimulatory effect of oleic acid on apolipoprotein secretion and (ii) mevinolin, another inhibitor of cholesterol synthesis also reversed the stimulatory effect of oleic acid on apolipoprotein B secretion. These results indicated that oleic acid may influence apolipoprotein B synthesis and secretion in hepatocytes probably by affecting cholesterol/cholesteryl ester formation which may be a critical component in the secretion of apolipoprotein B as lipoproteins     

Macrophages transfer [14C]-labelled fatty acids to pancreatic islets in culture

Macrophages are able to produce, export, and transfer fatty acids to lymphocytes in culture. The purpose of this study was to examine if labelled fatty acids could be transferred from macrophages to pancreatic islets in co-culture. We found that after 3 h of co-culture the transfer of fatty acids to pancreatic islets was: arachidonic > oleic > linoleic = palmitic. Substantial amounts of the transferred fatty acids were found in the phospholipid fraction; 87.6% for arachidonic, 59.9% for oleic, 53.1% for palmitic, and 36.9% for linoleic acids. The remaining radioactivity was distributed among the other lipid fractions analysed (namely polar lipids, cholesterol, fatty acids, triacylglycerol and cholesterol ester), varying with the fatty acid used. For linoleic acid, a significant proportion (63.1%) was almost equally distributed in these lipid fractions. Also, it was observed that transfer of fatty acids from macrophages to pancreatic islets is time-dependent up to 24 h, being constant and linear with time for palmitic acid and remaining constant after 12 h for oleic acid. These results lead us to postulate that in addition to the serum, circulating monocytes may also be a source of fatty acids to pancreatic islets, mainly arachidonic acid.     

Triglyceride metabolism in germinating Andropogon gayanus seeds

Seed triglycerides of Andropogon gayanus contained 17 fatty acid moieties, principally palmitic, oleic and linoleic acids. These were distributed in an essentially random manner amongst the triglycerides to form the following main types: POL, PLL, OOL, LLO and LLL. Triglycerides decreased during both light and dark germination but there was no evidence for selective hydrolysis. Free fatty acids appear to be derived from triglyceride hydrolysis but the free and triglyceride fatty acid composition differed. Less palmitic, oleic and linoleic acids and more stearic, linolenic and C₂₀-acids were found in the free state than combined in the triglycerides. Free fatty acids did not accumulate during germination.