Characterization of lipid-laden aortic cells from cholesterol-fed rabbits. II. Morphometric analysis of lipid-filled lysosomes and lipid droplets in aortic cell populations.
Explore the source record for details and available documents.
Animals↗
SEARCH · Search PubMed
Results for “Lipid”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Chemically induced lipid phase separation in model membranes containing charged lipids: a spin label study.
The lipid distribution in binary mixed membranes containing charged and uncharged lipids and the effect of Ca2+ and polylysine on the lipid organization was studied by the spin label technique. Dipalmitoyl phosphatidic acid was the charged, and spin labelled dipalmitoyl lecithin was the uncharged (zwitterionic) component. The ESR spectra were analyzed in terms of the spin exchange frequency, Wex. By measuring Wex as a function of the molar percentage of labelled lecithin a distinction between a random and a heterogeneous lipid distribution could be made. It is established that mixed lecithin-phosphatidic acid membranes exhibit lipid segregation (or a miscibility gap) in the fluid state. Comparative experiments with bilayer and monolayer membranes strongly suggest a lateral lipid segregation. At low lecithin concentration, aggregates containing between 25% and 40% lecithin are formed in the fluid phosphatidic acid membrane. This phase separation in membranes containing charged lipids is understandable on the basis of the Gouy-Chapman theory of electric double layers. In dipalmitoyl lecithin and in dimyristoyl phosphatidylethanolamine membranes the labelled lecithin is randomly distributed above the phase transition and has a coefficient of lateral diffusion of D = 2.8-10(-8) cm2/s at 59 degrees C. Addition of Ca2+ dramatically increases the extent of phase separation in lecithin-phosphatidic acid membranes. This chemically (and isothermally) induced phase separation is caused by the formation of crystalline patches of the Ca2+-bound phosphatidic acid. Lecithin is squeezed out from these patches of rigid lipid. The observed dependence of Wex on the Ca2+ concentration could be interpreted quantitatively on the basis of a two-cluster model. At low lecithin and Ca2+ concentration clusters containing about 30 mol % lecithin are formed. At high lecithin or Ca2+ concentrations a second type of precipitation containing 100% lecithin starts to form in addition. A one-to-one binding of divalent ions and phosphatidic acid at pH 9 was assumed. Such a one-to-one binding at pH 9 was established for the case of Mn2+ using ESR spectroscopy. Polylysine leads to the same strong increase in the lecithin segregation as Ca2+. The transition of the phosphatidic acid bound by the polypeptide is shifted from Tt = 47.5 degrees to Tt = 62 degrees C. This finding suggests the possibility of cooperative conformational changes in the lipid matrix and in the surface proteins in biological membranes.
Calcium↗
Lipid peroxidation of the microsomal fraction and extracted microsomal lipids from DAB-induced hepatomas.
NADPH- and ascorbic acid-induced microsomal lipid peroxidation was almost absent in subcutaneously implanted DAB-induced hepatomas D23, D30 and D192A, and present at greatly reduced levels in DAB-induced primary hepatomas when compared with normal liver controls. Fatty acid analysis of the microsomal lipid from passaged tumours demonstrated adequate levels of substrate in the phospholipid fractions to support lipid peroxidation. Lipid extracted from hepatoma microsomal fractions was shown to undergo ascorbic acid-induced lipid peroxidation, but to a lesser extent that the corresponding liver extract. This may be partially explained by a decrease in the phospholipid content of hepatoma microsomal membranes. However, phospholipid extracted from microsomal fractions of hepatoma and liver supported lipid peroxidation to a similar extent. The possible role of the non-lipid component of the membrane in the process of lipid peroxidation is discussed.
Animals↗
Membrane lipid metabolism in Acholeplasma laidlawii A EF 22. Influence of cholesterol and temperature shift-down on incorporation of fatty acids and synthesis of membrane lipid species.
1. Membrane lipid metabolism in Acholeplasma laidlowii A EF 22 has been studied under different conditions by applying three different techniques for changing membrane viscosity: fatty acid and cholesterol supplementation and temperature changes. 2. The molar relationship between the two dominating membrane lipids, monoglucosyldiglyceride and diglucosyldiglyceride, is to a large extent determined by membrane viscosity properties. This is shown by the varying metabolic responses occurring during incorporation of different fatty acids with and without cholesterol and by temperature shift-down experiments. Higher viscosity in membranes stimulates synthesis of monoglucosyldiglyceride at the expense of diglucosyldiglyceride. Synthesis of phospho and phosphoglucolipids is affected as well. 3. Temperature shift-down from 37 degrees C to 17 degrees C results in an immediate synthesis of monoglucosyldiglyceride accompanied by an increased incorporation of unsaturated fatty acids into this lipid. Synthesis of the other membrane lipid species (containing more unsaturated fatty acids) lags behind temporarily. 4. Incorporation from an equimolar mixture of palmitic and oleic acids together with cholesterol yields greater amounts of oleic acid in membrane lipids than incorporation in the absence of cholesterol, indicating that incorporation is viscosity dependent. 5. Studies of precursor relationships reveal that all main lipids have an active turnover which differs depending on membrane composition and conditions. Furthermore, this turnover proceeds with different intra-lipid pools. 6. Isolated membranes contain no detectable lipolytic enzymes capable of hydrolyzing membrane phospho or glycolipids. It is suggested that lipid turnover is partly mediated by enzymatic interlipid conversions, thus not allowing intermediates to accumulate.
Acholeplasma laidlawii↗
Cholesterol-lipid interactions in membranes. The saturation concentration of cholesterol in bilayers of various lipids.
1. The integration of cholesterol in a lipid bilayer can be visualized by changes in the fluorescence properties of the probe N-phenyl-1-naphthylamine (NPN). An increasing cholesterol content in the lipid phase corresponds to decreasing fluorescence intensity of NPN and a short wave shift of the emission spectrum. 2. Equilibrium constants for the partition of NPN between water and the various lipid phases are reported. An increasing cholesterol content in a bilayer decreases the solubility of NPN in the bilayer. 3. The saturation concentration of cholesterol in bilayers of various lipids prepared by ultrasonication is determined using the flourescence probe NPN. The maximal molar ratio of cholesterol : lipid is 2 : 1 for sphingomyelin or egg phosphatidylcholine and 1 : 1 for cerebroside, dipalmitoyl phosphatidylcholine, or dipalmitoyl phosphatidylethanolamine. 4. The comparison of the maximal molar ratio of cholesterol : lipid with the number of proton donor and proton acceptor sites in the lipid moiety is used for a discussion of the polar interactions of cholesterol within a lipid bilayer.
1-Naphthylamine↗
Replacement time for alveolar lipid removed by pulmonary lavage: effects of multiple lavage on lung lipids.
Daily washing in vivo of the lung with 0.15 M saline did not deplete the Beagle dog lung of surfactant lipids, but rather increased the quantity of surfactant lipid in the tissue. Replacement time for the lung lipids removed by the lavage was approximately 5 hours. This rate is one indication of the time required for movement of surfactant lipid from storage areas to the surface of the alveoli. The increase in tissue surfactant lipid following multiple lavage suggests that the rate of surfactant lipid synthesis is controlled in part by the level of surfactant lipid in the alveoli.
Animals↗
A method for microdetermination of major neutral lipids and an application of the procedure to tissue lipids.
Cholesterol, cholesteryl esters, triglycerides and fatty acids as major neutral lipids and phospholipids were examined in quantitative analysis. The method consisted of three steps: (1) separation of lipids by one-dimensional thin-layer chromatography on silica gel plates; (2) elution of neutral lipids from scraped silica gel with chloroform-methanol (4:1); and (3) colorimetric determination of individual neutral lipids in eluates and phospholipids in silica gel. The conditions were modified for chromotropic acid reaction for determining triglycerides. Laurell's method for determining fatty acids was also modified to apply to quantitative thin-layer chromatography. The accuracy of the modified methods was well-defined as the absorbance values were on a linear curve. A quantitative study was made of the recovery of triglycerides and fatty acids after chromatography. Combining these modified methods and colorimetry for determination of cholesterol cholesteryl esters and phospholipids, the author established a micromethod for determining the major neutral lipids and phospholipids by thin-layer chromatography. Lipids from HeLa, S-3 cells were analyzed to examine the applicability of this method to tissues. The results indicated that the new method permitted a reliable estimation of the major neutral lipids and phospholipids from small amounts of tissues.
Cholesterol↗
The binding of lipid to the lipid-free adenosine triphosphatase protein of sarcoplasmic reticulum.
Dinitrophenylated dipalmitoyl phosphatidylethanolamine and its lyso derivative have been shown to bind to the lipid-free ATPase protein derived from the sarcoplasmic reticulum. The binding of these lipids is accompained by the quenching of up to 95% of the tryptophyl fluorescence of the protein. This effect is reversed by 9-10 mM deoxycholate. The solubility of the lipid-free ATPase protein in the absence of deoxycholate and the solubility of submillimolar concentrations of the dinitrophenylated monopalmitoyl phosphatidylethanolamine anion in aqueous media allowed binding experiments using this lipid ligand to be carried out in a simple buffer system. It is shown that in the case of this lipid the initial phase of the binding process displays an apparent positive co-operatively. Data from the second phase in the saturation of the protein with this lipid is consistent with binding to independent, equivalent, non-interacting sites with a microscopic (intrinsic) association constant of 1.63 x 10(6) M-1, the fluorescence being quenched in the geometric fashion. Altogether a total of about 15 molecules of this lipid may be bound by the protein.
Adenosine Triphosphatases↗
Effects of force-feeding and dietary cellulose on liver lipid accumulation and lipid composition of liver and plasma in growing chicks.
Changes of content and composition of lipid in liver and plasma affected by force-feeding and dietary cellulose were investigated in 14-day old Single-Comb White Leghorn male chicks. They were given a purified high energy diet (starch-casein diet without fiber) supplemented with or without dietary cellulose. Chicks were fed ad libitum or force-fed the experimental diet. Force-feeding of excess food improved the growth rate of chicks and feed efficiency, but feeding of cellulose did not affect body weight gain and feed efficiency, though a slight improvement in nitrogen retention was observed. Liver weight and lipid content in liver and plasma were markedly elevated by force-feeding, and were markedly depressed by dietary cellulose in the force-fed chicks. It is suggested that changes of liver lipid by force-feeding and dietary cellulose are mainly originated from the changes of triglyceride in the liver lipid. No marked changes were observed in fatty acid composition of abdominal fat and liver lipid in the cellulose-fed chicks. These results suggest that dietary cellulose may affect lipid metabolism in growing chicks.
Animals↗
Remodeling of host lipid metabolism by Wolbachia strain wAlbB is associated with lipid accumulation and cardiolipin dysregulation in the Aedes aegypti fat body.
BACKGROUND: The intracellular symbiont Wolbachia, particularly the wAlbB strain, is a promising biocontrol agent against mosquito-borne diseases. Although Wolbachia infection is known to perturb host metabolism, the underlying mechanisms, especially those related to lipid metabolism, remain poorly understood. METHODS: We performed an integrated multi-level analysis of the Aedes aegypti fat body in uninfected and wAlbB-infected mosquitoes, combining histology, biochemistry, untargeted liquid chromatography-mass spectrometry (LC-MS) lipidomics, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis, reverse transcription quantitative PCR of key metabolic genes, and quantification of acetyl-coenzyme A (acetyl-CoA) and reduced nicotinamide adenine dinucleotide (NADH) levels. RESULTS: wAlbB infection increased fat body wet weight and thickness, accompanied by accumulation of triglyceride and of lipid droplets. Lipidomic analysis further revealed extensive lipidome remodeling, with elevated free fatty acid, diglyceride, and triglyceride, but broad depletion of glycerophospholipids, particularly cardiolipin. These changes were supported by transcriptional alterations: upregulation of fatty acid synthase 1 and glycerol-3-phosphate acyltransferase 1, and downregulation of adipose triglyceride lipase and carnitine palmitoyltransferase 1. Cardiolipin depletion correlated with downregulation of genes involved in its synthesis and remodeling, including phosphatidylglycerophosphate synthase and calcium-independent phospholipase A2γ. These lipid changes were also associated with accumulation of acetyl-CoA and NADH. CONCLUSIONS: Our findings suggest that wAlbB infection is associated with extensive lipid metabolic remodeling in the Aedes aegypti fat body, characterized by accumulation of neutral lipids and cardiolipin depletion, accompanied by transcriptional remodeling of key metabolic enzymes. This study establishes the fat body as a primary tissue-level hub for Wolbachia-associated lipid remodeling and provides a foundational framework for future mechanistic investigations into host-symbiont metabolic interactions.
Animals↗
Relation of fluorescence in lipid-containing red cell membrane extracts to in vivo lipid peroxidation.
Previous studies have demonstrated that the abnormal fluorescent peak consistent with the crosslinking of red cell membrane constituents by the lipid-peroxide decomposition product MDA is demonstrable in lipid-containing extracts of red cells obtained from patients treated with the oxidizing hemolytic agent diaminodiphenylsulfone. The present studies were primarily aimed at ascertaining the specificity of this fluorescence as an indicator of in vivo red cell lipid peroxidation. Repetitive injection of phenylhydrazine or acetylphenylhydrazine in rats resulted in gradually increasing levels of fluorescence despite the return in hematocrit toward normal. Chromatography on Sephadex LH-20 of lipid-containing extracts obtained from the red cells of rats and rabbits treated with phenylhydrazine revealed fluorescent peaks similar to those observed in red cell lipid incubated with MDA. In addition, increased levels of fluorescence were observed in the extracts of red cells obtained from three vitamin E--deficient premature infants. These findings suggest that the fluorescence assay is a sensitive indicator of in vivo red cell lipid peroxidation.
Animals↗
Lipid-protein interactions in membranes: effect of lipid composition on mobility of spin-labeled cysteine residues in yeast plasma membrane.
In order to gain direct evidence for lipid-dependent protein conformation in membrane, effects of modification of lipid composition on mobility of spin-labeled cysteine residues were investigated in the plasma membrane of the yeast Saccharomyces cerevisiae. Conversion of the bulk of phospholipids to diglycerides by treatment of the membrane with phospholipase C substantially enhanced spectral anisotropy. However, alterations of the viscosity of the lipid-bilayer by enriching the membrane with palmitelaidic or oleic acid had no effect on mobility of spin-labeled cysteine residues. These observations indicate that while the spin-labeled residues are not in direct contact with the lipid core of the membrane, there are lipid-protein interactions to the extent that removal of the polar portion of the bulk of phospholipids induces conformational changes in proteins, which in turn restrict mobility of these residues. It is concluded that conformation of membrane proteins on lipid structure and that phospholipids have a role in preserving the native conformation of proteins.
Adenosine Triphosphatases↗
Lipid chemotaxins isolated from culture filtrates of Escherichia coli and from oxidized lipids.
Lipid extracts of sterile culture filtrates of Escherichia coli were shown to contain approximately 75% of the chemotactic activity for human polymorphonuclear leukocytes and rabbit alveolar macrophages. Fractionation and purification of these lipids revealed the presence of many unknown lipids of widely different properties, but all were anionic and at very low concentrations, chemotactic. The only one of active molecules that could be identified was an unsaturated ultraviolet-absorbing hydroxy fatty acid, which, following catalytic reduction with hydrogen, was found to be hydroxyeicosanoic acid. This fatty acid's chromatographic behavior was very similar to that of 12-hydroxy-5,8,10,14-eicosatetraenoic acid (HETE), which is a potent chemotaxin for polymorphonuclear leukocytes and macrophages. Unknown chemotaxins could be generated by the oxidation of known unsaturated lipids. Prostaglandins A2 and E2 produced potent chemotaxins upon aerobic oxidation. Malonaldehyde, a peroxidation product of unsaturated lipids, when reacted with phosphatidylethanolamine in aerobic conditions, also produced strong chemotactic agents. The chemotactic activity of these products could be destroyed by catalytic reduction with hydrogen and by methylation with dry methanolic HCl. These data indicate that the nonenzymatic oxidation of unsaturated lipids generates some products that are potent chemotaxins for mammalian inflammatory cells.
Animals↗
Liquid chromatographic analysis of sebum lipids and other lipids of medical interest.
A technique is described for the high-pressure liquid chromatographic (HPLC) analysis of sebum lipid classes. The lipid present in sebum are separated by gradient elution HPLC from a microparticulate silica column and detected using a moving-wire detector. The system described can be linked to a computer. Quantitation can be carried out by comparing peak areas obtained with those of an internal standard. Peak trapping for further investigations of the separated components, for example by gas chromatography-mass spectrometry, is very easy. Sebum lipids are separated into the following lipid classes: hydrocarbons and squalene, cholesterol esters and wax esters, fatty acids as their methyl esters, triglycerides, 1,3-diglycerides, 1,2-diglycerides, free cholesterol, monoglycerides and other polar materials. Besides to sebum, the method has been successfully applied to other lipid mixtures, such as serum lipids. Examples of other applications are shown.
Chromatography, High Pressure Liquid↗
Diol lipids of rat liver. Quantitation and structural characteristics of neutral lipids and phospholipids derived from ethanediol, propanediols, and butanediols.
Specific enzymatic and chemical degradation of neutral lipid and phospholipid fractions from rat liver revealed the presence of novel types of lipid metabolites bearing a short-chain diol backbone. Diol-derived lecithin and cephalin analogs were readily cleaved by phospholipase C (EC 3.1.4.3) from Bacillus cereus, although the cephalin analogs required "carrier" lecithin to sustain hydrolysis. The products of phosphilipase hydrolyses as well as the neutral lipid fractions were subjected to alkaline and acidic methanolysis, and constituent short-chain diols were analyzed as long-chain cyclic acetals. Gas chromatographymass spectrometry confirmed that 1,2-ethanediol, 1,2-propanediol, 1,3-propanediol, and 1,3 butanediol can form the polyol backbone of neutral lipids and phospholipids. [1,1,2,2-2H]Ethanediol monohexadecanoate, dihexadecanoate, hexadecanoylphosphorylcholine, hexadecanoylphosphorylethanolamine were synthesized chemically and served as internal standards to assure accurate quantitation of the low levels of diol lipids (350 mug/g ot total lipid) present in rat liver.
Animals↗
Lipid deficient extender for bovine spermatozoa: its development and use in measuring freezing-induced lipid loss.
To facilitate the measurement of lipid losses from spermatozoa due to freezing, three low-lipid seminal extenders containing lactose, bovine serum albumin, or soybean protein were evaluated as potential cryoprotectants. All extenders were formulated to have an osmotic pressure within the range of 270 to 330 mosmol and a pH of 6.8 to 7.0. Soybean protein (Promine-D) maintained the highest post-thaw motility of spermatozoa with similar survival for spermatozoa frozen in ampules and straws. The extender derived from testing several components consisted of Tris(hydroxymethyl) aminomethane (245 mM), and citric acid monohydrate (78mM), as the buffering compounds; and fructose (69 mM), glycerol (7% vol/vol), and Promine-D (1.5% wt/vol). Post-thaw sperm motility of approximately 40% was not different from the Tris-egg yolk control. Fertility of fresh rabbit semen treated with the extender was normal. After freeze-thawing, protected spermatozoa contained more lipid (1.61 versus 1.20 mug/10-6 sperm) and lost less glutamic oxaloacetic transaminase enzyme (102 versus 108 Karmen units) than when Promine-D was not incorporated. However, even with protection by soybean protein, spermatozoa lipid content decreased from 2.43 to 1.61 mug/10-6 sperm after one freeze-thawing. The lipid status of spermatozoa frozen and thawed in conventional bull seminal extenders containing large amounts of lipids is unknown.
Animals↗
[Comparative study of lipid A from pertussis microbes differing in the toxicity of their O-antigens. I. Chemical composition and stability of the bond between lipid A and the specific polysaccharide in B. pertussis lipopolysaccharide].
Data presented in this work indicated that antigens, contrastive by toxicity, obtained by Boiven's method and O'Neill and Tood's method from two strains of Bordetella pertussis differed by stability of lipid A binding with the specific polysaccharide. The influence of duration of the lipopolysaccharide hydrolysis on the fatty acid content in lipid A, and of heptose in the specific polysaccharide was demonstrated. Lipid A fatty acid composition was studied. It is supposed that bound fatty acids are presented as C14 and C19--C22. There was a correlation between the antigen toxicity and the stability of lipid A bond with the specific polysaccharide. Stability of the lipopolysaccharide complex bond depended on heptose and lipid A content and on the composition and the amount of fatty acids in the lipid A preparations.
Antigens, Bacterial↗
Biosynthesis of lipid-linked oligosaccharides in cotton fibers. Stimulation by lipids from pig liver.
An acidic lipid fraction isolated from pig liver (Forsee, W. T. & Elbein, A.D. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 2574-2578) stimulated the incorporation of mannose from GDP-[14C]mannose into lipid-linked oligosaccharides using a particulate enzyme fraction from maturing cotton bolls. This lipid fraction did not stimulate mannose incorporation into the mannosylphosphorylpolyprenol. The 14C-oligosaccharides, formed in the presence or absence of the pig liver "acceptor" lipid, were released from the lipid-linked oligosaccharides by mild acid hydrolysis and were isolated by paper chromatography. Both sets of 14C-oligosaccharides had similar mobilities suggesting that they were qualitatively similar, except that those formed in the presence of "acceptor" lipid had much more radioactivity. The individual oligosaccharides were purified on a calibrated column of Sephadex G-25 and were then subjected to various treatments to obtain information about their structures. The molecular weights of the larger oligosaccharides ranged from about L1210 to 1720 indicating they probably contained from 6 to 10 sugar residues. Strong acid hydrolysis of the oligosaccharide with a molecular weight of 1350, followed by reduction with NaB3H4, gave a ratio of [3H]hexitol to [3H]hexosaminitol that was compatible with the molecular weight (about 5:2). However, the hexitol fraction contained small amounts of [3H]glucitol in addition to [3H]mannitol. Thus, this oligosaccharide may contain small amounts of glucose in addition to mannose and GlcNAc. When the smaller oligosaccharides (i.e. those having 5 or 6 sugars) were treated with alpha-mannosidase, essentially of all the 14C was released as mannose, but only 30 to 50% of the radioactivity could be released from the larger oligosaccharides by this enzyme. Acetolysis of the oligosaccharide with a molecular weight of 1720 released the radioactivity as mannobiose and mannotriose suggesting that these oligosaccharides contain di- and trisaccharide branches linked to the main chain in 1 leads to 6 linkages.
Animals↗