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PubMedDate not supplied

Lipids in blood-brain barrier models in vitro II: Influence of glial cells on lipid classes and lipid fatty acids.

Lipids of brain tissue and brain microvascular endothelial cells contain high proportions of long-chain polyunsaturated fatty acids (long PUFAs). The blood-brain barrier (BBB) is formed by the brain endothelial cells under the inductive influence of brain cells, especially perivascular glia, and coculture of endothelial cells and glial cells has been used to examine this induction. The objective of this study was to investigate whether C6 glioma cells are able to influence the lipid composition and shift the fatty acid (FA) patterns of the BBB model cell lines RBE4 and ECV304 toward the in vivo situation. Lipid classes of the three cell lines were analyzed by thin-layer chromatography and lipid FA patterns by high-performance liquid chromatography. Only ECV304 cells showed altered lipid composition in coculture with C6 cells. The fractions of triglycerides and cholesteryl esters (depending on the support filter) were about twice as high in coculture as when the cells were grown alone. Triglyceride fractions reached 13 to 15% of total lipids in coculture. The three cell lines showed an increase in the percentage of long PUFAs with respect to unsaturated FAs, mainly because of an increase in the percentages of arachidonic acid, all cis-7,10,13,16-docosatetraenoic acid, and all cis-7,10,13,16,19-docosapentaenoic acid. It is concluded that glioma C6 cells are able to induce a more in vivo-like FA pattern in BBB cell culture models. However, changes were not significant for the individual PUFAs, and their levels did not reach in vivo values.

Animals↗
PubMed1988

Pulmonary oxygen toxicity in rats: prevention by pyrogenic diphosphoryl lipid A and potentiation by nontoxic monophosphoryl lipid A and lipid X.

Pretreatment with gram-negative bacterial endotoxin protects rats from the lethal intrathoracic edema produced by continuous exposure to greater than 95% O2 for 3 days. To help elucidate structure-activity relationships of endotoxin, we administered lipid part-structures of endotoxin intraperitoneally (at dosages up to 5.0 mg/kg) to male Sprague-Dawley rats prior to exposure to greater than 99% O2 (at 760 mm Hg) for 72 hr. We found that Salmonella minnesota mutant Re595 diphosphoryl lipid A protected rats from O2 toxicity as effectively as the parent S. minnesota endotoxin molecule at equimolar concentration. Diphosphoryl lipid A exhibited less acute toxicity (2 hr post-treatment hypothermia) than did endotoxin, although both produced prolonged fevers (greater than 24 hr) and similar patterns of protection in O2. In contrast, nontoxic lipid X (2,3-diacylglucosamine 1-phosphate) and monophosphoryl (1-dephospho) lipid A potentiated pulmonary O2 toxicity. We conclude that within the range of dosages investigated, diphosphoryl lipid A is the minimal essential structure responsible for the protection of rats from O2 toxicity by endotoxin.

Animals↗
PubMed1993

Contribution of hydrogen bonding to lipid-lipid interactions in membranes and the role of lipid order: effects of cholesterol, increased phospholipid unsaturation, and ethanol.

It is proposed that increased phospholipid unsaturation in membranes and perturbation by agents such as ethanol weaken interlipid hydrogen bonding involving water and that the process is independent of effects on lipid order. To investigate this, the rates of phospholipid desorption, as a measure of the strength of interlipid interactions, from "donor" lipid vesicles was determined. This was accomplished using (7-nitrobenzo-2-oxa-1,3-diazole-4-yl)aminohexanoate (C6-NBD) labeled phospholipids, the rate of desorption being followed from changes in fluorescence with time. The rates of desorption of the NBD-phospholipids from phosphatidylcholine (PC) donor vesicles was in the order phosphatidylcholine (PC) > phosphatidylserine (PS) > phosphatidylethanolamine (PE), the slower rates in the PS and PE reflecting direct interlipid hydrogen bonding. For PC, the interlipid hydrogen bonding was restricted to the "hydration layer", the network of hydrogen-bonded water molecules extending between phospholipid head groups. The rate of C6-NBD-PC desorption was elevated with higher levels of donor PC sn-2 unsaturation, due the increased head group spacing weakening the lipid-lipid interactions that occur via the hydration layer. Ethanol also increased the rate of NBD-phospholipid desorption from donor PC vesicles in the order PC > PS > PE, showing that PC interactions, here limited to the weaker hydrogen-bonded water molecule network, were more susceptible compared to stronger, direct interlipid hydrogen bonds involving PE and PS. The relative magnitude of the ethanol-induced increase in the desorption rate was amplified with higher levels of donor lipid sn-2 unsaturation. Cholesterol had little effect on the rate of phospholipid desorption.(ABSTRACT TRUNCATED AT 250 WORDS)

4-Chloro-7-nitrobenzofurazan↗
PubMed1983

Protein-mediated lipid transfer. The effects of lipid-phase transition and of charged lipids.

The protein-mediated phospholipid exchange between small unilamellar vesicles was investigated by fluorescence polarization measurements with diphenylhexatriene as optical probe. Thermotropic phase-transition measurements were taken after mixing two vesicle preparations of distinct and different phase-transition temperatures or having different states of charge. From the heights of each phase-transition step, we were able to follow the lipid-exchange process in the presence, as well as in the absence (natural exchange), of so-called transfer protein isolated from beef liver. A strong enhancement of the lipid transfer was observed at the corresponding lipid-phase-transition temperature, which is explained by the presence of fluctuating fluid and ordered domains co-existing at the lipid-phase-transition temperature. A unidirectional lipid transfer of the neutral component was observed between negatively charged phosphatidic acid and neutral phosphatidylcholine vesicles. Fluorescence polarization measurements showed the disappearance of the phosphatidylcholine phase transition, whereas the phosphatidic acid phase transition broadened and its phase transition temperature became lower.

Androgen-Binding Protein↗
PubMed1990

Effects of dietary lipidic charge in the concentration of bovine plasmatic lipids and lipoproteins. Its influence on the saturation degree of fatty acids' storaged lipids.

To estimate physiological changes due to added fat in bovine diet, whole cottonseed (19% lipids) was furnished to half-breed zebu cattle during 7 months. Thirty animals had increased its dietary lipidic charge in 2.6% (lot S), thirty were used as controls (lot C) and other two bovines were hyper-dosed with 7.9% added fat (lot H). Weight, clinical signs and plasmatic values of total cholesterol (TC), triglycerides (TG), lipoproteins (LP) and other biochemical parameters were monthly controlled. At the end of the assay the steers were sacrificed and the quality of meat and fat was evaluated. The initial levels of TC were statistically homogeneous in all groups (mean = 1.0 g/l); they remained without modifications in C (1.53 g/l), but they had increased progressively in supplemented animals. At the end of the work TC was greater in H (3.95 g/l) than in S (2.44 g/l). Latest TG (initial mean = 0.30 g/l) also were greater in H (0.74 g/l) than in S (0.71 g/l). The lipoproteinogram stayed inalterable in C, meanwhile alpha LP were increased and beta LP were decreased in animals on which the dietary lipidic charge had been increased. The weight's gain/animal/day (X) was 130 g (C), 290 g (S) and 88 g (H). The general physiological condition was deteriorated in the last lot. After comparing lots C and S, results indicated that beefsteaks in the last group were heavier (2.45 vs. 2.33 kg), with more lipids (0.47 vs. 0.36 kg) and major fat veined proportion (1.40 vs. 1.25%). The stored lipids showed high saturation degree in S, with increment in the mean percentage of stearic acid (28.6%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

Characteristics and lipid distribution of a large, high-risk, hypertensive population: the lipid-lowering component of the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT).

The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) consisted of 42,418 participants randomized to one of four antihypertensive treatment groups: chlorthalidone, amlodipine, lisinopril, or doxazosin. A subset of these participants with fasting low-density lipoprotein cholesterol levels 100-189 mg/dL were randomized into a lipid-lowering component: 5170 to receive pravastatin (40 mg daily) and 5185 to receive usual care. This report describes the characteristics and lipid distribution of these participants. There were no important differences between the randomized treatment groups. Women had higher total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol than men. There was a similar finding for black participants compared with whites, except blacks had lower triglycerides. Diabetics had lower high-density lipoprotein cholesterol and higher triglycerides than nondiabetics, and patients with body mass index <25 kg/m(2) had higher high-density lipoprotein cholesterol but lower low-density lipoprotein cholesterol and triglycerides than patients with higher body mass index. The success of the randomization of this large, diverse population and the differences in the lipid distributions among its subgroups will allow further understanding of optimal lipid-lowering treatment.

Aged↗
PubMed1976

Lipid-lipid and lipid-protein interactions as studied with a novel type of fluorescent fatty acid and phospholipid probes.

A novel fluorescent-labelled group of fatty acids and phospholipids has been applied to determine phase transitions in liposomes by fluorescence intensity and polarisation measurements. The chromophore of these amphiphilic lipids proved to be very suitable to demonstrate temperature-dependent lipid-lipid interactions. Liposomes from 1,2-dipalmitoyl-3-sn-glycero-phosphoethanolamine and from lipids isolated from membranes of E. coli K 1062 mutant grown on elaidic acid were used in these studies. These probes also made it possible to observe conformational changes in membrane proteins in isolated plasma membranes from this mutant. The changes in protein conformation were dependent on structural changes in the lipid phase.

Binding Sites↗
PubMed2003

Regulation of lipases by lipid-lipid interactions: implications for lipid-mediated signaling in cells.

Lipases are extracellular peripheral proteins that act at the surface of lipid emulsions stabilized, typically, by phospholipids. At a critical composition lipase activity toward substrates in phospholipid monolayers is discontinuously switched on by a small increase in substrate mole fraction. This occurs in part because lipase binding is inhibited by phospholipids. Binding of the lipase cofactor, colipase, is also inhibited by phospholipids. The initial rate of colipase binding increases abruptly at a substrate mole fraction that is approximately half the critical composition for lipase activity and just above that in substrate-phospholipid complexes. Moreover, complex collapse areas show an approximately 1:1 correlation with phospholipid excluded areas determined from an analysis of colipase adsorption rates. Thus, complexes inhibit colipase binding rate. Additionally, the switching of lipase activity likely occurs when uncomplexed substrate becomes the majority species in the interface. Lipase substrates, e.g. diacylglycerols, are typically the same lipids generated in the cytoplasmic surface of the plasma membrane of stimulated cells. As colipase binding is nonspecific and complexes involving lipase substrates form on the basis of lipid-lipid interactions alone, complexes should form in the plasma membrane of stimulated cells and may regulate protein translocation to the membrane.

Kinetics↗
PubMed2003

Perspectives from the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial--Lipid Lowering Trial and the Anglo-Scandinavian Cardiac Outcomes Trial--Lipid Lowering Arm.

PURPOSE OF REVIEW: The design, process and outcomes are compared between two large clinical trials of LDL cholesterol reduction with statin treatment in patients with known high blood pressure. This new information is placed in the context of previous clinical trials of cholesterol reduction, which have provided analyses of sub-groups with high blood pressure. RECENT FINDINGS: The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial failed to find a significant reduction of total mortality (primary endpoint), cardiovascular mortality or major cardiovascular events. This differed sharply from the Anglo-Scandinavian Cardiac Outcomes Trial, Lipid Lowering Arm, which was stopped before the planned termination due to a marked reduction (36%) in coronary death or myocardial infarction (primary endpoint). This trial also found significant reductions in stroke (27%) and separately, all major vascular events (21%). The two studies were similar in that they each contained over 10 000 participants with documented high blood pressure requiring drug therapy and they both used a fixed dose of a single statin. Pravastatin (40 mg/day) was used in the former and atorvastatin 10 mg/day in the latter. The major difference was that the control group in the Anglo-Scandinavian trial was treated with placebo with a double blind design whereas antihypertensive and lipid-lowering trial was open label with controls receiving usual care. SUMMARY: The benefit of achieving and maintaining significant LDL cholesterol reduction in patients with high blood pressure was convincingly demonstrated in the Anglo-Scandinavian trial. The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial failed to achieve similar success due to use of a less effective drug and loss of the differential effect with increasing statin treatment in the usual care control group.

Aged↗
PubMed2006

Sensitivity and specificity of pediatric lipid determinations for adult lipid status: findings from the Princeton Lipid Research Clinics Prevalence Program Follow-up Study.

OBJECTIVE: The goal was to determine the diagnostic utility of the National Cholesterol Education Program pediatric guidelines. METHODS: With the use of pediatric lipid data from the Cincinnati Clinic of the Lipid Research Clinics Prevalence Study and lipid and cardiovascular disease data collected for the same subjects as adults in the Princeton Follow-up Study, the sensitivity and specificity of the National Cholesterol Education Program pediatric guidelines were calculated overall and according to age. Furthermore, whether use of parental cardiovascular disease history during childhood influenced the sensitivity and specificity was assessed. RESULTS: Overall sensitivities were 43% to 46% and specificities were 82% to 86% for total and low-density lipoprotein cholesterol levels. There was considerable variation in sensitivities according to age, with the lowest sensitivities at ages 14 to 16 years and the highest sensitivities at ages 5 to 10 years and 17 to 19 years. Results were similar whether or not the population was restricted to children with a positive parental history of cardiovascular disease. CONCLUSIONS: Results of our analyses suggest that the sensitivity and specificity for evaluating total cholesterol or low-density lipoprotein cholesterol levels that are elevated in adulthood are not improved by selecting children with a positive parental history. These data also show the strong role that age (particularly the pubertal years between 10 and 15 years of age) plays in lipid measurements for children and adolescents. Continued prospective and longitudinal studies designed with age as well as other risk parameters are needed to determine the best guidelines for clinical screening in the future.

Adolescent↗
PubMedDate not supplied

Lipid composition of aorta of Watanabe heritable hyperlipemic and comparably hypercholesterolemic fat-fed rabbits. Plasma lipid composition determines aortic lipid composition of hypercholesterolemic rabbits.

Aortic and plasma lipid compositions were compared during a 12-month period in Watanabe heritable hyperlipemic (WHHL), comparably hypercholesterolemic fat-fed, and age-matched control normolipidemic rabbits to determine whether exposure to equivalent degrees of endogenous or exogenous hypercholesterolemia led to differences in the composition and concentration of lipids deposited in the artery wall. Although there were marked differences in the distribution of cholesterol among the lipoproteins in the WHHL versus the fat-fed rabbits, the contents of both free and esterified cholesterol were elevated to an equivalent degree in the aorta and plasma. In contrast, the triglyceride content of both the plasma and aorta were elevated only in the WHHL rabbits. However, aortic total phospholipids were increased in both the WHHL and fat-fed animals as compared to control rabbits. In the control rabbits, there was an age-dependent enrichment in aortic relative to plasma cholesteryl-oleate consistent with low density lipoprotein (LDL) receptor-directed intracellular cholesteryl ester processing. In contrast, enrichment in cholesteryl-oleate in aortae relative to plasma was not detected in either WHHL or fat-fed groups, suggesting that the plasma cholesteryl ester composition was the primary determinant of the aortic cholesterol composition. Thus, during chronic hypercholesterolemia, deposition of lipids in the artery wall appears to be determined by plasma lipoprotein levels and composition if the LDL receptor is either absent on a genetic basis or suppressed due to a high-fat, high-cholesterol diet.

Animals↗
PubMed1998

Lipids, lipid domains and lipid-protein interactions in endocytic membrane traffic.

It has long been known that lipids are responsible for the structural integrity of biological membranes, and confer specific dynamic properties to the bilayer. However, they have often been considered as passive components of the membrane, beyond their well-known function as second messengers in signal transduction pathways. Evidence has been accumulating that lipids are sorted along the routes of membrane traffic, and that they are directly involved in the regulation of protein sorting and membrane flow. In this review, we will discuss recent studies on the organization and possible functions of lipids in the endocytic pathway.

ADP-Ribosylation Factors↗
PubMed2000

Lipids, lipid modification and lipid-protein interaction in membrane budding and fission--insights from the roles of endophilin A1 and synaptophysin in synaptic vesicle endocytosis.

Studies on the endocytosis of synaptic vesicles have provided two novel insights into the mechanism of vesicle formation from donor membranes, both of which concern lipids. One is the essential role of endophilin, a cytosolic protein converting lysophosphatidic acid by addition of the fatty acid arachidonate into phosphatidic acid. The other is the essential role of membrane cholesterol, which specifically interacts with synaptophysin, the major transmembrane protein of synaptic vesicles. These findings reveal novel modes of membrane lipid modification and lipid-protein interaction in vesicle biogenesis.

Adaptor Proteins, Signal Transducing↗
PubMed1991

Dietary fiber and lipid peroxidation: effect of dietary fiber on levels of lipids and lipid peroxides in high fat diet.

Effect of feeding coconut and blackgram fiber isolated as neutral detergent fiber (NDF) on the levels of lipids and lipid peroxides was studied in rats given a high fat diet. Concentration of cholesterol, free falty acid and phospholipids showed significant decrease in the serum, liver aorta and intestine of coconut and blackgram fiber groups. Concentration of malondialdehyde (MDA) and conjugated dienes was significantly decreased in liver and intestine of both fiber groups, while hydroperoxides showed significant increase in liver and heart of both the fiber groups. SOD and catalase activity was found to be increased in liver, intestine, heart proximal colon and distal colon of both the fiber groups. Serum ceruloplasmin levels showed a slight increase in animals fed coconut and blackgram fiber groups. Glutathione levels in liver, intestine proximal colon, distal colon and heart also showed a significant decrease in the animals of both the fiber groups.

Animals↗
PubMed1990

Organic pesticides modify lipid-lipid and lipid-protein domains in model membranes. A laser Raman study.

The effect of hexachlorocyclohexane (all isomers) on the thermal transition properties of phospholipid liposomes was determined by Raman spectroscopy. Raman spectra of liposomes with and without the presence of hexachlorocyclohexanes were recorded in the C-H stretching region which shows three major bands around 2850, 2880 and 2930 cm-1. Thermal transition properties were estimated from plots of I2880/I2850 and or I2930/I2850 vs. temperature, where I represents the intensity of the respective band. Our data on phospholipid liposomes reveal that delta- and gamma-hexachlorocyclohexanes drastically reduce and broaden the main thermal transitions of phospholipids at toxic level concentrations. These effects are more pronounced in liposomes containing 18 or more carbon atom long acyl chains. Alpha- and beta-isomers at similar concentrations show a minimum effect on the thermal transition properties of phospholipids. Raman analysis of phospholipid liposomes containing melittin, interestingly, reveal that the delta-isomer unlike the gamma-isomer strongly alters the transition properties of boundary lipids. These data suggest that the effect of hexachlorocyclohexanes on the thermal transition properties of membranes is stereo specific and that the delta-isomer preferably disrupts the lipid-protein domains. Results are explained on the basis of the dynamic flexibility owing to the equatorial and axial chlorine atoms of various hexachlorocyclohexane isomers.

Chemical Phenomena↗
PubMed2004

Relationship between sterol/steroid structure and participation in ordered lipid domains (lipid rafts): implications for lipid raft structure and function.

The formation and stability of ordered lipid domains (rafts) in model membrane vesicles were studied using a series of sterols and steroids structurally similar to cholesterol. In one assay, insolubility in Triton X-100 was assessed in bilayers composed of sterol/steroid mixed with dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylcholine, or a 1:1 mixture of these phospholipids. In a second assay fluorescence quenching was used to determine the degree of ordered domain formation in bilayers containing sterol/steroid and a 1:1 mixture of DPPC and a quencher-carrying phosphatidylcholine. Both methods showed that several single modifications of the cholesterol structure weaken, but do not fully abolish, the ability of sterols and steroids to promote ordered domain formation when mixed with DPPC. Some of these modifications included a shift of the double bond from the 5-6 carbons (cholesterol) to 4-5 carbons (allocholesterol), derivatization of the 3-OH (cholesterol methyl ether, cholesteryl formate), and alteration of the 3-hydroxy to a keto group (cholestanone). An oxysterol involved in atherosclerosis, 7-ketocholesterol, formed domains with DPPC that were as thermally stable as those with cholesterol although not as tightly packed as judged by fluorescence anisotropy. It was also found that 7-ketocholesterol has fluorescence quenching properties making it a useful spectroscopic probe. Lathosterol, which has a 7-8 carbon double bond in place of the 5-6 double bond of cholesterol, formed rafts with DPPC that were at least as detergent-resistant as, and even more thermally stable than, rafts containing cholesterol. Because lathosterol is an intermediate in cholesterol biosynthesis, we conclude it is unlikely that sterol biosynthesis continues past lathosterol in order to create a raft-favoring lipid.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1979

Lipid protein interactions in mitochondria. VII. A comparison of the effects of lipid removal and lipid perturbation of the kinetic properties of mitochondrial ATPase.

We investigated the kinetics of mitochondrial ATPase in bovine heart mitochondria and submitochondrial particles upon treatment with phospholipase A2, or upon addition of n-butanol to perturb the lipid protein interactions. The changes observed are the following: (1) Lipid removal or perturbation with butanol is accompanied by loss of ATPase activity with decrease of both V and of the KM for ATP. (2) There are changes of activation energy of ATPase activity at temperatures above the discontinuity normally observed for membrane-bound enzymes in mitochondria. In particular, butanol abolishes the discontinuity, and induces a constant activation energy of about 32 kcal/mol in the range 8--37 degrees C. (3) Butanol modifies the pH dependence of ATPase shifting the pH optimum from around 10 to less alkaline values. The optimum for Mg2+ concentrations is increased by the solvent. (4) Treatment with phospholipase A2 results in a removal of oligomycin-sensitive ATPase, whereas butanol addition prevents oligomycin inhibition of ATPase. (5) In beef heart mitochondria, a spin-labelled analog of the inhibitor, dicyclohexyl carbodiimide, did not show any change in environment upon butanol addition, unlike that found in mitochondria from Saccharomyces cerevisiae.

Adenosine Triphosphatases↗
PubMedDate not supplied

Interaction of lipid and carbohydrate metabolism after infusions of lipids or of lipid lowering agents: lack of a direct relationship between free fatty acid concentrations and glucose disposal.

The present work was planned to study the effects of changes in lipid metabolism irrespective of FFA concentrations (FFA) on the regulation of oxidative and nonoxidative disposal of a glucose infusion during hyperinsulinaemia. Fifteen normal volunteers participated in the 3 protocols, in which 1) Intralipid 2) beta-pyridylcarbinol or 3) isotonic saline were infused during 2 hours. Thereafter, these infusions were discontinued and a two-hour euglycaemic hyperinsulinaemic clamp was performed. All three studies were carried out in combination with indirect calorimetry to measure glucose uptake, and oxidative and nonoxidative glucose disposal (corresponding essentially to glucose storage). Plasma FFA concentrations were 508 +/- 34, 601 +/- 43 and 546 +/- 45 mumol/l in the basal state during the Intralipid, beta-pyridylcarbinol and control protocols. It increased to 960 +/- 71 mumol/l after the Intralipid infusion, fell to 246 +/- 17 mumol/l after the beta-pyridylcarbinol infusion, vs 600 +/- 48 mumol/l in the control. At the end of the glucose-insulin clamp the values were low in the 3 protocols: 263 +/- 17, 233 +/- 19 and 204 +/- 14 mumol/l. Intralipid infusion prior to the clamp protocol induced a suppression of both insulin-mediated glucose uptake (4.91 +/- 0.46 (Intralipid) vs 6.83 +/- 0.63 mg.kg-1.min-1 (saline)) and storage (1.61 +/- 0.34 vs 2.99 +/- 0.53 mg.kg-1.min-1) while beta-pyridylcarbinol infusion induced an increased insulin-mediated glucose uptake (8.58 +/- 0.37 mg.kg-1.min-1) and in glucose storage (4.29 +/- 0.31 mg.kg-1.min-1) (p less than 0.5 vs Intralipid). These changes occurred even though FFA plasma concentrations were similar in the 3 experimental conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗