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At least 1,081 records · Page 60Linked to original sources
PubMed1987

Purification of horse (Equus caballus) serum lecithin:cholesterol acyltransferase.

1. A method for the purification of horse serum lecithin:cholesterol acyltransferase has been established. 2. The method involves the adsorption of the enzyme from diluted horse serum on DEAE-Sephadex A-50, (NH4)2SO4 fractionation, 1-butanol treatment, and chromatographic techniques of DEAE-Sepharose CL-6B, DEAE-Sephadex A-50, Affi-Gel blue and hydroxylapatite. 3. The resultant enzyme preparation essentially formed a single main band when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. 4. The final purification of the enzyme was 20,000-fold with 7% yield. 5. The apparent mol. wt of the enzyme was 64,000. 6. The activity of the enzyme was stable for 3 days at 0 degree C.

Animals↗
PubMedDate not supplied

Influence of hypoxanthine and other purines on the lecithin:cholesterol acyltransferase reaction in the plasma of rat and other species.

1. Esterification of radiolabelled cholesterol in the plasma of rat, mouse, pig, ox and, to a lesser extent, guinea pig was partially inhibited by hypoxanthine, xanthine and guanine; esterification in human plasma and in plasma from 12 other vertebrate species was unaffected by purines. 2. Esterification of endogenous cholesterol and the formation of lysolecithin in rat plasma were decreased in the presence of purines indicating that it was the lecithin:cholesterol acyltransferase (LCAT) reaction that was inhibited rather than the isotopic equilibration of labelled cholesterol with the endogenous substrate lipoproteins. 3. Maximum inhibition of the LCAT reaction in rat plasma occurred at 1.4 mM hypoxanthine or xanthine; inhibition was not dependent upon the concentration of LCAT or plasma lipoproteins but increased with the amount of lipoprotein depleted rat plasma (LDRP) present in the incubation mixture. 4. Partial inhibition of the LCAT reaction in rat or mouse plasma by purines had no significant effect on the fatty acyl composition of the cholesteryl esters (CE) formed by LCAT. 5. In the presence of heated rat plasma, LDRP or, to a lesser extent, rat high density lipoproteins (HDL) prepared from heated plasma, the LCAT reaction in human plasma was inhibited by hypoxanthine. 6. Rat HDL and LDRP prepared from plasma pre-incubated at 37 degrees C for 4 hr before heating increased and decreased, respectively, the inhibitory effect of hypoxanthine on human plasma LCAT compared with HDL and LDRP prepared from unincubated rat plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Perinatal dietary supplementation with a soy lecithin preparation: effects on development of central catecholaminergic neurotransmitter systems.

Previous work has shown that exposure of developing rats to soy lecithin preparations (SLP) influences macromolecular constituents of immature brain cells and causes abnormal behavioral patterns. To determine if synaptic mechanisms are adversely affected by SLP, we examined the developmental characteristics of noradrenergic and dopaminergic pathways in discrete brain regions. Although transmitter levels were unaffected, the utilization rates of both catecholamines were profoundly disturbed in an age-dependent, regionally-selective manner. Utilization tended to be subnormal in the preweanling stage, but demonstrated a postweaning elevation in cerebellum and midbrain + brainstem. Enhanced utilization persisted in the latter region only, and cerebral cortex actually showed a lowered utilization rate in adulthood (60 days of age). Transmitter uptake capabilities were also affected by developmental exposure to SLP, as was tyrosine hydroxylase activity. The patterns of effects on these two variables indicated that the altered transmitter utilization rate probably reflected a change in impulse activity in the affected neuron populations, with promotion of activity in the midbrain + brainstem and reduced activity in the cerebral cortex. These data indicate that dietary supplementation with SLP throughout perinatal development alters synaptic characteristics in a manner consistent with disturbances in neural function.

Aging↗
PubMed1993

Baboon lecithin cholesterol acyltransferase (LCAT): cDNA sequences of two alleles, evolution, and gene expression.

Lecithin cholesterol acyltransferase (LCAT) is a key enzyme of cholesterol metabolism that catalyzes esterification of cholesterol for packaging in high-density lipoprotein (HDL) particles. In this study, we cloned and sequenced LCAT cDNA from baboon, a nonhuman primate model of atherosclerosis. LCAT sequences have been highly conserved over approximately 25 million years since the divergence of the baboon and human lineages. The baboon and human sequences are 97% identical at the nucleotide (nt) level and 98% identical at the amino acid (aa) level. Only 18% of the nt substitutions change the aa sequence (nonsynonymous substitutions). The substitutions between baboon and human LCAT do not alter key functional sites including the interfacial substrate active site, asparagine-linked glycosylation sites, or sites at which rare mutations cause human familial LCAT deficiencies. We also sequenced LCAT cDNA for a less common allele that is associated with higher LCAT activities and altered lipoprotein phenotypes. There were no sequence differences between the two alleles, which suggests that genotypic effects are most likely due to allelic differences in gene expression. The tissue specificity of LCAT expression was investigated using an RNase protection assay calibrated with known amounts of synthetic human LCAT RNA. In a survey of baboon tissues, the highest levels of LCAT mRNA were found in the cerebellum and liver and trace amounts in the ileum, spleen and cerebral cortex.

Alleles↗
PubMedDate not supplied

Changes in the levels of polyunsaturated fatty acids in the lung and lecithin cholesterol acyl transferase activity in plasma of monkeys exposed to ambient levels of ozone.

A group of 8 sub-adult bonnet monkeys (Macaca radiata) was exposed to 0.15 ppm ozone (O3) and another group of 8 to 0.3 ppm O3, 8 h/day, for 90 days. Four monkeys in the control group breathed filtered air during this period. Polyunsaturated fatty acids (PUFA) comprised about 22% of total fatty acids in the lungs of controls. The PUFA level decreased to about 90% and 6% in monkeys exposed to 0.15 ppm and 0.3 ppm O3, respectively. Lecithin cholesterol acyl transferase (LCAT) activity in the plasma of monkeys exposed to 0.15 ppm O3 was similar to that in the controls. However, the LCAT activity of plasma was significantly increased by the exposure of monkeys to 0.3 ppm O3.

Air↗
PubMed1994

Simultaneous microanalysis of biliary cholesterol, bile acids and fatty acids in lecithin using capillary column gas chromatography: an advantage to assess bile lithogenecity.

Simultaneous determination of biliary lipids was performed by alkaline hydrolysis, the formation of the methyl ester derivatives of fatty acids that are constituents of phospholipids and of the acetylated methyl ester derivatives of bile acids, and subsequent analysis by capillary column gas chromatography. Complete separation and satisfactory recovery of cholesterol, bile acids, and fatty acids were achieved. Also, the accuracy of the calculation of the bile cholesterol saturation index was enhanced by computation. Since the degree of acyl chain unsaturation affects the cholesterol-holding capacity in vesicles, this method provides a unique insight into bile metastability by the quantitative assessment of fatty acids in lecithin.

Bile↗
PubMed1986

Perinatal dietary exposure to soy lecithin: altered sensitivity to central cholinergic stimulation.

The effects of perinatal exposure to soy lecithin preparation (SLP) on the development of cholinergic responses in the rat brain were examined by assessing the ability of intracisternally administered carbachol to stimulate 33Pi incorporation into phospholipids in vivo, an effect of carbachol mediated by muscarinic cholinergic receptors. Maternal intake of SLP produced a suppression of the cholinergic response in the offspring, an effect which was specific in that basal (unstimulated) incorporation rates were not reduced (in fact, they eventually became elevated), nor was the response to another neurotransmitter (dopamine) compromised. The effect occurred early in the preweanling stage, a period in which SLP exposure also enhances development of cholinergic nerve terminals. These results suggest that SLP exposure has a major effect on cholinergic synaptic development and reactivity, followed by secondary changes in other neurotransmitter pathways and by more generalized effects on basal membrane phospholipid turnover.

Animals↗
PubMed1986

Effect of dietary carbohydrate type on lipoprotein lipase, hepatic lipase, and lecithin:cholesterol acyltransferase activities in cynomolgus monkeys.

The effects of dietary sucrose and starch with and without exogenous cholesterol on postheparin plasma lipoprotein lipase (PHLA) and hepatic lipase (HLA) were studied in cynomolgus monkeys. Serum triglyceride levels were higher in sucrose-fed animals than starch and exogenous cholesterol lowered serum triglyceride levels when added to sucrose diet but not starch diets. Sucrose markedly increased insulin levels, more so than starch; however, dietary cholesterol lowered insulin levels in sucrose diet but increased the levels in starch diet. PHLA activity was increased two- to threefold greater in sucrose than in starch diets. Exogenous cholesterol lowered PHLA activity in sucrose diet but increased PHLA activity in starch diet. HLA activity was increased with sucrose more than starch. Lecithin:cholesterol acyltransferase (LCAT) activity was significantly higher in sucrose diets than in the starch diet. Addition of cholesterol to either of these diets lowered the LCAT activity. These results indicate that PHLA, HLA, and LCAT activities not only are affected by the nature of carbohydrates, but also are related to triglyceride metabolism. The interaction of carbohydrates and cholesterol in the diet by influencing these selected enzymes plays an integrated role in lipoprotein particle interconversion processes.

Animals↗
PubMed1988

Effects of cold stress on serum lipids, lipoproteins, and the activity of lecithin:cholesterol acyltransferase in rabbits.

Effects of prolonged stress on lipid metabolism factors were studied for 9 weeks using four groups of young New Zealand rabbits. Two groups (A,B) were rendered atherosclerotic by administering 1% (w/w) cholesterol. One group (C) was subjected to cold stress together with one of the atherosclerotic groups (B); one group was used as control (N). At the end of treatment serum total cholesterol and total lipids of A and B increased significantly, while in stress group (C) a significant decrease was observed. HDL-C levels were reduced in all experimental groups. Triglycerides did not change in A, while they were reduced in both stress groups (C,B). Serum lecithin:cholesterol acyltransferase (LCAT) activity levels of B and C were decreased. Lipoprotein electrophoresis patterns showed a significant redistribution of percentage values in all experimental groups: %LDL-C increased and %VLDL-C decreased in all groups, %HDL-C declined in A and B and did not change in C. The combination of stress and atherosclerosis in rabbits elicits far greater alterations in lipid and lipoprotein profiles than stress or atherosclerosis alone. A stress and atherosclerotic diet combination may be a hazardous one in relation to CHD and atherosclerosis.

Animals↗
PubMed1992

The inhibiting effect of catecholamine-melanins on UV-induced lecithin peroxidation.

It was found that the yield of thiobarbituric acid reactive substances in UV-irradiated liposome membranes was significantly suppressed in the presence of catecholamine-melanins, indicating their ability to inhibit lipid peroxidation. The extent of inhibition depended on the type and concentration of melanin polymers. Melanin-copper complexes inhibited lecithin photooxidation less effectively than copper-free melanins derived from the same precursor. The antioxidant efficiency of melanins appears to be related to the levels of intrinsic and photo-induced free radical centers in the melanin polymer, as well as to accessibility of these centers for active species formed during irradiation of liposomes.

Catecholamines↗
PubMed2005

Comparison of the TDx-FLM II and lecithin to sphingomyelin ratio assays in predicting fetal lung maturity.

OBJECTIVE: We evaluated the usefulness of the TDx-FLM II and lecithin to sphingomyelin (L/S) ratio assays in predicting fetal lung maturity. STUDY DESIGN: We retrospectively reviewed 218 consecutive paired TDx-FLM II and L/S ratio results. Women who delivered viable infants within 72 hours of amniotic fluid collection (n=109) were included in the analysis of sensitivity and specificity. Concordance between tests was determined for all women tested during the study period, and in the subset of women who delivered viable infants within 72 hours of amniotic fluid collection. RESULTS: There were 9 respiratory distress syndrome (RDS)-affected infants born during the study period. Both the TDx-FLM II and L/S ratios had 100% sensitivity in detecting RDS at their best apparent cut-offs. There was a trend towards increased specificity of the L/S ratio compared with the TDx-FLM II (80% for L/S vs 73% for FLM II). The overall concordance between the TDx-FLM II and L/S ratio was approximately 75%. CONCLUSION: The TDx-FLM II and L/S ratios are both sensitive tests for RDS; however, there is not good concordance between the two. The results provide new insight into the optimal use, in sequential or reflex cascade testing, of the TDx-FLM II and L/S ratio.

Amniotic Fluid↗
PubMed2004

The effect of interaction between K(+) ions and gramicidin D on the lecithin membrane interfacial tension.

The effect of the presence of gramicidin D in a lecithin membrane on its interfacial tension has been studied. The studies have been carried out at various forming solution compositions and at various potassium ion concentrations in the electrolyte solution. Potassium chloride was used as the electrolyte. The complex was formed between the gramicidin molecule and K(+) ion. The following parameters describing the complex were determined: the surface area occupied by GK(+) complex (A(GK(+))), the interfacial tension of the GK(+) membrane complex (gamma(GK+)), and the stability constant of the gramicidin-K(+) complex (K). These values are 156 A(2), 1.89 mN m(-1) and 0.033 m(3) mol(-1), respectively.

Binding Sites↗
PubMed2005

Studies on direct electron transfer and biocatalytic properties of heme proteins in lecithin film.

Myoglobin (Mb), hemoglobin (Hb) and horseradish peroxidase (HRP) were incorporated in lecithin (PC) film on glassy carbon (GC) electrode by the method of vesicle-fusion. A pair of well-defined and quasi-reversible cyclic voltammetric peaks was obtained, which reflected the direct electron transfer of heme proteins. UV-Vis and reflectance absorption infrared (RAIR) spectroscopy showed that proteins in PC films remained at their secondary structure similar to their native states. Scanning electron microscopy (SEM) demonstrated the interaction between the proteins and PC would make the morphology of protein-PC films very different from the PC films alone. The immobilized proteins retained their biocatalytic activity to the reduction of NO and hydrogen peroxide, which provide the perspective to be the third generation sensors.

Calibration↗
PubMed2004

T13M mutation of lecithin-cholesterol acyltransferase gene causes fish-eye disease.

BACKGROUND: Lecithin-cholesterol acyltransferase (LCAT) esterifies free cholesterol (FC) in plasma and plays a crucial role in the maturation of prebeta1-HDL (lipid-poor HDL) into alpha-migrating HDL (spherical HDL). Natural mutations of LCAT gene cause familial LCAT deficiency (FLD) or fish-eye disease (FED). The relationship between mutations and their phenotypes gives important clues to the functions of specific regions of LCAT. We investigated the first homozygous case with a substitution of threonine to methionine at codon 13 (T13M) of LCAT gene. METHODS: We evaluated LCAT activity, LCAT distribution among HDL subfractions and conversion of prebeta1-HDL to alpha-migrating HDL by native two-dimensional gel electrophoresis (N-2DGE). RESULTS: The proband had corneal opacity, severe hypo-alpha-lipoproteinemia, half-normal LCAT activity and near normal cholesteryl ester/total cholesterol (TC) ratio in plasma. These features were characteristic of FED. Plasma prebeta1-HDL concentration was near normal, but not converted to alpha-migrating HDL during 37 degrees C incubation. As expected, alpha-migrating HDL (especially large particles) was markedly reduced. In the immunoblot against LCAT, the small alpha-migrating HDL from the proband had much less LCAT in this patient than in controls. CONCLUSION: T13M mutation of LCAT gene causes FED.

Adult↗
PubMed2005

The effect of the presence of valinomycin on the interfacial tension of lecithin membrane.

The effect of the presence of valinomycin in lecithin membrane on its interfacial tension has been studied. The experiments have been carried out at various forming solution compositions and at various potassium ion concentrations in electrolyte solution. Potassium chloride was used as the electrolyte. A complex was formed between the valinomycin molecule and K+ ion. The following parameters describing the complex were determined: K, the stability constant of the valinomycin-K+ complex and B, partition coefficient. These values are equal to 3.52 x 10(5) m3 mol(-1) and 6.0, respectively.

Anti-Bacterial Agents↗
PubMed2004

Comparative therapeutic efficacy of a novel lyophilized amphotericin B lecithin-based oil-water microemulsion and deoxycholate-amphotericin B in immunocompetent and neutropenic mice infected with Candida albicans.

The in vivo efficacy of a new amphotericin B (AmB) oil-in-water lecithin-based microemulsion delivery system (M-AmB) compared to deoxycholate-AmB (D-AmB) was studied in an immunocompetent and neutropenic murine model of systemic candidiasis. D-AmB was administered at the maximum tolerated dose of 1 mg/kg whereas M-AmB was given at the doses of 1, 2 and 3 mg/kg; doses were well tolerated due to their reduced toxicity. Both formulations were administered 24, 48 and 72 h after infection in immunocompetent mice, and 2, 6 and 24 h after infection in neutropenic mice. Kaplan-Meier survival curves showed that the M-AmB treated group had a better survival time than infected mice without treatment used as a control group (P = 4.66 x 10(-6)), and the Mann-Whitney W statistical test indicated that it reduced the percentage of mortality and fungal load in the most representative organs. This new formulation is a designed competitor which has proved to present better results than D-AmB in an established infection not only in immunocompetent but in neutropenic mice as well.

Amphotericin B↗
PubMed2005

Lecithin:retinol acyltransferase in ARPE-19.

The purpose of this study is to investigate if a readily available cell line (APRE-19) may be used to study in vitro function of visual cycle enzymes such as lecithin:retinol acyltransferase (LRAT). Cells incubated with exogenous retinol accumulated intracellular all-trans retinol and all-trans retinyl ester. Membrane proteins from ARPE-19 exhibited LRAT activity, which was inhibited by an LRAT inhibitor, retinyl bromoacetate (RBA). Gene microarray and Western blot results indicated that ARPE-19 cells expressed LRAT transcript and the LRAT protein. Therefore, our data show that ARPE-19 contains an active LRAT enzyme and suggest that it is an appropriate cell system to study visual cycle enzymes.

Acyltransferases↗
PubMed2004

Cloning, gene organization and identification of an alternative splicing process in lecithin:retinol acyltransferase cDNA from human liver.

Lecithin:retinol acyltransferase (LRAT) catalyzes the synthesis of retinyl esters in many tissues and is crucial for the transport and intracellular storage of vitamin A. LRAT expression is highly regulated in the liver. In this study, we have cloned and sequenced the full-length LRAT mRNA from human liver and identified its 5'- and 3'-ends. Full-length LRAT mRNA comprises 5023 nt with a predicted ORF of 230 amino acids, a short 5'UTR, and a relatively long 3'UTR of 4 kb containing several polyadenylation signals and AU-rich regions. Based on alignment of this mRNA with human genomic DNA in the GenBank database, the human LRAT gene spans about 9.1 kbp and consists of two exons and a relatively long 4-kbp intron. Further analysis of normal liver revealed a minor alternative splicing variant which lacks a 103 nt polynucleotide contained in the 5'UTR of the full-length LRAT transcript. This variant predicts that the LRAT gene is organized into three exons and two introns, as reported for LRAT cloned from retinal pigment epithelium (RPE) cells. These two LRAT mRNA variants are also present in testis, which is known to express LRAT and contain retinyl esters. Major and minor transcription start sites for human liver LRAT mRNA were identified and the sequence of the upstream proximal promoter region was retrieved from the GenBank database and physically analyzed for the presence of putative cis-acting elements essential for basal transcription. This region contains a TATA box, CCAAT box and Sp1 site, which are apparently conserved in mouse and rat LRAT genes. Our results provide evidence that multiple LRAT mRNA transcripts, which are expressed in a tissue-specific manner, may result from several mechanisms including differential splicing of the 5'UTR region and the use of multiple polyadenylation signals in the 3'UTR.

3' Untranslated Regions↗