Cholesteryl Ester Composition Research Articles

Inflammation and wound healing in cats with chronic gingivitis/stomatitis after extraction of all premolars and molars were not affected by feeding of two diets with different omega‐6/omega‐3 polyunsaturated fatty acid ratios

Feline chronic gingivitis/stomatitis (FCGS) is a painful inflammatory disease in cats. Extraction of teeth, including all premolars and molars, has been shown to be the therapy of choice in cats not responding sufficiently to home care (e.g. tooth brushing) and/or medical treatment (corticosteroids and/or antibiotics). In this study, we hypothesize that a cat food with an omega-6 polyunsaturated fatty acid (ω6 PUFA) to ω3 PUFA ratio of 10:1 reduces inflammation of the FCGS and accelerates soft tissue wound healing of the gingiva after dental extractions, compared to a cat food with a ω6:ω3 PUFA ratio of 40:1. The cats were fed diets with chicken fat and fish oil as sources of fatty acids. In one diet, part of the fish oil was replaced by safflower oil, resulting in two diets with ω6:ω3 PUFA ratios of 10:1 and 40:1. This double-blinded study in two groups of seven cats revealed that dietary fatty acids influence the composition of plasma cholesteryl esters and plasma levels of inflammatory cytokines. The diet with the 10:1 ratio lowered PGD(2) , PGE(2) and LTB(4) plasma levels significantly, compared to the diet with the 40:1 ratio (p = 0.05, p = 0.04, and p = 0.02 respectively). However, feeding diets with dietary ω6:ω3 PUFA ratios of 10:1 and 40:1, given to cats with FCGS for 4 weeks after extraction of all premolars and molars, did not alter the degree of inflammation or wound healing.

On the Presence and Role of Polar Lipids in Meibum

On the Presence and Role of Polar Lipids in Meibum

The fatty acid composition of plasma cholesteryl esters and estimated desaturase activities in patients with nonalcoholic fatty liver disease and the effect of long-term ezetimibe therapy on these levels

The aim of this study was to investigate the relationship between fatty acid composition of plasma cholesteryl esters (CEs) and estimated desaturase activity and the development and progression of nonalcoholic fatty liver disease (NAFLD). The study also assessed the effect of ezetimibe on CE levels. Plasma CEs fatty acid composition was analyzed in 3 groups: patients with a NAFLD activity score (NAS) ≤ 4 (n=31) or NAS ≥ 5 (n=32) and normal controls (n=25). The estimated desaturase activities were calculated using ratios of 16:1n-7/16:0 (D9-16D), 18:1n-9/18:0 (D9-18D), 18:3n-6/18:2n-6 (D6D) and 20:4n-6/20:3n-6 (D5D). Compared with controls, the levels of palmitate, palmitoleate, γ-linoleate, D9-16D and D6D were significantly increased, whereas levels of linoleate and D5D were significantly decreased. Patients with NAS ≥ 5 had significantly higher palmitate levels than patients with NAS ≤ 4. The levels of these fatty acids, especially palmitate and palmitoleate, correlated with NAFLD-related lipid, metabolic, and inflammatory parameters. Long-term therapy with ezetimibe caused significant improvements in the levels of these fatty acids, estimated desaturase activity index and NAFLD-related parameters. Our results suggest that fatty acids and desaturase activity associate with the development and progression of NAFLD, and that ezetimibe may be a novel treatment for this disorder.

Fatty acid composition of cholesteryl esters of human meibomian gland secretions

Very long chain cholesteryl esters (CE) are a major group of lipids found in meibomian gland secretions (MGS, also called meibum). MGS are produced by the meibomian glands of human and animal eyelids. They are a critical part of the tear film which covers the exposed ocular surface and serves various physiological roles. The composition of CE of MGS is complex, and still remains poorly understood. Here, a liquid chromatography–ion trap mass spectrometry (LC–MS) procedure developed to analyze CE is described, and a detailed composition of human meibomian CE is reported. MGS were collected from donors, analyzed without any modifications by LC–MS in positive and negative ion modes (PIM and NIM), and quantified using lipid standards where available. CE comprised about 30% of human meibum by mass. More than 40 individual CE species were found and characterized. In PIM, CE were observed as spontaneously in-source generated product ions m/ z 369. The signals of the proton adducts of intact CE (M+H) + were of very low intensity. In NIM, all tested CE spontaneously fragmented in-source producing signals of their respective FA. By combining the LC and MS information, the most abundant CE were found to be based on FA ranging from C 16 to at least C 32 in the following order C 26:0 > C 25:0 > C 24:0 > C 27:0 > C 24:1 = C 18:1 = C 20:0 > other CE. We conclude that the FA composition of CE can be successfully established in LC–MS experiments conducted in NIM. Meibomian CE have a large presence of both saturated and unsaturated FA with an average molar ratio of 4 to 1, respectively.

Plasma Fatty Acid Composition, Estimated Desaturase Activities, and Intakes of Energy and Nutrient in Japanese Men with Abdominal Obesity or Metabolic Syndrome

To examine predictive factors for abdominal obesity or metabolic syndrome, we investigated the association of plasma fatty acid composition, estimated desaturase activity, and nutrient intakes, with abdominal obesity or metabolic syndrome in Japanese males. Clinical characteristics, the fatty acid composition of plasma cholesteryl esters, and energy and nutrient intakes were analyzed in 3 groups: metabolic syndrome (MS, n=24), abdominal obesity (OB, n=43), and control (n=27). The estimated desaturase activities were calculated by the ratio of 16:1n-7/16:0, 18:3n-6/18:2n-6, and 20:4n-6/20:3n-6 in plasma cholesteryl esters as surrogates of the measure of the delta 9, delta 6, delta 5 desaturase (D9-16D, D6D and D5D) activities, respectively. Plasma fatty acid composition did not differ significantly between the OB group and the control group. The MS group had higher levels of palmitoleic, oleic, and gamma-linolenic acids, but a lower level of linoleic acid than the control. Stronger D6D activity and weaker D5D activity were observed in the OB group. A higher level of D9-16D activity as well as a higher level of D6D activity and a lower level of D5D activity was observed in the MS group. A logistic regression analysis showed that the low D5D activity and high D9-16D activity were predictive of the development of abdominal obesity from controls (odds ratio=0.39, p<0.05) and metabolic syndrome from abdominal obesity (odds ratio=2.44, p<0.05), respectively. In the multiple linear regression analysis, D5D activity positively correlated with the intake of eicosapentaenoic acid (EPA). In conclusion, the estimated D5D activity was a predictive factor for abdominal obesity and the estimated D9-16D activity was a predictive factor for developing metabolic syndrome from abdominal obesity in Japanese male subjects. Dietary intake of EPA would play an important role in preventing abdominal obesity and the development of metabolic syndrome.

Fatty Acid Profile of Sunshine Bass: II. Profile Change Differs Among Fillet Lipid Classes

Fatty acid (FA) profile of fish tissue mirrors dietary FA profile and changes in a time-dependent manner following a change in dietary FA composition. To determine whether FA profile change varies among lipid classes, we evaluated the FA composition of fillet cholesteryl esters (CE), phospholipids (PL), and triacylglycerols (TAG) of sunshine bass (SB, Morone chrysops x M. saxatilis) raised on feeds containing fish oil or 50:50 blend of fish oil and coconut, grapeseed, linseed, or poultry oil, with or without implementation of a finishing period (100% FO feed) prior to harvest. Each lipid class was associated with a generalized FA signature, irrespective of nutritional history: fillet PL was comprised largely of saturated FA (SFA), long-chain polyunsaturated FA (LC-PUFA), and total n-3 FA; fillet TAG was higher in MC-PUFA and total n-6 FA; and fillet CE was highest in monounsaturated FA (MUFA). Neutral lipids reflected dietary composition in a near-direct fashion; conversely, PL showed evidence of selectivity for MC- and LC-PUFA. Shorter-chain SFA were not strongly reflected within any lipid fraction, even when dietary availability was high, suggesting catabolism of these FA. FA metabolism in SB is apparently characterized by a division between saturated and unsaturated FA, whereby LC-PUFA are preferentially incorporated into tissues and SFA are preferentially oxidized for energy production. We demonstrated provision of SFA in grow-out feeds for SB, instead MC-PUFA which compete for tissue deposition, meets energy demands and allows for maximum inclusion of LC-PUFA within fillet lipids.

Separation and Determination of Fatty Acids from Lipid Fractions by High-Performance Liquid Chromatography: Cholesterol Esters of Umbilical Cord Arteries

ABSTRACTPreeclampsia is accompanied by an extensive remodeling of the extracellular matrix of umbilical cord. It is associated with an increase in collagen content in the umbilical cord artery. Furthermore, preeclampsia distinctly reduces proteolytic and gelatinolytic activity, especially after activation with various agents.We decided to develop a method for separation and determination of fatty acids from different tissues by high-performance liquid chromatography. That method allowed us to determine cholesteryl ester composition and content in umbilical cord arteries. Studies were performed on the umbilical cord arteries taken from 10 newborns delivered by healthy mothers and 10 newborns delivered by mothers with preeclampsia. Cholesteryl esters were isolated by thin layer chromatography. Fatty acids were liberated by basic hydrolysis and analyzed by HPLC of their p-bromophenacyl derivatives using detection at 254 nm. It was found that saturated fatty acids were the main group of fatty acids incorporated to cholesteryl esters in all control and preeclamptic umbilical cord arteries. Preeclampsia caused a significant increase in cholesteryl ester content in the umbilical cord arteries. An increase of neutral lipid content in vessel walls of newborns delivered by mothers with preeclampsia may be one of the factors that evoke the initiation of hypertension in utero and its amplification throughout childhood and adult life. The described method reduces time and cost consumption and allows us to determine almost all fatty acids forming cholesteryl esters contained in the tissue sample.

Functional LCAT deficiency in human apolipoprotein A-I transgenic, SR-BI knockout mice

Reduction of plasma LCAT activity has been observed in several conditions in which the size of HDL particles is increased; however, the mechanism of this reduction remains elusive. We investigated the plasma activity, mass, and in vivo catabolism of LCAT and its association with HDL particles in human apolipoprotein A-I transgenic, scavenger receptor class B type I knockout (hA-ITg SR-BI-/-) mice. Compared with hA-ITg mice, hA-ITg SR-BI-/- mice had a 4-fold higher total plasma cholesterol concentration, which occurred predominantly in 13-18 nm diameter HDL particles, a significant reduction in plasma esterified cholesterol-total cholesterol (EC/TC) ratio, and significantly lower plasma LCAT activity, suggesting a decrease in LCAT protein. However, LCAT protein in plasma, hepatic mRNA for LCAT, and in vivo turnover of 35S-radiolabeled LCAT were similar in both genotypes of mice. HDL from hA-ITg SR-BI-/- mice was enriched in sphingomyelin (SM), relative to phosphatidylcholine, and had less associated [35S]LCAT radiolabel and endogenous LCAT activity compared with HDL from hA-ITg mice. We conclude that the decreased EC/TC ratio in the plasma of hA-ITg SR-BI-/- mice is attributed to a reduction in LCAT reactivity with SM-enriched HDL particles.

Dietary Fat–Induced Alterations in Atherosclerosis Are Abolished by ACAT2-Deficiency in ApoB100 Only, LDLr −/− Mice

The enzyme acyl-coenzymeA (CoA):cholesterol O-acyltransferase 2 (ACAT2) in the liver synthesizes cholesteryl esters (CE) from cholesterol and fatty acyl-CoA, which get incorporated into apoB-containing lipoproteins that are secreted into the bloodstream. Dietary fatty acid composition influences the amount and fatty acid composition of CE within apoB-containing lipoproteins. We hypothesized that when ACAT2 activity is removed by gene deletion, hepatic CE synthesis and secretion would be minimal and, as a result, dietary fat-related differences in atherosclerosis would be eliminated. Groups of female apoB100 only, LDLr-/- mice with and without ACAT2 were fed diets enriched in either omega-3 or omega-6 polyunsaturated fat, saturated fat, and cis or trans monounsaturated fat. After 20 weeks on diet, mice fed diets enriched in monounsaturated or saturated fat exhibited significantly higher amounts of plasma cholesterol, larger LDL particles enriched in monounsaturated CE, and more atherosclerosis than mice fed polyunsaturated fat. The dietary fat-induced shifts in plasma cholesterol, LDL size, LDL CE composition, and atherosclerosis were not observed in ACAT2-/- mice. Regardless of the diet fed, the ACAT2-/- mice were protected from atherosclerosis. The results indicate that in apoB100 only, LDLr-/- mice, ACAT2 plays an essential role in facilitating dietary fat type-specific atherosclerosis through its various effects on plasma lipoprotein concentration and composition.

Fatty acids in serum cholesteryl esters in relation to asthma and lung function in children

Dietary fatty acid intake has been proposed to contribute to asthma development with n-6 polyunsaturated fatty acids (PUFA) having a detrimental and n-3 PUFA a protective effect. The aim of our analysis was to explore the relationship between fatty acid composition of serum cholesteryl esters as marker of dietary intake and prevalence of asthma, impaired lung function and bronchial hyper-responsiveness in children. The study population consisted of 242 girls and 284 boys aged 8-11 years, living in Munich, Germany. Data were collected by parental questionnaire, lung function measurement and skin prick test according to the International Study of Asthma and Allergies in Childhood phase II protocol. Confounder-adjusted odds ratios (OR) with 95% confidence intervals (CI) were calculated for the association between quartiles of fatty acid concentration and health outcomes with the first quartile as reference. n-3 PUFA: levels of eicosapentaenoic acid were not related to asthma and impaired lung function. Linolenic acid levels were positively associated with current asthma (OR for fourth quartile 3.35, 95% CI 1.29-8.66). Forced expiratory volume in 1 s (FEV(1)) values decreased with increasing levels of linolenic acid (p for trend=0.057). n-6 PUFA: there was a strong positive association between arachidonic acid levels and current asthma (OR(4th quartile) 4.54, 1.77-11.62) and a negative association with FEV(1) (P=0.036). In contrast, linoleic acid was negatively related to current asthma (OR(4th quartile) 0.34, 0.14-0.87) and FEV(1) values increased with increasing levels of linoleic acid (P=0.022). The ratio of measured n-6 to n-3 PUFA as well as levels of palmitic and oleic acid were not consistently related to asthma or lung function. Our data do not support the hypothesis of a protective role of n-3 PUFA. Elevated arachidonic acid levels in children with asthma may be because of a disturbed balance in the metabolism of n-6 PUFA or may be secondary to inflammation in these patients.

Lipoprotein-like Particles and Cholesteryl Esters in Human Bruch’s Membrane: Initial Characterization

To isolate and characterize cholesteryl ester-containing, lipoprotein-like particles (LLPs) from normal aged human Bruch's membrane (BrM)/choroid (Ch). From BrM/Ch of 20 eyes of 10 donors aged >60 years, LLPs were released by high-salt buffer, fractionated by density gradient ultracentrifugation, and characterized by determining cholesterol, triglyceride, and phospholipid concentration (by enzymatic colorimetry and fluorometry); cholesteryl ester composition (by electrospray ionization mass spectrometry, ESI/MS); and particle morphology (by negative stain electron microscopy). Apolipoprotein (apo) gene expression was determined with RT-PCR, Western blot analysis, and immunofluorescence of retinal-choroidal cryosections. In paraformaldehyde-preserved eyes (20 eyes of 20 donors), cholesteryl ester composition of BrM/Ch, cornea, and sclera was determined by ESI/MS. A pooled fraction of LLP released from BrM/Ch (concentrated total LLP, density [d] < 1.24 g/mL fraction) was fractionated into two peaks. A large Peak 1 (with plasma LDL-HDL density range), containing predominantly phospholipid and unesterified cholesterol, was morphologically heterogeneous. A small Peak 2 (with plasma VLDL density range), enriched with esterified cholesterol, contained approximately 100 nm diameter round electron-lucent particles. Both peaks contained apoB and apoA-I, RPE and retina contained apoA-I mRNA transcripts, and BrM and drusen contained apoA-I immunoreactivity. Peaks 1 and 2, native RPE, and fresh BrM/Ch were cholesteryl linoleate enriched and contained little cholesteryl docosahexaenoate. Preserved BrM/Ch was cholesteryl oleate-enriched, unlike sclera and cornea. BrM/Ch LLP do not resemble plasma lipoproteins in density profile, cholesterol distribution, or morphology. Peak 2 contains EC-rich LLP resembling BrM particles in situ. BrM/Ch cholesteryl esters respond to long-term storage differently than esters of plasma lipoprotein origin accumulated in other ocular tissues. Evidence of intraocular apoB and apoA-I expression supports an emerging hypothesis that the RPE assembles and secretes a large, possibly novel, lipoprotein particle.

Characteristics of Lipoprotein Peak x Eluted from a Column with the Eluent of High-magnesium Ion Concentration in Lipoprotein Analysis Using the Cation-exchange Chromatography

The new lipoprotein analysis method using a cation-exchange chromatography, which contains a sulfopropyl-ligand column and two magnesium ion-containing eluents was previously reported. This method can separate serum lipoproteins on the column gel with a magnesium ion concentration gradient and high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL) and an unspecified lipoprotein peak are eluted in order from the column. We have now characterized the unspecified lipoproteins, designated peak x, which is last eluted from the column with the eluent of high-magnesium ion concentration. The peak x was small size chylomicron fraction with a part of VLDL. Furthermore, the cholesterol values in the peak x were significantly correlated with remnant-like particle (RLP)-cholesterol values. The peak x separated from a hyperlipidemic patient included apolipoprotein B-100, B-48, E, A-1 and Cs (C-I, C-II, C-III) and its composition of free cholesterol, cholesteryl esters, triglyceride (TG) and phospholipids in total lipids were 6, 15, 66 and 13%, respectively. These results suggest that a major part of the lipoprotein peak X may be composed of remnants of chylomicron and VLDL, but it remains to be elucidated.

The influence of dietary fish oil vs. sunflower oil on the fatty acid composition of plasma cholesteryl-esters in healthy, adult cats.

The question addressed was whether the fatty acid composition of plasma cholesteryl esters (CEs) in cats reflects the intake of fatty acids. Diets containing either fish oil or sunflower oil were fed to six healthy, adult cats in a cross-over trial. The dry cat foods contained approximately 18.5% crude fat, of which two-third was in the form of the variable oil. Blood samples were collected at the end of each 4-week feeding period, and the fatty acid composition of plasma CEs and plasma concentrations of lipoproteins were determined. Consumption of the diet with fish oil was associated with significantly greater proportions of eicosapentaenoic acid, arachidonic acid, alpha-linolenic acid, oleic acid, palmitic acid and myristic acid in plasma CEs. The intake of fish oil instead of sunflower oil reduced the percentage of linoleic acid in CEs. The plasma concentrations of total cholesterol, high-density lipoprotein cholesterol, phospholipids and triglycerides were not affected by fish oil vs. sunflower oil feeding.

N−3 Fatty acids and 5-y risks of death and cardiovascular disease events in patients with coronary artery disease

Data on the association of n-3 fatty acid content in serum lipids with mortality in patients with coronary artery disease (CAD) are limited. We hypothesized that a high proportion of n-3 fatty acids in serum lipids would be associated with reduced risks of death and coronary events in patients with established CAD. We measured dietary intakes via food records and the fatty acid composition of serum cholesteryl esters (CEs) in 285 men and 130 women with CAD (x age: 61 y; range: 33-74 y). The patients participating in the EUROASPIRE (European Action on Secondary Prevention through Intervention to Reduce Events) study were followed up for 5 y. During the follow-up, 36 patients died, 21 had myocardial infarctions, and 12 had strokes. The relative risks (RRs) of death adjusted for cardiovascular disease risk factors for subjects in the highest tertile of fatty acids in CEs compared with those in the lowest tertile were 0.33 (95% CI: 0.11, 0.96) for alpha-linolenic acid, 0.33 (0.12, 0.93) for eicosapentaenoic acid, and 0.31 (0.11, 0.87) for docosahexaenoic acid (P for trend = 0.063, 0.056, and 0.026, respectively). A high proportion of eicosapentaenoic acid in CEs was associated with a low risk of CAD death. Compared with no consumption, consumption of fish tended to be associated with a lower risk of death [1-57 g/d, RR = 0.50 (0.20, 1.28); > 57 g/d, RR = 0.37 (0.14, 1.00); P for trend = 0.059]. High proportions of n-3 fatty acids in serum lipids are associated with a substantially reduced risk of death.

Removal of cholesteryl esters from diet-induced atherosclerotic lesions in two long-term studies in Rhesus monkeys

In two long-term studies of dietary-cholesterol-induced atherosclerosis lesions in rhesus monkeys, we determined the fatty acid composition of cholesteryl esters in the arterial intima-media preparations. In the first study (2-year study) monkeys were fed an atherogenic diet for 2 years; in the second study, (5-year study), monkeys were fed the atherogenic diet for about 5.4 years. Dietary cholesterol was removed from regressed animals and the regression periods in the 2-year study were 30 and 52 weeks and in the 5-year study were 20, 30, 52, 101, and 191 weeks. In both studies in thoracic and abdominal aortic segments, the percent removal of 18:2 cholesteryl ester (CE) was small, whereas percent removal of 18:1 CE and other CEs was much higher. We postulate that 18:2 CE is not hydrolyzed to the extent comparable to 18:1 and other CEs in lesions. Perhaps 18:2 CE is not a preferred substrate for arterial neutral cholesterol ester hydrolase.

Fatty acid composition of phospholipids and cholesteryl esters in maternal serum in the early puerperium

The fatty acid composition of serum phospholipids (PL) and cholesteryl esters (CE) in 26 healthy pregnant women at the end of term and 1 and 3 days after delivery was analysed in order to determine whether the maternal serum fatty acid composition changes in the early puerperium. The composition of the saturated fatty acids significantly changes in the PL fraction: 16:0 decreased and 18:0 increased. Both 20:4n-6 and 20:5n-3 significantly increased after parturition in serum PL while 22:6n-3 remained constant at the three sampling time points. The sum of HUFA was slightly higher 3 days postpartum compared to the prepartum data. The essential fatty acid index significantly increased after delivery. In the CE fraction too differences occurred during puerperium: 18:2n-6 and 20:4n-6 increased and 18:1n-9 decreased after parturition. The sum of the n-3 fatty acids in CE remained unaltered. The EFA index significantly improved both in PL as in CE after delivery. In conclusion, the previously reported changes in the fatty acid composition of PL and CE during normal pregnancy diminish shortly after delivery. In fact, very soon after delivery the maternal fatty acid composition returns to more normal values.

FA composition of cholesteryl esters and phospholipids in maternal plasma during pregnancy and at delivery and in cord plasma at birth.

The purpose of this study was to assess the FA composition of both cholesteryl esters (CE) and phospholipids (PL) in maternal plasma during pregnancy and at delivery and in umbilical plasma at birth. A longitudinal study of 32 normal pregnant women was carried out with three cutoff points during pregnancy (first, second, and third trimester) and at delivery. Few significant differences occurred in the FA profile of maternal CE: 18:1n-9 increased, 18:2n-6 dropped slightly, and 18:3n-3 decreased with progressing gestation. In maternal PL, long-chain highly unsaturated FA concentrations dropped and were replaced by saturated FA as gestation progressed. Additionally, changes in saturated FA in PL occurred: Shorter-chain 16:0 was higher whereas longer-chain 18:0 was lower at delivery compared to early pregnancy. The FA profile of umbilical venous plasma was strikingly different from that of maternal plasma at delivery. Cord plasma CE contained more saturated and monounsaturated FA than maternal CE. The polyunsaturates 18:2n-6 and 18:3n-3 are lower in umbilical CE than in maternal CE whereas 20:4n-6 and 22:6n-3 are twice as high in umbilical CE. Cord plasma PL have a higher content of long-chain highly unsaturated FA than maternal plasma PL at delivery. In contrast to maternal plasma PL, 16:0 was lower and longer-chain saturated FA were higher in cord plasma PL. The FA profile of umbilical plasma at birth shows preferential accumulation of 20:4n-6 and 22:6n-3, with low concentrations of 18:2n-6 and 18:3n-3 in CE and PL, indicating a preferential supply of the fetus with long-chain highly unsaturated FA needed for fetal development.

Correlation of the fatty acid composition and fluid property of the cholesteryl esters in the serum of Nigerian children with sickle cell disease and healthy controls

In a previous study conducted in Nigeria, we found that children with sickle cell disease (SCD) had exceedingly low total serum cholesterol levels (mean=100−102 mg/dl). The fact that significant reductions in the levels of certain polyunsaturated fatty acids (PUFA) have been documented in the serum phospholipids of these same SCD subjects led us to inquire as to the fatty acid composition of the cholesteryl esters (CE) in their serum. Lecithin:cholesterol acyl transferase (LCAT), the enzyme in blood that catalyzes the reaction in which tissue cholesterol is acylated prior to its removal from cell membranes, is relatively specific for certain PUFA. CE in blood serum from 43 male and 42 female children with SCD, ages 4–18 years, and equal numbers of age- and gender-matched controls were analyzed for their fatty acid composition. Relative to the non-SCD controls, the CE of the SCD subjects contained 9% less linoleic acid, 16% less arachidonic acid, 40% less α-linolenic acid, 50% less eicosapentaenoic acid, and 36% less docosahexaenoic acid, but 15% more palmitic acid and 10% more oleic acid. Overall, the acyl chains of the CE of the SCD subjects were less fluid than those of the controls, as determined by comparison of their mean melting points (MMP) and double bond indices (DBI). MMP and DBI were both estimated from the individual constituent fatty acids comprising the CE acyl chains. The strongest correlations between MMP and fatty acid mole percent were seen with palmitic acid and linoleic acid. These results show that the fatty acid composition of the serum CE of children with SCD is abnormal relative to controls who do not have this hematologic disorder. We speculate that suboptimal fatty acid nutrition in Nigerian children with SCD compromises their ability to remove cholesterol from their tissues due to preference of the LCAT enzyme for PUFA, thereby accounting, in part at least, for the low total serum cholesterol levels one finds in children with SCD.

Alteration of plasma HDL cholesteryl ester composition with transgenic expression of a point mutation (E149A) of human LCAT

We have previously identified a single amino acid mutation (hE149A) in human LCAT that increases its in vitro reactivity with phosphatidylcholine species containing sn-2 arachidonate (Wang et al. 1997. J. Biol. Chem. 272: 280–286). The purpose of the present study was to determine whether in vivo overexpression of hE149A compared with human wild-type LCAT (hLCAT-wt) would be sufficient to enrich the steady state composition of plasma HDL cholesteryl esters (CE) with long chain (>18 carbon) polyunsaturated fatty acyl species. Transgenic lines with 20-fold overexpression of hLCAT were created and studied between 12 and 16 weeks of age while consuming a chow diet. Transgenic overexpression of hE149A compared with hLCAT-wt significantly enriched HDL with CE species containing 20:4 (45%) and 22:6 n-3 (108%), at the expense of those containing 18:2, without a significant change in the plasma HDL concentration, particle size, or phospholipid fatty acyl composition. Removing the contribution of endogenous mouse LCAT by crossing the transgenic mice into the mouse LCAT knockout background resulted in even greater changes in HDL CE composition, with a 2.4-, 5-, and 5-fold increase in 20:4, 20:5 n-3, and 22:6 n-3 cholesteryl esters in the hE149A mice compared with hLCAT-wt Tg mice, respectively.Our results demonstrate that in vivo expression of hE149A significantly enriches HDL cholesteryl esters in 20- and 22-carbon fatty acyl species without affecting HDL concentration or size. Furthermore, the data suggest that endogenous mouse LCAT in hLCAT transgenic mice contributes to the plasma HDL CE pool out of proportion to its mass, presumably because the hLCAT transgene is poorly activated by mouse apolipoprotein A-I.

Total Fat Intake Modifies Plasma Fatty Acid Composition in Humans

Plasma fatty acid composition reflects dietary fatty acids. Whether the total fat content of the diet alters the fatty acid composition of plasma phospholipid, cholesteryl ester, triacylglycerol and free fatty acids is unknown. To evaluate the effects of low versus high fat diets on plasma fatty acids, a 12-wk, randomized, crossover, controlled feeding trial was conducted in healthy men and women with isoenergic low fat (20% energy) and high fat (45% energy) diets containing constant proportions of fatty acids. Ten subjects consumed one experimental diet for 28 d, their usual diet for 4 wk and the alternate experimental diet for 28 d. Endpoint measures of plasma fatty acids were determined at the end of each experimental period. The effects of the two diets were compared within subjects by analysis of variance. Plasma fatty acids (%) varied in response to total dietary fat with significantly greater total polyunsaturated fat, (n-6) and 18:2(n-6) levels in phospholipids and cholesteryl esters after high fat dietary consumption. The low fat diet was associated with significantly greater total (n-3) fatty acids, 20:5(n-3) and 22:6(n-3) levels in plasma phospholipid fatty acids and cholesteryl esters. Consumption of a low fat diet alters fatty acid patterns in a manner similar to that observed with feeding of (n-3) long-chain fatty acids. This change is likely related to decreased competition for the enzymes of elongation and desaturation, with reduced total intake of 18:2(n-6) favoring elongation and desaturation of available (n-3) fatty acids.