The effect of dietary n−3 polyunsaturated fatty acids on HDL cholesterol in Chukot residentsvs muscovites

Native Chukot Peninsula residents, in contrast to Muscovites, consume a diet rich in n−3 polyunsaturated fatty acids. This dietary peculiarity is reflected in differences in plasma lipid and apolipoprotein contents. The Chukot residents have lower contents of total cholesterol, triglyceride, LDL (low density lipoprotein) cholesterol and apolipoprotein B, but higher HDL (high density lipoprotein) cholesterol levels than do Muscovites. The apolipoprotein A-I levels were identical in both groups. A higher HDL cholesterol to apolipoprotein A-I ratio was determined in the coastline Chukot residents (0.52±0.01) than in Muscovites (0.43±0.01; p<0.01). In contrast to Muscovites, the coastline Chukot residents also had higher n−3 and lower n−6 polyunsaturated fatty acid percentages in plasma and erythrocyte lipids, and lower phosphatidylcholine and higher sphingomyelin or phosphatidylethanolamine levels in HDL_2b and HDL₃. The higher HDL cholesterol levels in the plasma of the coastline Chukot residents appears to reflect the higher cholesterol-scavenging capacity of their HDL. We conclude from this study that the regular consumption of dietary n−3 polyunsaturated fatty acids by the coastline Chukot residents decreased LDL cholesterol transfer from plasma to peripheral cells, and enhanced cholesterol efflux from cellular membranes toward HDL.     

Species variation in the atherogenic profile of monkeys: Relationship between dietary fats,lipoproteins, and platelet aggregation

Because lipoproteins and platelet aggregation have been implicated in atherogenesis, relative differences in the response of these variables to dietary fat saturation were compared in three species of monkeys differing in their susceptibility to atherosclerosis (cebus, rhesus, and squirrel monkeys). Both long-term (8–12 years) and short-term (8 weeks) responses to diets containing 31% fat calories were examined in the same monkeys. As expected, long-term feeding of coconut oil by comparison to corn oil produced significantly higher plasma concentrations of total cholesterol, LDL cholesterol, apoB, and triglycerides, as well as higher ratios of LDL/HDL cholesterol and apo B/apo A-I. These responses were characteristic of all species with cebus being most responsive and rhesus the least. The shortterm plasma cholesterol response to animal fats (butter, lard, beef tallow) was significantly less than that to coconut oil. When fish oil was substituted for two-thirds of either corn oil or coconut oil, exceptional decreases occurred in plasma cholesterol and triglycerides, as well as in HDL cholesterol and apo A-I concentrations despite the fact that the fish oil diets contained more saturated fat and less polyenes than the corn oil diet. Platelet aggregation tended to increase with saturated fat consumption and greatly decreased with fish oil intake in all monkeys, although cebus monkeys were ten-fold more resistant to platelet aggregation than the other two species. The molecular species of platelet phosphatidylcholine (PC) varied with both the dietary fat fed and species of monkey. An inverse correlation (r=−0.60; p<0.001) was found between changes in one such PC molecular species (18∶0−20∶4) induced by diet and the platelet aggregation threshold. These results demonstrate that the lipemic and platelet responses to dietary saturated fat depend upon both the type of fat (i.e., the specific combination of dietary fatty acids, including the chain length of saturated fatty acids and the degree of polyunsaturation) and the species of monkey (genetic component) in which the response is elicited.     

Compositional changes and apoprotein A-I metabolism of plasma high density lipoprotein in estrogenized chicks

The effect of estrogen on compositional changes, apolipoprotein (apo) A-I metabolism and the morphology of plasma high density lipoprotein (HDL) were investigated in chicks. The administration of 17β-estradiol (25 mg/kg body weight) to growing male chicks (8-week-old) markedly reduced the concentrations of plasma HDL components, except for triglyceride (TG). At the same time, levels of TG, total cholesterol (TC) and phospholipid (PL) in plasma were greatly elevated. The respective values for TG, TC, PL and protein in HDL were 13.9, 89.3, 154.1 and 231.7 (mg/dL) in the control, and 39.0, 35.1, 113.8 and 160.0 (mg/dL) in chicks upon estrogen treatment for one day.In vivo kinetic studies showed that the fractional catabolic rate of HDL apo A-I was significantly higher (p<0.05) in estrogen-treated chicks than in control birds, indicating an increased efficiency of HDL removal in the former. The production rate of HDL apo A-I also was significantly lower (p<0.05) in estrogen-treated chicks. Sodium dodecyl sulfate-acrylamide gel electrophoresis followed by laser scanning densitometry of HDL apolipoproteins in estrogen-treated chicks revealed a reduction of apo A-I and the occurrence of new apolipoproteins which had been absent in HDL of untreated birds. The HDL particles showed that the mean particle size of HDL became larger upon estrogen treatment. Particles with diameters between 70 and 123 Å were predominant in HDL of control chicks, while particles with diameters between 97 and 143 Å were most abundant in HDL of estrogen-treated chicks.     

Serum apolipoproteins B and A-I and naturally occurring fatty liver in dairy cows

Serum lipids and apolipoprotein (apo) B and A-I concentrations were determined in 164 dairy cows which had undergone liver biopsy in early lactation. The animals were divided into groups according to fatty liver severity on the basis of hepatic triglyceride content. The serum free fatty acid (FFA) concentration was higher in cows that developed fatty livers than in normal cows, and it correlated highly with liver triglycerides. Serum total cholesterol and triglyceride levels did not correlate with hepatic triglycerides. Both apo B and apo A-I levels were significantly decreased in fatty liver cows. In particular, apo B levels showed a strongly negative correlation with liver triglycerides. The present results suggest that hepatic apolipoprotein synthesis is impeded in fatty liver cows.     

LXR Agonist Increases the Lymph HDL Transport in Rats by Promoting Reciprocally Intestinal ABCA1 and apo A-I mRNA Levels

Liver and intestine are major sites of apo A-I synthesis in mammals. ABCAI is reported to be involved in the secretion of nascent HDL from cultured intestinal cells. However, whether ABCA1 participates in the secretion of nascent HDL from the intestine has not been assessed directly in vivo. This study examined the effect of a synthetic LXR-agonist “TO” on the lymphatic transport of HDL in thoracic duct-cannulated rats. The feeding of a TO-containing diet resulted in an increased transport of cholesterol and apo A-I in the lymph d > 1.063 g/ml lipoprotein fraction than did the feeding of a control diet without TO. The transport of cholesterol in whole lymph was lower, whereas the transport of apo A-I was higher, in the TO group. The abundance of mRNAs for ABCAI and apo A-I in the intestine was increased in the TO group. Furthermore, although the TO-containing diet reportedly increased the serum HDL concentration in intact mice and rats, no such effect was observed in the cannulated rats. The LXR agonist stimulated in vivo the synthesis of nascent HDL by increasing reciprocally the mRNA for ABCAI and apo A-I in the intestine, thereby contributing to an increase in the circulating HDL.     

Plasma high density lipoprotein subgroup distribution in rats fed diets with varying amounts of sucrose and sunflower oil

The effect of varying the dietary sunflower oil/sucrose (SO/SU) ratio on rat plasma lipid concentration and lipoprotein distribution was studied. Four groups of 10 rats were fed for 4 weeks diets with varying SO/SU ratios. Lipoprotein components were then estimated in whole plasma and after cumulative density ultracentrifugation. Whole plasma triacylglycerol (TG), total cholesterol (TC) and free cholesterol (FC) decreased with increasing SO/SU ratio; the CE/FC ratio increased, because CE remained virtually unaltered. Plasma TG-lowering was due to a decrease in VLDL and LDL-TG. Protein, CE and FC in d=1.063–1.100 g/ml (HDL_2b) and d=1.100–1.125 g/ml (HDL_2a) lipoproteins decreased upon increasing the SO/SU ratio. In contrast, in d=1.125–1.200 g/ml (HDL₃) lipoproteins, there was a concomitant increase in these components. Although increasing the SO/SU ratio effected more protein and CE transportation in HDL₃ and less in HDL₂, the total amount of these components in high density lipoproteins (d=1.063–1.200 g/ml) remained constant. Apo A-I and apo C-III decreased in HDL₂ but increased in HDL₃ upon increasing the SO/SU ratio. Also, HDL₂ apo E, and the apo C-II/apo C-III and small apo B/large apo B ratios in VLDL and LDL were lowered by increasing the SO/SU ratio. The hepatic VLDL-TG output during isolated liver perfusion was lowest in rats fed the diet with the highest SO/SU ratio. In perfusate, like in plasma, the VLDL and LDL apo C-II/apo C-III ratio, as well as the small apo B/large apo B ratio, decreased upon increasing the dietary SO/SU ratio. The results indicate that there can be appreciable diet-dependent variations in plasma HDL subgroup distribution in spite of unchanged total HDL levels.     

HDL3-mediated cholesterol efflux from cultured enterocytes: The role of apoproteins A-I and A-II

High density lipoproteins (HDL) were recently demonstrated in an enterocyte model (CaCo-2 cells) to mediate reverse cholesterol transport by retroendocytosis. The present study was carried out to define the role of the major HDL apoproteins (apo) A-I and apo A-II in this pathway. HDL₃ was fractionated by heparin affinity chromatography into the two main fractions containing either apo A-I only (fraction A) or both apo A-I and apo A-II (fraction B). In addition, liposomes were reconstituted from purified apo A-I or apo A-II and dimyristoyl phosphatidylcholine. The cell binding properties and cholesterol efflux potential were studied in the lipoprotein fractions and the liposomes. Both fractions exhibited similar maximal binding capacities of 4427 (A) and 5041 (B) ng/mg cell protein, but their dissociation constants differed (40.5 and 167.7 μg/mL, respectively). Fraction A induced cholesterol efflux and stimulated cholesterol synthesis more than did fraction B. Fraction A mobilized both cellular free and esterified cholesterol, whereas fraction B preferentially mobilized cholesteryl esters. Liposomes, containing either apo A-I or apo A-II, showed specific binding, endocytosis and endosomal transport, and were released as intact particles. Apo A-I liposomes also mediated cholesterol efflux. In conclusion, there is evidence that the HDL₃ subfractions A and B, as well as reconstituted liposomes containing either apo A-I or apo A-II, were specifically bound and entered a retroendocytosis pathway which was directly linked to cholesterol efflux. Quantitatively, the apo A-I subfraction appeared to play the dominant role in normal enterocytes. The apo A-II content of fraction B was related to the mobilization of cholesteryl esters.     

Quantitation of high density lipoproteins

The demand for high density lipoprotein (HDL) quantitation has dramatically increased with the renewed awareness of the importance of HDL as a negative risk factor for coronary heart disease. HDL is usually estimated by specific precipitation of the non-HDL apoB-containing lipoproteins by polyanions and divalent cations followed by measurement of cholesterol in the supernatant. A common procedure involves precipitation with sodium heparin at 1.3 mg/ml and MnCl₂ at 0.046 M (final concentrations). This method is appropriate for serum but less than ideal for plasma because of incomplete precipitation and sedimentation of the apoB-containing lipoproteins. A two-fold increase in Mn²⁺ to 0.096 M improves precipitation of the apoB-associated lipoproteins from plasma without excessive precipitation of HDL. This modified heparin-Mn²⁺ procedure gives results nearly identical to the results with the ultracentrifugal reference method (cholesterol in the d>1.063 fraction corrected for losses and the presence of apoB-associated cholesterol). The dextran sulfate 500-Mg²⁺ and the sodium phosphotungstate-Mg²⁺ procedures give results consistently 2–4 mg/dl lower than does the reference method. In contrast, a heparin-Ca²⁺ method gives results 5–8 mg/dl higher than does the reference method. Immunochemical analysis of apoA-I in the precipitate and apoB in the supernatant indicates that lower values for the phosphotungstate-Mg²⁺ procedure is due to partial precipitation of the A-I-containing lipoproteins, while higher values by the heparin-Ca²⁺ method are due to incomplete precipitation of the apoB-containing lipoproteins. Quantitation of the principal apoproteins of HDL, A-I and A-II, represent an important additional index of HDL concentrations and composition. Quantitation of plasma A-I and A-II concentrations by radial immunodiffusion indicates that women generally have higher HDL concentrations than men (women, A-I, 135±25, A-II, 36±6; men, A-I, 120±20, A-II, 33±5; mean±S.D., in mg/dl). A-I and A-II do not increase with age in men but show a slight increase with age in women. Estrogen increases HDL cholesterol and protein and may in part account for the higher HDL in women. The lighter density HDL subclass has a higher A-I/A-II ratio than the denser HDL subclass, with women generally having significantly more of the lighter HDL subclass. Density-gradient ultracentrifugation in CsCl₂ gradients indicates that HDL contains subpopulations of differing hydrated density which vary in the A-I/A=II ratio. Immunoassay of A-I and A-II when used in combination with HDL cholesterol analysis is a powerful tool for studies of HDL structure, epidemiology and metabolism.     

Regulation of intestinal apolipoprotein A-I synthesis by dietary phosphatidylcholine in newborn swine

Phospholipid (PL) from both dietary sources and biliary secretions may be important in the regulation of intestinal apolipoprotein (apo) synthesis. We previously demonstrated the up-regulation of apo A-I secretion by phosphatidylcholine (PC) in a newborn piglet intestinal epithelial cell line. We hypothesized that dietary PC increases small intestinal apo A-I synthesis in vivo in the newborn piglet. Two-day-old female swine were fed by gavage for 48 h. Diets consisted of a formula containing 51% of calories as triacylglycerol providing 180 kcal/kg/24 h. The experimental group (+PC, n=7) received 1 g/L added soybean PC, and the control group (−PC, n=7) received no added PC. At the end of the study period, jejunal apo A-I, B, and A-IV synthesis was measured, and apo A-I mRNA levels were quantitated. Jejunal mucosal PI content and serum lipids and apo B and A-I levels were measured. Jejunal apo A-I synthesis was almost twice as high in the +PC group as compared to the −PC group with no difference in apo A-I mRNA levels. Jejunal content of PL was higher in the +PC group than in the −PC group. There were no differences in jejunal apo B and A-IV synthesis or serum levels of lipids and apo-lipoproteins between the two groups. Dietary PC supplementation in newborn swine up-regulated jejunal apo A-I synthesis. Apo A-IV synthesis, which is sensitive to fatty acid flux, was not significantly increased, which suggests a specific effect of PC on apo A-I synthesis. Lumenal PC may be important in the regulation of intestinal apo A-I synthesis in the neonate.     

Serum Opacity Factor Enhances HDL-Mediated Cholesterol Efflux,Esterification and Anti Inflammatory Effects

Serum opacity factor (SOF) is a streptococcal protein that disrupts the structure of human high density lipoproteins (HDL) releasing lipid-free apo A-I while forming a large cholesteryl ester-rich particle and a small neo HDL. Given its low cholesterol and high phospholipid contents, we tested the hypotheses that neo HDL is a better substrate for cholesterol esterification via lecithin:cholesterol acyltransferase (LCAT), better than HDL as an acceptor of THP-1 macrophage cholesterol efflux, and improves reduction of oxidized LDL-induced production of inflammatory markers. We observed that both cholesterol efflux and esterification were improved by recombinant (r)SOF treatment of whole plasma and that the underlying cause of the improved cholesterol esterification in plasma and macrophage cholesterol efflux to rSOF-treated plasma was due to the rSOF-mediated conversion of HDL to neo HDL. Moreover, the reduction of secretion of TNF-α and IL-6 by THP-1 cells by neo HDL was twice that of HDL. Studies in BHK cells overexpressing cholesterol transporters showed that efflux to neo HDL occurred primarily via ABCA1 not ABCG1. Thus, rSOF improves two steps in reverse cholesterol transport with a concomitant reduction in the release of macrophage markers of inflammation. We conclude that rSOF catalyzes a novel reaction that might be developed as a new therapy that prevents or reverses atherosclerosis via improved reverse cholesterol transport.     

Dietary fat in the prevention of cardiovascular disease — a review

In developed countries atherosclerotic cardiovascular disease is the reason for about 50% of all deaths. With growing prosperity coronary heart disease (CHD) is becoming the major cause of premature death. Most complications of atherosclerosis occur unexpectedly and more than 50% of the patients developing a myocardial infarction do not survive more than one year. Because of the severe morbidity and high mortality primary prevention is likely to be the only solution. Epidemiological studies show a strong, positive relationship between plasma cholesterol concentrations and the incidence of CHD. People who immigrate from low‐risk to high‐risk areas usually acquire similar plasma cholesterol levels as the native population and a similar CHD risk. This demonstrates that environmental rather than genetic factors lead to the differences in cardiovascular risk and supports the notion that nutrition and lifestyle play a major role. The association between dietary intake of fat and cholesterol and the extent of atherosclerosis and CHD has been recognized in previous studies. The amount of saturated fat in the diet correlates stronger with the incidence of CHD than with total fat intake. The consumption of unsaturated fatty acids, however, appears to be beneficial, since it is inversely correlated with the plasma cholesterol concentration and risk of myocardial infarction. Lately additional nutritional factors like trans fatty acids with a negative influence on risk as well as positive factors like linolenic acid have attracted much attention. In conclusion, as a challenge to public health, preventive medicine needs to focus on changes in dietary habits with priority, particularly fat modification. A nutrition low in total fat primarily avoiding saturated and trans fatty acids, but rich in fruit and vegetables should be recommended.     

Xanthophylls, Phytosterols and Pre-β1-HDL are Differentially Affected by Fenofibrate and Niacin HDL-Raising in a Cross-Over Study

Fenofibrate and extended‐release (ER) niacin similarly raise high‐density lipoprotein cholesterol (HDL‐C) concentration but their effects on levels of potent plasma antioxidant xanthophylls (lutein and zeaxanthin) and phytosterols obtained from dietary sources, and any relationship with plasma lipoproteins and pre‐β1‐HDL levels, have not been investigated. We studied these parameters in 66 dyslipidemic patients treated for 6 week with fenofibrate (160 mg/day) or ER‐niacin (0.5 g/day for 3 week, then 1 g/day) in a cross‐over study. Both treatments increased HDL‐C (16 %) and apolipoprotein (apo) A‐I (7 %) but only fenofibrate increased apoA‐II (28 %). Lutein and zeaxanthin levels were unaffected by fenofibrate but inversely correlated with percentage change in apoB and low‐density lipoprotein cholesterol and positively correlated with end of treatment apoA‐II. ApoA‐II in isolated HDL in vitro bound more lutein than apoA‐I. Xanthophylls were increased by ER‐niacin (each ~30 %) without any correlation to lipoprotein or apo levels. Only fenofibrate markedly decreased plasma markers of cholesterol absorption; pre‐β1‐HDL was significantly decreased by fenofibrate (?19 %, p < 0.0001), with little change (3.4 %) for ER‐niacin. Although fenofibrate and ER‐niacin similarly increased plasma HDL‐C and apoA‐I, effects on plasma xanthophylls, phytosterols and pre‐β1‐HDL differed markedly, suggesting differences in intestinal lipidation of HDL. In addition, the in vitro investigations suggest an important role of plasma apoA‐II in xanthophyll metabolism.     

Apolipoprotein A-I,A-IV and E synthesis in the liver of copper-deficient rats

Copper deficiency induces hypercholesterolemia in the rat. This hypercholesterolemia is mainly due to an increase in apo E-rich high density lipoproteins (HDL₁). The present study was undertaken to determine whether the HDL increase could be explained by altered low-molecular weight apolipoprotein (apo) synthesis in the liver. The effect of copper deficiency on apo A-I, apo A-IV and apo E concentrations in plasma, as well as on respective mRNA levels and synthesis in the liver, were therefore investigated. We observed that the increased HDL₁ levels in the plasma of copper-deficient rats were associated with a significant rise in plasma apo E concentrations; however, plasma apo A-I and apo A-IV concentrations remained unchanged. Liver apo synthesis and respective apo mRNA levels were not significantly altered in copper-deficient animals when compared to control rats. No changes in apo E mRNA levels in various tissues from copper-deficient, as compared to control rats, were noted. Based on the data obtained, it was concluded that the observed changes in plasma lipoprotein and apo concentrations are not related to changes in low-molecular weight apo synthesis in the liver. The mechanisms of the impaired catabolism of HDL₁ should be further evaluated to possibly explain the observed increase in this fraction in copper-deficient rats.     

Low fat-monounsaturated rich diets containing high-oleic peanuts improve serum lipoprotein profiles

Postmenopausal hypercholesterolemic women are at risk for cardiovascular disease and are encouraged to follow low-fat (LF) (≤30% energy) diets. However, these diets may have undesirable effects on high density lipoprotein cholesterol (HDL-C), apolipoprotein A-I (apo A-I) and triglycerides, whereas diets high in monounsaturated fats do not. Twenty postmenopausal hypercholesterolemic women previously consuming high-fat diets (34% energy) were placed on a low fat-monounsaturated rich diet (LFMR: 26%, 14% energy, respectively) for 6 mon. Sixteen women already eating LF diets (24% energy) were also followed to monitor variations in serum lipids due to seasonal variations. Twenty-five women successfully completed the study (LFMR=12, LF=13). Serum cholesterol decreased 10% (264 to 238 mg/dL, P≤0.01) and low density lipoprotein cholesterol (LDL-C) decreased 12% (182 to 161 mg/dL, P≤0.01) in the LFMR group, but did not change in the LF group. The reduction in serum cholesterol in the LFMR group was greater than estimated by predictive formulas. Serum triglycerides and apo A-I did not change in the LFMR group. A modest decrease in HDL-C, HDL₃-C, and apolipoprotein B (apo B) occurred in both groups, but only the LFMR group showed a trend toward beneficial changes in LDL-C/HDL-C and apo A-I/apo B ratios. Overall, the LFMR diet was well tolerated and resulted in an improved serum lipid and apolipoprotein profile. A portion of this material was presented earlier at the annual meeting of the American Oil Chemists’ Society and in abstract from (O’Byrne, D.J., Shireman, R.B., and Knauft, D., 1993. The effects of a low-fat/high-oleic acid diet on lipoproteins in postmenopausal hypercholesterolemic women. INFORM 4(4), 553, #SS7).     

Effect oftrans fatty acids on plasma lipids,platelet function and systolic blood pressure in stroke-prone spontaneously hypertensive rats

To investigate the effect oftrans fatty acids on plasma lipid levels and systolic blood pressure, hydrogenated corn oil was fed to SHRSP (stroke-prone spontaneously hypertensive rats) and WKY (Wistar-Kyoto) rats for 30 days. Significantly lower systolic blood pressure and plasma total cholesterol were observed in SHRSP rats fedtrans fatty acids when compared with rats fedcis fatty acids from olive oil. In addition, higher HDL cholesterol and lower VLDL plus chylomicron cholesterol levels were found in SHRSP rats fedtrans fatty acids. Although no significant changes of systolic blood pressure and plasma total cholesterol levels were observed in WKY rats aftertrans fatty acids treatment, WKY rats fedtrans fatty acids had lower plasma LDL cholesterol and higher HDL cholesterol levels. In addition, platelet aggregation induced by collagen was decreased in WKY rats fedtrans fatty acids. It is interesting thattrans fatty acids increased the activity of plasma lecithin:cholesterol acyltransferase (LCAT) in both SHRSP and WKY rats. The observed influence oftrans fatty acids on plasma lipid levels, systolic blood pressure and platelet aggregation suggests thattrans fatty acids might prevent thrombotic disorders in SHRSP rats.     

Effects of diet and high density lipoprotein subfractions on the removal of cellular cholesterol

The effects of isocaloric substitutions of dietary polyunsaturated and saturated fat on the composition and function of plasma high density lipoproteins (HDLs) were studied in 3 normal subjects who were fed saturate-rich and polyunsaturate-rich diet programs. Compared to the saturated diets (P/S=0.4), polyunsaturated fat diets (P/S=4 or 2) reduced both plasma cholesterol and triglyceride levels. In 2 of the subjects, HDL cholesterol concentrations increased with polyunsaturated fat caused a reduction in HDL fatty acyl content of oleate and an increase in linoleate. To determine whether the altered composition affected the removal of cell membrane cholesterol, HDL and their subfractions, HDL₂ and HDL₃, which were isolated from each of the diets, were incubated with Ehrlich ascites cells in vitro. The cells were prelabeled with [³H] cholesterol, and the release of labeled cholesterol from the cells into the medium containing the various HDL fractions was determined. HDL, irrespective of the type of dietary fat, caused a release of [³H] cholesterol from the cells into the medium. The amount of [³H] cholesterol recovered in the medium was dependent on the absolute concentration of HDL cholesterol added to the cells and was independent of the type of diet. These results indicate that HDL facilitates the removal of cholesterol from cells, but that the amount and rate of removal are independent of the changes in HDL composition that can be obtained by dietary perturbations.     

Liver and serum lipids and lipoproteins of rats fed 5% L-lysine

Soybean protein and casein supplemented with 1% Arg were compared for their ability to prevent fatty livers caused by excess dietary Lys. The concentrations of serum lipids and lipoproteins of rats fed 5% Lys and having vatty livers were also compared with those of rats fed the identical diet but lacking fatty livers when killed. The total liver lipids, triglycerides and cholesterol of rats fed 15% casein +5% Lys were 3.9, 12.4 and 2 times control values, respectively. Rats fed 5% Lys +1% Arg or 5% Lys with 15% soybean protein had liver lipid concentrations similar to controls fed no supplemental Lys. Serum total lipids, triglycerides, cholesterol, phospholipids and free fatty acids also did not change, and serum ketone bodies were slightly elevated with Lys feeding whether the rats had fatty livers or not. The concentrations of circulating HDL were slightly depressed in all rats fed 5% Lys while LDL were significantly elevated, particularly in rats without fatty livers. Serum VLDL did not change with 5% dietary Lys. Overall, excessive dietary Lys caused fatty livers which were prevented by varying the diet or length of feeding. Excess Lys feeding altered lipoprotein metabolism shown by decreased serum HDL and a substantial elevation in LDL. The latter was more apparent when the fat accumulation in liver was less severe or absent. The data suggest that the fatty liver from Lys excess is probably unrelated to increased fat mobilization from storage, decreased fat oxidation or to a major block in the transport of triglycerides from the liver to the circulation.     

Determination of HDL2 cholesterol by precipitation with dextran sulfate and magnesium chloride: Establishing optimal conditions for rat plasma

Optimal conditions for analyzing HDL₂ cholesterol in small amounts of rat plasma have been studied using different concentrations of dextran sulfate and MgCl₂ to precipitate lipoproteins containing apolipoprotein B and/or apo E. When the MgCl₂ level was 91 mM, the supernate cholesterol was rather constant at a level of about 50–60% of the total plasma cholesterol concentration. Immunochemical determination of the apo A-I content indicated that no major losses of the HDL₂ fraction took place under these conditions. The recovery of about 96% of HDL₂ lipoproteins after the precipitation of rat plasma and the almost complete absence of lipoproteins belonging to the VLDL, LDL and HDL₁ fractions was demonstrated by agarose gel electrophoresis. Thus, the method should be suitable for screening the HDL₂ cholesterol content in small volumes of rat plasma.     

Higher Lipophilic Index Indicates Higher Risk of Coronary Heart Disease in Postmenopausal Women

Fatty acids (FAs) are essential components of cell membranes and play an integral role in membrane fluidity. The lipophilic index [LI, defined as the sum of the products between FA levels and melting points (°C), divided by the total amount of FA: \({\text{LI}} = \frac{{\mathop \sum \nolimits_{k} [{\text{fatty acid}} \times {\text{melting point}}]}}{{\mathop \sum \nolimits_{k} {\text{fatty acid}} }}\)] is thought to reflect membrane and lipoprotein fluidity and may be associated with the risk of coronary heart disease (CHD). Therefore, we examined the associations of dietary and plasma phospholipid (PL) LI with CHD risk among postmenopausal women. We determined dietary LI for the cohort with completed baseline food frequency questionnaires and free of prevalent cardiovascular diseases in the Women’s Health Initiative (WHI) observational study (N = 85,563). We additionally determined plasma PL LI in a matched case-control study (N = 2428) nested within the WHI observational cohort study. Cox proportional hazard regression and multivariable conditional logistic regression were used to calculate HRs/ORs for CHD risk between quartiles of LI after adjusting for potential sources of confounding and selection bias. Higher dietary LI in the cohort study and plasma PL LI in the case-control study were significantly associated with increased risk of CHD: HR = 1.18 (95% CI 1.07–1.31, P for trend <0.01) and OR = 1.76 (95% CI 1.33–2.33, P for trend <0.01) comparing extreme quartiles and adjusting for potential confounders. These associations still persisted after adjusting for the polyunsaturated to saturated fat ratio. Our study indicated that higher LI based on either dietary or plasma measurements, representing higher FA lipophilicity, was associated with elevated risk of CHD among postmenopausal women.     

A preliminary study on platelet aggregation in postmenopausal women consuming extra-virgin olive oil and high-oleic acid sunflower oil

The purpose of the present study was to examine the effects of two monounsaturated fatty acid-rich oils, extravirgin olive oil (EVOO) and high-oleic sunflower oil (HOSO), on platelet aggregation in 14 postmenopausal women (aged 62.9 ± 1.8 yr) with high-fat dietary habits. Both oils contained oleic acid as the major compound (≈76% of total fatty acids), but the content of palmitic and linoleic acids and many minor compounds was significantly different. These oils were used as the only culinary fats during two 28-d periods, and represented ≈62% of the total lipid intake (≈46% of total energy consumption). Other dietary components were matched. The daily energy contribution of saturated, monounsaturated, and polyunsaturated fatty acids to the total energy consumption was 11.8, 28.5, and 2.8%, respectively, during the EVOO dietary period and 10.3, 27.8, and 4.6%, respectively, with HOSO. Aggregation in platelet-rich plasma was measured after addition of ADP. Platelet aggregation (expressed as cm/5 min) was significantly lower after the EVOO diet than after HOSO (2.1 ± 1.1 and 3.0 ± 1.4, respectively; P<0.05). Although maximal aggregation time was 40.2% higher in HOSO than in EVOO, the difference was not significant. Independent of serum cholesterol level, platelet aggregation tended to be different on the EVOO diet when women were classified according to cholesterol levels: <220 mg/dL or ≥220 mg/dL. Results suggest that other compounds present in the oils aside from the fatty acids may play an important role in modulating platelet aggregation in these postmenopausal women.