Jean T. Snook

Mechanisms mediating lipoprotein responses to diets with medium-chain triglyceride and lauric acid

Medium-chain triglycerides (MCT) are often used in specialized formula diets or designer fats because of their special properties. Yet their influence on lipid metabolis is not completely understood. In this two-period cross-over study, the effects of MCT (8∶0+10∶0) in contrast to a similar saturated fatty acid (12∶0) were compared. Eighteen healthy women ate a baseline diet [polyunsaturated (PUFA)/saturated fat=0.9] fer 1 wk. Then, they consumed test diets (PUFA/saturated fat=0.2) for 4 wk. Monounsaturated fat and cholesterol were constant in baseline and treatment diets. MCT and 12∶0, substituted for part of the PUFA, provided 14 energy (en)% of the test diets. In comparison to the PUFA baseline diet, a 16% increase in mean serum low density lipoprotein (LDL)-cholesterol (C) on the 12∶0 diet was accompanied by a 21% decrease in mean receptormediated degradation of LDL by freshly isolated mononuclear cells (MNC) in vitro. The MNC assay theoretically gives an indication of receptor-mediated degradation of LDL. In contrast, the MCT diet raised mean receptor-mediated degradation of LDL by 42%, a finding out of line with the mean 11% increase in serum LDL-C. Perhaps MCT, by increasing the rate of LDL-C production, overcame the rate of LDL-C clearance. The 12∶0 diet enhanced some factors involved in reverse cholesterol transport (e.g., high density lipoprotein fractions) while MCT had a different of less pronounced effect. The overall effects of MCT on cholesterol metabolism may or may not be desirable, whereas those of 12∶0 appear largely undesirable as previously reported.

Effect of apolipoprotein E polymorphism on serum lipoprotein response to saturated fatty acids

This report summarizes two studies which investigated the effects of apolipoprotein E (apoE) polymorphism on the serum total cholesterol (TC) and lipoprotein cholesterol responses to 8∶0+10∶0 and 12∶0 diets (Study I) and 14∶0, 16∶0, and 18∶0 diets (Study II). Eighteen healthy premenopausal women (3 apoE 3/2, 12 apoE 3/3, 3 apoE 4/3) in study I and another 18 healthy premenopausal women (4 apoE 3/2, 10 apoE 3/3, 3 apoE 4/3, 1 apoE 4/2) in study II consumed a baseline diet providing 40 en% total fat, 11 en % 18∶2, 15 en% 18∶1, 11.5 en% saturated fat for the first week of each 5-wk period. The experimental diets for both studies provided 40 3n% total fat, 13–14 en% as one of five test saturated fatty acids (SFA), 14–16 en% 18∶1, and 3–4 en% 18∶2. Analysis by apoE phenotypes showed that both the 8∶0+10∶0 diet and the 12∶0 diet in Study Linduced significant increases in serum TC in subjects with different apoE phenotypes with the exception of apoE 3/2 in the medium-chain triglyceride group. In contrast, in Study II, individuals with apoE 4/3 consuming the 14∶0 diet showed significant increases in serum TC, high density lipoprotein-cholesterol (HDL-C), and HDL2-C, but the same subjects consuming the 16∶0 diet showed significant increases in serum TC and low density lipoprotein-cholesterol. The findings from both studies indicated serum lipoprotein responses to SFA were different and the variation of responsiveness may be regulated, at least in part, by apoE polymorphism, especially when 14∶0, 16∶0, or 18∶0 was consumed.

Effect of ethanol use and other lifestyle variables on measures of selenium status

Relationships between blood levels of selenium (SE) and SE-dependent glutathione peroxidase (GPX) activity and selected lifestyle variables including ethanol ingestion, smoking behavior, nutrient intake, and nutrient supplement use were studied in 124 male and female subjects, half of whom drank alcoholic beverages lightly or moderately. Among the 19 independent lifestyle variables included in correlation and multiple regression analysis, ethanol intake was most strongly and consistently associated with levels of plasma and whole blood SE and plasma GPX activity, r = .32-.34, p less than 0.01. Light to moderate drinkers had higher, p less than 0.05, whole blood and plasma SE and GPX than subjects abstaining from alcohol. SE intake was not different. This positive association was in contrast to some previous reports in which alcoholics were shown to have lower blood SE levels than control subjects. A possible explanation could be the adequate SE intake and the light to moderate ethanol consumption of drinkers in this study.

Effect of Chronic Ethanol Consumption on Selenium Status and Utilization in Rats

Effects of chronic ingestion of 2 levels of alcohol on selenium (Se) utilization were determined in initially Se-depleted rats. Male weanling rats were fed ad lib a Se deficient (0.012 mg/kg) basal diet for 4 weeks and then were meal-fed low or marginally adequate Se in the form of high Se yeast for 4 weeks. During Se repletion, ethanol, which replaced medium-chain triglycerides in the diet, provided 10 or 20 percent of food energy. The basal diet provided 80% of food energy as well as adequate protein, vitamins and minerals. In rats given adequate Se moderate chronic ethanol consumption did not influence Se absorption or retention, but increasing ethanol level raised Se in liver and whole blood in a linear fashion and in kidney in a quadratic manner. In this rat model measures of Se status were reduced by low Se intake, not chronic moderate ethanol ingestion.

Relative effects of high saturated fatty acid levels in meat, dairy products, and tropical oils on serum lipoproteins and low-density lipoprotein degradation by mononuclear cells in healthy males

To determine the effects of three saturated fatty acid combinations on lipoprotein metabolism, we fed 18 21- to 32-year-old men three diets in a crossover design for 28-day periods separated by washout periods of 4 to 6 weeks. The men self-selected a prescribed diet at home emphasizing saturated fat as the visible fat for 1 week. Then, they ate experimental diets providing 40%, 15%, 17%, and 7% of food energy, respectively, as total, saturated, monounsaturated, and polyunsaturated fatty acids, levels representing amounts available in the US diet. Different test fatty acid combinations, given at 4 to 6 energy% (en%) each, were incorporated into food products: 12:0 + 14:0, 14:0 + 16:0, and 16:0 + 18:0. Test fatty acids were equalized by giving free myristic acid (14:0) with palm kernel oil or butter and sheanut butter (high in 18:0) with lard. The diet highest in 12:0 + 14:0 also provided 4.2 en% 16:0, the most common saturated fatty acid in the US diet. Mean apparent absorption of all fatty acids was at least 90%. The three diets produced similar concentrations of serum total and low-density lipoprotein (LDL) cholesterol and apolipoprotein (apo) B-100 regardless of the apo E phenotype of the subjects. Compared with baseline, the experimental diets affected serum high-density lipoprotein (HDL) concentrations (P < .06), with the highest values occurring on diet 12:0 + 14:0. When the change from baseline in receptor-mediated degradation of 125I-LDL in freshly isolated mononuclear cells (MNC) was stratified by apo E phenotype, diet 16:0 + 18:0 produced a 30% increase, compared with a 9% decrease on diet 12:0 + 14:0 and a 6% increase on diet 14:0 + 16:0 in subjects with the apo E3/3 phenotype. These results suggested that different saturated fatty acid combinations, consumed at levels typical of availability in the United States and with diets providing ample unsaturated fat, had similar cholesterolemic properties in healthy males despite some subtly different effects on lipoprotein metabolism.

The cholesterolaemic effects of dietary fats in cholesteryl ester transfer protein transgenic mice.

In order to investigate the role of cholesteryl ester transfer protein (CETP) in the cholesterolaemic response to dietary fats, we analysed plasma lipid profiles of CETP-transgenic and control C57BL/6 mice fed standard chow (AIN-93G; AIN), a low-fat diet, and diets high in butter (saturated fatty acids; SFA), high-oleic acid safflower oil (monounsaturated fatty acids; MUFA), and safflower oil (polyunsaturated fatty acids; PUFA) for 5 weeks. Each group contained four or five mice. There were significant diet and dietxgenotype effects on plasma total cholesterol (TC; and respectively), liver TC ( and respectively), and esterified cholesterol (EC; and respectively); diet effects on plasma triacylglycerol liver free cholesterol and body weight a genotype effect on body-weight gain and a dietxgenotype effect on energy intake In transgenic mice the SFA diet caused significantly higher plasma TC than the PUFA diet In control mice MUFA and PUFA diets, but not the SFA diet, caused significantly higher plasma TC than the low-fat and AIN diets Transgenic mice fed PUFA had lower plasma TC while transgenic mice fed MUFA had lower LDL+VLDL-cholesterol than controls in the same dietary groups. Transgenic mice fed MUFA and PUFA diets also had significantly higher liver TC and respectively) and EC and respectively) than controls fed the same diets. In the present study we showed that: (1) CETP transgenic mice had a cholesterolaemic response to dietary fats similar to that in human subjects; (2) CETP transgenic mice fed PUFA showed significantly lower plasma TC, while those fed MUFA had lower LDL+VLDL-cholesterol than controls; (3) hepatic accumulation of cholesterol, possibly resulting from the combination of the enhanced cholesteryl ester transfer to apolipoprotein B-containing lipoproteins and increased hepatic uptake of cholesterol, may contribute to the cholesterol-lowering effect of MUFA and PUFA in CETP-transgenic mice; (4) CETP may play a role in appetite and/or energy regulation.

Cholesteryl ester transfer and cholesterol esterification in type 1 diabetes: relationships with plasma glucose.

Abstract The activities of two crucial enzymes of reverse cholesterol transport, cholesterol ester transfer protein (CETP) and lecithin:cholesterol acyltransferase (LCAT), and their relationships with lipid profile and fasting plasma glucose were examined in 35 type 1 diabetic children. The CETP and LCAT activities were significantly lower (p<0.05) in the 4 subjects with normal fasting plasma glucose levels (<6.39 mmol/l) than in the 28 with high plasma glucose levels (CEPT activity, 10.63±3.81 vs. 32.18±13.94 nmol/ml h; LCAT activity, 25.52±4.53 vs. 39.52±12.52 nmol/ml h; both p<0.05). The subjects with high plasma glucose levels also had higher total and LDL-cholesterol than those with normal glucose levels. CETP activity was positively correlated with fasting plasma glucose, CETP concentration, LCAT activity, total cholesterol, free cholesterol, LDL-C, and LDL-cholesterol ester, while negatively correlated with cholesteryl ester to free cholesterol ratio, LDL triglyceride to protein ratio, and LDL triglyceride to cholesteryl ester ratio. LCAT activity was found to positively correlate with CETP activity, total cholesterol, free cholesterol. LDL-C, CETP concentration, and LDL-cholesteryl ester, while it negatively correlated with cholesteryl ester to free cholesterol ratio. The results observed in type 1 diabetic subjects suggest that (1) accelerated LCAT and CETP activities may result in the accumulation of LDL-cholesteryl ester; and (2) fasting plasma glucose may be a major determinant of CETP activity.

Sandwich enzyme-linked immunosorbent assay for plasma cholesteryl ester transfer protein concentration

OBJECTIVES Cholesteryl ester transfer protein (CETP) mediates the transfer of HDL cholesterol to apoB-containing lipoproteins. Its mass and activity are increased in several pro-atherogenic conditions. The objective of this study is to develop a cost- and time-effective sandwich ELISA for plasma CETP concentration. DESIGN AND METHODS Monoclonal anti-CETP, TP20, was used as the capture antibody, while the other biotinylated monoclonal anti-CETP, TP2, was used for detection. The results were expressed in an arbitrary unit, ng biotin-TP2 bound per microl plasma. Plasma CETP concentrations, activities and their relationship were assessed in 35 IDDM children. RESULTS The assay had an intra-assay CV of 8.75% and an inter-assay CV under 10%. Plasma CETP concentration of these subjects ranged from 0.36-1.89 ng biotin-TP2/microL. CETP concentration was significantly correlated with CETP activity (r = 0.51, p < 0.01). CONCLUSION The sandwich ELISA we have developed carried sufficient sensitivity for assaying plasma CETP concentration in human.

Does a diet high in corn oil lower LDL cholesterol levels in women via an effect on LDL receptor activity

Abstract To determine if a high intake of corn oil alters LDL receptor activity, a model emphasizing freshly isolated mononuclear cells (MNC) was used with 12 women consuming 40 energy% fat diets based on corn oil or butter in a randomized crossover design. Each phase included 1 week of a prescribed but selfselected high saturated fat (SFA) diet at home followed by 4 weeks of a designated fat-based diet consumed in a metabolic kitchen. There was a 7-week washout period between phases. LDL degradation through and binding to LDL receptors were 63 and 100% higher, respectively, on the corn oil diet than the butter diet ( P r = −0.4 to −0.6). Compared with the butter group, total unsaturated and saturated fatty acid contents of MNC were 166% higher and 150% lower, respectively, in the corn oil group. Changes in saturated and unsaturated fatty acids and receptor-mediated LDL degradation were associated ( r = −0.6 and 0.6, respectively). These findings suggest that the change in fatty acid composition in MNC may enhance the LDL receptor-mediated pathway. Receptor-mediated LDL degradation may account for only part of the effect of corn oil on serum LDL cholesterol concentrations in healthy women.

Effect of moderate to very low fat defined formula diets on serum lipids in healthy subjects

Serum total cholesterol, high density lipoprotein (HDL) cholesterol, and triglyceride levels were studied in healthy male and female subjects consuming for one-week periods a diet of conventional food (CF) providing 42% of energy as fat, principally butter fat, and then in random order nutritionally complete, defined formula diets of moderate (32%) to very low (1%) fat content. Compared to CF, the formula with 32% of energy as corn oil lowered serum cholesterol by 25% and the ratio of total to HDL-cholesterol by 13%. Low (9%) and very low (1–3%) fat formulas reduced HDL-cholesterol by as much as 40%, raised the total: HDL-cholesterol ratio by about 20% and raised serum triglyceride levels by as much as 100%. When low and very low fat formulas were ingested for three weeks, these effects persisted although maximal responses occurred during the first week. These results demonstrated that a moderate fat formula diet with a high P/S ratio had a more favorable effect on serum lipid levels than various low fat formulas. Low fat conventional food diets should be studied in long-term controlled metabolic experiments before such diets are recommended to the general population for coronary heart disease or cancer prevention.