Effects of finishing diets containing wet distillers grains plus solubles on beef quality attributes and fatty acid profile.

The objective of this study was to evaluate the effects of feeding wet distillers grains plus solubles (WDGS) on quality attributes of three beef muscles (longissimus lumborum, psoas major, and infraspinatus). Ninety-six, yearlings crossbred steers were randomly assigned to one of three dietary treatments (Corn, 15%, or 30% WDGS - DM basis) and fed for 133 d. No significant differences were observed in marbling score (P=0.89), marbling texture (P=0.70), and marbling distribution (P=0.36). Greater concentrations of PUFA and lower levels of 18:1(n-7) were observed in beef from steers fed 30% WDGS when compared to other treatments. Lipid oxidation was also greater in beef from steers fed 30% WDGS (P≤0.05). No significant differences were observed in sensorial attributes and Warner-Bratzler shear force (WBSF) for all muscles (P>0.05). Feeding WDGS increased PUFA and lipid oxidation, which may lead to shorter shelf life.

Effects of feeding modified distillers grains plus solubles on marbling attributes, proximate composition, and fatty acid profile of beef.

Wet distillers grains contain approximately 65% moisture. A partially dried product [modified distillers grains plus solubles (MDGS)] contains about 50% moisture. However, both have similar nutrient composition on a dry matter basis. The objective of this study was to investigate the effects of finishing diets varying in concentration of MDGS on marbling attributes, proximate composition, and fatty acid profile of beef. Yearling steers (n = 268) were randomly allotted to 36 pens, which were assigned randomly to 0, 10, 20, 30, 40 and 50% MDGS (DM basis) and fed for 176 d before harvest. The 48-h postmortem marbling score, marbling texture, and marbling distribution were assessed by a USDA grader and 1 ribeye slice (longissimus thoracis) 7 mm thick was collected from each carcass for proximate and fatty acid analyses. Treatments did not significantly alter marbling score or marbling distribution (P ≥ 0.05). United States Department of Agriculture Choice slices had coarser marbling texture when compared with USDA Select. Although dietary treatment affected marbling texture, no consistent pattern was evident. Diets did not influence fat content, moisture, or ash of the ribeye (P ≥ 0.05). For treatments 0, 10, 30, 40 and 50%, there were positive linear relationships between marbling score and fat percentage in the ribeye (P ≤ 0.05), and all slopes were similar (P = 0.45). Feeding MDGS linearly increased stearic, linoelaidic, linoleic, linolenic, PUFA, and n-6 fatty acids. As dietary MDGS increased, linear decreases were observed in all n-7 fatty acids and cubic relationships were detected for the 18:1 trans isomers [trans-6-8-octadecenoic acid (6-8t), elaidic acid (9t), trans-10-octadecenoic acid (10t), and trans vaccenic acid (11t)]. No effects were observed for saturated fatty acids containing 6 to 14 carbons. Feeding MDGS resulted in increased PUFA, trans, and n-6 fatty acids, minimal effects on marbling texture, and no effects on the relationship of marbling to intramuscular fat content relationship.

Beef quality of calf-fed steers finished on varying levels of corn-based wet distillers grains plus solubles.

Calf-fed crossbred steers (n = 94) were randomly allocated to 3 dietary treatments (0%, 15%, or 30% wet distillers grains plus solubles, WDGS; DM basis) and fed for 167 d to determine the effects on quality attributes of beef. At 48 h postmortem, marbling score, marbling texture, and marbling distribution were assessed by a USDA grader. After grading, one rib eye slice (longissimus thoracis) ¼7 mm thick was excised from each carcass, trimmed of subcutaneous fat, and analyzed for fatty acid profile and lipid content. At 7 d postmortem, 48 top blades (infraspinatus), strip loins (longissimus lumborum), and tenderloins (psoas major) (16 per treatment) were removed from shoulder clods and short loins and 2 steaks were obtained to measure mineral content, fatty acid profile (except strip loins), trained sensory analysis, objective color, and lipid oxidation. Finishing diet did not influence the content of total lipid (P = 0.19) or marbling, marbling texture, or marbling distribution (P = 0.46, 0.84, 0.40, respectively). Feeding WDGS created a linear increase (P < 0.01) of PUFA in all three muscles (longissimus thoracis showed 4.90%, 5.91%, and 6.23% PUFA for 0%, 15%, and 30% WDGS, respectively). Similar responses were observed for 18:2(n-6) and total n-6 fatty acids. Conversely, lower proportions of 18:1(n-7) fatty acid were observed in beef from animals fed 30% WDGS (P < 0.01). Total trans fatty acids increased linearly in strip loin and top blade steaks (P < 0.01), whereas proportions of 16:0 and 14:1(n-5) fatty acids decreased in all muscles (P < 0.01) as WDGS increased. Diet did not affect mineral content of top blades or strip loins. For tenderloin steaks, S concentration was lower when 30% of WDGS was fed (P = 0.05). No effects on sensory attributes and Warner-Bratzler shear force were observed (P > 0.50), except a minimal effect on strip loin juiciness (5.32, 4.86, and 5.52 for 0%, 15%, and 30% WDGS, respectively; P = 0.02). Top blade and tenderloin steaks from cattle fed 30% WDGS were significantly less red (lower a* values) on d 3 of simulated retail display (P < 0.04). Inclusion of 30% WDGS in the diet resulted in higher levels of oxidation after 7 d of retail display for top blade and strip loin steaks (P < 0.01). Feeding WDGS to calf-fed steers altered fatty acid profile, increased oxidation, and decreased color stability during retail display.

Chemical and sensory properties of beef of known source and finished on wet distillers grains diets containing varying types and levels of roughage.

Beef knuckles (n = 160) were obtained from source-verified cattle finished on 30% wet distillers grains plus solubles enriched with varying levels of alfalfa hay (4 or 8%), corn silage (6 or 12%), or corn stalks (3 or 6%) based on NDF. Proximate analysis, pH, oxidation-reduction potential, fatty acid composition, and sensory analysis were conducted on the rectus femoris muscle to determine if roughage inclusion, in conjunction with wet distillers grains plus solubles and cattle source, affects beef flavor with particular interest in liver-like off-flavor. Proximate analysis, fat content, and oxidation-reduction potential were unaffected (P ge;0.129) by diet or source. For s.c. adipose tissue, cattle from Nebraska (NE) had greater amounts of MUFA (P = 0.048) and unsaturated fatty acids (P = 0.068) but less SFA (P = 0.065) when compared with cattle from South Dakota. Diet affected s.c. adipose tissue levels of 15:0, 17:0, and n-3 fatty acids in which cattle from NE finished on the low corn stalk diet had (P < or =0.050) lower levels. Cattle from NE had (P < or = 0.049) greater i.m. adipose proportions of 13:0 and CLA. Dietary effects (P < or = 0.050) were observed for i.m. adipose tissue proportions of 16:0, 18:1(n-9), 18:2(n-6), 20:4(n-6), 22:5(n-3), MUFA, PUFA, and n-6 fatty acids. Sensory analysis revealed that cattle from NE were (P < or = 0.023) less juicy and had less bloody notes when compared with cattle from South Dakota. Cattle finished on the low alfalfa diet were (P < or = 0.014) more tender and juicy but had more bloody notes. No (P ge; 0.670) dietary or source effects were noted for liver-like off-flavor. Subcutaneous amounts of 18:2(n-6 trans) (r = -0.17) were inversely related to the incidence of liver-like off-flavor, whereas 20:1(n-9) (r = 0.21), CLA cis-9, trans-11 (r = 0.16) were directly related. Data from this study indicate that type and level of roughage inclusion and cattle source have minimal effects on fatty acid profiles and sensory properties of the musculus rectus femoris. However, individual fatty acids of s.c. and i.m. adipose tissue were significantly correlated with liver-like off-flavor.

Soy isoflavones improve plasma lipids in normocholesterolemic, premenopausal women.

BACKGROUND: Soy consumption is known to reduce plasma total cholesterol and LDL cholesterol in hypercholesterolemic subjects, but the responsible soy components and the effects in normocholesterolemic subjects remain unclear. OBJECTIVE: The effects of soy isoflavone consumption on plasma total cholesterol, HDL-cholesterol, LDL-cholesterol, triacylglycerol, apolipoprotein A-I, apolipoprotein B, and lipoprotein(a) concentrations and on LDL peak particle diameter were examined in normocholesterolemic, premenopausal women. DESIGN: Thirteen healthy, normocholesterolemic, free-living, premenopausal female volunteers took part in this randomized, crossover-controlled trial. Each subject acted as her own control. Three soy isoflavone intakes (control: 10.0 +/- 1.1; low: 64.7 +/- 9.4; and high: 128.7 +/- 15.7 mg/d), provided as soy protein isolate, were consumed for 3 menstrual cycles each. Total cholesterol, HDL cholesterol, LDL cholesterol, and triacylglycerol were measured over the menstrual cycle. Apolipoprotein A-I, apolipoprotein B, lipoprotein(a), and LDL peak particle diameter were evaluated in the midluteal phase. RESULTS: Total cholesterol, HDL-cholesterol, and LDL-cholesterol concentrations changed significantly across menstrual cycle phases (P < 0.005). During specific phases of the cycle, the high-isoflavone diet lowered LDL cholesterol by 7.6-10.0% (P < 0.05), the ratio of total cholesterol to HDL cholesterol by 10.2% (P < 0.005), and the ratio of LDL to HDL cholesterol by 13.8% (P < 0.002). CONCLUSIONS: Isoflavones significantly improved the lipid profile across the menstrual cycle in normocholesterolemic, premenopausal women. Although of small magnitude, these effects could contribute to a lower risk of developing coronary heart disease in healthy people who consume soy over many years.

Dietary stearic acid reduces cholesterol absorption and increases endogenous cholesterol excretion in hamsters fed cereal-based diets.

The observation that dietary stearic acid does not raise plasma cholesterol concentration is well documented, although the regulating mechanisms are not completely understood. Therefore, we examined the effect of dietary stearic acid on cholesterol absorption and sterol balance using male Syrian hamsters fed modified NIH-07 cereal-based diets selectively enriched in palmitic acid (16:0), stearic acid (18:0), trans fatty acid (18:1t), cis oleic acid (18:1c) or linoleic acid (18:2). All diets contained 17 g/100 g total fat and 0.05 g/100 g cholesterol; the five fat blends were enriched 30% with the fatty acid of interest above a constant fatty acid background. Cholesterol absorption efficiency was 50-55% in all treatment groups except for the 18:0 group, in which cholesterol absorption was significantly reduced to 21%. Plasma total cholesterol concentration was significantly lower in the 18:0 group compared to the 16:0 group. Fecal neutral steroid excretion was significantly greater in hamsters fed the high 18:0 diet compared to the other treatment groups. After accounting for unabsorbed dietary cholesterol, endogenous cholesterol excretion was about 100% higher in the 18:0 group. Consequently, the calculated rate of whole body cholesterol synthesis was significantly increased by dietary 18:0. Bile acid excretion accounted for only 12-20% of total sterol output by the hamsters in this study. Thus, the data suggest that reduced plasma cholesterol concentration in hamsters fed high 18:0 diets may be influenced by reduced cholesterol absorption and increased excretion of endogenous cholesterol.

Cholesteryl ester enrichment of plasma low-density lipoproteins in hamsters fed cereal-based diets containing cholesterol.

Male Syrian hamsters were fed 0.02, 0.03, or 0.05% cholesterol to test the hypothesis that moderate cholesterol intake increases the cholesteryl ester content of the plasma low-density lipoproteins (LDL). Dietary cholesterol levels of 0.02%-0.05% were chosen to reflect typical human intakes of cholesterol. Hamsters were fed ad libitum a cereal-based diet (modified NIH-07 open formula) for 15 weeks. Increasing dietary cholesterol from 0.02% to 0.05% resulted in significantly increased plasma LDL and high-density lipoprotein cholesterol concentration, increased liver cholesterol concentration, and increased total aorta cholesterol content. The cholesteryl ester content of plasma LDL was determined as the molar ratio of cholesteryl ester to apolipoprotein B and to surface lipid (i.e., phospholipid + free cholesterol). Increasing dietary cholesterol from 0.02% to 0.05% resulted in significantly increased cholesteryl ester content of LDL particles. Furthermore, cholesteryl ester content of LDL was directly associated with increased total aorta cholesterol, whereas a linear relationship between plasma LDL cholesterol concentration and aorta cholesterol was not observed. Thus, the data suggest that LDL cholesteryl ester content may be an important atherogenic feature of plasma LDL.

Biliary cholesterol and bile acid excretion do not increase in hamsters fed cereal-based diets containing cholesterol.

The major compensatory responses to increased cholesterol consumption are decreased cholesterol synthesis and increased cholesterol excretion through the bile either as free cholesterol or bile acids. The objective of this study was to test the hypothesis that biliary cholesterol excretion is increased in hamsters fed low levels of cholesterol reflecting normal human intake. The hypothesis was based on observations that hamsters generally resist changes in bile acid synthesis when fed large amounts of cholesterol; therefore, increased biliary cholesterol excretion represents a potentially significant pathway for elimination of excess cholesterol in this species. Hamsters were fed modified NIH-07 cereal-based diets containing 0.02%, 0.03%, and 0.05% cholesterol (0.04, 0.06, and 0.10 mg cholesterol/kcal, respectively). The primary response to increasing amounts of dietary cholesterol was downregulation of whole-body cholesterol synthesis, reduced from 3.93+/-0.14 micromol x d(-1) x 100 g(-1) body weight in hamsters fed 0.02% cholesterol to 0.52+/-0.14 micromol x d(-1) x 100 g(-1) in the 0.05% cholesterol group. Biliary cholesterol excretion was also slightly reduced in hamsters fed 0.05% cholesterol, whereas bile acid excretion was not altered by dietary cholesterol. Despite a pronounced downregulation of whole-body cholesterol synthesis, liver and plasma cholesterol concentrations increased in hamsters fed 0.05% cholesterol. The data indicate that increased biliary cholesterol excretion is not a major compensatory route of cholesterol excretion in hamsters consuming cholesterol. Furthermore, cholesterol added to the diet at 0.05% appears to be the approximate threshold at which compensatory mechanisms can prevent increases in liver and plasma cholesterol in male Syrian hamsters. Consequently, this species may be an appropriate animal model for "hyperresponding" individuals in the human population.

Hepatic origin of cholesteryl oleate in coronary artery atherosclerosis in African green monkeys. Enrichment by dietary monounsaturated fat.

Relationships among plasma lipoprotein cholesterol, cholesterol secretion by the isolated, perfused liver, and coronary artery atherosclerosis were examined in African green monkeys fed diets containing cholesterol and 35% of calories as fat enriched in polyunsaturated, monounsaturated, or saturated fatty acids. The livers of animals fed monounsaturated fat had significantly higher cholesteryl ester concentrations (8.5 mg/g wet wt) than the livers of the other diet groups (3.65 and 3.37 mg/g wet wt for saturated and polyunsaturated fat groups, respectively) and this concentration was highly correlated with plasma cholesterol and apoB concentrations in each diet group. Cholesteryl oleate was 58 and 74. 5% of the liver cholesteryl ester in the saturated and monounsaturated fat groups. In each diet group, perfusate cholesteryl ester accumulation rate was highly correlated to liver and plasma cholesterol concentrations, and to plasma LDL cholesteryl ester content. Cholesteryl oleate was 48 and 67% of the cholesteryl esters that accumulated in perfusate in the saturated and monounsaturated fat animals, and this percentage was very highly correlated (r = -0.9) with plasma apoB concentration. Finally, in these two diet groups, liver perfusate cholesteryl ester accumulation rate was well correlated (r >/= 0.8) to coronary artery cholesteryl ester concentration, a measure of the extent of coronary artery atherosclerosis that occurred over the five years of diet induction in these animals. These data define an important role for the liver in the cholesteryl oleate enrichment of the plasma lipoproteins in the saturated and monounsaturated fat groups, and demonstrate strong relationships among hepatic cholesteryl ester concentration, cholesteryl ester secretion, and LDL particle cholesteryl ester content. The high correlation between liver cholesteryl ester secretion and coronary artery atherosclerosis provides the first direct demonstration of the high degree of importance of hepatic cholesteryl ester secretion in the development of this disease process. The remarkable degree of enrichment of cholesteryl oleate in plasma cholesteryl esters of the monounsaturated fat group may account for the relatively high amount of coronary artery atherosclerosis in this group.

Cholesterol-lowering effects of modified animal fats in postmenopausal women.

OBJECTIVE: In an attempt to improve the nutritional value of animal fats (including milkfat and lard), two technological approaches (i.e., cholesterol removal by steam distillation and linoleic acid enrichment by addition of safflower oil) were tested for cholesterolemic effects in a cohort of 29 older women (age 68 +/- 7 years). METHODS: Test fat sources were incorporated into crackers, cookies, cheese, ice cream, whipped topping, sour cream, baking shortening, and table spreads. Subjects were permanent residents of a convent where meals were prepared in a centralized kitchen, allowing test fats to be provided in daily food menu items. The foods containing test fats were introduced into three sequential dietary treatment periods, each lasting 4 weeks, in the following order: cholesterol-reduced animal fat (CRAF): fatty-acid modified, cholesterol-reduced animal fat (FAMCRAF); and-unaltered animal fat (AF). Subjects were offered menu items cafeteria style and encouraged to make food selections consistent with their habitual diets, which were recorded daily. RESULTS: Fasted blood lipid profiles determined at the end of each treatment period showed that FAMCRAF reduced mean plasma total cholesterol, LDL cholesterol, and apolipoprotein B concentrations relative to AF (p < 0.05). Mean HDL cholesterol concentrations were not influenced by diet. DISCUSSION: Relative to native products, animal fats modified by cholesterol removal and linoleic acid enrichment reduced plasma total and LDL cholesterol concentrations in a predictable manner similar to that based on studies of men.

Increased intestinal contents viscosity reduces cholesterol absorption efficiency in hamsters fed hydroxypropyl methylcellulose.

This study was conducted to test the hypothesis that increased intestinal contents viscosity lowers plasma cholesterol concentrations by decreasing cholesterol absorption. Male Golden Syrian hamsters were fed for 4 wk diets containing 0.12% cholesterol, and either 4% cellulose or four different viscosity grades of hydroxypropyl methylcellulose (HPMC). Dietary HPMC confers viscosity in the small intestine but is resistant to fermentation. Cholesterol absorption efficiency was measured using the dual isotope ratio method, and plasma and liver cholesterol concentrations were determined enzymatically. Ex vivo viscosity of intestinal contents supernatants was measured using a Wells-Brookfield cone/plate viscometer, and the means of treatment groups ranged from 6 to 6532 mPa.s. Relative to dietary cellulose, all viscosity grades of HPMC resulted in significantly lower cholesterol absorption efficiency, lower plasma cholesterol concentration, and lower liver cholesteryl ester content. The logarithm of intestinal contents ex vivo viscosity was inversely correlated with dietary cholesterol absorption (r2 = 0.84, P = 0.028). Furthermore, dietary cholesterol absorption was positively correlated with plasma cholesterol concentration (r2 = 0.89, P = 0.017) and liver cholesteryl ester content (r2 = 0.96, P = 0.0031). Thus, the data suggest an independent role of intestinal contents viscosity in lowering plasma cholesterol concentration and liver cholesteryl ester content by reducing cholesterol absorption efficiency.

ACAT inhibitors decrease secretion of cholesteryl esters and apolipoprotein B by perfused livers of African green monkeys.

To test the hypothesis that newly synthesized cholesteryl esters are required for hepatic lipoprotein assembly and secretion, isolated livers of African green monkeys were perfused with medium containing an ACAT inhibitor. Three ACAT inhibitors with different structural and chemical properties were used: CI-976 (Parke-Davis), CP-113818 (Pfizer), or PD-138142-15 (Parke-Davis). Each compound produced variable effects on secretion of lipids and apolipoproteins. A significant decrease in the secretion rates of cholesteryl ester and apoB was common to all three inhibitors, indicating that fewer lipoprotein particles were secreted during ACAT inhibition. Triacylglycerol secretion was decreased in the presence of CP-113818 and PD-138142-15 but no decrease in triacylglycerol secretion was observed with CI-976. Particles secreted in the presence of CI-976 were enriched in triacylglycerol relative to apoB. Effects on secretion of other apolipoproteins (apoA-I, apoA-II, and apoE) were variable. When all data were combined, the percent inhibition of cholesteryl ester secretion and apoB secretion in the presence of ACAT inhibitors was positively correlated (r = 0.84), whereas a similar relationship was not observed between triacylglycerol and apoB. While the results demonstrate some lack of specificity for these ACAT inhibitors, the complimentary results using the three different ACAT inhibitors suggest that secretion of cholesteryl ester and apoB are coordinately regulated. The data suggest that newly synthesized cholesteryl esters may participate in and promote the assembly and secretion of hepatic apoB-containing lipoproteins. The availability of triacylglycerol during lipoprotein assembly, while also important, would appear to serve a role separate from that of cholesteryl ester.

Atherogenic and anti-atherogenic factors in the human diet.

New atherosclerosis causative factors and preventive modalities have been identified. Atherogenic factors include lipid oxidation products, such as cholesterol oxidation products, malonaldehyde and other aldehydes; trans-fatty acids; some saturated fatty acids (lauric, myristic and possibly palmitic acids); and myristic acid plus cholesterol. Lipid oxidation products are well suited to induce arterial damage, based on their known cytotoxic effects; evidence also indicates the possibility of plaque promotion and stimulation of thrombogenesis. Anti-atherogenic factors include antioxidants, fish oils and other polyunsaturates (if protected from oxidation), fibre and trace minerals such as copper, manganese, selenium and zinc. Iron is unique, being considered as both a potential promoter of atherosclerosis (component of ferritin, conceivably inducing lipid oxidation) and a possible anti-atherogenic component (of antioxidant enzyme catalase). It is apparent that an entire new series of research challenges has been uncovered.

Interrelationships of alpha-tocopherol with plasma lipoproteins in African green monkeys: effects of dietary fats.

The distributions of plasma lipoprotein alpha-tocopherol and lipids were studied in African green monkeys consuming diets enriched in saturated, monounsaturated, or polyunsaturated fatty acids. Plasma total alpha-tocopherol concentrations were not different among the animals fed the three diets, whereas plasma total cholesterol concentrations were significantly different among the diet groups. The alpha-tocopherol: total lipid molar ratio in plasma high density lipoproteins (HDL) was significantly higher compared to low density lipoproteins (LDL) and very low plus intermediate-sized low density lipoprotein (VLDL + ILDL) of each diet group, suggesting that HDL may exhibit a greater affinity for alpha-tocopherol. The presence of a positive correlation between HDL alpha-tocopherol and plasma apoA-I concentration and the absence of a correlation between HDL alpha-tocopherol and total lipid in HDL suggested that alpha-tocopherol associates with the protein moiety of HDL on the surface of the particle. A direct relationship between the plasma apoA-I: apoB molar ratio and the percentage of alpha-tocopherol found in the HDL fraction indicated that a greater proportion of alpha-tocopherol associates with HDL as the number of HDL particles in plasma increases relative to LDL particles. LDL from monkeys fed diets high in saturated fat contained 40% and 33% fewer alpha-tocopherol molecules per particle than LDL from monkeys fed polyunsaturated and monounsaturated fats, respectively. The phase transition temperature of LDL cholesteryl esters, indicative of the physical state of the lipids in the particle core, was well above body temperature in LDL from saturated fat-fed monkeys and was significantly higher compared to the other diet groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Enzymatic determination of triglyceride, free cholesterol, and total cholesterol in tissue lipid extracts.

Triglyceride, free cholesterol, and total cholesterol were quantified in lipid extracts of liver and thoracic aorta from nonhuman primates using commercially available enzymatic reagents. Lipids were solubilized in water by the addition of Triton X-100. Results of the enzymatic assays compared favorably with chemical assays of lipids separated by thin-layer chromatography. In addition to saving time, the present method has the advantage of measuring each lipid class from a single sample preparation. Furthermore, the procedure has been adapted for use with microtiter plates that conserve both sample and reagent.

Hepatic ACAT activity in African green monkeys is highly correlated to plasma LDL cholesteryl ester enrichment and coronary artery atherosclerosis.

Previous studies and this study of African green monkeys show a strong positive correlation between plasma low density lipoprotein (LDL) size and the extent of coronary artery atherosclerosis (CAA). Increased LDL size was principally due to the accumulation of cholesteryl oleate molecules within the particle core, suggesting that many of these cholesteryl esters were of tissue origin, i.e., from the acyl-coenzyme A:cholesterol acyltransferase (ACAT) reaction instead of the lecithin:cholesterol acyl-transferase (LCAT) reaction. The current study was conducted to test the hypothesis that ACAT in the liver is the source of the increased numbers of cholesteryl oleate molecules in plasma LDL particles that appear to increase the atherogenic potential of LDL. Monkeys were fed diets rich in fat (lard, safflower oil, or fish oil) and cholesterol for 3-6 years before liver perfusion, ACAT assay, and evaluation of CAA. Hepatic ACAT activity was positively correlated with hepatic cholesteryl ester secretion (r = 0.61, p < 0.001), plasma LDL cholesteryl ester content (r = 0.60, p < 0.0001), and the extent of CAA (r = 0.62, p < 0.0001). The number of cholesteryl oleate molecules within LDL increased proportionally with LDL size in each of the diet groups. Hepatic cholesteryl oleate concentration was correlated with the accumulation of cholesteryl oleate in liver perfusate (r = 0.72, p < 0.01) and with plasma LDL cholesterol oleate content (r = 0.73, p < 0.0001). Our interpretation is that these data, obtained in a relevant primate model of CAA, suggest that hepatic ACAT increases the atherogenicity of LDL by augmenting both the secretion by the liver and accumulation in plasma LDL of cholesteryl oleate.

Incorporation of tritiated water into sterol in copper-deficient rats.

The effect of copper deficiency on absolute rates of cholesterol synthesis was investigated in the rat. Male weanling rats were fed semi-purified diets containing adequate (7.13 ppm) or deficient (0.621 ppm) levels of copper for 6 weeks. Copper-adequate and -deficient animals (n = 6/group) were injected intraperitoneally with 50 mCi 3H-labelled water and killed 1 h post-injection. Copper-deficient animals had elevated heart weights and reduced body and spleen weights. Plasma cholesterol levels were significantly elevated and hematocrits were reduced. Absolute rates of carcass cholesterol synthesis per organ were 1.9-fold higher in copper-deficient rats (P less than 0.025). Previous studies have indicated that hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (E.C.1.1.1.34) activity is increased by copper deficiency; however, de novo synthesis of cholesterol from 3H-labelled water was not significantly elevated in the liver. The present data indicate that newly synthesized cholesterol exported to the plasma was increased 2.1-fold (P less than 0.01) in copper-deficient rats. Since it has been demonstrated that hepatic export of cholesterol is increased with copper deficiency and that the major tissue for export of newly synthesized cholesterol is the liver, we hypothesize that the origin of radiolabeled cholesterol in the plasma was the liver. These data support the hypothesis that elevated levels of hepatic HMG-CoA reductase seen with copper deficiency are associated with an increased rate of whole body and hepatic cholesterol synthesis.

High-density lipoprotein cholesteryl ester and protein catabolism in hypercholesterolemic rats induced by copper deficiency.

High-density lipoprotein (HDL) catabolism induced by copper deficiency was examined in vivo in hypercholesterolemic Sprague-Dawley rats. Doubly labeled HDL was used to trace the catabolic pathways of both cholesteryl ester and protein moieties of HDL particles. The catabolic rate of removal from the plasma, as well as uptake by various tissues, was determined for each HDL component. Copper-deficient rats exhibited a 30% increase in HDL cholesterol concentration, confirming hypercholesterolemia. In addition, plasma volume was enlarged 38% in deficient animals, resulting in a significantly increased intravascular pool of all HDL components of at least 60%. These data emphasize the importance of determining plasma volume and total pool size of pertinent plasma components in this hypercholesterolemic model. The absolute catabolic rate (ACR) of HDL protein removal from the plasma was 369 +/- 22 and 278 +/- 12 micrograms/h in copper-deficient and control rats, respectively. The ACR of HDL cholesteryl ester was 647 +/- 37 micrograms/h in deficient animals and 321 +/- 13 micrograms/h in controls, suggesting that the mechanisms of selective clearance of HDL cholesteryl ester (compared with protein) were increased threefold by copper deficiency. Virtually all of the increased removal of HDL cholesteryl ester in deficient rats occurred in the liver. Since previous studies indicate that increased hepatic cholesterol excretion may not occur in copper deficiency, the present results suggest that cholesterol delivered to the liver as HDL cholesteryl ester is possibly reassembled into new HDL particles at an increased rate in copper-deficient rats.

In vivo apoprotein catabolism of high density lipoproteins in copper-deficient, hypercholesterolemic rats.

High density lipoprotein (HDL) apoprotein catabolism was examined in male Sprague-Dawley rats deficient in dietary copper. Twenty-four rats were randomly divided into two groups: copper-adequate (control, 5 mg of copper/kg diet) and copper-deficient (0.6 mg of copper/kg diet). After 5 weeks, animals were administered a tracer dose of iodinated HDL protein previously isolated from donor rats that were subjected to the same dietary treatments as the test animals. Copper-deficient rats exhibited a 54% increase in plasma volume and a 26% increase in HDL protein concentration above controls. Consequently, the intravascular pool of total HDL protein was increased 2-fold. The fractional catabolic rate of total HDL protein was similar between groups. However, because of the increased intravascular HDL pool in copper-deficient animals, the absolute catabolic rate was greater (640 +/- 49 micrograms/hr vs 316 +/- 12 micrograms/hr in controls). Tissue uptake of total HDL protein in copper-deficient rats tended to be greater in the kidneys, spleen, and testes compared with controls; the heart exhibited a significant 2.3-fold increase. In contrast, the catabolic rate of HDL protein in the liver and adrenal gland were not different between treatment groups. That an obligatory increase in HDL protein uptake was not observed in the liver and adrenal gland (organs which are sensitive to and can further metabolize cholesterol) suggests that these organs may be regulated, possibly contributing to the observed hypercholesterolemia in this model. These data imply that total HDL apoprotein catabolism is increased in response to the increased intravascular pool of HDL in copper-deficient rats.

Alterations in lipid composition and fluidity of liver plasma membranes in copper-deficient rats.

In view of the importance of membrane fluidity on cell functions, the influence of phospholipid acyl groups on membrane fluidity, and the changes in lipid metabolism induced by copper (Cu) deficiency, this study was designed to examine the influence of dietary Cu on the lipid composition and fluidity of liver plasma membranes. Male Sprague-Dawley rats were divided into two dietary treatments, namely Cu deficient and Cu adequate. After 8 weeks of treatment, liver plasma membranes were isolated by sucrose density gradient centrifugation. The lipid fluidity of plasma membranes, as assessed by the intramolecular eximer fluorescence of 1,3-di(1-pyrenyl) propane, was significantly depressed by Cu deficiency. In addition, Cu deficiency significantly reduced the content of arachidonic and palmitoleic acids but increased the docosatetraenoic and docosahexaenoic acids of membrane phospholipids. This alteration in unsaturated phospholipid fatty acid composition, especially the large reduction in arachidonic acid, may have contributed to the depressed membrane fluidity. Furthermore, Cu deficiency also markedly altered the fatty acid composition of the triacylglycerols associated with the plasma membranes. Thus, the lipid composition and fluidity of liver plasma membranes are responsive to the animal's Cu status.