Alfalfa seeds: effects on cholesterol metabolism

Plasma cholesterol concentrations were reduced in 3 human volunteers during ingestion of diets containing alfalfa seeds (AS) for 3 weeks. No signs of toxicity were detected through serum determinations of multiple parameters. The ingestion of AS in rats decreased the concentration of plasma cholesterol, reduced intestinal absorption of exogenous and endogenous cholesterol, and increased fecal biliary excretion.     

Hepatic cholesterol and bile acid metabolism in subjects with gallstones: comparative effects of short erm feeding of chenodeoxycholic and ursodeoxycholic acid

The activity of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase and 7alpha-hydroxylase, the enzymes controlling the rate of hepatic synthesis, respectively, of cholesterol and bile acids, and the microsomal cholesterol content were evaluated in 25 patients with cholesterol gallstones and 17 subjects without gallstones. The same quantities were estimated in 16 additional patients with gallstones given chenodeoxycholic (CDCA) or ursodeoxycholic acid (UDCA) at a dose of 15 mg/kg per day in order to investigate the comparative effect of a short term (7 days) administration of the two bile acids on the hepatic sterol metabolism. As compared to the controls, subjects with gallstones exhibited a 36% decrease of 7alpha-hydroxylase (26.8 +/- 6.2 versus 41.7 +/- 4.2 pmol/min per mg protein) and a 24% increase of the microsomal cholesterol (78.7 +/- 15.3 versus 63.1 +/- 18.1 nmol/mg protein). Although higher in the gallstone patients, the activity of HMG-CoA reductase did not differ significantly in the two groups of subjects. Administration of CDCA and UDCA changed the bile acid pool composition so that the fed bile acid predominated in the bile (mean CDCA 73% and mean UDCA 54%). Bile lipid composition did not appreciably change. In the eight subjects treated with CDCA the activity of HMG-CoA reductase was reduced to 45% of the value of untreated subjects (27.9 +/- 14.5 versus 63.5 +/- 25.3 pmol/min per mg protein) whereas in the eight subjects treated with UDCA the same enzyme showed a twofold increase (123.5 +/- 20.9). In the treated groups 7alpha-hydroxylase activity was somewhat decreased but the values did not differ significantly from those of the untreated subjects. Microsomal cholesterol content decreased with CDCA (64.8 +/- 11.6 nmol/mg protein) as well as with UDCA (59.1 +/- 10.1) treatment; however in the latter the difference attained statistical significance (P < 0.05). Altogether the results would suggest that in the liver of patients with gallstones the conversion of cholesterol to bile acids is somewhat reduced, and that changing the bile acid pool composition, by exogenous bile acid feeding, has disparate effects on hepatic cholesterol synthesis. The findings could represent the acute changes produced by bile acid feeding, however they could imply that the effects of two bile acids in dissolving cholesterol gallstones might not be related only to the changes in hepatic sterol metabolism.-Carulli, N., M. Ponz De Leon, F. Zironi, A. Pinetti, A. Smerieri, R. Iori, and P. Loria. Hepatic cholesterol and bile acid metabolism in subjects with gallstones: comparative effects of short term feeding of chenodeoxycholic and ursodeoxycholic acid.     

Resistant starch decreases serum total cholesterol and triacylglycerol concentrations in rats

Rats were meal-fed semipurified diets containing a low (0.8 g/MJ) and a high (9.6 g/MJ) amount of resistant starch (RS) or various amounts of RS (0.8 to 9.6 g/MJ) and guar gum (0 to 8.8 g/MJ). In one experiment, rats were fed the low and high RS diets in three dietary regimens (ad libitum consuming, 12 h ad libitum/12 h food deprived, and meal fed). Effects of RS and guar gum on serum postprandial and postabsorptive concentrations of total cholesterol (TC) and triacylglycerol (TAG), growth, hydrogen excretion, tissue weights and contents of small intestine and cecum, and pH of cecal contents were investigated. In addition, effects of RS on food intake, de novo hepatic synthesis of fatty acids and neutral sterols, and on lipoprotein lipase activity and weight of epididymal fat pads were investigated. Compared with feeding the low RS diet, the high RS diet reduced the serum TC and TAG concentrations, with these effects observed after 1 and 2 wk of feeding, respectively. The dietary regimen did not influence the effect of RS on the serum TC and TAG concentrations, but it did affect the serum TAG concentration. Resistant starch had no effect on the hepatic synthesis of fatty acids and neutral sterols or on the lipoprotein lipase activity in epididymal fat pads. Guar gum also reduced the serum TC concentration, but it had no effect on serum TAG concentration. The tissue weights and contents of small intestine and cecum as well as hydrogen excretion increased with increasing amounts of dietary RS and guar gum, whereas the pH of cecal contents decreased. No effects of RS on food intake and total body weight gain were found, whereas guar gum decreased weight gain. Feeding the high RS diet also led to a lower weight of the epididymal fat pads. We conclude that dietary RS can reduce serum TC and TAG concentrations and fat accretion.     

Acute inflammation, acute phase serum amyloid A and cholesterol metabolism in the mouse

We describe a patient, RM, who suddenly became amnesic for premorbid autobiographic events in the absence of any known precipitating event. Learning abilities as well as semantic knowledge were normal. Knowledge of famous facts and persons was good, although not perfect. Whether RM suffered from organic or psychogenic isolated retrograde amnesia (IRA) could not be established on the basis of available clinical and neuropsychological elements. Regardless of its aetiology, RM's case respects the boundaries between semantic and episodic memory and so gives further support to the distinction between these two memory systems.     

Fat-modified diets influence serum concentrations of cholesterol precursors and plant sterols in hypercholesterolemic subjects

Serum noncholesterol sterols, cholesterol precursors and plant sterols, are indicators of cholesterol absorption and synthesis. Serum plant sterol concentrations correlate positively with cholesterol absorption, but have also been found to correlate with dietary unsaturated to saturated fatty acid ratios. We studied the concentration of serum noncholesterol sterols during four different fat-modified diets, (1) high-fat, saturated fat-enriched (control), (2) reduced-fat, sunflower oil-enriched (SO-enriched), (3) rapeseed oil-enriched (RO-enriched), and (4) reduced-fat, saturated fat-enriched (reduced-fat), followed for 6 months in hypercholesterolemic subjects in a parallel design. The proportion of lathosterol (micrograms per 100 mg cholesterol), a precursor of cholesterol synthesis, increased significantly (P < .05) in both SO-enriched (mean +/- SD 147 +/- 57 v 167 +/- 76, 0 v 6 months) and RO-enriched (147 +/- 54 v 157 +/- 52) groups, where the reduction in low-density lipoprotein (LDL) cholesterol was also significant. The proportion of sitosterol, a plant sterol, decreased significantly in the control group (137 +/- 48 v 122 +/- 42), and the proportion of another plant sterol, campesterol, increased in the RO-enriched group (280 +/- 141 v 333 +/- 162), reflecting changes in the use of vegetable oils in these two groups rather than increased cholesterol absorption. In the whole study population, the proportion of linoleic and alpha-linolenic acid (a marker of the use of RO) in cholesterol esters (CEs) correlated (P < .001) with the proportion of sitosterol (r = .43) and campesterol (r = .36) in serum at the end of the study. In conclusion, serum cholesterol precursors were found to be useful indicators of cholesterol metabolism, but changes in serum plant sterols reflected dietary changes rather than cholesterol metabolism during long-term dietary intervention with fat-modified diets.     

Dietary cholesterol affects serum lipids, lipoproteins and LDL metabolism in cynomolgus monkeys in a dose-dependent manner

To examine the mechanism(s) underlying the cholesterolemic response to dietary cholesterol and saturated fatty acids, low density lipoprotein (LDL) metabolism was studied in two groups of cynomolgus monkeys fed diets containing 30 or 36% of total energy as fat. At each dietary fat level, the same group of monkeys was sequentially fed three dietary cholesterol concentrations as egg yolk in the following sequence: low (0.01 mg/kJ), medium (0.03 mg/kJ) and high (0.05 mg/kJ) for 30, 32 and 24 wk, respectively. Dietary polyunsaturated and monounsaturated fatty acids were the same in the two groups; the 6% difference in fat was due to the saturated fatty acids, 12:0 and 14:0. Serum total cholesterol, LDL cholesterol and LDL apolipoprotein B concentrations increased (P < 0.05) with dietary cholesterol in a dose-dependent manner in both fat groups. These elevations were the result of generally increasing LDL apolipoprotein B production rates, concomitant with reduced LDL apolipoprotein B fractional clearance at the high cholesterol intake. Serum HDL cholesterol and HDL apolipoprotein A-I concentrations were not affected in a consistent manner. These results demonstrate that cynomolgus monkeys are hyperresponsive to dietary cholesterol compared with humans, suggesting that this model may be useful in identifying metabolic and genetic predictors for hyperresponsiveness to dietary cholesterol in humans as well as assessing the metabolic heterogeneity of responses to dietary cholesterol.     

Milk type during mixed feeding: contribution to serum cholesterol ester fatty acids in late infancy

OBJECTIVE: To evaluate the contribution of the type of milk on serum cholesterol ester fatty acids in infants receiving mixed feeding, we analyzed 3-day dietary records and serum cholesterol ester fatty acid composition of 397 seven-month-old infants. STUDY DESIGN: The infants received, in addition to solid food, only one type of milk: human milk (n = 218), a ready-to-use liquid formula (n = 139), a powdered formula (n = 33), or soy formula (n = 7). RESULTS: Mean fat intakes were low and varied from 28% to 31% of energy; the milks provided 43% to 64% of the fat. The mean polyunsaturated/saturated fat ratios of solid foods were from 0.52 to 0.63 and of milks from 0.20 to 0.45. Breast-fed infants' relative serum linoleic acid (18:2n-6) concentration was low (51.2%), whereas infants fed liquid formula had low serum oleic acid (18:1n-9) in accordance with low oleic acid content in that formula. The breast-fed infants had markedly higher serum concentrations of arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) than the infants fed any of the formulas. CONCLUSION: The typical fatty acid patterns of breast- or formula-fed infants were still evident in 7-month-old infants who already received 60% to 70% of their energy from solid food. Marked differences were seen also in the relative concentrations of docosahexaenoic acid and arachidonic acid despite their small contribution in cholesterol esters.     

Clinical characteristics and coronary risk factors of patients with low concentrations of serum low-density lipoprotein cholesterol and total cholesterol

We investigated the clinical characteristics and coronary risk factors of Chinese patients with suspected coronary artery disease (CAD) having low serum concentrations of both low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC). Of 1,450 patients with suspected CAD (age range, 30-92 years; 948 men and 502 women), 760 had established CAD. The patients were divided into three groups according to lipid profile patterns. Group 1 patients (n = 138) had low LDL-C concentrations (< 100 mg/dL) and low TC concentrations (< 160 mg/dL). They were characterized by lower triglyceride concentrations, lower frequencies of high TC/high-density lipoprotein cholesterol (HDL-C) ratios (> 5) and LDL-C/HDL-C ratios (> 5), and lower frequencies of a family history of CAD and obesity. Group 3 patients (n = 610) had LDL-C concentrations of 130 mg/dL or above and TC concentrations of 200 mg/dL or above, much higher than in group 1. The prevalence of CAD was 41.3% (57/138) in group 1. 46.7% (328/702) in group 2, and 61.5% (375/610) in group 3. Groups with higher TC and LDL-C concentrations had a higher CAD prevalence. Coronary risk factors of group 1 patients appeared to be low HDL-C concentration, high TC/HDL-C ratio, advanced age, cigarette smoking, hypertension, and diabetes mellitus. Among these risk factors, HDL-C and hypertension were independent predictors of CAD. Unlike in the other two groups, hypertension was the only independent nonlipid risk factor. We conclude that in therapy or prevention of CAD, the goals should be to reduce LDL-C concentration to below 100 mg/dL and the TC concentration to below 160 mg/dL. However, other risk factors should also be considered.     

Regulation of classic and alternative bile acid synthesis in hypercholesterolemic rabbits: effects of cholesterol feeding and bile acid depletion

The effect of cholesterol feeding (3 g/day) on bile acid synthesis was examined in 10 New Zealand white rabbits (NZW), 8 Watanabe heterozygous and 10 homozygous rabbits with partial and complete deficiencies of LDL receptors. After 10 days of cholesterol feeding, bile fistulas were constructed and bile acid pool sizes were measured. Cholesterol feeding increased plasma and hepatic cholesterol levels in all rabbit groups. Baseline bile acid pool sizes were smaller (P < 0.01) in heterozygotes (139 +/- 3 mg) and homozygotes (124 +/- 30 mg) than NZW rabbits (254 +/- 44 mg). After feeding cholesterol, bile acid pool sizes doubled with increased cholic acid synthesis in NZW and, to a lesser extent, in Watanabe heterozygous rabbits but not in homozygotes. Baseline cholesterol 7alpha-hydroxylase activity in NZW and heterozygotes declined 69% and 53% (P < 0.001), respectively, after cholesterol feeding. Sterol 27-hydroxylase activity reflecting alternative bile acid synthesis increased 66% (P < 0.01) in NZW and 37% in Watanabe heterozygotes but not in homozygotes after feeding cholesterol. Bile fistula drainage stimulated cholesterol 7alpha-hydroxylase activity but not sterol 27-hydroxylase activity in all three rabbit groups. These results demonstrated that dietary cholesterol increased hepatic sterol 27-hydroxylase activity and alternative bile acid synthesis to expand the bile acid pool and inhibited cholesterol 7alpha-hydroxylase in NZW and in Watanabe heterozygous rabbits but not in homozygotes with absent hepatic LDL receptor function. Thus, in rabbits, sterol 27-hydroxylase is up-regulated by the increased hepatic cholesterol that enters the liver via LDL receptors whereas cholesterol 7alpha-hydroxylase is controlled by the circulating hepatic bile acid flux.     

Absorption and metabolism of nivalenol in pigs

Effects of chronic concentrations of linuron (0, 0.5, 5, 15, 50, and 150 micrograms/L) were studied in indoor, macrophyte dominated, freshwater microcosms. The concentrations were kept at a constant level for 4 weeks. This paper is the first in a series of two and summarizes the course of the linuron concentrations in time and its effects on macrophytes, periphyton, and phytoplankton. These endpoints were studied from 3 weeks before the start of the treatment until 11 weeks after the start. The degradation of linuron in the water was lower at higher treatment levels, probably due to a decrease in pH. Linuron treatment resulted in a decrease in biomass of the macrophyte Elodea nuttallii and a clear decrease in abundance of the algae Cocconeis, Chroomonas, and Phormidium foveolarum. It was found that Cocconeis first decreased in biovolume and after 2 weeks also in abundance. The alga Chlamydomonas increased in abundance at the two highest doses, resulting in higher chlorophyll-a levels. The NOECs of 0.5 micrograms/L for the inhibition of the growth and photosynthesis of Elodea nuttallii, the abundance of Cocconeis and Chroomonas, and the oxygen and pH levels were the lowest recorded in the microcosms. The safety factors adopted by the EU in the Uniform Principles appeared to ensure adequate protection for the ecosystem in the case of chronic exposure to linuron.     

The effects of simvastatin and cholestyramine, alone and in combination, on hepatic cholesterol metabolism in the male rat. off

The influence of dietary simvastatin, cholestyramine, and the combination of simvastatin plus cholestyramine on hepatic cholesterol metabolism has been investigated in male rats. Recovery from the effects of the drugs was also investigated by refeeding normal chow for 24 h. Both drugs, alone and in combination, increased 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase activity in vitro, but activity returned toward control values, after drug withdrawal. Acyl-CoA:cholesterol acyltransferase (ACAT) was significantly reduced (P < 0.001) by simvastain (-75%), cholestyramine (-71%), and by the drug combination (-81%), due both to a decrease in microsomal cholesterol and to nonsubstrate-dependent modulation of enzyme activity. Refeeding control diet increased ACAT activity but not to control levels. The enhanced activity arose partly from higher microsomal cholesterol and partly from increases in total enzyme activity. Cytosolic neutral cholesteryl ester hydrolase (CEH) activity was substantially elevated by simvastatin (3-fold) and by the drug combination (6-fold), whereas the effect of cholestyramine was smaller (1.5-fold). Normal chow for 24 h only partially returned cytosolic CEH activity to control values. Microsomal CEH activity was increased by simvastatin, alone and in combination with cholestyramine (1.4 to 1.7-fold), and was also enhanced, in the cholestyramine-treated animals, following drug withdrawal. Removal of simvastatin did not allow recovery of this enzyme activity, while withdrawal of the drug combination led to values 29% below controls. The results indicate that in the rat, simvastatin and cholestyramine alter both ACAT and CEH activity, as well as inhibiting HMG-CoA reductase activity.     

The effects of cholesterol/fat feeding on lipid levels and morphological structures in liver, kidney and spleen in guinea pigs

Guinea pigs were fed a semisynthetic diet containing 10 per cent (by weight) cottonseed oil with or without 1 per cent cholesterol. In the animals fed fat, the lipid levels and the morphology remained normal in all tissues studied. Concomitantly with a marked accumulation of cholesteryl ester (CE) in the liver, however, many microscopical changes occurred in guinea pigs fed cholesterol/fat. A prominent deposition of lipids in vacuoles, mostly without delimitating membranes, where observed at centrilobular sites. Multivacuolated, secondary lysosomes, membrane bound lipid vacuoles (lipolysosomes) and myelin figures were found both in hepatocytes and Kupffer cells. Myelin figures and crystalline clefts were observed more often in Kupffer cells than in hepatocytes. The granular endoplasmic reticulum in the Kupffer cells was grossly dilated and filled with an amorphous material. Both the biochemical and the morphological findings in hepatocytes and Kupffer cells are very similar to those observed in cholesteryl ester storage disease and in Wolman's disease. These two lipid storage diseases are both related to deficiency of an acid lipase in the liver. Measurement of the acid liver CE hydrolase in guinea pigs fed fat and in those fed cholesterol/fat showed similar activity. A relative deficiency of this enzyme activity could be the reason for the development of the enormous CE storage in guinea pig livers. These findings suggest that guinea pigs fed cholesterol/fat, in some respects, can be used as a model for Wolman's disease and cholesteryl ester storage disease. We did not find any microscopical changes in the kidneys from animals fed cholesterol/fat, thus indicating that the experimental condition it not useful as a model for studies of the kidney changes in lecithin:cholesterol acyltransferase (LCAT) deficiency.     

Independence of the effects of cholesterol and degree of saturation of the fat in the diet on serum cholesterol in man

The effect on the fasting serum lipid levels of adding daily 291 mg of cholesterol to diets containing 3 mg of cholesterol and equal fat content, but different fatty acid composition, was tested on 12 young men. The saturated diet provided 97 g/day of a staurated oil made up of 2 parts of palm oil and 1 part of coconut oil. The polyunsatured diet provided 97 g/day of safflower oil. The cholesterol was dissolved in 40 g of either oil incorporated into a spread. A similar spread, devoid of cholesterol, was fed during the cholesterol free periods. Duration of dietary periods was 14 days. Addition of cholesterol produced a mean elevation of serum cholesterol of 9 mg/dl (SE +/- 2.1) in the presence of the saturated diet, and of 8 mg/dl (SE +/- 1.6) in the presence of the polyunsaturated diet. Both cholesterol elevations were significant (P less than 0.01) but not significantly different from each other. Substitution of the saturated diet for the polyunsatured diet caused a significant elevation of serum cholesterol which was the same when the substitution was made in the presence or in the absence of added dietary cholesterol.     

Fractionation of serum cholesterol: a critique

A procedure for fractionation of serum cholesterol is developed by employing digitonin for the precipitation of free cholesterol in isopropanol. This method is based on the ferric acetate color reaction which is unaffected by traces of digitonin. It is influenced only negligibly by the presence of stanols in serum and is shown to be equally chromogenic with respect to both free and ester forms of cholesterol. The proposed procedure shows a three-fold improvement in precision (3.1 percent coefficient of variation [CV]) compared to that of the procedure by Leffler and McDougald (10.5 percent C.V.) which is based on the ferric chloride color reaction. The proposed free cholestrol procedure showed a mean recovery of 99.5% percent (98.1 to 102.5 percent) when cholesterol in 40 to 200 mg per dl concentrations was added to serum. The analytical performance of the proposed fractionation of serum cholesterol is critically reviewed with respect to its potential application in the diagnosis of liver diseases and in basic or experimental research.     

Dietary soluble fiber lowers plasma LDL cholesterol concentrations by altering lipoprotein metabolism in female guinea pigs

This experiment was designed to evaluate the effects of pectin (PE), guar gum (GG) and psyllium (PSY) intake on VLDL and LDL metabolism in female guinea pigs fed high dietary cholesterol. Guinea pigs were fed a 15 g/100 g fat diet containing 0.25 g/100 g cholesterol with 12.5 g/100 g PE, 12.5 g/100 g GG, 7.5 g/100 g PSY or 12.5 g/100 g cellulose (control diet) for 4 wk. Plasma cholesterol concentrations were 29, 43 and 39% lower in guinea pigs fed PE, GG or PSY, respectively, compared with the control group (P < 0.0001). Plasma apolipoprotein (apo) B concentrations were 16-22% lower in the groups fed soluble fiber compared with the control group (P < 0.01). In contrast, hepatic cholesterol and triglyceride concentrations were not different among the PE, GG, PSY and control groups. No differences in triacylglycerol (TAG) or apo B secretion rates, measured by blocking VLDL catabolism by triton (WR 1339) injection, were observed, whereas plasma LDL apo B fractional catabolic rates (FCR), determined by injection of radiolabeled LDL, were higher in guinea pigs fed GG or PSY than in those from the control group. All sources of dietary soluble fiber reduced LDL apo B flux (P < 0.05). These results suggest that the mechanisms of plasma LDL cholesterol lowering by dietary soluble fiber are distinctive for each fiber source and result in specific alterations in lipoprotein metabolism in female guinea pigs. Differences between male and female guinea pigs in response to these diets are discussed.     

Linkage between cholesterol 7alpha-hydroxylase and high plasma low-density lipoprotein cholesterol concentrations

Interindividual differences in plasma low-density lipoprotein cholesterol (LDL-C) levels reflect both environmental variation and genetic polymorphism, but the specific genes involved and their relative contributions to the variance in LDL-C are not known. In this study we investigated the relationship between plasma LDL-C concentrations and three genes with pivotal roles in LDL metabolism: the low-density lipoprotein receptor (LDLR), apolipoprotein B (APOB), and cholesterol 7alpha-hydroxylase (CYP7). Analysis of 150 nuclear families indicated statistically significant linkage between plasma LDL-C concentrations and CYP7, but not LDLR or APOB. Further sibling pair analyses using individuals with high plasma LDL-C concentrations as probands indicated that the CYP7 locus was linked to high plasma LDL-C, but not to low plasma LDL-C concentrations. This finding was replicated in an independent sample. DNA sequencing revealed two linked polymorphisms in the 5' flanking region of CYP7. The allele defined by these polymorphisms was associated with increased plasma LDL-C concentrations, both in sibling pairs and in unrelated individuals. Taken together, these findings indicate that polymorphism in CYP7 contributes to heritable variation in plasma LDL-C concentrations. Common polymorphisms in LDLR and APOB account for little of the heritable variation in plasma LDL-C concentrations in the general population.