Serum lipids, lipoprotein composition and liver cholesterol in genetically obese Zucker rats fed semipurified diets containing either casein or soy protein

The effect of semipurified diets containing either casein or soy protein on serum lipids, lipoprotein composition and liver cholesterol was studied in genetically obese Zucker rats. The ingestion of a cholesterol-enriched semipurified diet containing casein resulted in elevated levels of serum cholesterol and phospholipids compared to the feeding of a soy protein diet. No differences in serum triglycerides were observed. Differences in serum cholesterol and phospholipids were mainly reflected in the very low density lipoproteins and low density lipoproteins and to a minor extent in the high density lipoproteins. Liver cholesterol paralleled the levels of cholesterol in the serum, the rats fed casein exhibited markedly higher levels of liver cholesterol than those fed soy protein. Furthermore, the rats fed casein also had enlarged livers. Thus, this study clearly shows the differential cholesterolemic effect of dietary casein and soy protein in genetically obese Zucker rats.     

Influence of fish protein as compared to casein and soy protein on serum and liver lipids, and serum lipoprotein cholesterol levels in the rabbit

Serum and hepatic cholesterol and triglyceride levels, and serum lipoprotein cholesterol were investigated in rabbits fed fish protein as compared to casein and soy protein as part of a 20% protein, low fat, cholesterol-free, semi-purified diet. A nonpurified diet was used as a control. After a 28-day experimental period, rabbits fed casein developed hypercholesterolemia compared to those fed the soy protein diet. Serum cholesterol levels of rabbits fed fish protein was intermediate and not different from that of the casein or the soy protein group. However, serum triglycerides were higher in the fish group than in the casein group. Feeding of fish protein resulted in a reduction of hepatic cholesterol compared to casein, indicating no direct relationship between serum and hepatic cholesterol. In addition, fish protein induced a decrease of cholesterol in the low density lipoproteins (LDL) compared to casein and an increase of cholesterol in the high density lipoproteins (HDL) compared to casein and soy protein. Reduction in LDL-cholesterol (LDL-C) and elevation in HDL-cholesterol (HDL-C) caused a 10-fold decrease in the LDL-C/HDL-C ratio of fish protein fed rabbits compared to those fed casein. This ratio was similar to that observed with soy protein which was also lower than the ratio of the casein group. Thus, since the LDL-C/HDL-C ratio has been shown to be a good indicator of the atherosclerosis risk, these results suggest that fish protein, as well as soy protein, may reduce the risk of atherosclerosis in rabbits, compared to casein.     

Accumulation of cholesteryl esters and triglycerides in cultured macrophages incubated with plasma very low density lipoproteins from rats fed on casein or soybean protein diets containing moderate levels of cholesterol

Even when administered at a comparatively low level, dietary cholesterol produces significant changes in the properties of plasma lipoproteins in rats, particularly the d less than or equal to 1.006 g/ml fraction (VLDL). The occurrence of these changes is promoted by dietary casein. To test the hypothesis that these dietary-induced perturbations might include properties influencing lipoprotein-cell interactions of relevance to atherogenesis, cultured mouse peritoneal macrophages were incubated with VLDL isolated from male rats fed on diets containing either 0, 0.25 or 0.5% cholesterol with casein or soybean protein, respectively, as the sole source of protein. No increase in cholesteryl ester content, and a comparatively small rise in triglyceride content, was observed in macrophages incubated with VLDL from rats fed on cholesterol-free diets. In contrast, a significant and apparently saturable cellular accumulation of cholesteryl esters as well as triglycerides was produced by VLDL from cholesterol-fed rats. The curves showing cellular lipid accumulation versus VLDL-protein (or VLDL-cholesterol) content in the cell medium indicated different cellular affinity for VLDL from casein-fed rats in comparison with VLDL from soybean protein-fed rats. The apoprotein composition of VLDL differed between groups of rats fed on different types of dietary protein with higher proportions of apo C's in the casein-fed rats. In addition, cholesterol feeding resulted in increased proportions of apo A-I and apo A-IV in the plasma VLDL fraction.     

Lipid metabolism in pregnancy. VIII. Effects of dietary fat versus carbohydrate on lipoprotein and hepatic lipids and tissue triglyceride lipases

We have asked, is hypertriglyceridemia in the fed state in pregnancy due to intolerance to exogenous fat, accumulation of endogenous triglycerides, or accumulation of remnants of d < 1.006 lipoprotein metabolism? To answer these questions, we fed rat-free diets high in starch or sucrose, or diets containing fat or fat plus cholesterol to pregnant and nonpregnant rats for 12 days until gestational day 21 (term = 22 days). Blood was obtained 0, 4, or 8 hr after removal of food from the cages. Lipid concentrations were determined in chylomicrons and very low, low, and high density lipoproteins. Hypertriglyceridemia in pregnancy exists on both starch and sucrose containing fat-free diets and is exaggerated 4 and 8 hr after food is removed from the cage. The triglyceride rise occurs in d < 1.006 lipoproteins. With fat feeding, chylomicron triglyceride concentrations are not significantly elevated in pregnant rats, 0 or 8 hr postabsorptively despite greater food intake in pregnancy. In contrast, very low density lipoprotein (VLDL) triglyceride concentrations are elevated at all times following fat feeding in pregnant compared to nonpregnant animals. A significant contribution of lipoprotein remnants to the triglyceride rise in d < 1.006 lipoproteins seems unlikely since an isolated increase in VLDL cholesterol is not observed. No statistically significant accumulation of hepatic triglycerides occurs on any diet in pregnancy. Diet induced shifts in adipose tissue and muscle lipoprotein lipase activity are exaggerated in pregnancy while hepase in unaffected. Fetal weight is similar on all diets except sucrose where weight is reduced. Conclusions: Hypertriglyceridemia in fed pregnant rats is due to an increase in endogenous triglycerides. Remnant lipid accumulation does not appear to contribute to the endogenous hypertriglyceridemia. There is no intolerance to exogenous (dietary) fat. The results are compatible with an unimpaired delivery of exogenous fat to fat oxidizing tissues thereby maximizing glucose availability for fetal growth.     

Effects of soy components on blood and liver lipids in rats fed high-cholesterol diets

AIM: To assess the effects of soy protein, isoflavone, and saponin on liver and blood lipid in rats that consumed high-cholesterol diets. METHODS: High-cholesterol diets (1%) with or without soy material were fed to 6-wk-old male Sprague-Dawley rats for 8 wk. Blood lipids, liver lipids, glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) levels were measured. The in vitro bile acid-binding ability of soy materials was analyzed. RESULTS: The results of in vitro studies showed that soy protein isolate had a significantly higher bile acid-binding ability (8.4±0.8%) than soy saponin (3.1±0.7%) and isoflavone (1.3±0.4%, P<0.05). On the other hand, at the end of the experimental period, rats that consumed soy protein diets had lower GOT and GPT levels than rats that consumed casein under high-cholesterol diets. Rats that consumed soy protein also had lower total cholesterol (TC) levels in the liver than those that consumed casein under high-cholesterol diets. Rats that consumed the soy protein diet containing both saponin and isoflavone had lower hepatic TC level than those that consumed the soy protein diet without isoflavone alone. The effect of different types of proteins on triglyceride was not significant. CONCLUSION: Consumption of soy provided benefits to control lipid levels under high-cholesterol dieting conditions in this rat model of hypercholesterolemia. The major component that reduced hepatic TC was not saponin, but possibly isoflavone.     

Varied effects of dietary sucrose and cholesterol on serum lipids, lipoproteins and apolipoproteins in rhesus monkeys.

Serum lipid, lipoproteins, apolipoproteins and plasma insulin and glucose were studied in rhesus monkeys (Macaca mulatta) fed high sucrose diets (69%, w/w), with and without added cholesterol. When compared to basal diet, a high sucrose diet with no added cholesterol fed for 6 weeks increased serum total cholesterol and triglycerides by factors of 1.2 and 2.8, respectively. Cholesterol supplementation of sucrose diets increased the serum total cholesterol levels by a factor of 2.2 and decreased the serum triglycerides by 0.47. The serum cholesterol response to experimental diets was reflected predominantly in beta-lipoprotein and to a lesser extent in alpha-lipoprotein. Sucrose diets without cholesterol enriched the beta- and pre-beta-lipoproteins with triglycerides and protein at the expense of cholesterol. On the same diet, the protein content of alpha-lipoprotein increased at the expense of cholesterol and triglycerides. In contrast, dietary cholesterol decreased the triglyceride content and increased the cholesterol content of all the lipoprotein classes. Sucrose feeding seems to increase ApoB more than non-ApoB proteins. The proportion of ApoC-II relative to ApcoC-III increased in each animal on a sucrose diet; exogenous cholesterol further increased this trend. While sucrose diet decreased ApoA-I/ApoA-II ratios, cholesterol supplementation reversed this trend. Dietary sucrose increased the plasma glucose, insulin, and insulin-glucose ratios. The addition of cholesterol also tended to decrease plasma glucose and insulin levels. These observations indicate varied responses of serum lipoproteins and apoproteins to dietary sucrose with and without cholesterol supplementation.     

Plasma lipid and lipoprotein cholesterol levels in swine. Modification of protein-induced response by added cholesterol and soy fiber

Piglets, aged 8 weeks and weighing 12-18 kg, were fed semi-purified casein or soy protein diets, with or without cholesterol and soy hull fiber, for 2 months. In addition to observing the effects of the dietary treatments on growth, the modification of the primary hypocholesterolemic action of soy protein by cholesterol and soy fiber was studied. Pigs fed the soy protein or casein diets grew normally with no difference in weight gain. Plasma triglyceride and phospholipid levels, as well as several plasma metabolic indices examined, were not significantly affected by dietary treatment. However, plasma total cholesterol was higher (but not significantly) in pigs fed casein than in those fed soy protein alone. Cholesterol feeding induced markedly significant (P less than 0.05) hypercholesterolemia with either protein source, compared to feeding the proteins without added cholesterol. Dietary soy fiber fed simultaneously with cholesterol decreased the cholesterol-induced hypercholesterolemia, but the reduction was significantly greater (P less than 0.05) with soy protein than with casein in the diet. Analyses of the lipoprotein cholesterol indicated that LDL cholesterol was much more sensitive to the changes induced by feeding cholesterol and soy fiber than either HDL or VLDL cholesterol. These findings suggest a beneficial role of dietary soy fiber in hypercholesterolemia.     

Methionine-induced elevation of plasma homocysteine concentration is associated with an increase of plasma cholesterol in adult rats

BACKGROUND AND AIMS: Dietary methionine affects cholesterol metabolism in growing rats. Methionine effects on adult rats and mechanisms by which methionine alters the lipid metabolism are not fully elucidated. We investigated possible mechanisms by which dietary methionine acts on lipid metabolism of adult rats. METHODS: Male adult rats were divided into three groups (n=10) and were fed casein-based diets differing in methionine concentration (low-methionine diet: 0.96 g/kg; adequate-methionine diet: 2.22 g/kg, high-methionine diet: 6.82 g/kg) for 4 weeks. Concentrations of triacylglycerols and cholesterol in plasma and lipoproteins, concentration of homocysteine in plasma, concentration of cholesterol in liver, fecal lipid excretion, expression of hepatic HMG-CoA reductase, phosphatidylethanolamine N-methyltransferase 2 (PEMT-2) and of LDL receptor were measured. RESULTS: Rats fed the high-methionine diet had higher plasma homocysteine concentrations than rats fed the low-methionine diet (p<0.05). Although concentrations of cholesterol in plasma and lipoproteins were not different between the groups, there was a distinct positive correlation between circulating plasma homocysteine and plasma cholesterol (R(2)=0.55, p<0.001). The fecal excretion of cholesterol and bile acids was not altered by dietary methionine. The relative mRNA concentration of HMG-CoA reductase and of LDL receptor remained unaffected by dietary methionine. Gene expression of PEMT-2 was higher in rats fed the high-methionine diet than in rats fed the other diets (p<0.05). CONCLUSION: The results demonstrate that dietary methionine contributes to a rise in circulating homocysteine concentration which positively correlates with the concentration of plasma cholesterol. However, the effects of methionine on cholesterol metabolism of adult rats were relatively weak.     

Effects of soy protein and saponins on serum, tissue and feces steroids in rat

Four groups of rats were fed, for 45 days, one of the following semipurified diets containing sucrose 55% (w/w) and (a) casein 25%, (b) casein 24%, saponins (from Saponaria officinalis) 1%, (c) isolated soy protein 25%, (d) soy protein 24%, saponins 1%. The soy protein diet, compared to the casein one, produced an increase in the fecal excretion of neutral sterols on the 29th and 42nd days, without any modification in the liver, aorta and serum cholesterol concentrations. The effect of soy protein cannot be attributed to its saponin content but other substances associated to soy protein may interfere. With the casein diet, added saponins increased the fecal excretion of neutral sterols and bile acids and decreased liver and aorta cholesterol levels. Serum cholesterol was found unchanged. The effects of saponins were suppressed or greatly reduced with the soy protein diet. These results could be explained by binding of the sterols in insoluble forms.     

Dietary protein, fat, and minerals in nephrocalcinosis in female rats.

Young female rats fed semipurified diets containing casein or a soy protein isolate had extensive nephrocalcinosis at the junction between the outer and inner stripe of the outer medullary zone after 5 wk on the diets, whereas rats fed a diet containing a lactalbumin concentrate did not. Although the percentages of actual protein and of total ash were similar in all three diets, the concentrations of individual minerals were not, owing to methods used in isolating the proteins. Comparison of the individual mineral contents of these diets with those in other laboratories as compiled from the literature suggested that factors other than minerals, including protein, are also implicated. Dietary fat appeared to be another such factor in a series of experiments in which saturated medium-chain triglycerides and corn oil were included in diets containing soy protein isolate. Although these diets had identical mineral compositions, the rats fed medium-chain triglycerides had less severe lesions.     

Compositional changes in serum lipoproteins during developing hypercholesterolemia induced in rabbits by cholesterol-free, semipurified diets

Changes in the concentration and composition of serum very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) were studied in rabbits transferred from chow diet to cholesterol-free, semipurified diets containing casein or isolated soy protein. The fat and fibre content of these diets was similar to that of chow but a higher protein level was used to enhance the hypercholesterolemia. During the first week on the casein diet, there was a marked increased in LDL-cholesterol, protein and phospholipids, and these higher levels were maintained during the subsequent 3 weeks of the study. Similar but less marked changes were obtained with the soy protein diet. The components of VLDL showed relatively little change after introduction of the diets to the animals. In both VLDL and LDL, the proportion of cholesterol increased and that of triglycerides decreased after 1 week on the casein diet and a similar trend was seen in HDL. The concentration of HDL-cholesterol showed little change but triglycerides, protein and phospholipids all tended to decline on both casein and soy protein diets.     

Experimental atherosclerosis in rabbits fed cholesterol-free diets. 13. Interaction of proteins and fat

The atherogenic and cholesterolemic effects of animal protein vis-a-vis plant protein are well documented. Virtually all the studies were carried out using diets high in saturated fat, such as coconut oil. In order to determine if the same effects were seen with less saturated fat, we have compared atherogenic effects of an animal protein (casein) with those of a plant protein (soybean protein isolate) fed with partially hydrogenated soybean oil (PHS) (iodine value 72) or soybean oil (iodine value 134) as part of a cholesterol-free semipurified diet. After 6 months only rabbits fed casein-PHS exhibited elevated levels of plasma and liver cholesterol and triglycerides and atherosclerosis. Rabbits fed soy protein-PHS had slightly higher plasma cholesterol and triglycerides than did those fed soy protein and soybean oil, but values in both groups were in the normal range. The different effects of animal and plant protein on lipidemia and atherosclerosis can be influenced by dietary fat and appear to be dependent on fat saturation.     

Melatonin effect on plasma adiponectin,leptin, insulin,glucose, triglycerides and cholesterol in normal and high fat–fed rats

Abstract: Melatonin effect on body weight progression, mean levels and 24‐hr pattern of circulating adiponectin, leptin, insulin, glucose, triglycerides and cholesterol were examined in rats fed a normal or a high‐fat diet. In experiment 1, rats fed a normal diet were divided into two groups: receiving melatonin (25 μg/mL drinking water) or vehicle for 9 wk. In experiment 2, animals were divided into three groups: two fed with a high‐fat diet (35% fat) and melatonin (25 μg/mL) or vehicle in drinking water for 11 wk, while a third group was given a normal diet (4% fat). At the end of experiments, groups of eight rats were killed at six different time intervals throughout a 24‐ hr period. Melatonin administration for 9 wk decreased body weight gain from the 3rd wk on without affecting food intake. A significant reduction in circulating insulin, glucose and triglyceride mean levels and disrupted daily patterns of plasma adiponectin, leptin and insulin were observed after melatonin. In high fat–fed rats, melatonin attenuated body weight increase, hyperglycemia and hyperinsulinemia, as well as the increase in mean plasma adiponectin, leptin, triglycerides and cholesterol levels. The high‐fat diet disrupted normal 24‐ hr patterns of circulating adiponectin, insulin and cholesterol, the effects on insulin and cholesterol being counteracted by melatonin. Nocturnal plasma melatonin concentration in control and obese rats receiving melatonin for 11 wk attained values 21–24‐fold greater than controls. The results indicate that melatonin counteracts some of the disrupting effects of diet‐induced obesity in rats.     

Intestinal apolipoprotein B-48 synthesis and lymphatic cholesterol transport are lower in swine fed high fat, high cholesterol diet with soy protein than with casein

Effects of dietary proteins on intestinal lipoproteins were studied in 8 Yorkshire swine fed a high fat, high cholesterol diet with either casein or soy protein. After 5 weeks of feeding, the casein group exhibited moderately elevated levels of serum cholesterol (334 +/- 46 mg/dl). The soy protein group showed significantly less hypercholesterolemia as compared to the casein group (122 +/- 8 mg/dl). Swine were subjected to cannulation of mesenteric lymph duct under halothane anesthesia. A single dose of 250 microCi [14C]cholesterol and 10 mCi [3H]leucine was infused into the upper jejunum 2 h after one-fifth of daily food was given. The 3-h lymphatic transport of cholesterol in casein-fed swine was significantly higher than in those fed soy protein. Triglyceride transport values were similar in the 2 groups. The [3H]leucine incorporation study revealed that transport of apo B-48 bore a significant positive relationship to transport of cholesterol in both chylomicron and VLDL fractions of mesenteric lymph. A greater apo B-48 secretion with higher specific activity was probably responsible for the greater transport of cholesterol in chylomicrons in casein-fed than in soy protein-fed swine. Similarly, the transport of lymph VLDL cholesterol in swine fed casein or soy protein paralleled the amount of accompanying apo B-48. Dietary proteins probably influence the intestinal synthesis of apo B-48 which in turn affects cholesterol transport into the lymphatics.     

Relationship between dietary proteins, their in vitro digestion products, and serum cholesterol in rats

A relationship was assessed between the amino acid composition of 9 protein sources or of their in vitro digestion products and total serum cholesterol in rats. Three animal proteins (casein, beef, fish) and 6 vegetable proteins (soy, pea, peanut meal, rapeseed, oatmeal, wheat gluten) were tested. The intact protein sources were submitted to an enzymatic proteolysis according to a new in vitro digestion method. Each protein source was hydrolyzed for 30 min with pepsin at pH 1.9, then with 10 mg pancreatin at basic pH in a dialysis cell. The digestion products diffused through the dialysis membrane of the cell and were collected by a circulating sodium phosphate buffer over a 6-h period. They were likely to correspond to end products luminal in vivo digestion. The aromatic and the basic amino acids were present in higher proportions in the digestion products than in the intact protein sources, reflecting the specificity of the proteolytic enzymes. Total serum cholesterol was measured on male Sprague-Dawley rats fed cholesterol-free or cholesterol-enriched (1% cholesterol, 0.5% cholic acid) semipurified diets containing protein sources. Total serum cholesterol ranged from 70 mg/dl with the pea diet to 98 mg/dl with the peanut meal diet in rats fed cholesterol-free diets and from 163 mg/dl with the wheat gluten diet to 313 mg/dl with the casein diet in rats fed the cholesterol-enriched diets. These results suggested no specific effect of protein from animal or vegetable origin on total serum cholesterol in rats. In rats fed cholesterol-enriched diets, significant correlations were observed between total serum cholesterol and tyrosine content or leucine/isoleucine ratio of digestion products. These correlations were stronger than those observed with intact protein sources.     

Plasma and lipoprotein cholesterol in rats fed L-amino acid-supplemented diets

The effect of feeding amino acid-supplemented diets on plasma cholesterol concentration and distribution in the lipoproteins was studied on adult rats. A control diet (without any amino acid addition), and experimental diets supplemented with one of the following L-amino acids: lysine (10%), cystine (5%), methionine (1%), tryptophan (10%), valine (5%), and histidine (5%), were given for 2-4 months. Rats fed the lysine-, cystine- and tryptophan-added diets exhibited constant weights throughout the experiment, whereas those fed the other amino acid-added diets showed body weight gains quite similar to control rats. Two amino acids were shown to lower plasma cholesterol concentration: lysine (by 30%) and tryptophan (by 35%); one amino acid increased it: cystine (by 46%). The cholesterol distribution in the lipoproteins was significantly modified, principally when rats ingested cystine-enriched diets: as compared to control rats, the cholesterol concentration in lipoproteins of density between 1.040 and 1.063, and in high density lipoproteins (HDL), was increased by 174 and 58%, respectively.     

Effects of soy protein hydrolysate on blood pressure and angiotensin-converting enzyme activity in rats with chronic renal failure

We investigated the effects of soy protein and soy protein hydrolysate on blood pressure control, angiotensin-converting enzyme (ACE) activity, and renal function in a rat chronic renal failure model. Rats that had undergone a 5/6 nephrectomy were separated into three groups and fed different experimental diets for 14 weeks. At the end of the study, rats that fed a diet containing soy protein as the protein source had better blood pressure control and renal function, as well as lower circulating ACE activity and renal tumor-necrosis factor-alpha (TNF-alpha) concentration than rats fed a casein protein diet. Soy protein hydrolysate was shown to be as effective as soy protein in preventing the elevation of blood pressure, the progression of renal failure, and decreases in kidney TNF-alpha level, plasma ACE activity, and insulin concentration. In conclusion, the beneficial effects of consuming soy protein on blood pressure and renal function may be mediated mostly by its pepsin-digested hydrolysate through its ACE inhibitory activity.     

Casein-induced hypercholesterolemia in rabbits: distribution of cholesterol, triglycerides and phospholipids between serum and liver

Replacement of soy protein by casein in the cholesterol-free, semipurified diet of rabbits caused hypercholesterolemia within 7 days. After 36 days, the serum of casein-fed rabbits displayed elevated levels of free and esterified cholesterol and phospholipids, but not of triglycerides. Most of the excess of serum cholesterol in the casein group was localized in the LDL fraction, but there were marked variations in the density profile of the serum lipoproteins between individual rabbits. Dietary casein induced an increased content in liver of free and esterified cholesterol, but not of phospholipids and triglycerides. The molar ratio of free to esterified cholesterol in the liver was decreased by casein. In contrast, feeding casein resulted in an increase of this ratio in the serum.     

Effects of cholesterol-free, semipurified diets containing different levels of casein or soy protein on distribution of cholesterol and protein among serum lipoproteins of rabbits

Rabbits fed cholesterol-free, low-fat, semipurified diets have more cholesterol and protein in serum low density lipoprotein (LDL) relative to high density lipoprotein (HDL) than rabbits fed Chow diet. This difference was accentuated by a casein semipurified diet but was also observed with a soy protein diet even though the latter did not produce an elevation of serum cholesterol. To investigate the reason for these differences, the formulation of the semipurified diets was altered by reducing the level of protein from 27 to 16%, increasing the fat from 1 to 4% and the fiber from 5 to 13%, to correspond more closely to the proportions in Chow. With this formulation, the soy protein diet gave a lipoprotein pattern similar to that of Chow, whereas the casein diet produced a moderately elevated serum cholesterol level with more cholesterol in LDL than in HDL. When the protein in the newly formulated diets was increased back to 27%, the lipoprotein patterns reverted to those obtained with the original formula. In this case, soy protein-fed rabbits had moderately elevated serum cholesterol whereas casein-fed animals showed hypercholesterolemia. These results indicate that the altered lipoprotein pattern observed previously in rabbits fed semipurified diets is related to the high level of protein in those diets.     

The effect of dietary lysine to arginine ratio on cholesterol kinetics in rabbits

The effect of varying the compositions of dietary proteins on the relative cholesterolaemic effects of animal and vegetable proteins was investigated in rabbits. In experiments using high fat diets, the amino acid compositions of dietary proteins (soya or casein) were altered by blending them 1:1 (w/w) with gelatin. This reduced the differences in amino acid compositions and also made soya more and casein less hypercholesterolaemic. In experiment 2a, soya protein was compared with dried skim milk in low fat diets and in experiment 2b, these proteins were supplemented with lysine or arginine, respectively, so that the lysine:arginine ratio of soya was similar to dried skim milk and vice-versa. Serum cholesterol was significantly higher in milk-fed than soya-fed rabbits and was not influenced by reversing the lysine:arginine ratio. In the three experiments, parameters of cholesterol kinetics were estimated from the die-away curve of injected [4 14-C]cholesterol. There were no significant effects of diet on the parameters of cholesterol kinetics. It was concluded that the lysine:arginine ratio of the diet is not the major determinant of the cholesterolaemic properties of proteins, but that the overall amino acid composition is primarily concerned.