Cholesterol-free Diet Research Articles

Rhesus monkey model of familial hypercholesterolemia: relation between plasma Lp[a] levels, apo[a] isoforms, and LDL-receptor function.

We previously described a family of rhesus monkeys in which three out of six members had a spontaneous hypercholesterolemia related to a decrease in number of low density lipoprotein receptors (LDL-R) (Scanu et al. 1988. J. Lipid Res. 29: 1671-1681). During the current work an additional female normocholesterolemic offspring was generated from the mating of the original dam and sire. Moreover, from the breeding of one of the affected male offspring with six unrelated normocholesterolemic female monkeys, eight offspring were generated of which three were hypercholesterolemic on a cholesterol-free diet and exhibited the same degree of LDL-R deficiency as shown by studies in skin fibroblast cultures. All of the animals studied had levels of plasma lipoprotein[a] protein ranging between 1.0 mg/dl and 57.5 mg/dl that were only weakly correlated with total plasma cholesterol, LDL cholesterol, and apoB. LDL-R deficiency correlated with plasma LDL but not Lp[a]. A 7 week fat challenge (16.5% lard, 0.64% cholesterol) that raised the plasma LDL levels markedly had no effect on plasma Lp[a]. Animals with the single band apo[a] phenotype moving on SDS-PAGE faster than apoB-100 exhibited a tendency for high plasma Lp[a] levels which, however, varied widely. Wide variations in Lp[a] levels were also noted with the other apo[a] phenotypes. Taken together our results demonstrate a successful transmission to second generation animals of the LDL-R deficiency phenotype and provide evidence that this phenotype correlates well with plasma LDL levels but not Lp[a]. Our data also suggest that the apo[a] gene is only partially involved in the regulation of the plasma Lp[a] levels.

Some Metabolic Aspects of the Hypocholesterolemic Effect of Soybean Protein in Rats Fed a Cholesterol-Free Diet

The present paper shows a series of experiments carried out to elucidate the possible mechanism by which soybean protein isolate (SPI) produces a lower level of plasma cholesterol than casein in rats fed a cholesterol-free diet. When the plasma cholesterol level was in a steady state characteristic of casein and SPI, SPI in the diet was substituted for casein and vice versa. Within 3 days after substitution of dietary protein, the plasma cholesterol level in each group reached a steady state level similar to that in its previous counterpart. The inherent responses of plasma cholesterol to casein and SPI were not changed by the resection of the jejunum or the ileum and the administration of cholestyramine or beta-sitosterol. The rates of the sterol synthesis in vivo of the liver and the small intestine were significantly higher in SPI-fed rats than in casein-fed rats. The hypocholesterolemic effect of SPI disappeared when Met was supplemented at a level equivalent to casein. The effects of casein and SPI were reproduced by their equivalent amino acid mixtures. The ratio of the postprandial increment of Met concentration to that of Gly concentration in the portal plasma was significantly higher when casein and its amino acid mixture were fed than when SPI and its amino acid mixture were fed. The casein-induced increase in the level of plasma cholesterol was attributed to an increase in high density lipoprotein (HDL)-cholesterol. HDL-cholesterol concentration showed the positive correlation with the lecithin:cholesterol acyltransferase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Hydrophobic Surfactant Inhibits Hypercholesterolemia in Pair-fed Rabbits on a Cholesterol-Free, Low-Fat Diet

Poloxalene, a hydrophobic surfactant, is known to prevent hypercholesterolemia in animals fed a high-fat, high-cholesterol diet. It has not been demonstrated, however, whether this agent is of benefit when hypercholesterolemia is induced in animals by means other than the feeding of a high-fat diet. In this study, hypercholesterolemia was produced in rabbits by feeding a low-fat, cholesterol-free diet with dietary protein supplied by casein for a period of 8 weeks. Controls were given this diet without poloxalene and experimentals were given the diet with poloxalene. Total serum cholesterol levels increased in both groups, but the rise was greater for the control group. Lipoprotein analysis performed at the conclusion of the study showed significantly greater low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol levels in the control group as compared to the experimental group. Total protein and apolipoprotein B (apo B) were also greater in control LDL. It was concluded that poloxalene favorably affects this model of hypercholesterolemia as total serum cholesterol, LDL cholesterol, and LDL apo B were all less and the HDL cholesterol to LDL cholesterol ratio was higher in surfactant-treated rabbits.

Hypolipidemic principle of the inflorescence stalk of plantain (Musa sapientum)

A pectin present in the juice of the inflorescence stalk of plantain(Musa sapientum) has been isolated. The material contained 32.4% hexoses and 52.5% uronic acid. On administration to rats fed both cholesterol free and cholesterol diet, this material showed significant lowering of cholesterol and triglycerides in the serum, liver and aorta. There was decreased cholesterogenesis in the liver as was evident from decreased activity of hydroxymethylglutaryl coenzyme A reductase and decreased incorporation of labelled acetate into hepatic cholesterol. Hepatic bile acids showed significant increase and there was increased fecal excretion of neutral sterols and bile acids. Release of lipoproteins into the circulation was lower. The material also caused increase in the activity of lipoprotein lipase in the heart and adipose tissue and also of plasma lecithin: cholesterol acyl transferase

Predominant conjugation with glycine of biliary and lumen bile acids in rats fed on pectin

1. Bile acids were analysed in the bile and lumen samples of rats which received a cholesterol-free or cholesterol-enriched (5 g/kg) diet free from fibre, or containing cellulose or citrus pectin at the level of 100 g/kg. 2. Dietary pectin but not cellulose increased biliary bile acid concentration and excretion. Dietary cholesterol did not affect biliary bile acids quantitatively. 3. Biliary bile acids were almost exclusively conjugated with glycine or taurine in the various experimental situations. The predominant portion of bile acids in rats fed on the cholesterol-free diet was conjugated with taurine when the diet was either free from fibre or contained cellulose; the ratio of bile acids conjugated with glycine: those conjugated with taurine (G:T) was less than 0.2. In contrast, with pectin as a fibre source, the conjugation with glycine increased enormously (G:T increased to approximately 4). Cholesterol enrichment of the diet also increased the glycine conjugation in all groups of rats. Even in this situation, the G:T was highest in rats fed on pectin. 4. Pectin, but not cellulose, increased the bile acid content of the small intestine and caecum, both in rats fed on the cholesterol-free and cholesterol-enriched diets. Cholesterol feeding doubled the bile acid content of the caecum in rats fed on a fibre-free diet or a cellulose diet, but not in those fed on pectin. No such effect of cholesterol was observed in the small intestine, except for the ileal bile acid content in rats fed on cellulose. 5. A considerable portion of the bile acids in the small intestine was deconjugated. The extent of the deconjugation was higher in the ileum than in the jejunum. As in the bile, G:T in rats fed on pectin (3-5.5) were higher than those in the other groups (0.05-1.05) in various situations. Also, cholesterol feeding considerably increased the ratio in all groups of rats. 6. The observed dietary alteration of the partition of bile acids between glycine and taurine may be of physiological significance in regulating bile acid and lipid metabolism in rats.

Oxysterol Regulators of 3-Hydroxy-3-methylglutaryl-CoA Reductase in Liver: Effect of Dietary Cholesterol

Hepatic regulatory oxysterols were analyzed to determine which oxysterols were present in livers of mice fed a cholesterol-free diet and whether repression of 3-hydroxy-3-methylglutaryl-CoA reductase following cholesterol feeding was accompanied by an increase in one or more oxysterols. Analysis of free and esterified sterols from mice fed a cholesterol-free diet resulted in the identification and quantitation of six regulatory oxysterols: 24-hydroxycholesterol, 25-hydroxycholesterol, 26-hydroxycholesterol, 7 alpha-hydroxycholesterol, 7 beta-hydroxycholesterol, and 7-ketocholesterol. Following the addition of cholesterol to the diet for 1 or 2 nights, hepatic 3-hydroxy-3-methylglutaryl-CoA reductase activity declined and the levels of oxysterols, especially those of the side-chain-hydroxylated sterols, increased. Total 3-hydroxy-3-methylglutaryl-CoA reductase repressor units attributable to identified free oxysterols increased 2.5- and 6-fold after 1 and 2 nights, respectively, of cholesterol feeding. The amounts of esterified 24-, 25-, and 26-hydroxycholesterol also increased, with the increase in esterified 24-hydroxycholesterol being the greatest. The 24-hydroxycholesterol was predominantly the 24S epimer and the 26-hydroxycholesterol was predominantly the 25R epimer, indicating enzymatic catalysis of their formation. The observed correlation between increased levels of regulatory oxysterols and repression of 3-hydroxy-3-methylglutaryl-CoA reductase in cholesterol-fed mice is consistent with a hypothesis that intracellular oxysterol metabolites regulate the level of the reductase.

Effects of addition of sulfur-containing amino acids and glycine to soybean protein and casein on serum cholesterol levels of rats.

The effects of the supplementation of methionine (Met), cystine (Cys), and glycine (Gly) to soybean protein or casein on serum and liver lipid levels were studied in rats. Rats were fed cholesterol-free diets containing 25% soybean protein or casein supplemented with 0.75% Met, 2.5% Gly, or a combination of these two for 4 weeks. The addition of Met to soybean protein caused a significant increase in serum cholesterol and this was slightly ameliorated when Gly was given simultaneously. In rats fed casein diets, serum cholesterol tended to decrease when Gly, or Met and Gly were added. A simultaneous supplementation of Met and Gly to casein resulted in a reduction of hepatic cholesterol. Cystine added at the 0.6% level did not cause demonstrable changes in lipid concentrations except for a drop in serum triglyceride of the casein group. When 2.0% Gly was added to cholesterol-enriched diets containing 20% protein, serum cholesterol decreased significantly only when the protein source was casein and the level attained was comparable to that observed in rats fed soybean protein. Liver cholesterol was also markedly decreased by the addition of Gly to casein. The results suggest a possible role of Gly in the regulation of serum cholesterol levels by dietary protein.

Mechanism of hypercholesterolemia produced by biotin deficiency

The effect of biotin deficiency on the metabolism of cholesterol was studied in rats fed cholesterol-free and cholesterol-containing diet. Biotin deficiency induced by feeding raw egg-white resulted in higher cholesterol in the serum and aorta, and higher high density lipoprotein cholesterol and low density lipoprotein + very low density lipoprotein cholesterol. In the liver, cholesterol increased only in the cholesterol diet group but not in the cholesterol-free diet group. Levels of triglycerides were lower in the biotindeficient, cholesterol-free diet group, but triglycerides were elevated in the cholesterol diet group. Concentration of bile acids in the liver and activity of lipoprotein lipase in the heart and adipose tissue were significantly decreased in the biotin-deficient rats. Release of lipoproteins into the circulation, incorporation of [1,2-14C] acetate into cholesterol, and activity of plasma lecithin: cholesterol acyl transferase were higher.

The effects of cholesterol supplementation on plasma lipoprotein-cholesterol levels in rats fed diets deficient in vitamin e and/or selenium

Rat plasma lipoprotein-cholesterol levels were measured in male Fischer-344 rats fed a vitamin E and selenium deficient basal diet containing 1% cholesterol (B+cho) or identical diets to which vitamin E (B+E+cho), selenium (B+Se+cho) or both micronutrients were added (B+E+Se+cho). In each of these dietary groups, cholesterol supplementation caused an increase in plasma-cho and low density lipoprotein-cho (LDL-cho) but a decrease in high density lipoprotein-cho (HDL-cho) compared with values previously measured (1) in rats red corresponding cholesterol-free diets. Selenium deficiency (B+cho and B+E+cho) was found to be an important factor (P < 0.001) associated with increased levels of LDL-cho and plasma-cho. In contast, vitamin E did not significantly influence either plasma-cho or LDL-cho. Rats deficient in both vitamin E and selenium (B+cho group) were, however, found to develop elevated very low density lipoprotein-cho (VLDL-cho) and to have decreased HDL-cho. Cholesterol supplementation was found to significantly interact with selenium deficiency in producing elevated LDL-cho levels. Our results are discussed with respect to the antioxidant properties of both vitamin E and selenium and the role of cholesterol in modulating the plasma clearance rate of lipoproteins.

Effects of a daisaikoto extract on the biliary constituents in mice and rats.

The effects of an extract of Daisaikoto (a traditional Chinese medicine) on biliary constituents was studied in mice fed with a lithogenic diet containing 0.5% cholesterol and 0.25% sodium cholate (the control diet) and in rats fed with a cholesterol-free diet. The Daisaikoto extract was added to the control diet at a level of 0.5%. A high incidence of cholesterol gallstones were found in the control mice, but not in the mice given the Daisaikoto extract. This difference was concluded to have been due to the absolute concentration of bile acid in the bile being significantly higher in the mice given the Daisaikoto extract than in the control mice. The result from rats fed with the cholesterol-free diet also demonstrated that the Daisaikoto extract caused an increase in the absolute concentration of bile acid in the bile.

On the source of sterols in the green peach aphid, Myzus persicae, reared on holidic diets

Cultures of Myzus persicae were maintained for five generations on holidic diet lacking sterol. A method to incorporate cholesterol into the aqueous diet is described. Addition of cholesterol to the diet (at 2.5 μg ml −1) reduced the larval development time of apterous M. persicae raised to the fourth generation on cholesterol-free diet by 12–16%, but had no significant effect on their teneral weight, the number of offspring deposited or the birth weight of their offspring. The symbiotic microorganisms in the aphids' mycetocytes were disrupted by treatment with the antibiotic chlortetracycline. These “aposymbiotic” aphids produced viable offspring on the diet containing cholesterol, but on the cholesterol-free diet development of the embryos was halted prior to germ band formation. These data suggest that the symbionts may synthesize sterols, which are required for embryogenesis of the aphid on diets. The possible involvement of microbial contaminants in the diet or gut microflora of the aphid in the sterol nutrition of M. persicae is excluded by rigorous microbiological studies.

Serum and liver lipids of rats fed rubber seed oil.

Crude rubber seed oil (RSO) was incorporated at the 5% level in diets free of cholesterol or containing 1% cholesterol, respectively. These studies were initiated for the purpose of evaluating the palatability and potential toxicity of RSO. Corn oil was used as a control. A considerable amount of unsaponifiable matter was detected in RSO. In addition, RSO was found to possess a fatty acid profile which was fairly different from that of corn oil, primarily due to a much higher content of linolenic acid and a lower content of linoleic acid in RSO. No adverse effects on food intake and average daily gain were observed in rats fed RSO in both cholesterol-free and cholesterol diets. The presence of RSO in cholesterol-free diets results in lower (p less than 0.05) serum and liver total cholesterol levels than in control animals. A relative hypercholesterolemic effect compared to corn oil was observed however, when RSO was added to diets containing 1% cholesterol. In summary, the physicochemical properties of RSO as well as the presence of cholesterol in the diet are important factors in evaluating the cholesterolemic effect of RSO.

Serum and Liver Lipids of Rats Fed Rubber Seed Oil

Crude rubber seed oil (RSO) was incorporated at the 5% level in diets free of cholesterol or containing 1% cholesterol, respectively. These studies were initiated for the purpose of evaluating the palatability and potential toxicity of RSO. Corn oil was used as a control. A considerable amount of unsaponifiable matter was detected in RSO. In addition, RSO was found to possess a fatty acid profile which was fairly different from that of corn oil, primarily due to a much higher content of linolenic acid and a lower content of linoleic acid in RSO. No adverse effects on food intake and average daily gain were observed in rats fed RSO in both cholesterol-free and cholesterol diets. The presence of RSO in cholesterol-free diets results in lower (p<0.05) serum and liver total cholesterol levels than in control animals. A relative hypercholesterolemic effect compared to corn oil was observed however, when RSO was added to diets containing 1% cholesterol. In summary, the physicochemical properties of RSO as well as the presence of cholesterol in the diet are important factors in evaluating the cholesterolemic effect of RSO.

Monoclonal antibody characterization of hepatic and extrahepatic cytochrome P-450 activities in rats treated with phenobarbital or methylcholanthrene and fed various cholesterol diets

Monoclonal antibodies (MAb) against 3-methylcholanthrene (MC)- and phenobarbital (PB)-inducible forms of cytochrome P-450 isozyme were used to characterize changes in aryl hydrocarbon hydroxylase (AHH) and ethoxycoumarin O-deethylase (ECDE) activities modulated by dietary cholesterol. Rats were induced by MC or PB, and immunochemical inhibition of AHH and ECDE activities was studied as an indication of changes in cytochrome P-450 isozyme patterns. Feeding of a cholesterolfree diet markedly decreased enzyme activities both in liver and in small intestinal mucosa, and the highest activities were observed after feeding rats a high (2%)-cholesterol diet for one month. As a control, a normal pelleted diet (0.1% cholesterol) was used; in rats fed this diet, intermediate levels of monooxygenase activities were present. Although no diet-dependent change in total AHH and ECDE activities was observed in kidneys and lungs, diet apparently modulated isozyme composition in the lungs, as indicated by a change in the immunochemical inhibition pattern with MAb; no such shift was observed in the kidneys. In liver and intestine, in addition to changes in total activity, isozyme composition was also altered, as indicated by inhibition of the catalytic activities of cytochrome P-450 by MAb. Our data infer that dietary cholesterol can: (i) modulate total monooxygenase activities, especially in the intestine; (ii) change the cytochrome P-450 isozyme composition in liver and intestine; (iii) change isozyme composition without changing overall enzyme activity, e.g. in lungs; and (iv) have no effect in a tissue (e.g. kidney) that lacks constitutionally the P-450 isozyme responsive to cholesterol.

Intestinal cholesterol absorption in the chyluria model

Isotopic methods for the measurement of dietary cholesterol absorption were compared with the lymph cholesterol balance procedure in filarial chyluria patients. After a single intravenous injection of radioactive cholesterol, absorption was found to be 746 +/- 136 mg/day by method I, which is based upon the fecal endogenous neutral steroid mass measurement, and 471 +/- 135 mg/day by the simultaneously measured lymph/plasma ratio of cholesterol specific activity (dpm/mg). The corresponding value, determined as the difference between lymph cholesterol transport on a cholesterol-containing diet (1500 mg) and on a cholesterol-free diet, was 622 mg/day. When radioactive cholesterol (1487 mg/day) was fed daily to a second patient, absorption determined by isotopic fecal recovery (353 mg/day) matched that obtained by the lymph balance procedure (326 mg/day). Transudation of plasma cholesterol into the intestinal lymph, estimated by the single intravenous injection of radioactive beta-sitosterol, was independent of both the luminal content of plant sterols and the absorption of dietary cholesterol. The absorption of endogenous cholesterol was calculated by: 1) subtracting the cholesterol originating from plasma (transudation) together with the absorbed dietary cholesterol found in lymph from the total mass of cholesterol transported in lymph, and 2) the lymph balance method, i.e., after interrupting the endogenous cholesterol mucosal uptake by beta-sitosterol feeding (9 g/day) while on a cholesterol-free diet. Endogenous cholesterol was preferentially absorbed compared to dietary cholesterol, but there was no competition for absorption. The major portion of dietary cholesterol found in lymph was esterified, but esterification was not a prerequisite for absorption.

Fecal steroid excretion in chickens with hereditary hyperlipidemia.

Plasma lipids (cholesterol, triglycerides, and phospholipids; mg/dl) and the fecal excretion (mg/day) of neutral steroids and bile acids were studied in layers (L), hereditary nonlayer hens (NL), and roosters (R) fed a basal cholesterol-free grain diet ad libitum. Each group had significantly (P less than 0.05) different levels of plasma cholesterol, triglycerides, and phospholipids when compared to the other groups. The highest lipid values were found in the NL group (cholesterol, 798 +/- 89; triglycerides, 8914 +/- 679; phospholipids, 2458 +/- 112). There was no difference in the fecal excretion of neutral steroids between L and NL; however, fecal bile acid excretion by these two groups was significantly different (P less than 0.05) (L, 13.1 +/- 1.7 vs NL, 26.9 +/- 3.4). Fecal neutral steroid excretion by R was significantly greater (P less than 0.05) than that by either L or NL (L, 6.4 +/- 1.3; NL, 6.0 +/- 1.4; R, 14.4 +/- 1.2). While fecal excretion of bile acids by R (36.1 +/- 4.0) was also greater than that by either L or NL, only the difference between R and L was statistically significant (P less than 0.05). Since, in the steady state, fecal bile acid excretion is equal to its synthesis, these results suggest that bile acid metabolism in these animals can be affected by both sex and egg-laying status.

Dietary acetate and cholesterol metabolism in rats.

Rats were fed either cholesterol-free or high-cholesterol (1%, w/w) semipurified diets containing acetate (0.5%) or cholestyramine (0.44%) or both compounds for 29 days. The bile-acid binding resin, cholestyramine, did not affect serum and liver cholesterol, irrespective of whether the diet was cholesterol-free or contained cholesterol. In the cholesterol-free diets, acetate tended to lower the concentration of serum cholesterol, but did not influence liver cholesterol. When the diets contained cholesterol, acetate lowered liver cholesterol concentrations by about 20% both in the presence and absence of cholestyramine. Acetate did not affect the excretion of bile acids in feces. The mechanism by which acetate may partly counteract the increase in liver cholesterol seen after cholesterol feeding of rats, remains to be established.

Effects of pure capsaicinoids (capsaicin and dihydrocapsaicin) on plasma lipid and lipoprotein concentrations of turkey poults

The effects of capsaicin and dihydrocapsaicin on blood lipid and lipoprotein concentrations were determined in two groups of turkeys. The first group was maintained on a cholesterol-free diet, while the second received a diet supplemented with 0.2% cholesterol. Daily administration of capsaicinoids occurred at a dose of 4 mg per animal. Neither drug had an effect on serum triglyceride concentrations in the animals receiving the cholesterol-free diet. However, total cholesterol, LDL-cholesterol and HDLcholesterol concentrations were increased significantly, while VLDL cholesterol concentrations were decreased significantly by both drugs relative to controls. In the cholesterol-fed group triglycerides, total cholesterol and LDL-cholesterol decreased significantly with dihydrocapsaicin treatment. Both compounds reduced VLDL-cholesterol and increased HDL-cholesterol in the cholesterol-fed animals. Dihydrocapsaicin had a greater efficacy in producing benificial anti-hyperlipidemic effects in the cholesterol-fed animals.

Hydrophobic surfactant effects on aortic cholesterol accumulation and atherosclerosis in hypercholesterolemic rabbits

Studies were performed in hypercholesterolemic rabbits to determine whether the hydrophobic surfactant, Poloxalene 2930 (Pol), is of benefit under these conditions. Lipoprotein analyses plus chemical and morphologic studies of the aorta were performed to evaluate the results. In one study, rabbits were made hypercholesterolemic by dietary means and then divided into two groups and given a cholesterol-free diet with one group additionally given Pol with treatment continued for 10 weeks. Pol treatment resulted in less atherosclerosis but the mechanism for this effect was not apparent from lipoprotein analysis. In the other study 3 groups of rabbits were given a cholesterol-rich diet for 16 weeks. Two groups received Pol supplement with one of these groups receiving a dose that was too small to prevent hypercholesterolemia. In this group plus the group on diet alone comparable degrees of hypercholesterolemia were maintained throughout the study. Lipoprotein abnormalities were similar in these two groups except that those on Pol had a more normal cholesterol to apolipoprotein B ratio. The amount of atherosclerosis in both groups was mild but aortic cholesterol content was much less for the Pol group. It is concluded that Pol limits cholesterol accumulation in the aortic wall of hypercholesterolemic rabbits and can retard the development of atherosclerosis.

Flow cytometric quantification of cholesteryl ester-containing “foam” cells: I. Analysis of aortas from normolipidemic swine

We have quantified using flow cytometry foam cells of aortas from normolipidemic swine varying in age from 6 months to 12 years. These swine were maintained throughout their lives on a low-fat, cholesterol-free diet. Intimal-medial tissues removed from the swine aortas were enzymatically dissociated to prepare Formalin-fixed cell suspensions. Foam cells were labeled by specific staining of their intracellular cholesteryl ester using the fluorescent dye filipin. This was carried out by first removing cellular unesterified cholesterol with ethanol, then enzymatically hydrolyzing cellular cholesteryl ester, and finally staining with filipin the unesterified cholesterol derived from hydrolysis of cholesteryl ester. Results of flow cytometric analysis indicated that thoracic foam cell densities of female swine became more variable and tended to increase with age. It appeared that there were two sub-groups of female swine with either high or low levels of thoracic foam cells. A similar finding was not observed for abdominal foam cells in female swine nor for thoracic or abdominal foam cells in the smaller number of older male swine. Abdominal foam cell densities in younger males, however, also appeared to be comprised of low and high foam cell groups. Foam cell densities did not correlate with serum cholesterol or triglyceride levels. However, a genetic basis for some of the variability in foam cell densities among these animals was suggested by the observation that females with high thoracic foam cell densities had greater commonality among ancestors than did females with low thoracic foam cell densities.