bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes of high density lipoprotein subfraction concentration and composition by intralipid in vivo and by lipolysis of intralipid in vitro.
Authors: Taskinen M R , Nikkilä E A , Kuusi T , Tulikoura I
Published: June 2011

Serum lipoproteins were measured during a single infusion of intralipid and during parenteral nutrition with intralipid and glucose. Postheparin plasma lipolytic enzymes and plasma LCAT activity were assayed before and after the parenteral nutrition. Both single and repeated infusions of intralipid were followed by a significant rise of HDL2 concentration (p less than 0.01), whereas the HDL3 decreased. The composition of HDL subclasses altered. The HDL2 triglyceride and phospholipids increased, while the HDL3 esterified cholesterol and protein decreased. In vitro incubation of serum with intralipid alone caused no changes in the zonal profile of HDL subclasses, but hydrolysis of intralipid by lipoprotein lipase was followed by conversion of HDL3 into lighter particles floating in the density range of HDL2. The present results provide additional evidence for a precursor-product relationship between the HDL2 and HDL3. During 4 days of parenteral nutrition with intralipid, the basal (morning) values of serum total and VLDL triglyceride did not change. The LDL phospholipids increased progressively (from 67 to 98 mg/dl, p less than 0.05). The total HDL cholesterol decreased and this change was due to the fall of HDL3 cholesterol esters (from 19 to 12 mg/dl, p less than 0.05). Also the basal values of apo A-I and A-II in HDL3 decreased. The basal level of the HDL2 remained constant. Postheparin plasma LPL activity increased by 52% (p less than 0.01) but hepatic lipase activity fell by 49% (p less than 0.05). These changes may account for the maintenance of plasma HDL2, whereas the progressive fall of the basal HDL3 is probably due to the lack of intestinal apoprotein synthesis during absent intestinal absorption.
Biochemical changes in endothelial cell monolayers induced by fibrin deposition in vitro.
Authors: Schleef R R , Birdwell C R
Published: June 2011

Endothelial cells in vivo come into contact with fibrin during numerous physiological and pathological processes, but the nature of this interaction is poorly understood. The interaction of fibrin and endothelial cells was examined by incubating bovine aortic endothelial cell monolayers in vitro with fibrinogen (100 micrograms/ml) in medium supplemented with 10% calf serum. This treatment produced a layer of fibrin above the endothelial cell monolayer, which induced disorganization of the monolayer and remained associated with the monolayer even after washing. Endothelial cell pinocytosis of 14C-sucrose was elevated by 25% and 60% after incubation with fibrin for 8 hours and 2 days, respectively. Fibrin deposition on endothelial cell monolayers increased the rate of DNA synthesis (3H-thymidine incorporation) for 1 to 3 days by as much as 60% over controls. Concomitant with the lysis of fibrin above endothelial cells and the reorganization of the monolayer at Days 4 or 5 was a reduction of endothelial cell pinocytosis and DNA synthesis to control values. In contrast, there was no effect on the cell density at any time after incubation with fibrin. These results indicate that fibrin interacts with endothelial cells and induces a disorganization of the monolayer that is accompanied by changes in the rate of pinocytosis and in the rate of DNA synthesis.
Relationship between cortisol and cholesterol in men with coronary artery disease and type A behavior.
Authors: Schwertner H A , Troxler R G , Uhl G S , Jackson W G
Published: June 2011

To further understand the hormonal mechanisms linking behavior pattern and coronary artery disease (CAD), we investigated the relationship between 0930-hour plasma cortisol and cholesterol in relatively young males who had undergone coronary angiography and in a subgroup of individuals who had undergone the structured interview for classification of behavior pattern. A statistically significant association (p less than 0.05) was found between cortisol and cholesterol for individuals who had either minimal CAD (20% to 49% narrowing) or significant CAD (greater than or equal to 50% narrowing), but not for subjects without CAD. An association between cortisol and cholesterol was also found to be significant for the subgroup of individuals with Type A-1 behavior pattern, but not for those with Type A-2, X, or B behavior patterns. The findings suggest that hormonal mechanisms involving cortisol and cholesterol metabolism may be operative in individuals with CAD as well as in individuals with Type A-1 behavior.
Family resemblance for plasma lipids and lipoprotein concentrations in blacks. Cincinnati Lipid Research Clinic Family Study.
Authors: Laskarzewski P M , Glueck C J , Rao D C
Published: June 2011

Our specific aim in this study was to assess the family resemblance for lipids and lipoproteins in blacks randomly selected from the Princeton School District's Family Study cohort, and compare the family resemblance of lipids and lipoproteins between the blacks and whites from the same cohort. There were 160 white and 59 black nuclear families ascertained through randomly selected family study probands. Familial correlations were estimated by the method of maximum likelihood. Father-child and sib-sib correlations were of larger magnitude in whites than blacks for each lipid and lipoprotein, with the exception of high density lipoprotein cholesterol, where the correlations in blacks were higher than in whites. Estimates of genetic heritability were larger in whites than in blacks for each lipid and lipoprotein, with the exception of high density lipoprotein cholesterol. Whereas environment has a substantial effect on high density lipoprotein cholesterol levels in both blacks and whites, there may be a greater genetic effect accounting for higher levels of high density lipoprotein cholesterol in blacks. This is highlighted in our current study by the consistent observation in blacks of increased measures of within-family resemblance for high density lipoprotein cholesterol alone, of all lipids and lipoproteins.
Effect of weight loss in moderate obesity on plasma lipoprotein and apolipoprotein levels and on high density lipoprotein composition.
Authors: Zimmerman J , Kaufmann N A , Fainaru M , Eisenberg S , Oschry Y , Friedlander Y , Stein Y
Published: June 2011

Plasma lipids, lipoprotein and apolipoprotein levels were determined in seven women and seven men with moderate obesity before, during 7 weeks of continuous weight loss (10.4% to 9.6% of body weight, 1000 kCal/day diet), and after 3 months at a stable, reduced weight. Plasma triglyceride levels decreased by 30.4% in men and by 39.4% in women (p less than 0.0001) after 1 week of caloric restriction and remained at this level throughout the study period. The plasma cholesterol decreased by 19.0% in men (p less than 0.001) and by 10.9% in women (p less than 0.01) in the period of active weight loss, but returned to prediet values after stabilization at a leaner body mass. Similar changes were observed in LDL cholesterol levels. No change in high density lipoprotein (HDL) cholesterol levels occurred during active weight reduction, but after 3 months at a reduced weight, a significant increase in HDL cholesterol was evident, and the ratio of HDL cholesterol to plasma cholesterol increased over prediet values (p less than 0.001), women). Separation of HDL subpopulations by zonal ultracentrifugation before and after weight reduction revealed that HDL2 increased slightly in men and decreased slightly in women. In both genders, HDL3 tended to decrease after weight reduction. Plasma levels of apolipoprotein A-I decreased during active weight loss, but this was significant only in women (p less than 0.05). After 3 months of reduced weight, plasma apo A-I increased to prediet levels. No significant changes in plasma apo A-II or apo E were noted.(ABSTRACT TRUNCATED AT 250 WORDS)
Electron microscopic structure of serum lipoproteins from patients with fish eye disease.
Authors: Forte T M , Carlson L A
Published: June 2011

The structure and composition of lipoprotein fractions from two patients with fish eye disease were examined. The composition of very low density lipoproteins (VLDL) was normal, although total mass was greatly elevated. The mean particle sizes of the VLDL were 44.6 +/- 22.2 and 42.8 +/- 19.8 nm for Patients 1 and 2, respectively. Intermediate density lipoprotein (IDL) concentrations in patients were elevated and contained increased triglyceride content; particle sizes for Patients 1 and 2 were 29.4 +/- 3.5 nm and 28.0 +/- 4.1 nm, respectively. In both patients, the triglyceride/cholesteryl ester ratio in LDL was approximately tenfold higher than in normal individuals; however, the LDL particles were somewhat smaller in diameter (23.5 +/- 3.0 nm for Patient 1 and 23.3 +/- 3.8 nm for Patient 2) than those of controls (25.8 +/- 3.0 nm and 24.9 +/- 3.4 nm). In both patients, large vesicular structures were occasionally encountered in the LDL region. The high density lipoprotein (HDL) fraction of fish eye disease patients showed the greatest abnormalities. Not only was the total HDL concentration extremely low (approximately 10% of control levels), but unesterified cholesterol was increased relative to cholesteryl ester. Particle morphology was heterogeneous; the major HDL species was a small spherical particle with a diameter of 7.6 nm. Discoidal particles with a thickness of 4.4 nm and diameters between 17.4 and 20.8 nm were also present, together with large (40-90 nm) vesicles.(ABSTRACT TRUNCATED AT 250 WORDS)
In vitro metabolism of LDL labeled with a nondegradable cholesteryl ester analogue.
Authors: Higgs D W , van der Westhuyzen D R , Gevers W , Coetzee G A , Stein O , Stein Y
Published: June 2011

Low density lipoprotein (LDL) metabolism by human skin fibroblasts was studied using LDL labeled in the neutral lipid fraction with a nonhydrolysable cholesteryl ester analogue 3H-cholesteryl linoleyl ether (3H-CLE-LDL). LDL uptake could be quantitated accurately using 3H-CLE-LDL, since the label accumulated intracellularly due to its resistance to hydrolysis. 3H-CLE-LDL was taken up via apo B,E receptor-mediated endocytosis in a manner similar to 125I-labeled LDL. This was demonstrated by similar rates of uptake of the two differently labeled LDL preparations, saturation kinetics of uptake with respect to 3H-CLE-LDL concentration, regulation of 3H-CLE-LDL uptake by procedures that up-regulate or down-regulate the number of apo B,E receptors, and negligible uptake of 3H-CLE-LDL by receptor-negative cell strains. The major difference between the handling of 125I-LDL and 3H-CLE-LDL was the finding that, while the amount of trypsin-releasable (surface) 3H-CLE increased progressively over a 24-hour experimental period, trypsin-releasable 125I-LDL reached a maximum within 30 minutes. After 24 hours of incubation, the 3H radioactivity released by brief trypsinization was three to four times higher than could be accounted for by 125I radioactivity released by a similar treatment. Possible reasons for this behavior are discussed.
LDL, scavenger, and beta-VLDL receptors on aortic endothelial cells.
Authors: Baker D P , Van Lenten B J , Fogelman A M , Edwards P A , Kean C , Berliner J A
Published: June 2011

Primary and first passage aortic endothelial cells were shown to possess a high affinity receptor for beta-migrating very low density lipoproteins (beta-VLDL) distinct from the low density lipoprotein (LDL) receptor and scavenger receptor on these cells. In bovine aortic endothelial cells, 125I-rabbit beta-VLDL was taken up and degraded by a high affinity process that was competed for by unlabeled rabbit beta-VLDL and unlabeled postprandial VLDL from a fat-fed normal subject. However, unlabeled human or rabbit LDL, human LDL modified by malondialdehyde (MDA-LDL), or VLDL from a fasted normal human or a rabbit were not effective competitors for the degradation of 125I-rabbit beta-VLDL. In contrast to the receptor-mediated degradation of 125I-human or rabbit LDL and 125I-human-MDA-LDL, cell density did not affect the receptor-mediated degradation of 125I-rabbit beta-VLDL. Endothelial cells from a Watanabe heritable hyperlipidemic (WHHL) rabbit virtually did not degrade rabbit LDL, but degraded rabbit beta-VLDL at a rate equal to that seen in normal rabbit endothelial cells. It was concluded that the beta-VLDL receptor on endothelial cells is genetically distinct from the LDL receptor. Incubation of cells for 3 days with 100 micrograms/ml protein of unlabeled beta-VLDL caused an 88% increase in cellular cholesterol content, even though the beta-VLDL receptor activity was down-regulated by 60%. Endothelial cells and monocyte-macrophages are thus far the only cells known to possess the LDL receptor, the scavenger receptor, and the beta-VLDL receptor.
Effect of a diet low in saturated fatty acids on plasma lipids, lipoproteins, and HDL subfractions.
Authors: Ehnholm C , Huttunen J K , Pietinen P , Leino U , Mutanen M , Kostiainen E , Iacono J M , Dougherty R , Puska P
Published: June 2011

The effect on serum high density lipoprotein subfractions of a low fat diet with a high ratio of polyunsaturated-to-saturated fatty acids was studied in 38 middle-aged volunteers (19 men and 19 women) in North Karelia, Finland. The mean serum HDL2 cholesterol decreased from 32 +/- 2 mg/dl (mean +/- SE) to 28 +/- 2 mg/dl (p less than 0.001) during the experimental diet and returned to 33 +/- 2 mg/dl (p less than 0.001) after a return to the original diet. No changes were observed in the concentration of HDL3 cholesterol. A highly significant decrease was observed in serum apoprotein A-I concentration, but not in apoprotein A-II concentration during the experimental diet. It is concluded that a low-fat, high-P/S ratio diet lowers LDL and HDL2 cholesterol in healthy volunteers, but does not influence the level of HDL3 subfraction.
Inhibition of low density lipoprotein synthesis by dietary omega-3 fatty acids in humans.
Authors: Illingworth D R , Harris W S , Connor W E
Published: June 2011

Diets rich in omega-3 fatty acids derived from fish oils lower the plasma concentrations of low density lipoproteins (LDL) and very low density lipoproteins in humans. The present study was designed to examine the mechanism(s) by which diets enriched in omega-3 fatty acids reduce plasma LDL cholesterol levels in normal subjects. Seven healthy volunteers with normal plasma lipid levels consumed two metabolically controlled diets for a period of 4 weeks each. The control diet contained predominantly saturated and monounsaturated fatty acids, whereas the fish-oil diet contained 24 gm of omega-3 fatty acids per day. The total fat and cholesterol content of the two diets were similar for each subject. Total and LDL cholesterol levels decreased from 162 +/- 26 mg/dl and 103 +/- 27 mg/dl on the control diet to 124 +/- 26 mg/dl and 82 +/- 27 mg/dl on the omega-3-rich diet. Triglyceride levels fell from 91 +/- 34 mg/dl to 52 +/- 19 mg/dl. Kinetic studies of 125I-LDL metabolism disclosed a significantly lower rate of synthesis of LDL apoprotein B on the omega-3-rich diet (9.5 +/- 1.3 mg/kg/day) as compared to the control diet (13.6 +/- 3.7 mg/kg/day; p less than 0.05). In contrast, the fractional catabolic rate was similar on both diets. We conclude that dietary omega-3 fatty acids lower plasma LDL levels in normal human subjects by reducing the rate of synthesis of apoprotein B.
Studies of hypercholesterolemia in the nonhuman primate. I. Changes that lead to fatty streak formation.
Authors: Faggiotto A , Ross R , Harker L
Published: June 2011

Morphologic studies resulting from events that occur during the development of the lesions of atherosclerosis were studied in chronic, diet-induced hypercholesterolemia in a series of nonhuman primates. Within 12 days of hypercholesterolemia in Macaca nemestrina, monocytes became adherent to the surface of the endothelium. These monocytes appeared to migrate subendothelially, accumulate lipid, and become lipid-laden macrophages (foam cells). Within a month, a "serofibrinous insudate" formed together with variable numbers of subendothelial lipid-laden macrophages. By the second month, foam cells increased in number, often in multilayers, to form a fatty streak. Concomitantly, the luminal surface of the arteries became increasingly irregular due to the subendothelial accumulation of foam cells. Numerous monocytes continued to attach to the endothelial surface over the fatty streaks, and many of them appeared to enter the intima and participate in the growth of the fatty streaks. Lipid-laden smooth muscle cells appeared in small numbers and formed two to four layers between the macrophages and the internal elastic lamella at 2 to 3 months. During the third month of hypercholesterolemia, endothelial cell continuity over the lipid-laden macrophages became interrupted, exposing the underlying foam cells to circulating blood. Foam cells were then readily observed in whole blood smears, suggesting that many of the lipid-laden macrophages leave the intima and enter the circulation. After 4 months, significant endothelial denudation was found in the iliac artery and many exposed macrophages were covered by adherent platelets in the form of a mural thrombus. Thus, the early components of atherosclerosis induced by chronic hypercholesterolemia centered around the monocyte-macrophage and its interaction with endothelium in the induction of the fatty streak. Subsequent changes that lead to macrophage-smooth muscle interactions, platelet-macrophage interactions, and platelet-endothelial interactions appeared to set the stage for the development of more advanced proliferative lesions.
Studies of hypercholesterolemia in the nonhuman primate. II. Fatty streak conversion to fibrous plaque.
Authors: Faggiotto A , Ross R
Published: June 2011

This report presents the second portion of the morphologic studies on chronic, diet-induced hypercholesterolemia in nonhuman primates (Macaca nemestrina) examined sequentially between 5 and 13 months. A direct relationship was observed between the rate of cholesterol increase, the level and duration of hypercholesterolemia, and the changes in the artery wall that led to the formation of fatty streaks and their conversion to fibrous plaques. A loss of endothelial continuity was first observed in the iliac arteries between 3 and 4 months of atherogenic diet and appears to be a critical step in the conversion of many fatty streaks to fibrous plaques. With breaks in endothelial junctions and exposure of some of the macrophages in a fatty streak, many of the lipid-filled macrophages appeared to detach and enter the circulation. The number of circulating foam cells increased precipitously between 3 and 4 months, the time when increased sites of endothelial dysjunction and macrophage egress were observed. Exposure of subendothelial macrophages also permitted adherence of platelets to these macrophages and to exposed connective tissue. Fibrous plaques were found at similar anatomic sites where endothelial denudation had been observed at earlier time points but were more prevalent in the abdominal aorta and iliac arteries. These changes subsequently occurred at every level of the aortic tree and appeared to progress in a cephalad fashion with increasing rate, level, and duration of hypercholesterolemia. The results of these studies stress the importance of following cholesterol levels of each animal throughout the entire period of the study and of sampling the entire arterial tree at every level with time. This helped us to understand the complicated interrelationships between the various cells in atherogenesis, provided further support for the "Response to Injury Hypothesis of Atherosclerosis," and helped to explain how hypercholesterolemia may be involved in the different stages of atherogenesis in nonhuman primates and possibly in humans.
Kinetics of low density lipoprotein interactions with rabbit aortic wall following balloon catheter deendothelialization.
Authors: Alavi M , Moore S
Published: June 2011

The kinetics of the interaction of circulating lipoproteins with the balloon catheter-injured aortic wall of rabbits were investigated. The fate of radioiodinated low density lipoprotein (LDL) was followed up to 48 hours after intravenous injection and compared with radioiodinated albumin injected simultaneously. The accumulation of LDL was calculated from the radioactivity in the aortic tissue divided by the average specific activity, measured from the area below the plasma radioactivity versus the time curve, and expressed as cpm/ml/hr. It was observed that the aortic area of endothelial regeneration (AER), which is relatively impermeable to Evans blue dye and shows very little albumin concentration, accumulated considerably more LDL than control (uninjured) tissues. Furthermore, removal of labeled lipoprotein from the intima during a 24-hour efflux period following the initial 48-hour loading period indicated that there was greater removal from normal and deendothelialized aortic (DEA) tissues than from neointima covered by endothelium. These findings are consistent with previous observations indicating changes in the morphological and chemical composition of AER. There is evidence that greater neointimal lipid accumulation in response to endothelial injury parallels the concentration of glycosaminoglycans (GAG). The relatively increased accumulation of label in AER, while plasma-specific activity is rapidly decreasing, indicates that there may be an ionic interaction of GAG with LDL in the AER.
Endothelial healing in the rabbit aorta and the effect of risk factors for atherosclerosis. Hypercholesterolemia.
Authors: Walker L N , Bowyer D E
Published: June 2011

The effect of diet-induced moderate hypercholesterolemia on endothelial healing has been investigated in the rabbit following a narrow superficial injury to aortic endothelium without damage to the media of the vessel. The healing process was compared with that observed in normocholesterolemic animals. The degree of platelet involvement was similar in both normo- and hypercholesterolemic animals. Reendothelialization occurred within 48 hours in both groups of animals, showing that hypercholesterolemia did not delay endothelial healing. It was found that esterase-positive cells, which morphologically resembled monocyte-macrophages, adhered to and penetrated regenerated endothelium only in hypercholesterolemic animals. After reendothelialization in normocholesterolemic animals, there was no increase in the number of cells within the intima of the vessel and no evidence of lipid accumulation. In hypercholesterolemic animals, cells accumulated in the intima in areas of regeneration, and lipid accumulation occurred within both the intima and the media in areas of regeneration.
Smooth muscle cells produce an inhibitor of endothelial cell growth.
Authors: Schumacher B L , Grant D , Eisenstein R
Published: June 2011

A potent inhibitor of endothelial cell growth and neovascularization has previously been isolated from both bovine and human aorta. In an attempt to determine if the major cellular component of that tissue, smooth muscle cells, synthesized molecules with this activity, we studied the effects of serum-free conditioned medium from bovine aortic smooth muscle on the growth of cultured endothelial cells from bovine aorta. The smooth muscle conditioned medium was found to contain both growth inhibitory and stimulating activities that could be distinguished by their heat stability. The inhibitor is precipitable by ammonium sulfate, heat stable (80 degrees C), and inactivated by dithiothreitol, trypsin, and 2-mercaptoethanol. It has an estimated molecular weight of approximately 35,000 to 40,000 daltons (by column chromatography). It therefore appears that smooth muscle cells produce an inhibitor of endothelial cell growth that belongs to a class of natural endothelial cell growth inhibitors derived from avascular tissues we tentatively term "avasculins." Proof of its identity with the inhibitor isolated from intact aorta (or other tissues) must await purification.
Platelet activation.
Authors: Zucker M B , Nachmias V T
Published: June 2011

Platelets are discoidal cytoplasmic particles that respond to a variety of stimuli by developing filopodia and rounding up (shape change), developing the ability to bind fibrinogen from the medium, and, with strong stimuli such as thrombin and PAF-acether, secreting the contents of several types of granules. Arachidonic acid is cleaved from phospholipids by phospholipase A2 and converted by the platelets to endoperoxides, and then to thromboxane A2. The bound dimeric fibrinogen molecules probably cause aggregation by forming bridges between platelets. Aggregation is reinforced by secreted fibrinogen and thrombospondin, which binds the platelets, and by thromboxane A2 and endoperoxides, as well as secreted ADP, which cause additional receptor-mediated activation. The responses to these stimuli are initiated when the agonists bind to specific receptors on the plasma membrane. Subsequent steps resemble those in other types of responsive cells: breakdown of phosphatidylinositol bisphosphate into diacylglycerol, a stimulator of protein kinase C, and inositol-1,4,5-trisphosphate, recently shown to be a potent calcium ionophore. The response of shape change results from increased cytoplasmic Ca2+ which permits phosphorylation of one of the light chains of myosin by a calcium-calmodulin-dependent kinase, with resulting enhanced actin-myosin interaction. Secretion is associated with phosphorylation of a 40,000 to 47,000 dalton protein by the diacylglycerol-activated protein kinase C. These recent findings have increased our understanding of the mechanisms of platelet activation, but much remains to be learned. How do agonist-receptor complexes influence PIP2 breakdown? Is this indeed the first step in activation? What mediates adhesion of platelets to the injured blood vessel wall? Does transduction of this stimulus occur by the same mechanism as transduction of commonly used soluble stimuli? What is the role of the phosphorylated 40-47 K protein in secretion? What change in GP IIb-IIIa promotes their ability to bind fibrinogen? What is the role of calcium-activated protease? Of the phosphorylation of actin-binding protein? Progress is being made rapidly, and these questions may be answered within a few years.
Retro-endocytosis of low density lipoprotein by cultured human skin fibroblasts.
Authors: Aulinskas T H , Oram J F , Bierman E L , Coetzee G A , Gevers W , van der Westhuyzen D R
Published: June 2011

A fraction of the low-density lipoprotein (LDL) internalized by cells via receptor-mediated endocytosis follows a short-circuit pathway, termed "retro-endocytosis," that results in the rapid exocytosis of ligand. Results from the current study suggest that retro-endocytosis of LDL in human fibroblasts is caused by resurfacing of endocytotic vesicles that contain both free and receptor-bound ligand, resulting in discharge of vesicular contents and in spontaneous dissociation of LDL from its receptor. The bulk of the released LDL particles had the same size, density, and immunogenic properties as native LDL, indicating that they were discharged intact. Some of the retro-endocytosed LDL was larger than native LDL, and some exhibited altered sedimentation properties. When fusion of endosomes with lysosomes was inhibited by chilling cells to 18 degrees C, the proportion of intracellular LDL subsequently released was unaffected, suggesting that retro-endocytosis does not require lysosomal participation. Furthermore, the shorter the internalization phase the greater was the proportion of LDL subsequently released, suggesting that LDL was discharged from compartments formed early in endocytosis. Retro-endocytosis of LDL was stimulated by agents that neutralize acid intracellular compartments, such as ionophores (monensin) and weak bases (chloroquine and methylamine). Monensin increased the proportion of intracellular LDL released, suggesting that it had a direct effect on retro-endocytosis. The effect of weak bases appeared to be secondary to their ability to promote cellular accumulation of undegraded LDL. Thus, retro-endocytosis of LDL becomes a major pathway when intracellular compartments fail to maintain a low pH or where the intracellular concentration of LDL reaches abnormal levels.
Apolipoprotein B-48 and B-100 very low density lipoproteins. Comparison in dysbetalipoproteinemia (type III) and familial hypertriglyceridemia (type IV).
Authors: Tercé F , Milne R W , Weech P K , Davignon J , Marcel Y L
Published: June 2011

A protein band having the same migration as apolipoprotein (apo) B-48 was observed by SDS electrophoresis in the plasma very low density lipoprotein (VLDL) from 14 Type IV and three Type III hyperlipoproteinemic subjects and from six normal fasting subjects. The VLDL from five Type IV, three Type III, and one normal subject were separated into two subfractions, retained and nonretained, by immunoaffinity chromatography on monoclonal anti-apo B-100 Sepharose. Based on results of electrophoresis and radioimmunoassay, we have concluded that these two fractions represent apo B-48 and apo B-100 lipoproteins that we have named apo B-48 and apo B-100 VLDL. When compared to their respective apo B-100 VLDL, the apo B-48 VLDL from either Type III or Type IV was principally enriched in total lipids, in apo E, and had an electrophoretic migration similar to chylomicrons. This suggests that apo B-48 VLDL has the same origin (i.e., intestinal) in the two disorders. Both apo B-48 and apo B-100 VLDL were enriched in cholesteryl ester (CE) and depleted in triglyceride (TG) in Type III; however, both fractions were rich in TG and poor in CE in Type IV and in normal subjects. In addition, compared to their respective apo B-100 VLDL, the apo B-48 fraction was enriched in CE in Type III and in TG in Type IV. We conclude that, despite a possible similar origin for apo B-48 VLDL in Type III and in Type IV subjects, the composition of apo B-48 VLDL is variable and the CE/TG ratio is more characteristic of the type of hyperlipidemia than of the particular VLDL subfractions.
Molecular biology in arteriosclerosis research.
Authors: Williams D L
Published: June 2011

The topics discussed in this article illustrate how molecular biology will have a dramatic impact on arteriosclerosis research. DNA clones for a small number of relevant proteins have been isolated, and studies are underway in numerous laboratories to extend these initial studies. The techniques of molecular biology will provide major advances in our understanding of numerous proteins directly or indirectly involved in the atherogenic process. Cloning technology will solve the primary structures of many proteins that can not be purified in quantities sufficient for classical methods of analysis. Studies of regulation will benefit from the availability of DNA probes, the ability to generate site-directed antibodies, and the use of reverse genetics to identify nucleic acid sequences involved in the regulation of gene expression. Studies of gene structure and genetic polymorphisms will unravel the genetic basis for defects in lipid and lipoprotein metabolism and should provide valuable reagents for clinical screening and diagnosis. The reverse genetics approach will permit the systematic analysis of structure-function relationships at the protein level in a manner not previously possible. Each of these will contribute to our understanding of the atherogenic process and should provide insight into ways of preventing and treating arteriosclerosis.
Histochemical characterization of low density lipoprotein receptors in internalization-defective familial hypercholesterolemia.
Authors: Takaichi S , Tajima S , Miyake Y , Yamamoto A
Published: June 2011

We studied the localization of low density lipoprotein (LDL) bound to the receptors on the cultured fibroblasts from a patient (MN) with homozygous familial hypercholesterolemia and a defect in internalization of LDL and compared the localization with normal fibroblasts and with those from another internalization-defective cell, GM2408A. Monolayers of cells were cultured with lipoprotein-deficient human serum, and the cells were incubated with 125I- or ferritin-labeled LDL. The LDL binding was observed by autoradiography or by an electron microscope. Autoradiographs of bound 125I-LDL confirmed that MN's cells could not internalize LDL inside the cell at 37 degrees C. In these cells, ferritin LDL was found mainly in noncoated regions at 4 degrees C; it was not found in endocytic vesicles after incubation at 37 degrees C. Ferritin cores that bound on the surface of the cells from the normal subject, from GM2408A, from MN, and from MN's parents, were counted and quantitatively analyzed at 4 degrees C. In normal cells, 62% of the ferritin cores were bound in the coated pits; in MN's cells, only 11% of the ferritin LDL was found in the coated pits; in the GM2408A cells, 12% of the ferritin LDL was found in the coated pits; in the cells from MN's parents, 40% of the ferritin LDL was found on the coated pits. The results indicate that the internalization defect in MN's cells is the same as that in the GM2408A cells; neither can localize the LDL and LDL receptor complex in coated pits.
Antiatherosclerotic effects of nicardipine and nifedipine in cholesterol-fed rabbits.
Authors: Willis A L , Nagel B , Churchill V , Whyte M A , Smith D L , Mahmud I , Puppione D L
Published: June 2011

Dutch-belted rabbits fed a 2% cholesterol diet for 8 weeks developed atherosclerotic lesions that covered 37.2% +/- 3.5% of the aortic luminal surface. In samples of aortic arch, accumulation of cholesterol and triglyceride was also observed. Oral administration of nicardipine or nifedipine at dosages of 40 mg/kg twice daily for 8 weeks reduced plaque area by 49.2% and 58.7%, respectively. Nicardipine and nifedipine reduced cholesterol accumulation in the aortic arch by 74.5% and 69%, respectively. Triglyceride accumulation was totally abolished. Neither drug significantly altered cholesterol concentration of plasma low density lipoprotein or high density lipoprotein, although nicardipine produced a 42% reduction in cholesterol concentration of the d less than 1.006 g/ml fraction. The above results suggest potential therapeutic utility of nicardipine in atherosclerosis.
Lipoprotein levels in genetically selected mice with increased susceptibility to atherosclerosis.
Authors: Breckenridge W C , Roberts A , Kuksis A
Published: June 2011

Male mice of two inbred strains, C57BR/cdJ and CBA/J, were maintained either on an atherogenic diet rich in cholesterol (5%) and saturated fat (30%) or on a control Purina Laboratory Chow diet. After 3 weeks on a 90% atherogenic/10% chow diet, the C57BR/cdJ mice showed a fivefold increase in plasma total cholesterol levels, while the CBA/J strain indicated only a twofold increase. On the atherogenic diet, both groups of animals showed marked increases in a lipoprotein of a broad density spectrum (1.006 greater than d less than 1.063) with prebeta and beta mobility on agarose gel electrophoresis, but the C57BR/cdJ strain showed a greater increase than the CBA/J strain. The very low density lipoprotein (VLDL) contained predominantly lipoprotein of prebeta mobility, while intermediate density lipoprotein (IDL) and low density lipoprotein (LDL) contained lipoprotein of beta-mobility. In both strains, the major lipid component in this lipoprotein was cholesteryl ester and both strains showed large amounts of an apoprotein (apo) with E mobility as defined for known apoproteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis or by isoelectric focusing on polyacrylamide-urea disc gels. The phosphatidylcholine/free cholesterol ratio of the VLDL, IDL, and LDL fractions isolated from the C57BR/cdJ mice on the atherogenic diet (0.76 to 0.95) was noticeably lower than that (1.10 to 1.19) from the CBA/J mice maintained on the same diet. There was a marked decrease in the high density lipoprotein (HDL) fraction in both strains of mice on the atherogenic diet, but a greater reduction in the HDL level of the C57BR/cdJ strain with a 50% decrease in the phosphatidylcholine/free cholesterol ratio.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of chronic smoking on fibrinolysis.
Authors: Allen R A , Kluft C , Brommer E J
Published: June 2011

The aim of the study was to investigate the long-term chronic effects of smoking on the fibrinolytic enzyme system by comparing two groups of healthy male volunteers (aged 30 to 40 years). One group consisted of 15 habitual smokers who consumed 20 or more cigarettes a day; the other consisted of 15 nonsmokers. Fibrinolysis was studied at rest (baseline) and after infusion of 1-desamino-8-D-arginine vasopressin (DDAVP; 0.4 micrograms/kg body weight). Smokers had significantly lower baseline blood fibrinolytic activity as determined by the overall assays: dilute blood clot lysis (p less than or equal to 0.05) and euglobulin-fibrin plate assay (p less than or equal to 0.05). Further analysis showed that these low activities could be attributed to a lower baseline level of extrinsic tissue-type plasminogen activator (t-PA) activity (p less than or equal to 0.05) in the smokers. There were no significant differences between the groups in various fibrinolytic inhibitors or in the intrinsic fibrinolytic activation pathways. The increased levels of t-PA activity and factor VIII R:Ag in response to DDAVP were also reduced in the smokers (p less than or equal to 0.01). The relative increase (ratio of post-DDAVP activity/baseline activity) for these parameters was not significantly different for the two groups. Smokers also had significantly higher levels of the acute phase reactants, alpha 1-antitrypsin (p less than or equal to 0.02) and plasminogen (p less than or equal to 0.02) and C-reactive protein (p less than or equal to 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of plasma proteins on endothelial binding and vesicle loading of anionized ferritin in rabbit aorta.
Authors: Baldwin A L , Chien S
Published: June 2011

Endothelial binding and vesicle loading of anionized ferritin (AF, isoelectric point, 3.8 to 4.2) in plasma and in Tyrode's solution were investigated in the rabbit aorta. After 2 minutes of in situ perfusion, binding to vesicle necks was significantly higher than elsewhere on the membrane and was independent of AF concentration in the range of 0.05 g/ml to 0.2 g/ml for both perfusates. After 30 minutes, no particles were seen in abluminal vesicles and few had reached the basement membrane. Compared to AF in plasma, AF in Tyrode's solution showed lower endothelial binding and greater vesicle loading (p less than 0.005). The average number of particles per loaded vesicle was equal to the average particle density at vesicle necks in Tyrode's solution, but not in plasma, for 0.1 g/ml AF. Gel chromatography and electrophoresis demonstrated no detectable difference between AF particles incubated in plasma or Tyrode's solution. Our experiments indicate that adherence to, or entrapment in, the glycocalyx is an important step for molecules that are transported by vesicles. We hypothesize that plasma proteins adsorb to the endothelial surface and partially shield its net negative charge so that more AF may bind. Plasma proteins also inhibit vesicle loading either sterically or by modifying the electrical potential at the vesicle neck compared to that within the vesicle cavity.
Apolipoprotein E is the determinant that mediates the receptor uptake of beta-very low density lipoproteins by mouse macrophages.
Authors: Innerarity T L , Arnold K S , Weisgraber K H , Mahley R W
Published: June 2011

Beta very low density lipoproteins (beta-VLDL) from cholesterol-fed animals from patients with Type III hyperlipoproteinemia are internalized by a receptor-mediated process in mouse macrophages. Once internalized, the cholesteryl esters of beta-VLDL are hydrolyzed in lysosomes, and the released cholesterol is re-esterified, resulting in a massive accumulation of cholesteryl esters. In the present study, competitive binding experiments demonstrated that canine apo E HDLc (lipoproteins that contain almost exclusively apolipoprotein E) inhibited the receptor-mediated uptake of 125I-beta-VLDL. The incorporation of human apo E into beta-VLDL was also shown to modulate binding. Reductively methylated beta-VLDL (methyl beta-VLDL) were not taken up by macrophages and did not stimulate cholesteryl ester synthesis. When unmodified human apo E-3 was incorporated into the lipoprotein in place of the canine methyl apo E, these hybrid beta-VLDL (methyl beta-VLDL [E-3]) were internalized and degraded and were as effective as native beta-VLDL in stimulating cholesteryl ester synthesis in macrophages. In the reverse experiment, the incorporation of methyl apo E-3 into native canine beta-VLDL (beta-VLDL [methyl E-3]) reduced the binding activity of the beta-VLDL and abolished their ability to stimulate cellular cholesteryl ester synthesis. Canine beta-VLDL into which apo E-2(Arg158----Cys) had been incorporated had less ability to stimulate cholesteryl ester synthesis (20%) than did native beta-VLDL, but they were more active than beta-VLDL [methyl E-2] or beta-VLDL [methyl E-3], which had virtually no activity. These results demonstrate that apo E is the determinant mediating the receptor binding and uptake of beta-VLDL by mouse macrophages.
Cigarette smoke impairs endothelial cell prostacyclin production.
Authors: Reinders J H , Brinkman H J , van Mourik J A , de Groot P G
Published: June 2011

Production of prostacyclin by endothelial cells is considered to be important in rendering the vessel wall nonthrombogenic. Cigarette smoking is an important risk factor in the pathogenesis of atherosclerosis. Here we show that the incubation of cultured human endothelial cells with a cigarette smoke condensate impaired the basal prostacyclin release. Also, the enhanced release of prostacyclin provoked by phorbol myristate acetate was inhibited by cigarette smoke condensate. Furthermore, cigarette smoke condensate impaired the thrombin-induced prostacyclin production. The production of prostacyclin from exogenous arachidonate was not affected by cigarette smoke condensate, indicating that cigarette smoke condensate constituents exert their inhibitory properties on the level of arachidonate mobilization from cellular phospholipids, rather than on cyclooxygenase or prostaglandin synthetase. The effects noted for cigarette smoke condensate could not be attributed to the cigarette smoke constituents nicotine and cadmium. While inhibiting the endothelial cell prostacyclin production significantly, cigarette smoke condensate did not cause cell death or impairment of secretory function, as measured by the release of von Willebrand factor. This in vitro study shows that impairment of an endothelial cell function is related to a risk factor for atherosclerosis.
Local variation in arterial wall permeability to low density lipoprotein in normal rabbit aorta.
Authors: Stemerman M B , Morrel E M , Burke K R , Colton C K , Smith K A , Lees R S
Published: June 2011

Normal rabbits were injected intravenously with horseradish peroxidase (HRP) and 125I-labeled human low density lipoprotein (LDL), and the aortas were perfusion-fixed. Subsequent visualization of HRP in the aortas was produced by reaction of the tissue with diaminobenzidine and hydrogen peroxide. The luminal surface of the aortas showed many small punctate foci of brown reaction product to the HRP, which represented penetration of the HRP into the vessel wall. The foci were scattered over the luminal surface, and most of the focal areas were less than 1 mm in diameter. The concentration of LDL was up to 47 times greater in these focal areas than in surrounding noncolored regions not showing increased permeability to HRP. Small circumscribed foci of heightened permeability to LDL may predispose to the local accumulation of lipid and ultimately to the formation of atherosclerotic plaques.
Cholesteryl ester efflux from extracellular and cellular elements of the arterial wall. Model systems in culture with cholesteryl linoleyl ether.
Authors: Stein O , Halperin G , Stein Y
Published: June 2011

Cholesteryl ester (CE) accretion in human atheroma is a slow process during which lipoproteins infiltrate the arterial extracellular space and then gain entry into the cellular components. The present aim was to simulate this process in model systems in culture to learn whether cholesteryl ester transfer protein (CETP) may promote CE efflux at different stages of atheroma formation. To simulate CE efflux from arterial interstitium, cationized LDL labeled with 3H-cholesteryl linoleyl ether (3H-CLE) was added to fixed aortic smooth muscle cells (SMC) or to extracellular matrix. To study efflux of 3H-CLE taken up by cells via receptor-mediated endocytosis of LDL, the SMC cultures were fixed and permeabilized prior to the determination of CE efflux. The cellular model included macrophages, which had ingested acetylated LDL labeled with 3H-CLE. Efflux of 3H-CLE and 14C-CE was studied during postincubation of the labeled cultures with human lipoprotein deficient serum (LPDS) or partially purified CETP. As controls, we used SMC cultures incubated with albumin. In all systems, a 3- to 12-fold increase of 3H-CLE or 14C-CE efflux was found in the postincubation medium containing human LPDS or partially purified CETP when compared to controls. Permeabilization of the cells with saponin enhanced cellular 3H-CLE and 14C-CE efflux in the presence of human LPDS. The findings indicate that CETP may promote CE efflux from aortic interstitium or disintegrating cells. We propose that CETP may play an important role in aortic CE homeostasis under physiological and pathological conditions.
Development of coronary atherosclerosis in swine with severe hypercholesterolemia. Lack of influence of von Willebrand factor or acute intimal injury.
Authors: Griggs T R , Bauman R W , Reddick R L , Read M S , Koch G G , Lamb M A
Published: June 2011

The development of coronary atherosclerosis in response to acute intimal injury and severe hypercholesterolemia was studied in 18 swine, nine normal and nine with von Willebrand's disease, an inherited disorder affecting platelet-vessel wall interactions. The left anterior descending coronary artery was denuded of endothelium by balloon catheterization, while the circumflex and right coronary arteries served as nonballooned controls. All swine were maintained on a 2% cholesterol diet for 4 months. The extent of atherosclerotic involvement was evaluated from four indices: percent intimal area, percent luminal narrowing, ratio of intimal to medial area, and luminal form. No differences in coronary atherosclerosis were observed between phenotypes in either ballooned or nonballooned vessels, nor were there any differences between ballooned and nonballooned vessels within either phenotype (p greater than 0.05). The major variable affecting coronary atherosclerosis was serum cholesterol. There was a significant positive relationship between serum cholesterol concentration and the extent of intimal lesions (r = 0.62, p = 0.006) that was independent of plasma von Willebrand factor concentration. These findings suggest that severe hypercholesterolemia promotes the development of porcine coronary atherosclerosis through a mechanism(s) that is independent of acute intimal injury or von Willebrand factor-mediated platelet response to injury.