bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of serotonin-receptor blockade on angioplasty-induced vasospasm in an atherosclerotic rabbit model.
Authors: Sigal S L , Gellman J , Sarembock I J , LaVeau P J , Chen Q S , Cabin H S , Ezekowitz M D
Published: June 2011

Vasospasm occurs both in patients and animal models after angioplasty and may be associated with early closure of the dilated vessel. To investigate the mechanism of angioplasty-induced vasospasm, the effect of serotonin-receptor blockade with two serotonin2 (S2) antagonists, LY53857 and sergolexole, was examined in rabbits with focal femoral artery atherosclerosis. In preliminary studies, local infusion of 1-100 micrograms serotonin caused significant femoral artery vasoconstriction (p less than 0.05) in both normal and atherosclerotic rabbits. There was no significant difference in the degree of vasoconstriction induced by equal doses of serotonin in normal and atherosclerotic animals. Infusion of 10 micrograms serotonin produced a 23 +/- 5% decrease in luminal diameter in atherosclerotic femoral arteries. This was blocked by pretreatment with both S2 inhibitors given separately in different animals before serotonin infusion (p less than 0.002). In contrast, LY53857 (sergolexole was not tested) had no significant effect on phenylephrine-induced vasoconstriction, confirming its specificity as an S2-receptor antagonist. Balloon angioplasty of atherosclerotic vessels caused a significant increase in vessel diameter at the angioplasty site (45% increase from baseline diameter, p less than 0.05). This was associated with significant luminal narrowing both proximal (21% reduction from baseline, p less than 0.05) and distal (17% reduction from baseline, p less than 0.03) to the angioplasty site. These proximal and distal changes are most likely due to vasospasm, as there was no histological evidence of thrombus or dissection at these sites to explain the luminal narrowing. Pretreatment of animals with 10 mg LY53857 or 20 mg sergolexole blocked the proximal vasospasm (2.6 +/- 0.4 before versus 2.2 +/- 0.1mm after angioplasty for LY53857, 2.1 +/- 0.4 before versus 2.1 +/- 0.4 mm after angioplasty for sergolexole; p = NS). Treatment with 20 mg LY53857 inhibited both proximal (2.3 +/- 0.1 before versus 2.2 +/- 0.2 mm after angioplasty, p = NS) and distal (1.7 +/- 0.1 before versus 1.6 +/- 0.2 mm after angioplasty, p = NS) vasospasm after angioplasty. Proximal (2.3 +/- 0.5 before versus 2.5 +/- 0.3 mm after) and distal (1.7 +/- 0.2 before versus 1.7 +/- 0.4 mm after) vasospasm was also prevented by pretreatment with 40 mg sergolexole.(ABSTRACT TRUNCATED AT 400 WORDS)
Lipoprotein(a) and apolipoprotein(a) in a New World monkey, the common marmoset (Callithrix jacchus). Association of variable plasma lipoprotein(a) levels with a single apolipoprotein(a) isoform.
Authors: Guo H C , Michel J B , Blouquit Y , Chapman M J
Published: June 2011

In an earlier report (Chapman et al, Biochemistry 1979;18:5096-5108), we suggested that the common marmoset may represent an important model for the study of human plasma lipoprotein metabolism. We now extend the interest of this monkey model to the study of lipoprotein(a) (Lp[a]) and apolipoprotein(a) (apo[a]). Density gradient ultracentrifugal fractionation of marmoset plasma revealed a bimodal distribution of Lp(a), with one peak of concentration occurring in association with very low density lipoproteins (VLDLs) and a second in the density range 1.040-1.080 g/ml. The dense Lp(a) subspecies displayed physicochemical properties (chemical composition, particle size, and electrophoretic mobility) that closely resembled those of its counterpart in humans and baboons but that were distinct from those of low density lipoprotein (LDL). Furthermore, the particle size of marmoset Lp(a) was invariant (31 nm) over the density interval 1.040-1.080 g/ml, whereas that of LDL decreased progressively with an increase in density (approximately 26-25.2 nm). Use of polyclonal and monoclonal antibodies to human apo(a) and of a polyclonal antibody to marmoset Lp(a) allowed immunologic identification of a single apo(a) isoform in the marmoset whose size was similar to that of apo B-100; apo(a) and apo B-100 were associated in Lp(a) particles by a disulfide linkage. The total protein mass of apo-Lp(a) was estimated to be 800,000 or more by electrophoresis in sodium dodecyl sulfate-polyacrylamide-agarose gels. The amino acid compositions of marmoset and human apo(a) resembled each other but were distinct from those of the corresponding forms of apo B-100. Immunologic evidence is provided for a high degree of cross reactivity between apo(a) in marmosets, baboons, and humans, supporting the idea of the existence of a marked degree of structural homology between these proteins. In addition, electroimmunoblotting of marmoset apo(a) and marmoset plasminogen showed that these proteins shared certain epitopes in common, suggesting that marmoset apo(a) may possess kringle-like structural features. Finally, despite possession of a single apo(a) isoform, marmoset Lp(a) levels varied over a 100-fold range (0.5-49 mg/dl plasma). Considered together, our present findings suggest that the common marmoset monkey constitutes a unique model in which to study the regulation of apo(a) gene expression and the posttranslational processing of apo(a), as well as factors that modulate the synthesis, intravascular metabolism, and cellular catabolism of Lp(a).
The mutation causing the common apolipoprotein A-IV polymorphism is a glutamine to histidine substitution of amino acid 360.
Authors: Tenkanen H , Lukka M , Jauhiainen M , Metso J , Baumann M , Peltonen L , Ehnholm C
Published: June 2011

Apolipoprotein (apo) A-IV is a protein involved in the metabolism of chylomicrons and high density lipoproteins. This protein displays genetic polymorphism due to two main codominant alleles, A-IV1 and A-IV2. We have identified the mutation that leads to this polymorphism. It is caused by a single-base substitution of guanine for thymine in the third base of codon 360. This substitution leads to a glutamine to histidine change. Direct sequencing of amplified DNA from eight subjects in a three-generation pedigree has demonstrated that the guanine to thymine substitution can explain the apo A-IV polymorphism. In 32 unrelated individuals, a correspondence between apo A-IV phenotype determined by isoelectric focusing and genotype determined with Fnu4HI digestion of amplified DNA could be demonstrated. The enzyme lecithin:cholesteryl acyltransferase (LCAT) is activated by apo A-IV. Under our in vitro conditions, the isoprotein apo A-IV 1-1 is a better LCAT activator than is the isoprotein apo A-IV 2-2. A knowledge of the molecular mechanism underlying the apo A-IV polymorphism will help to elucidate the mechanisms involved in LCAT activation.
Effects of tumor necrosis factor on prostacyclin production and the barrier function of human endothelial cell monolayers.
Authors: Langeler E G , Fiers W , van Hinsbergh V W
Published: June 2011

The endothelium controls the influx of macromolecules into the tissues, a process that may be disturbed at sites of inflammation and in atherosclerotic plaques. In this article, we report our evaluations of the effects of the inflammatory mediator, tumor necrosis factor-alpha (TNF-alpha), on the production of prostacyclin and the barrier function of human endothelial cell monolayers in an in vitro model. TNF-alpha (500 units/ml) had no direct effect on the passage of sucrose, peroxidase, and low density lipoprotein through monolayers of human aortic endothelial cells. On the other hand, during the first hours after addition 500 units/ml TNF-alpha induced a reduction of the permeability of umbilical artery endothelial cell monolayers. Within 10 minutes TNF-alpha induced an increase in prostacyclin production by primary cultures of umbilical artery endothelial cells. However, the reduction in permeability was not caused by a change in prostacyclin production or by a change in cyclic AMP concentration because 1) the effect of TNF-alpha on permeability was not prevented by aspirin, 2) no change in the cellular cyclic AMP concentration could be observed after addition of TNF-alpha, and 3) TNF-alpha was still able to reduce the passage rate in the presence of 25 microM forskolin. The reduction in permeability was accompanied by a decrease of F-actin in stress fibers. With prolonged incubation with TNF-alpha, the permeability of umbilical artery endothelial cell monolayers increased, and F-actin was found again in stress fibers. However, these effects of TNF-alpha were only significant at high concentrations of TNF-alpha.(ABSTRACT TRUNCATED AT 250 WORDS)
A nifedipine-sensitive smooth muscle cell population is present in the atherosclerotic rabbit aorta.
Authors: Pauletto P , Scannapieco G , Borrione A C , Zanellato A M , Tonello M , Giuriato L , Pessina A C , Palù C D , Sartore S
Published: June 2011

We evaluated the ability of the Ca2+ channel blocker nifedipine to influence the severity of atherosclerotic lesions and the pattern of aortic smooth muscle cell (SMC) differentiation in cholesterol-fed New Zealand White rabbits. The animals were fed a 1% cholesterol-enriched diet for 12 weeks. After 4 weeks of the diet, some rabbits were given nifedipine (20 mg b.i.d.) for another 8 weeks without discontinuation of the cholesterol-enriched diet (experiment 1). Another group of rabbits was treated with nifedipine from the beginning of the cholesterol-enriched diet for the entire 12 weeks (experiment 2). The severity of ahterosclerotic lesions was determined by computerized planimetry, and qualitative effects of nifedipine on SMCs were studied by monoclonal antibodies specific for smooth muscle and nonmuscle myosins. In the aortic media of normal rabbits, these antibodies can identify an SMC population with an "immature" type of myosin pattern; a marked increase in the number of these cells is observed during atherogenesis. In experiment 1, we observed a marked decrease of medial SMCs with the immature type of myosin pattern, without any significant reduction in atherosclerosis severity. In experiment 2, disappearance of the previously mentioned medial SMC population was accompanied by a dramatic slowing of intimal lesion development. These results indicate that nifedipine treatment is effective in reducing atherosclerotic lesions only when given from the beginning of a cholesterol-enriched diet. Delay of nifedipine administration until the fourth week of the cholesterol-enriched diet fails to halt progression of the disease. The observed antiatherosclerotic activity can be attributable to a direct effect of the drug on the medial SMC population, which increases during the course of experimental atherogenesis.
Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase induce reductase accumulation and altered lamellar bodies in rat forestomach keratinocytes.
Authors: Singer I I , Kawka D W , Scott S , Bailey P , Kloss M W , Majka J , MacDonald J S
Published: June 2011

Lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and a potent hypocholesterolemic agent, induces a hyperplastic thickening of the rat forestomach mucosa after oral administration of its active form, a hydroxyacid. We studied the effects of lovastatin on the intracellular accumulation of HMG-CoA reductase immunostaining and the accompanying morphological changes in rat forestomach keratinocytes by immunofluorescence microscopy and transmission electron microscopy (TEM). Administration of lovastatin hydroxyacid induced increases in HMG-CoA reductase levels within forestomach keratinocytes that were dose and time dependent and reversible. The adjacent glandular stomach epithelium did not exhibit induction of reductase. A pharmacologically inactive epimer of lovastatin hydroxyacid did not increase keratinocyte reductase accumulation, and lovastatin lactone induced minimal forestomach reductase. TEM of forestomachs from rats given lovastatin hydroxyacid demonstrated profound alterations in epidermal lamellar bodies (organelles that transport lipids and steroids to the intercellular spaces of the stratum corneum). Treated cells lacked internal lipid lamellae and failed to secrete sheets of lipid material into the intercellular spaces of the stratum corneum. We hypothesize that sustained inhibition of HMG-CoA reductase in rat forestomach keratinocytes induces accumulation of HMG-CoA reductase and hyperplasia by inhibiting sterol synthesis, assembly of lamellar bodies, and formation of intercellular lipid sheets.
Waist to hip ratio in middle-aged women. Associations with behavioral and psychosocial factors and with changes in cardiovascular risk factors.
Authors: Wing R R , Matthews K A , Kuller L H , Meilahn E N , Plantinga P
Published: June 2011

Waist to hip ratio (WHR) was measured in 487 middle-aged women participating in the Healthy Women Study. Upper body fat distribution was found to be associated with numerous behaviors that affect cardiovascular risk, including smoking, low exercise levels, weight gain during adulthood, and higher caloric intake. Moreover, WHR was also associated with higher levels of anger, anxiety, and depression and lower levels of perceived social support. Women with upper body fat obesity had higher systolic blood pressure, total cholesterol, low density lipoprotein cholesterol, triglycerides, and apolipoprotein B and lower levels of high density lipoprotein (HDL) and the HDL subfractions 2 and 3. These associations remained significant after adjusting for body mass index. Among 108 women who had repeat measurements of WHR, changes in WHR over a 3-year period were significantly correlated with changes in activity and with decreases in HDL2. Thus, WHR appears to be an integral component of the cardiovascular risk profile. WHR is related to those behaviors and psychosocial attributes that influence cardiovascular risk.
Aortic intimal thickening and myosin isoform expression in hyperthyroid rabbits.
Authors: Giuriato L , Borrione A C , Zanellato A M , Tonello M , Scatena M , Scannapieco G , Pauletto P , Sartore S
Published: June 2011

The potential effect of thyroid hormones on the expression of cytoskeletal and cytocontractile proteins of vascular smooth muscle cells (SMCs) was examined by a panel of monoclonal antibodies and immunocytochemical procedures. L-Thyroxine was administered to adult New Zealand White rabbits for as long as 26 days, and the aortic SMC composition was studied at days 1, 2, 7, 15, and 26 from the beginning of hormonal treatment. A diffuse intimal thickening of the aorta became visible after 7 days of thyroxine administration. Histological and histochemical examination of intimal tissues from hyperthyroid rabbits revealed the presence of a homogeneous Sudan black-negative cell population. In immunofluorescence tests the intimal cells were found to be negative for antibodies specific for monocyte/macrophage or desmin and homogenously reactive (positive) for antibodies to vimentin and smooth muscle (SM) alpha-actin, thus indicating that cells present in the thickened intima were of the SM type. In addition, intimal SMCs from aortas of hyperthyroid rabbits showed a myosin isoform content similar to that found in normal developing aortic SM and in a specific medial SMC subpopulation of aortas from adult euthyroid animals. In the media underlying the intimal thickening, almost all the SMCs switched their myosin isoform expression toward the "immature" phenotype after 2 days of thyroxine treatment. When the level of thyroid hormones was reduced by propylthiouracil treatment, the medial SMC subpopulation with the immature myosin isoform content present in euthyroid rabbits completely disappeared. The study of DNA synthesis-related bromodeoxyuridine incorporation in aortas from hyperthyroid rabbits showed the presence of labeled nuclei in medial SMCs before the appearance of the intimal thickening as well as in the thickened intima and in the underlying media at days 7 and 15. These results are consistent with a specific role for thyroid hormones in inducing proliferation/migration of medial SMCs into the intima. Moreover, the switch in the expression of myosin isoforms induced by thyroid hormones appears to precede the accumulation of medial SMCs in the intima.
Relation of fasting plasma insulin concentration to high density lipoprotein cholesterol and triglyceride concentrations in men.
Authors: Laws A , King A C , Haskell W L , Reaven G M
Published: June 2011

Low plasma high density lipoprotein (HDL) cholesterol concentration is a risk factor for coronary heart disease (CHD) and is frequently associated with high triglyceride concentration. Both of these abnormalities have been related to insulin resistance as estimated by plasma insulin concentrations and to measures of obesity, regional adiposity, and physical fitness. To determine which of these variables (fasting plasma insulin, obesity as measured by body mass index [BMI], or regional adiposity as measured by waist to hip ratio [WHR]) best identifies men with low HDL cholesterol and high triglyceride concentrations, we divided 83 men, aged 50-65 years, who were free of CHD or diabetes, into tertiles based on BMI, WHR, or fasting plasma insulin concentration. Only for plasma insulin tertiles were there statistically significant differences in HDL cholesterol (tertile 1, mean +/- SEM, 1.34 +/- 0.08 mmol/l; 2, 1.16 +/- 0.05 mmol/l; 3, 1.10 +/- 0.06 mmol/l; p less than 0.03) and triglyceride (tertile 1, 1.05 +/- 0.08 mmol/l; 2, 1.48 +/- 0.12 mmol/l; 3, 1.82 +/- 0.17 mmol/l; p less than 0.005) concentrations. In forward stepwise regressions with HDL cholesterol and triglyceride as dependent variables, fasting insulin concentration but not BMI, WHR, or maximal oxygen uptake (VO2max), a measure of physical fitness, predicted HDL cholesterol (R2 = 0.07, p less than 0.02) and triglyceride (R2 = 0.20, p less than 0.001) concentrations. The data suggest that plasma insulin concentration is an important predictor of HDL cholesterol and triglyceride concentrations independent of BMI, WHR, or VO2max.
Collagen synthesis in cultured aortic smooth muscle cells. Modulation by collagen lattice culture, transforming growth factor-beta 1, and epidermal growth factor.
Authors: Schlumberger W , Thie M , Rauterberg J , Robenek H
Published: June 2011

We investigated the effects of transforming growth factor-beta 1 (TGF-beta 1) and epidermal growth factor (EGF) on the protein synthesis and production of collagen in cultured smooth muscle cells (SMCs) from the aortic media of pigs. SMCs were cultured as monolayers on plastic as well as in three-dimensional collagen lattices to gain some information about the influence of a preexisting collagenous matrix on the growth factor-induced effects. A 48-hour exposure of SMCs to TGF-beta 1 at concentrations of 5 ng/ml in the presence of 1% serum caused a marked enhancement of the production of collagen and noncollagen proteins. The rate of net collagen production by SMCs exposed to TGF-beta 1 was approximately threefold higher than that of control cells. Moreover, TGF-beta 1 specifically stimulated collagen synthesis, resulting in a greater proportion of collagen in total proteins synthesized compared with controls. The preexisting matrix of collagen lattices affects the response of SMCs to TGF-beta 1 and EGF. In monolayer cultures the collagen proportion increased twofold under the influence of TGF-beta 1, whereas in collagen lattices the specific stimulation of collagen synthesis was lower. We found that EGF enhanced TGF-beta 1-induced protein production in collagen lattices but not in monolayer cultures. In addition, the protein production by SMCs was influenced differently by EGF in these culture systems. Taken together, these data show a mutual influence of growth factors and extracellular matrix components on collagen production in SMCs, thus indicating that TGF-beta 1 may be an important pathophysiological regulator of collagen metabolism in the vessel wall.
The endothelium of advanced arteriosclerotic plaques in humans.
Authors: Bürrig K F
Published: June 2011

The functional morphology of the endothelial cells (ECs) covering advanced but uncomplicated sclerotic plaques in humans was studied in carotid endarterectomy specimens and in coronary arteries from hearts explanted because of advanced ischemic heart disease. The endothelial layer was nearly always intact, and the endothelial patterns reflected the anticipated local flow patterns along the narrowed arteries, with the majority of flow irregularities downstream from the stenosis. Large (giant) ECs (defined as ECs with a surface area of greater than or equal to 800 microns 2) were frequently found on the plaque surface, probably indicating accelerated EC senescence attributable to sustained nondenuding injury in the region of disturbed flow. Ultrastructurally, activation of ECs with hyperplasia of organelles was frequent. In addition, as a sign of immunological activation, about 5% of ECs express class II antigens (HLA-DR and rarely focal HLA-DQ), as demonstrated by double immunofluorescence with von Willebrand factor to identify the ECs. EC activation may be responsible for adherence to the intact luminal surface by activated platelets and monocytes, which were always present (in contrast with nonsclerotic artery segments). Furthermore, an increase in myo-endothelial contacts to subendothelial modified smooth muscle cells was a regular feature of the sclerotic lesions; this feature represents an unknown process of EC and smooth muscle cell interaction in the sclerotic lesion and may be a compensatory process for EC control of smooth muscle cell proliferation. In advanced plaques the ECs are altered without denudation but with changed properties, which may contribute to plaque growth and which are consistent with the postulated EC dysfunction in the pathogenesis of arteriosclerotic lesions.
Reduced proteoglycan binding of low density lipoproteins from monkeys (Macaca fascicularis) fed a fish oil versus lard diet.
Authors: Edwards I J , Gebre A K , Wagner W D , Parks J S
Published: June 2011

We examined the effect of isocaloric substitution of dietary fish oil for lard on the properties of low density lipoproteins (LDL) important in binding to arterial proteoglycans (PG). Cynomolgus monkeys (n = 10) were fed atherogenic diets enriched in fish oil or lard in a crossover study consisting of two 15-week phases of atherogenic diet separated by a 6-week monkey chow "wash-out period." LDL were isolated from plasma during each dietary phase, characterized for chemical and physical properties, and assessed for their ability to interact in vitro with arterial PG. Plasma LDL cholesterol was similar during fish oil and lard consumption (356 +/- 34 and 331 +/- 17 mg/dl, mean +/- SEM), but during fish-oil feeding relative to that of lard, LDL size was smaller (4.2 +/- 0.1 versus 4.9 +/- 0.1 g/mumol) and LDL particles differed in chemical composition. When animals were fed fish oil, significantly fewer (p less than 0.05) LDL particles bound to PG in both dietary phases: 1.00 +/- 0.27 (x10(12)) versus 5.31 +/- 0.83 (x10(12)) particles/micrograms PG in phase 1 and 3.56 +/- 0.67 (x10(12)) versus 6.00 +/- 0.52 (x10(12)) in phase 2 for LDL from animals fed fish oil and lard, respectively. These studies indicate that dietary fat-induced changes in LDL particles lead to altered in vitro interactions with artery wall PG and suggest a novel mechanism for the protective effect of fish oil against atherosclerosis.
Relation of extent of extracranial carotid artery atherosclerosis as measured by B-mode ultrasound to the extent of coronary atherosclerosis.
Authors: Wofford J L , Kahl F R , Howard G R , McKinney W M , Toole J F , Crouse J R
Published: June 2011

The extent of carotid artery atherosclerosis as measured by B-mode ultrasound has been shown to be strongly and independently correlated with the presence or absence of coronary atherosclerotic disease (CAD), but no studies to date have used carotid B-mode ultrasound to compare the extent of atherosclerotic disease in the two arterial circulations. We used data from a registry of patients undergoing cardiac catheterization and B-mode ultrasound of the carotid arteries to compare the extent of CAD (number of major coronary vessels with 50% or greater stenosis as judged by a consensus interpretation) with the extent of extracranial carotid atherosclerosis. Four hundred thirty-four patients (234 men, 200 women) greater than 40 years of age were stratified by gender and then divided into quartiles on the basis of a B-mode score that was derived by summing arterial wall thickness at nine sites in the left and nine sites in the right carotid arteries. Evaluation of extent of CAD for the four B-mode quartiles showed that men in the lowest B-mode quartile were over six times more likely to have normal coronary arteries than three- to four-vessel CAD, while men in the highest B-mode quartile were over 10 times more likely to have three- to four-vessel CAD than normal coronary arteries. The findings were similar for women but not as dramatic. Gender-specific discriminant function models using traditional risk factors alone or in combination with B-mode score were developed to predict the extent of CAD.(ABSTRACT TRUNCATED AT 250 WORDS)
Stimulating effect of oxidized low density lipoproteins on plasminogen activator inhibitor-1 synthesis by endothelial cells.
Authors: Latron Y , Chautan M , Anfosso F , Alessi M C , Nalbone G , Lafont H , Juhan-Vague I
Published: June 2011

Oxidized low density lipoproteins (ox-LDL) are thought to accelerate atherogenesis. It was recently demonstrated that patients with coronary heart disease have defects in plasma fibrinolysis due to increased plasminogen activator inhibitor-1 (PAI-1) levels. Investigation of PAI-1 synthesis by endothelial cells may allow insight into the effect of native LDL (N-LDL) and ox-LDL on endothelial cells. In the present study, secretion of PAI-1 by human umbilical vein endothelial cells (HUVEC) in culture was evaluated after incubation with N-LDL and ox-LDL. Ox-LDL were obtained by peroxidation under ultraviolet radiation, which induced compositional changes in LDL, namely, a decrease in the levels of arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, and alpha-tocopherol and an increase in the malondialdehyde content. Ox-LDL induced a dose-dependent increase in PAI-1 secretion by HUVEC as assayed by an enzyme-linked immunosorbent assay. After a 24-hour incubation, a twofold increase in the PAI-1 content was observed with 50 micrograms/ml ox-LDL protein. Studies with inhibitors of protein synthesis and metabolic labeling with [35S]methionine confirmed that PAI-1 synthesis was stimulated by ox-LDL. N-LDL had no detectable effect on PAI-1 secretion. Binding studies with radiolabeled lipoproteins showed that the effect of ox-LDL was independent of the B/E receptor. Our experiments indicate that ox-LDL stimulate PAI-1 secretion from HUVEC and that this effect may involve a scavenger receptor.
Hypertriglyceridemia and regulation of fibrinolytic activity.
Authors: Mussoni L , Mannucci L , Sirtori M , Camera M , Maderna P , Sironi L , Tremoli E
Published: June 2011

A relation between elevated triglyceride (TG) levels and alterations of the fibrinolytic system has been recognized in studies of patients with coronary heart disease. In this investigation, the total fibrinolytic activity and the levels of specific components of the fibrinolytic system were evaluated in plasma samples from a highly selected group of patients with type IV hyperlipoproteinemia before and after a dietary treatment aimed at reducing TG levels. The fibrinolytic response of type IV patients was comparable to that of normolipidemic subjects, whereas tissue-type plasminogen activator antigen levels before and after venous occlusion (p less than 0.01) and resting plasminogen activator inhibitor-1 (PAI-1) antigen (p less than 0.01) and activity (p less than 0.01) were significantly higher in hypertriglyceridemic subjects compared with controls. After dietary treatment, a 22% reduction in TG levels was attained in type IV patients, with no appreciable modification of fibrinolytic parameters. The analysis of the single-patient data revealed a tendency toward normalization of PAI-1 levels only in those patients who showed a TG reduction greater than or equal to 20%. Very low density lipoproteins (VLDLs) from both normal and type IV patients concentration-dependently stimulated PAI-1 release by endothelial cells and HepG2 cells, with the effect of VLDL from type IV patients being more pronounced on HepG2 cells. The release of PAI-1 induced by VLDL in competent cells may thus account for the elevated levels of this antifibrinolytic protein that occur in hypertriglyceridemic patients.
Role of apolipoprotein E on cholesteryl ester-enriched low density lipoprotein particles in coronary artery atherosclerosis of hypercholesterolemic nonhuman primates.
Authors: Stevenson S C , Sawyer J K , Rudel L L
Published: June 2011

Significant differences among individuals occur in the lipoprotein response to atherogenic diets in cynomolgus and African green monkeys. The range of concentrations of total plasma cholesterol (TPC) was 100-600 mg/dl and of apolipoprotein (apo) E (quantified by enzyme-linked immunosorbent assay) was 3-20 mg/dl in the animal groups of this study. The correlation between the concentrations of TPC and of apo E was r = 0.89 in these animals. To determine which lipoprotein classes contained the majority of apo E, agarose gel-filtration chromatography was used to subfractionate whole plasma. In hypercholesterolemic monkeys, the majority of the apo E and apo B-100 coeluted within the region of low density lipoprotein (LDL). In normocholesterolemic monkeys, the majority of apo E coeluted with apo A-I and high density lipoproteins. A strong positive correlation was seen between the concentrations of plasma apo E and LDL cholesterol (r = 0.9), but there was no significant correlation between high density lipoprotein apo E and either TPC or plasma apo E concentrations. A positive correlation of r = 0.8 was found between the LDL apo E to apo B-100 molar ratio and the average LDL particle size, suggesting an increase in the number of apo E molecules on the larger LDL particles. Within individual animals, LDL were heterogeneous and the LDL subfractions of lower density (1.02 less than d less than 1.03 g/ml) had the highest proportion of apo E, although apo E was present on LDL of all densities. A strong positive correlation between plasma apo E concentration and coronary artery atherosclerosis was identified, and in stepwise regression analysis, plasma apo B concentration and the apo E to apo B molar ratio of LDL together accounted for more than 90% of the variation in the atherosclerosis end point of coronary artery intimal area. These data strongly suggest that the enrichment of LDL with cholesteryl esters and apo E, which occurs in hypercholesterolemic primates, is an atherogenic feature of the plasma lipoproteins.
Regulation of the urokinase-type plasminogen activator receptor on vascular smooth muscle cells is under the control of thrombin and other mitogens.
Authors: Reuning U , Bang N U
Published: June 2011

The urokinase-type plasminogen activator receptor (u-PAR) was demonstrated on cultured smooth muscle cells (SMCs) of bovine aorta. Binding of 125I-urokinase-type plasminogen activator (u-PA) was concentration dependent and saturable within 45-60 minutes. A similar concentration and time dependence was found in functional plasminogen activation studies. Human two-chain high-molecular-weight u-PA and its proenzyme (pro-u-PA) bound specifically with identical affinity (Kd). Activation of pro-u-PA was strongly accelerated on binding to SMCs and occurred only in the presence of plasminogen on the cell surface. A 100-fold molar excess of unlabeled high-molecular-weight u-PA effectively blocked binding of the radiolabeled ligands; tissue-type plasminogen activator, plasminogen, low-molecular-weight u-PA, and unrelated proteins did not. 125I-u-PA binding was abolished by a monoclonal antibody against the specific u-PA sequence responsible for u-PAR binding. Binding of u-PA sharply decreased on SMC exposure to phosphatidylinositol-specific phospholipase C, confirming the glycan phospholipid cell anchorage of u-PAR. Bovine and human alpha-thrombin (240 nM) increased the binding of 125I-u-PA fivefold, translating into an increase in the number of sites per cell from about 10(5) to 5 x 10(5) without significant change in the Kd (1.29 +/- 0.39 nM). Active site blockade of thrombin by D-Phe-Pro-Arg-chloromethyl ketone resulted in the total loss of stimulatory activity, as did the use of the inactive active site thrombin mutant, S205A. Hirugen (100 microM), which blocks the anion-binding exosite of thrombin, blocked u-PAR stimulating activity. Thus, both the catalytic activity and integrity of the exosite are important for thrombin's stimulatory activity. Other SMC mitogens (epidermal growth factor, transforming growth factor-beta 1, basic fibroblast growth factor, platelet-derived growth factor, and phorbol 12-myristate 13-acetate) increased u-PAR expression on SMCs six- to 20-fold while concomitantly increasing Kd four- to 10-fold. In all cases the induction of u-PAR was dependent on de novo protein synthesis. These observations assign a possible role for thrombin and other mitogens in u-PAR regulation, thereby influencing the pericellular proteolysis that is important in SMC migration and atheromatous plaque development.
Delayed clearance of postprandial chylomicrons and their remnants in the hypoalphalipoproteinemia and mild hypertriglyceridemia syndrome.
Authors: Ooi T C , Simo I E , Yakichuk J A
Published: June 2011

Hypoalphalipoproteinemia (HPAL) with mild hypertriglyceridemia (HTG) is associated with increased coronary artery disease (CAD) risk. The aim of this study was to examine the metabolism of postprandial lipoproteins in HPAL/HTG subjects (n = 21). They had a fasting plasma high density lipoprotein (HDL) cholesterol level < 0.9 mmol/l, a triglycerides (TG) level of 2.0-7.1 mmol/l, and a normal low density lipoprotein (LDL) cholesterol level (< 3.7 mmol/l). They were either homozygous for apoprotein E3 (n = 13) or heterozygous for apoprotein E4 (n = 5) or E2 (n = 3). After ingestion of a vitamin A fat load, plasma and chylomicron (CM) retinyl palmitate (RP) response (areas under curves) was three times and non-CM RP response 2.5 times greater than in normolipidemic control subjects (n = 13). There was close correlation between fasting plasma TG level and postprandial RP response in HPAL/HTG subjects (plasma, r = 0.87; CM, r = 0.89; and non-CM, r = 0.84). In control subjects this correlation was present for plasma RP (r = 0.80) and CM RP (r = 0.61) but not for non-CM RP (r = 0.53). In contrast, postprandial RP response was not correlated with fasting plasma HDL cholesterol levels for both groups. There was also no correlation between fasting TG and fasting HDL cholesterol. Postheparin lipoprotein lipase and hepatic lipase activities were slightly higher in HPAL/HTG subjects. The pattern of postprandial change in HDL composition was similar to that in control subjects. These data indicate enhanced postprandial lipemia in the HPAL/HTG syndrome, and this may account for their increased CAD risk.(ABSTRACT TRUNCATED AT 250 WORDS)
Long-term nicotine exposure increases aortic endothelial cell death and enhances transendothelial macromolecular transport in rats.
Authors: Lin S J , Hong C Y , Chang M S , Chiang B N , Chien S
Published: June 2011

Repeated endothelial cell injury has been suggested as an initiating factor in atherogenesis. Dying or dead endothelial cells have been shown to make significant contributions to the local enhancement of transendothelial macromolecular transport. Since cigarette smoking is one of the major risk factors for atherosclerosis, we examined the hypothesis that smoking accelerates atherogenesis by increasing the frequency of endothelial cell death and hence transendothelial macromolecular transport. Sixteen male Sprague-Dawley rats were given nicotine at a weight-adjusted dose of 5 mg/kg body wt per day in their drinking water over a period of 6 weeks. A group of 16 age-matched male Sprague-Dawley rats not exposed to nicotine and maintained over the same time period served as the control group. In en face preparations of thoracic aorta, immunoglobulin G-containing dying or dead endothelial cells were identified by the indirect immunoperoxidase method, and endothelial leakage to Evans blue-albumin (EBA) complexes (5 minutes after intravenous injection) was visualized by fluorescence microscopy. The results showed that in nicotine-treated rats, 51% of dead endothelial cells were associated with EBA leakage, which was responsible for 57% of total EBA leaky foci. Both the frequency of endothelial cell death (0.94 +/- 0.11% versus 0.40 +/- 0.04%, p < 0.0001 by two-tailed, unpaired Student's t test) and the number density of EBA leaky foci (6.45 +/- 1.23/mm2 versus 3.30 +/- 0.49/mm2, p < 0.05 by two-tailed, unpaired t test) were significantly greater in nicotine-treated rats than in control rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Etoposide treatment suppresses atherosclerotic plaque development in cholesterol-fed rabbits.
Authors: de la Llera-Moya M , Rothblat G H , Glick J M , England J M
Published: June 2011

To study the mechanisms by which monocytes/macrophages and smooth muscle cells contribute to atherosclerotic lesions, we studied atherosclerotic plaque formation in cholesterol-fed rabbits treated with etoposide, a drug that has been shown to have several effects that could interfere with the proposed interactions between these two cell types (M.W. Aarnoudes et al, Virchows Arch B 1984;47:211-216 and M. Rozencweig et al, Cancer 1977;40:334-342). Our results show that long-term etoposide treatment of New Zealand White rabbits maintained on a high-cholesterol diet decreases the extent of fatty streak formation in the aortic intima. Moreover, the plaques formed in the presence of etoposide are thinner and at least focally have less fibrous tissue and fewer smooth muscle cell-derived foam cells than do plaques in control rabbits. These effects are independent of the extent of the diet-induced hyperlipemia or an effect of etoposide on blood cell count and may be related to the inhibition of intimal cell proliferation by etoposide.
Evidence for an altered lipid metabolic state in circulating blood monocytes under conditions of hyperlipemia in swine and its implications in arterial lipid metabolism.
Authors: Bell F P , Gerrity R G
Published: June 2011

Circulating blood monocytes were isolated from normal and hypercholesterolemic swine, and the monocyte lipid compositions and lipid biosynthesis profiles were assessed. The data indicate that monocytes freshly isolated from hyperlipemic swine have increased phospholipid and cholesterol contents and have increased biosynthetic capability for synthesizing phospholipids, triglycerides, and cholesteryl esters, but not cholesterol. The profile of the stimulated lipid synthesis capability is similar to that of the swine aortic intima undergoing atherogenic change. These studies indicate that circulating blood monocytes in hyperlipemic swine, which are known to give rise to intimal foam cells in the early fatty streak lesion, can contribute to altered vessel lipid metabolism without a requirement for in situ modification by wall factors.
Apolipoprotein (a) alleles determine lipoprotein (a) particle density and concentration in plasma.
Authors: Kraft H G , Sandholzer C , Menzel H J , Utermann G
Published: June 2011

The distribution of Lp(a) lipoprotein (Lp[a]) and genetic apolipoprotein(a) (apo[a]) isoforms in plasma samples from 29 healthy normolipidemic subjects of known apo(a) phenotype was evaluated by density gradient ultracentrifugation. The density of Lp(a) was directly related to the size of the apo(a) isoform, ranging from 1.043 g/ml for the LpF phenotype to 1.114 g/ml for the LpS4 phenotype. Heterozygotes had two distinct Lp(a) particles, each containing one of the respective isoforms in plasma. In each heterozygote, the concentration of the lighter Lp(a) species was higher than that of the denser Lp(a) population. These data suggest that apo(a) alleles determine the density and the metabolism and thereby also the concentrations of Lp(a) particles in plasma.
Low density lipoproteins inhibit endotoxin activation of monocytes.
Authors: Weinstock C , Ullrich H , Hohe R , Berg A , Baumstark M W , Frey I , Northoff H , Flegel W A
Published: June 2011

Human serum and low density lipoproteins (LDLs) were shown to inactivate endotoxin (lipopolysaccharide [LPS]) by testing the effect of LPS interactions with serum or LDL on the activation of human monocytes. Sera and LDL preparations from four patients with familial hypercholesterolemia were used to demonstrate the inhibition of LPS from inducing interleukin-1 release. Before LDL removal by immunoapheresis, the patients' sera were able to inactive approximately fivefold more LPS than after LDL removal. The LPS-inactivating capacity lost during apheresis could essentially be retrieved in the LDL-rich eluate from the immunoadsorption columns. Because patients were treated frequently with immunoapheresis, their LDL levels before LDL removal were not markedly elevated. These patients' sera before LDL removal were shown to inactivate amounts of LPS comparable to those inactivated by the sera from three healthy volunteers. LDL prepared by ultracentrifugation showed similar LPS inactivation as LDL prepared by immunoapheresis. We conclude that the inhibition of LPS-induced monocyte activation by human serum is dependent to a large extent on the LDL fraction. LDLs were demonstrated to inhibit LPS from inducing interleukin-1 release by human monocytes.
Effect of various blood-derived and semisynthetic nutrient media on in vitro uptake of 125I-albumin across the intact porcine aortic endothelial surface in an organ-support system.
Authors: Fry D L , Pap J M
Published: June 2011

The effects on porcine arterial structure and permeability of a 4-hour in vitro incubation at 37 degrees C in eight different blood-derived and synthetic nutrient media were examined. Changes in arterial permeability were inferred from the normalized, 1-hour, pressurized, transendothelial uptake (M/c0 cm) of porcine 125I-albumin in 60 porcine aortic tissue preparations using an organ-support system. The organ-support system provided experimental control of ambient gas composition, temperature, transmural pressure, flow (stirring), and nutrient media at a number of sites along the vessel. Light and electron microscopic (scanning and transmission) structural correlations with the observed permeability changes were examined. The M/c0 from the autogenous serum (AS) medium was used as the "control" measurement in each vessel preparation. (Grand mean M/c0 for AS from all studies was 0.312 +/- 0.011 [x10(-3)] [mean +/- SEM] cm, n = 60.) For brevity, M/c0 values from the other media are expressed below as a percentage of the corresponding paired M/c0 from the AS. Uptake from heparinized autogenous blood was 113 +/- 9% of that from AS (p = 0.119); from heparinized autogenous plasma was 135 +/- 10% (p = 0.048); from AS+heparin was 97 +/- 5% (p = 0.498); from pooled porcine serum was 113 +/- 9% (p = 0.037); from a synthetic medium was 131 +/- 8% (p = 0.004); and from a physiological hetastarch solution was 532 +/- 8% (p = 0.0002). Associated light microscopic structural changes and ultrastructural changes were not found. We conclude that 1) incubation with AS and heparinized blood (both of which are autogenous blood substances containing platelet products or platelets) provided the best support for the endothelial barrier function, whereas heparin plasma, pooled serum, a synthetic medium, and particularly hetastarch provided poorer support; 2) arterial permeability can increase significantly without discernible endothelial ultrastructural changes; and 3) AS and to a lesser extent heparin blood should provide a suitable nutrient medium for short-term (less than 4-hour) metabolic support of the endothelial surface and subjacent tissues.
Effect of induced hypercholesterolemia in rabbits on functional responses of isolated large proximal and small distal coronary arteries.
Authors: Simonsen U , Prieto D , Mulvany M J , Ehrnrooth E , Korsgaard N , Nyborg N C
Published: June 2011

We studied the effects of hypercholesterolemia on the vascular responses of proximal and distal parts of the rabbit coronary circulation in two consecutive studies. For 12 weeks, New Zealand White rabbits were fed a control diet or a diet with 1% cholesterol dissolved in either 3% coconut oil (study A) or ether (study B). Isolated proximal epicardial and distal intramyocardial coronary arteries from control and hypercholesterolemic rabbits were mounted for isometric tension recording in a double myograph. In study A for hypercholesterolemic rabbits (n = 12), the maximal relaxation and sensitivity to acetylcholine (ACh) were significantly decreased in proximal coronary segments contracted with 30 mmol/l potassium solution compared with segments from control rabbits (n = 13). The only change observed in distal coronary segments was a slight decrease in relaxation in response to low ACh concentrations (10(-8) and 3 x 10(-8) mol/l). However, in study B for proximal coronary and distal coronary segments from hypercholesterolemic rabbits (n = 13), the area under the ACh relaxation curve was increased compared with that of control rabbits (n = 12). Other parameters that were similarly affected in studies A and B include the following: 1) proximal coronary segments from hypercholesterolemic rabbits were more sensitive to sodium nitroprusside (SNP) than were those from control rabbits, but this was not true for distal coronary segments; 2) endothelial removal from arterial segments of control rabbits induced a significant increase in sensitivity and maximal relaxation to SNP of proximal coronary and distal coronary arteries; 3) in segments from hypercholesterolemic rabbits, the absence of endothelium did not alter the response of proximal coronary segments to SNP but did augment the relaxation of distal coronary segments to SNP; 4) the maximal response to 5-hydroxytryptamine in proximal coronary arteries from hypercholesterolemic rabbits was increased compared with those from control rabbits, whereas such changes were not observed in distal coronary arteries; and 5) histological examination showed the presence of atheromatous plaques in proximal coronary but not in distal coronary segments from treated animals. In conclusion, the present investigation demonstrates that induced hypercholesterolemia alters both the structure and function of proximal parts of the coronary circulation. In distal coronary arteries of hypercholesterolemic rabbits, the only change observed was an impaired endothelium-dependent cholinergic relaxation, but even this change appeared to be dependent on the manner in which cholesterol was added to the diet, although parallel studies are required to confirm this.