Atherosclerosis - Articles in Press

Response to: Animal models of vascular calcification (by O. Ivanovski and G. Nikolov) - Corrected Proof

Jacques Genest, on behalf of the Authors — 2012-05-14

We thank Dr. Ivanovski and Dr. Nikolov for their kind comments on our manuscript entitled: The LDL-R deficient mouse as a model for aortic calcification and quantification by micro-computed tomography. We agree fully that medial calcifications (Monkenberg's mecial sclerosis) and sub-intimal calcifications – as observed in our study differ in risk factors, causality and pathogenesis. Current imaging studies in man makes it difficult to differentiate. However, the clinical context, as mentioned by Dr. Ivanovski and Dr. Nikolov differs significantly. It is also likely that while both forms of aortic calcifications carry a poor long-term prognosis, preventing the calcification process has, to date, not been associated with changes in measurable outcomes.

Regulatory T cells and IL-10 levels are reduced in patients with vulnerable coronary plaques - Corrected Proof

2012-05-10

Highlights: ► Patients with stable atherosclerotic plaques have a higher level of Tregs and IL-10 than vulnerable patients. ► We found no difference in other chemokines studied between the two patient groups. ► We found no difference in EPCs, antibodies to β2GPI, antibodies to ox-LDL, adiponectin or resistin between the two groups.Abstract: Background: Despite having a similar large extent of atherosclerotic coronary affliction, some patients suffer of recurrent cardiac events, whereas others remain asymptomatic.Hypothesis: We hypothesized the existence of a systemic “signature” that could distinguish “vulnerable” patients with preexisting coronary atherosclerosis from those having similar risk factors and atheromatous burden, but no history of clinically evident plaque rupture/erosion.Methods: Twenty three patients who had at least two prior myocardial infarctions (“vulnerable group”) were matched in respect to their background and coronary atherosclerosis extent with twenty one patients without a history of previous myocardial infarction who underwent routine coronary angiography before valvular surgery. We studied a panel of cytokines, antibodies and hormones including IL-6, IL-10, IL-12, antibodies to β2 glycoprotein I (β2GPI), antibodies to oxidized-LDL, adiponectin and resistin, along with levels of circulating EPCs and Tregs.Results: A significantly higher level of Treg cells was present in the control (73.4%±4) than in the “vulnerable patient” group (62.2%±10.7), p<0.001. IL-10 level was also significantly higher in the control than in the vulnerable patients (2.6±1.2pg/ml versus 0.9±0.1pg/ml respectively, p=0.03). There was no significant difference in the circulating levels of the other cytokines, hormones or EPCs between the two groups.Conclusion: Regulatory T cells and serum IL-10 may discriminate “vulnerable” versus stable patients and may have a protective role against plaque rupture in patients with coronary atherosclerosis.

YKL-40 is elevated in patients with peripheral arterial disease and diabetes or pre-diabetes - Corrected Proof

2012-05-09

Highlights: ► The role of YKL-40 in atherosclerosis was investigated in a cohort study. ► YKL-40 was higher in patients with peripheral arterial disease and/or diabetes. ► YKL-40 is associated with age, metabolism, liver function and inflammation. ► YKL-40 is associated with albuminuria and mediasclerosis. ► Thus, YKL-40 may be involved in the specific diabetic form of atherosclerosis.Abstract: Objective: YKL-40 is secreted by macrophages in atherosclerotic lesions and involved in plaque rupture. YKL-40 is elevated in coronary artery disease, and predicts cardiovascular mortality. Experimental in vivo and in vitro data suggest a role of YKL-40 in tissue remodeling. A disease modulating potency of YKL-40 was not investigated in peripheral arterial disease (PAD).Methods: We measured YKL-40 in 460 subjects: 316 PAD: 71 normal glucose metabolism (PAD-NGM), 90 pre-diabetes (PAD-PREDM) and 155 diabetes (PAD-DM); 20 diabetes with atherosclerosis but without PAD (AS-DM); 85 diabetes without macro-vascular complications (DM) and 39 healthy controls (CO).Results: YKL-40 is higher in PAD vs. CO (median [25–75 percentile]: 103 [69–159] vs. 43 [30–80]ng/ml; p<0.001). In addition, YKL-40 is elevated in DM (p<0.001), PAD-NGM (p=0.001), PAD-PREDM (p<0.001), PAD-DM (p<0.001) and AS-DM (p=0.002) compared to CO. Among PAD, YKL-40 is increased in PAD-PREDM (p=0.001) and PAD-DM (p=0.01) vs. PAD-NGM. By multivariate regression YKL-40 is significantly associated with age (beta=0.272), triglycerides (beta=0.216), aspartate-amino-transferase (beta=0.177) and c-reactive-protein (beta=0.178). Underpinning its role YKL-40 was found to be associated with micro-/macroalbuminuria (p=0.014/p=008) – a strong remodeling inducer. In addition, YKL-40 was elevated in existence of mediasclerosis (p=0.008), a remodeling process.Conclusion: We are first to show that YKL-40 is higher in subjects with peripheral arterial disease. YKL-40 was higher in PAD patients with pre-/diabetes. In addition, YKL-40 was associated with the “severity” of generalized atherosclerosis estimated by affected vascular beds. All our findings point towards a role of YKL-40 in the progression/prognosis of patients with PAD and concomitant diabetes.

Leonurine: A new comer in the natural compounds affecting atherosclerosis - Corrected Proof

Giuseppe Danilo Norata, Alberico Luigi Catapano — 2012-05-07

Historically, pharmacology has relied upon the use of compounds extracted from natural resources to treat many common human disorders. Although most of these were demonstrated to be ineffective in the years of modern pharmacology, several molecules present in plants and adapted for human uses, such as aspirin, constitute the pillars of medicine. Over the last decades in the western world, the interest in compounds of natural origin was somewhat reduced, and was superseded by approaches that were more focused on lab-designed and chemically synthesized molecules, and the success of this approach has of course greatly improved disease management and life expectancy. However, in order to identify new and innovative drugs, with chemical features completely unrelated to the known molecules, many large pharmaceutical companies have started ambitious programs aimed at screening for new compounds in natural environments such as virgin forests. In addition, in collaboration with several academic institutions, programs have looked more closely at products used in eastern medicine, some of which could be highly effective.

A simple multiplier to calculate the impact of HDL cholesterol on cardiovascular risk estimation using SCORE - Corrected Proof

Olivier S. Descamps, Marie Therese Cooney, Guy De Backer, Ian Graham — 2012-05-07

Abstract: The 2011 ESC–EAS guidelines on the management of dyslipidaemias use four separate charts to illustrate the impact of differing HDL cholesterol levels on risk of cardiovascular disease. We developed an easy way to calculate the effects of differing HDL-C levels on risk by deriving HDL and sex specific multipliers and applying these to various reference charts. Of three strategies explored, one based on a low HDL (0.8mmol/l) reference chart was the simplest and was acceptably accurate. Such an approach simplifies risk estimation by avoiding the need for multiple charts.

Atherosclerosis and arterial stiffness in obstructive sleep apnea—A cardiovascular magnetic resonance study - Corrected Proof

2012-05-07

Highlights: ► We assessed whether OSA is independently associated with atherosclerosis and vascular dysfunction using CMR. ► Patients with OSA were compared to matched control subjects without OSA and the correlation between OSA severity and CMR markers was determined. ► We demonstrated an independent association between the presence and severity of OSA and increased carotid and aortic atheroma burden. ► We report for the first time an association between OSA and the presence of advanced and high risk carotid atheroma. ► We detected an association between OSA and central aortic stiffening, which appears to be dependent on an increase in blood pressure.Abstract: Objective: Obstructive sleep apnoea (OSA) has been linked to cardiovascular risk factors, such as hypertension, and clinical cardiovascular endpoints. Our aim was to assess whether OSA is independently associated with atherosclerosis and vascular dysfunction as assessed by cardiovascular magnetic resonance (CMR).Methods: 58 patients with OSA and 39 matched control subjects without OSA underwent CMR of the aorta and carotid arteries. Carotid and aortic wall thickness and aortic distensibility were measured. Multi-weighted, high resolution CMR imaging was used for carotid atheroma characterization according to the American Heart Association (AHA) atheroma classification, modified for CMR.Results: Carotid [1.47±0.03mm vs. 1.26±0.05mm, (P<0.01)] and aortic wall thickness [2.95±0.09mm vs. 2.05±0.07mm, (P<0.001)] were increased in patients with OSA compared to controls. Aortic distensibility was decreased in patients with OSA [3.62±0.3 vs. 4.75±0.2mmHg−1×10−3, (P<0.05)]. Prevalence of carotid plaque, average carotid atheroma class, and prevalence of high risk features of carotid atheroma were increased in patients with OSA (P<0.005 for all). On multivariate analysis, Oxygen desaturation index (ODI) emerged as an independent predictor of carotid and aortic wall thickness, but not of aortic stiffness.Conclusions: OSA is associated with increased carotid and aortic atheroma burden and with advanced, high risk carotid atherosclerotic plaques, but not with aortic stiffening.

Diastolic function parameters are improved by the addition of simvastatin to enalapril-based treatment in hypertensive individuals - Corrected Proof

2012-05-03

Highlights: ► Diastolic dysfunction is frequent and increases mortality . ► Pre-clinical data suggest that statins improve diastolic function. ► In this randomized controlled trial, the addition of simvastatin to enalapril in hypertensive patients improves parameters of diastolic function.Abstract: Objective: Diastolic dysfunction (DD) is a frequent condition in hypertensive patients whose presence increases mortality and whose treatment remains unclear. The aim of this study was to investigate in a prospective, double-blinded, placebo-controlled randomized design the additive effect of simvastatin on DD in enalapril-treated hypertensive patients with average cholesterol levels.Methods: Hypertensive patients with DD and LDL-cholesterol <160mg/dL underwent a run-in phase to achieve a systolic blood pressure (SBP) <135mmHg and diastolic blood pressure (DBP) <85mmHg with enalapril. Hydrochlorothiazide was added when need to achieve blood pressure control. Four weeks after reaching the optimum anti-hypertensive regimen patients were randomized to receive 80mg simvastatin (n=27) or placebo (n=28) for a period of 20 weeks. Echocardiograms were performed before and after treatment with measurement of maximum left atrial volume (LAV), conventional and tissue Doppler velocities in early diastole (E, e′) and late diastole (A, a′).Results: After 20 weeks, the simvastatin group presented reduction in SBP (−4±2mmHg, p=0.02), increase in E/A ratio (1.0±0.05 to 1.2±0.06, p=0.03) and decrease of LAV indexed to body surface area (24.5±0.9 to 21.1±0.8ml/m2, p=0.048), as compared with placebo arm. No change in systolic function and no correlation between the E/A ratio, LAV and changes in blood pressure or lipid profile were observed.Conclusions: The addition of simvastatin to enalapril in hypertensive patients with average cholesterol levels improves parameters of diastolic function independently of changes in blood pressure or cholesterol.

Resveratrol inhibits neointimal formation after arterial injury through an endothelial nitric oxide synthase-dependent mechanism - Corrected Proof

2012-05-01

Highlights: ► We investigated the mechanism whereby resveratrol, a red wine polyphenol, decreases restenosis. ► We used two models of restenosis, arterial balloon injury in rats and arterial wire injury in mice. ► We found that the effect of resveratrol to decrease neointimal growth after arterial injury was abolished in rats treated with an inhibitor of nitric oxide synthase and also in mice genetically deficient of endothelial nitric oxide synthase.Abstract: Revascularization procedures used for treatment of atherosclerosis often result in restenosis. Resveratrol (RSV), an antioxidant with cardiovascular benefits, decreases neointimal formation after arterial injury by a mechanism that is still not fully clarified. Our main objective was to address the role of nitric oxide synthases (NOSes) and more specifically the endothelial-NOS (eNOS) isoform as a mediator of this effect.RSV (4mg/kg/day, s.c.) alone or in combination with the NOS inhibitor N-nitro-l-arginine methyl ester (l-NAME) (2mg/kg/day, s.c.) was given to Sprague-Dawley rats beginning at 3 days before arterial (carotid or aortic) injury. RSV reduced neointimal formation by 50% (P<0.01), decreased intimal cell proliferation by 37% (P<0.01) and reduced inflammatory markers such as PECAM and MMP-9 mRNA. These effects of RSV were all abolished by coadministration of l-NAME. Oral RSV (beginning at 5 days before arterial injury) reduced neointimal thickness after femoral wire injury in mice, however this effect was not observed in eNOS knockout mice. This is the first report of RSV decreasing neointimal cell proliferation and neointimal growth through an eNOS-dependent mechanism.

Dietary fatty acids and peripheral artery disease in adults - Corrected Proof

Asghar Z. Naqvi, Roger B. Davis, Kenneth J. Mukamal — 2012-05-01

Highlights: ► In a national sample, higher dietary linolenic acid was associated with higher ABI. ► Higher dietary saturated fatty acid was positively associated with prevalence of PAD. ► Other fatty acids were not substantially associated with ABI or prevalence of PAD.Abstract: Background: Peripheral artery disease (PAD) is a debilitating condition involving atherosclerosis. Although saturated, monounsaturated and polyunsaturated fatty acids have strong associations with atherosclerosis, it is unclear if diets high in these fatty acids affect PAD.Methods: We studied 6352 adults aged 40 years and older who participated in the U.S. National Health and Nutrition Examination Survey between 1999 and 2004. Ankle brachial index (ABI) was assessed by standardized blood pressure measurements, and we defined PAD as an ABI<0.9. Fatty acid intake was assessed by validated 24-h dietary recall. We used multivariable linear and logistic regression to estimate associations between intakes of dietary saturated fatty acids (SFAs), monounsaturated fatty acids (MFAs), marine omega-3 fatty acids (N-3), linolenic acid (LNA), and omega-6 fatty acids (N-6) and ABI/PAD.Results: The prevalence and 95% confidence interval (CI) of PAD was 5.2% (95% CI 4.6–5.8).There were no associations between ABI and intakes of marine N-3 (p=0.83) or N-6 (p=0.19) in adjusted models. In contrast, LNA was associated with higher ABI (p=0.04) and SFA tended to be associated with lower ABI (p=0.06) in adjusted models. In addition, higher SFA was associated with a higher prevalence of PAD: adjusted odds ratio 1.30 (95% CI 1.01–1.67; p=0.04) and a trend toward slower gait speed (p=0.08).Conclusion: In this nationally representative sample, higher dietary intakes of LNA and SFAs were associated with higher and lower ABI, respectively. Prospective studies are needed to confirm the potential protective effects of dietary LNA and detrimental effects of dietary SFAs on PAD.

RAGE signaling mediates post-injury arterial neointima formation by suppression of liver kinase B1 and AMPK activity - Corrected Proof

2012-05-01

Highlights: ► RAGE deficiency inhibits post-injury vascular remodeling by activating AMPK pathway. ► Carotid ligation decreases p-AMPK and increases RAGE and ligands, which promotes SMCs proliferation and migration. ► RAGE/ligands mediates vascular remodeling by regulation of AMPK activity via LKB1 and STAT3.Abstract: Objective: Intima formation involves smooth muscle cell (SMC) proliferation and migration that ultimately drives arterial stenosis, thrombosis, and ischemia in atherosclerosis, hypertension, and arterial revascularization. Receptor for advanced glycation endproducts (RAGE) is a transmembrane signaling receptor implicated in diabetic renal and vascular complications, and post-injury intima formation, partly via Signal transducer and activator of transcription 3 (STAT3) activation. The metabolic super-regulator Adenosine monophosphate kinase (AMPK) inhibits SMC proliferation and intima formation. AMPK activation is promoted by liver kinase B1 (LKB1), and LKB1 inhibits STAT3 activation. Here, we tested the hypothesis that RAGE promotes arterial intima formation by modulating both LKB1 and AMPK.Methods and results: RAGE ligands (the calgranulin S100A11, and glycated albumin) suppressed AMPK activation in conjunction with increased proliferation and migration of cultured SMCs. These effects were inhibited both by RAGE deficiency and by prior AMPK activation. In SMCs, RAGE ligands decreased LKB1 activity. Moreover, knockdown of both LKB1 and AMPK were associated with increased STAT3 phosphorylation levels. In response to murine carotid artery ligation, expression of RAGE and S100A11 increased, whereas AMPK and LKB1 activity decreased in situ. Conversely, LKB1 and AMPK activity increased in situ, and neointima formation was attenuated in Rage−/− mice.Conclusion: The linkage of decreased LKB1 and AMPK activity with increased STAT3 in SMCs mediates the capacity of RAGE ligand-induced signaling to promote neointima formation in response to arterial injury.

Multiple factors and pathways involved in hepatic very low density lipoprotein-apoB100 overproduction in Otsuka Long-Evans Tokushima fatty rats - Corrected Proof

2012-04-30

Highlights: ► We examine the relationship between systemic/hepatic inflammation associated with insulin resistance and VLDL-apoB100 production. ► Inflammation, insulin resistance and dysregulated lipogenesis may be important underlying factors in apoB100 production. ► Hepatic insulin resistance related genes, proteins expression and phosphorylation. ► Hepatic lipogenesis related gene and protein expression. ► Decreased sirt 1, 2 and 3 mRNA expression in OLETF liver.Abstract: Aims: Overproduction of hepatic very low-density lipoprotein (VLDL) particles is a major abnormality of lipoprotein dysregulation in type 2 diabetes (T2D). We sought to examine the relationship between systemic/hepatic inflammation associated with insulin resistance and apolipoprotein (apo)B100-containing VLDL production.Methods and results: At the age of 19 wks, Otsuka Long-Evans Tokushima Fatty (OLETF) rats showed systemic inflammation (plasma TNF-α and interleukin (IL)-6 levels increased), insulin resistance (plasma retinol binding protein 4 and soluble CD36 levels were higher), dyslipidemia and fatty liver (plasma and liver triglyceride and cholesterol levels were higher as well as total VLDL-, VLDL1-, VLDL2-apoB100 and VLDL-triglycerides were overproduced), compared with the control rats. In livers of OLETF rats, mRNA levels of tnf, il1b and il6 were increased, but an anti-inflammatory protein, zinc finger protein 36, and its mRNA expression were decreased. We also found that the liver mRNA, protein levels, and tyrosine phosphorylation (pY) of insulin receptor (InsR) substrate (IRS) 2, but not IRS1, were decreased in OLETF rats; pY of InsR and Akt protein and phospho-Akt (ser437) were also reduced; but protein tyrosine phosphatase-1B protein was overexpressed. The gene expressions of glucose transporters 1 and 2, and glycogen synthase were decreased, but phosphatase and tensin homolog deleted on chromosome ten and glycogen synthase kinase 3β mRNAs were overexpressed, compared with the controls. Sterol regulatory element binding protein-1c mRNA, ATP-binding cassette transporter A1 mRNA, microsomal triglyceride transfer protein mRNA/protein, and CD36 mRNA/protein levels were increased and lipoprotein lipase and Niemann-Pick c1-like1 mRNA levels were decreased, which are all involved in lipogenesis. Decreased sirtuins1–3 mRNA levels were also observed in OLETF rats.Conclusions: These abnormal genes, proteins expression and phosphorylation of multiple pathways related to inflammatory, insulin signaling and lipogenesis may be important underlying factors in VLDL-apoB100 particles overproduction observed in T2D. Our data contribute to the further understanding of an association of dyslipoproteinemia with systemic metabolic disorders, fatty liver and dysregulated hepatic metabolic pathways.

Pancreatic β-cell function relates positively to HDL functionality in well-controlled Type 2 diabetes mellitus - Corrected Proof

Robin P.F. Dullaart, Wijtske Annema, Jan Freark de Boer, Uwe J.F. Tietge — 2012-04-30

Highlights: ► Abnormalities in HDL functionality may affect β-cell function. ► HDL functionality was assessed as its anti-oxidative capacity and cholesterol efflux to plasma. ► β-cell function was related to these HDL functionality measures in well-controlled Type 2 diabetes. ► Such relationships were not observed in subjects with normal and impaired fasting glucose. ► Better HDL functionality may contribute to maintenance of β-cell function in Type 2 diabetes.Abstract: Background: High density lipoproteins (HDLs) have been implicated in glucose homeostasis. Among subjects with normal fasting glucose (NFG), impaired fasting glucose (IFG) and Type 2 diabetes mellitus (T2DM) we tested whether pancreatic β-cell function relates to HDL functionality, as determined by HDL anti-oxidative capacity and cellular cholesterol efflux to plasma.Subjects and methods: HDL anti-oxidative capacity (inhibition of LDL oxidation in vitro), cellular cholesterol efflux (the ability of plasma to stimulate cholesterol efflux out of cultured fibroblasts obtained from a single human donor), glucose and insulin were determined in fasting plasma samples from 37 subjects with NFG, 36 with IFG and 22 with T2DM (no glucose lowering drug or insulin treatment; HbA1c 6.0±1.0%). Homeostasis model assessment was used to estimate pancreatic β-cell function (HOMA-β) and insulin resistance (HOMAir).Results: HOMA-β was lowest, whereas HOMAir was highest in T2DM (P<0.01 and P<0.001 vs. NFG). HDL anti-oxidative capacity and cellular cholesterol efflux did not differ significantly according to glucose tolerance category. In univariate analysis and after controlling for HOMAir both HDL anti-oxidative capacity (P<0.05) and cellular cholesterol efflux (P<0.01) were positively correlated with HOMA-β in T2DM, but not in NFG and IFG. In age-, sex- and HOMAir-adjusted analyses, T2DM status interacted positively with HDL anti-oxidative capacity (P=0.001) and cellular cholesterol efflux (P=0.042) on HOMA-β. HbA1c interacted similarly with HDL functionality measures on HOMA-β.Conclusions: Pancreatic β-cell function relates to pathophysiologically relevant measures of HDL function in T2DM, but not in NFG and IFG. Better HDL functionality may contribute to maintenance of β-cell function in subjects with well-controlled T2DM.

Reducing lifetime cardiovascular risk - Corrected Proof

Jane Stock — 2012-04-27

A proactive approach aimed at preventing cardiovascular (CV) risk factors to reduce lifetime risk of cardiovascular disease (CVD) is needed, according to the authors of a meta-analysis of 18 studies involving over 250,000 men and women . Among men aged 55 years or more, those with an optimal risk factor profile (nonsmokers without diabetes and optimal cholesterol and blood pressure) had a lower lifetime risk:

Aortic stiffness and calcification in men in a population-based international study - Corrected Proof

2012-04-26

Highlights: ► Aortic stiffness is an independent risk factor of cardiovascular disease. ► We examined the association of aortic stiffness with aortic calcification. ► We examined it in a population-based multi-ethnic cohort of 906 healthy men. ► Aortic stiffness was measured as carotid-femoral pulse wave velocity (cfPWV). ► cfPWV had a significant positive association with aortic calcification.Abstract: Objectives: Aortic stiffness, a hallmark of vascular aging, is an independent risk factor of cardiovascular disease and all-cause mortality. The association of aortic stiffness with aortic calcification in middle-aged general population remains unknown although studies in patients with end-stage renal disease or elderly subjects suggest that aortic calcification is an important determinant of aortic stiffness. The goal of this study was to examine the association of aortic calcification and stiffness in multi-ethnic population-based samples of relatively young men.Methods: We examined the association in 906 men aged 40–49 (81 Black Americans, 276 Japanese Americans, 258 White Americans and 291 Koreans). Aortic stiffness was measured as carotid-femoral pulse wave velocity (cfPWV) using an automated waveform analyzer. Aortic calcification from aortic arch to iliac bifurcation was evaluated using electron-beam computed tomography.Results: Aortic calcium score was calculated and was categorized into four groups: zero (n=303), 1–100 (n=411), 101–300 (n=110), and 401+ (n=82). Aortic calcification category had a significant positive association with cfPWV after adjusting for age, race, and mean arterial pressure (mean (standard error) of cfPWV (cm/s) from the lowest to highest categories: 836 (10), 850 (9), 877 (17) and 941 (19), P for trend <0.001). The significant positive association remained after further adjusting for other cardiovascular risk factors. The significant positive association was also observed in each race group.Conclusions: The results suggest that aortic calcification can be one mechanism for aortic stiffness and that the association of aortic calcification with stiffness starts as early as the 40s.

Carotid extra-medial thickness in childhood: Early life effects on the arterial adventitia - Corrected Proof

2012-04-25

Highlights: ► Extra-medial thickness is a noninvasive measure of arterial adventitial thickness. ► We measured carotid extra-medial thickness in 389 non-diabetic 8-year old children. ► Extra-medial thickness is associated with cardiovascular risk factors in childhood. ► The arterial adventitia may be involved in early atherosclerosis.Abstract: Objective: Structural modification of the arterial adventitia may be an early event in atherosclerosis. Carotid extra-medial thickness is a new measure of arterial adventitial thickness. We examined the association of cardiovascular risk factors with extra-medial thickness, in childhood.Methods: Carotid extra-medial thickness was assessed by high-resolution ultrasound in 389 non-diabetic children aged 8-years. A non-fasting blood sample was collected from 314 participants. Associations of gender, age, lipoproteins, blood pressure, BMI z-score, waist:height ratio and parental history of early vascular disease, with extra-medial thickness were examined.Results: Carotid extra-medial thickness was lower in girls (r=−.163, P=.001) and directly associated with systolic (r=.128, P=.009), diastolic blood pressure (r=.130, P=.009), and height (r=.170, P=.0006). These associations remained after adjustment for carotid intima-media thickness. In multivariable analysis including carotid intima-media thickness, only gender and height were significantly associated with carotid extra-medial thickness. In gender-stratified analysis, the strongest associations with extra-medial thickness were BMI z-score (r=.181, P=.01), height (r=.210, P=.003) and diastolic blood pressure (r=.167, P=.02) for boys; and systolic blood pressure (r=.153, P=.03) and parental history of premature cardiovascular disease (r=.139, P=.05) for girls. The association of BMI z-score with extra-medial thickness differed by gender (P-interaction=.04).Conclusions: Carotid extra-medial thickness is independently associated with gender and height in childhood. Extra-medial thickness may provide important information concerning early arterial health, particularly related to the arterial adventitia.

Reduced metal ion concentrations in atherosclerotic plaques from subjects with Type 2 diabetes mellitus - Corrected Proof

2012-04-23

Highlights: ► Elevated levels of metal ions have been proposed as catalysts in atherosclerosis. ► Lesions from people with and without Type 2 diabetes were analysed for metal ions. ► Levels of lipid and protein oxidation products were quantified. ► Lower levels of iron were detected in subjects with Type 2 diabetes than without. ► The data do not support a role for metal ions in diabetes-enhanced atherosclerosis.Abstract: Aims: Transition metal ions have been implicated in atherosclerosis. The goal of this study was to investigate whether metal ion levels were higher in people with diabetes, in view of their increased risk of aggravated atherosclerosis.Methods and results: Absolute concentrations of iron, copper, zinc and calcium, and products of protein and lipid oxidation were quantified in atherosclerotic lesions from subjects with (T2DM, n=27), without Type 2 diabetes (nonDM, n=22), or hyperglycaemia (HG, n=17). Iron (P<0.05), zinc (P<0.01) and calcium (P=0.01) were lower in T2DM compared to nonDM subjects. Copper levels were comparable. A strong correlation (r=0.618; P<0.001) between EPR-detectable and total iron in nonDM patients was not seen in T2DM. X-ray fluorescence microscopy revealed “hot spots” of iron in both T2DM and nonDM. Calcium and zinc co-localised and levels correlated strongly. F2-isoprostanes (P<0.05) and di-Tyr/Tyr ratio (P<0.025), oxidative damage markers were decreased in T2DM compared to nonDM, or HG.Conclusion: Advanced atherosclerotic lesions from T2DM subjects unexpectedly contained lower levels of transition metal ions, and protein and lipid oxidation products, compared to nonDM and HG. These data do not support the hypothesis that elevated metal ion levels may be a major causative factor in the aggravated atherosclerosis observed in T2DM patients.

Assessment of the value of a genetic risk score in improving the estimation of coronary risk - Corrected Proof

2012-04-23

Highlights: ► A multi-locus genetic risk score (GRS) was directly associated with risk of CHD events. ► This GRS improved risk reclassification in the population at intermediate coronary risk. ► These results indicate the potential value of genetic information in risk assessment.Abstract: Background: The American Heart Association has established criteria for the evaluation of novel markers of cardiovascular risk. In accordance with these criteria, we assessed the association between a multi-locus genetic risk score (GRS) and incident coronary heart disease (CHD), and evaluated whether this GRS improves the predictive capacity of the Framingham risk function.Methods and results: Using eight genetic variants associated with CHD but not with classical cardiovascular risk factors (CVRFs), we generated a multi-locus GRS, and found it to be linearly associated with CHD in two population based cohorts: The REGICOR Study (n=2351) and The Framingham Heart Study (n=3537) (meta-analyzed HR [95%CI]: ∼1.13 [1.01–1.27], per unit). Inclusion of the GRS in the Framingham risk function improved its discriminative capacity in the Framingham sample (c-statistic: 72.81 vs.72.37, p=0.042) but not in the REGICOR sample. According to both the net reclassification improvement (NRI) index and the integrated discrimination index (IDI), the GRS improved re-classification among individuals with intermediate coronary risk (meta-analysis NRI [95%CI]: 17.44 [8.04; 26.83]), but not overall.Conclusions: A multi-locus GRS based on genetic variants unrelated to CVRFs was associated with a linear increase in risk of CHD events in two distinct populations. This GRS improves risk reclassification particularly in the population at intermediate coronary risk. These results indicate the potential value of the inclusion of genetic information in classical functions for risk assessment in the intermediate risk population group.

Association between traditional cholesterol parameters, lipoprotein particle concentration, novel biomarkers and carotid plaques in retired National Football League players - Corrected Proof

2012-04-23

Highlights: ► We assessed the association between lipid parameters and carotid plaques in retired NFL players. ► Carotid artery plaques were common and seen in 41% of retired NFL players. ► Plaques were more strongly associated with LDL particle concentration than LDL cholesterol.Abstract: Objectives: We assessed whether low-density lipoprotein particle concentration (LDL-P) and high-sensitivity C-reactive protein [hs-CRP] can identify subclinical atherosclerosis better than traditional cholesterol parameters in retired National Football League (NFL) players.Background: It is not known whether LDL-P and the biomarker hs-CRP can identify subclinical atherosclerosis better than low-density lipoprotein cholesterol (LDL-C) or non-high-density-lipoprotein cholesterol (non-HDL-C) in retired NFL players, given high prevalence of metabolic syndrome in these players.Methods: Carotid artery plaque screening was performed with traditional lipids, LDL-P, and hs-CRP in 996 retired players. Logistic regression analyses comparing highest with the lowest quartile were performed.Results: Carotid artery plaques were seen in 41%. LDL-C (odds ratio [OR] 1.66, 95% confidence interval [CI] 1.06–2.59), non-HDL-C (OR 1.67, 95% CI 1.04–2.67), and LDL-P (OR 2.21, 95% CI 1.35–3.62) were associated with plaques in adjusted models. Among 187 retired players with metabolic syndrome, LDL-C (OR 1.40, 95% CI 0.53–3.72) was not associated with carotid plaques, whereas LDL-P (OR 3.71, 95% CI 1.16–11.84) and non-HDL-C (OR 2.63, 95% CI 0.91–7.63, p=0.07; borderline significant) were associated with carotid plaques. hs-CRP (OR 1.13, 95% CI 0.71–1.79) was not associated with carotid plaques.Conclusion: Carotid artery plaques were common in retired NFL players and were strongly associated with LDL-P, especially among those with metabolic syndrome. hs-CRP was not associated with carotid plaques in this cohort.

Associations between cellular growth factors and ischemic and hemorrhagic strokes - Corrected Proof

Yukihiko Momiyama — 2012-04-19

Accumulating evidence indicates that insulin-like growth factors I and II (IGF-I and -II) promote the growth of arterial cells and mediate the development of cardiovascular diseases. IGFs promote the proliferation and migration of vascular smooth muscle cells (SMCs) and suppress the apoptosis of SMCs . In atherosclerotic plaques, IGF-I expression was shown to be low or even absent in those with macrophage infiltration . These findings suggest that IGFs may play a protective role against plaque weakening and rupture. Although many cells in the body synthesize IGFs, the systemic IGFs levels are determined mainly by the production in the liver. Several population-based prospective studies have shown that low IGF-I levels were associated with increased risks of ischemic heart disease (IHD) , mortality from IHD , and myocardial infarction (MI) . Regarding the association between the IGF-I levels and strokes, Johnsen et al. reported that low IGF-I levels were associated with an increased risk of ischemic stroke, whereas Kaplan et al. found no such association. In this issue of Atherosclerosis, Iso et al. show that there is no association between the IGF-I levels and mortality from ischemic stroke in a Japanese population. However, it is interesting to note that they reported that the IGF-I levels were lower in subjects who died from cerebral hemorrhage than in controls, and that there was an inverse association with the risk of mortality from cerebral hemorrhage, independent of atherosclerotic risk factors.

The LDL-cholesterol to HDL-cholesterol ratio and the severity of coronary and aortic atherosclerosis - Corrected Proof

2012-04-19

The LDL-cholesterol to HDL-cholesterol (LDL/HDL-cholesterol) ratio is recognized as a stronger risk predictor of cardiovascular diseases than LDL-cholesterol and HDL-cholesterol levels . The close correlation between the LDL/HDL-cholesterol ratio and the severity of coronary artery stenosis is also reported . However, the association between the LDL/HDL-cholesterol ratio and aortic atherosclerosis has not been elucidated yet. Magnetic resonance imaging (MRI) is a useful tool for non-invasively evaluating atherosclerotic plaques in both the thoracic and abdominal aorta . Regarding this MRI method, we and others showed good correlations for the aortic plaque extent between the in vivo and ex vivo MRI findings and histopathology in animal models. In humans, we reported that MRI evaluations of the thoracic aorta closely correlated with TEE findings .

IL-1, quo vadis? - Corrected Proof

Alena Grebe, Eicke Latz — 2012-04-18

The build up of lipids and immune cells in the subintimal space of arteries is termed atherosclerosis. This slow, but progressive process occurs in the majority of humans. Atherosclerosis remains undetected until changes in the atherosclerotic vessel cause decreased blood flow, or result in luminal thrombotic occlusion.

Estimation of lipoprotein profile in patients with type II diabetes and its relevance to remnant lipoprotein cholesterol levels - Corrected Proof

2012-04-18

Abstract: Background: Remnant lipoprotein (RLP), associated with atherosclerosis progression, is often elevated in diabetes mellitus. The RLP level is estimated by immune-separation method and agarose-gel electrophoresis (AGE).Methods: The patients were grouped into three groups according to tertile of RLP-cholesterol (RLP-C) levels. The lipoprotein profiles of type II diabetic patients (T2DM) (n=194) were measured by an anion-exchange liquid chromatography (AEX-HPLC) and an AGE with lipid-staining or cholesterol-staining.Results: IDL- and VLDL-cholesterol by the AEX-HPLC, and VLDL-levels by the AGE with lipid-staining and with cholesterol-staining were significantly different in the three groups. In all the subjects, IDL-cholesterol (r=0.531) and VLDL-cholesterol (r=0.880) by the AEX-HPLC method were strongly correlated with RLP-C, but only VLDL levels were correlated with RLP-C in AGE, respectively.Conclusion: These results suggest that the AEX-HPLC, which can provide cholesterol levels of not only VLDL but also IDL, is helpful for estimation of lipid profiles in T2DM with high RLP-C.

PAPP-A negatively regulates ABCA1, ABCG1 and SR-B1 expression by inhibiting LXRα through the IGF-I-mediated signaling pathway - Corrected Proof

2012-04-16

Highlights: ► In the present study, we first found that PAPP-A inhibits ABCA1, ABCG1, SR-BI expression and reduces cholesterol efflux in THP-1 macrophage-derived foam cells. ► LXRα is involved in PAPP-A-induced down-regulation of ABCA1, ABCG1 and SR-BI in cells. ► PAPP-A promotes the IGF-1/PI3-K/Akt signaling pathway in cells. ► The IGF-1/PI3-K/Akt signaling pathway may be related to down-regulated expression of LXRα, ABCA1, ABCG1 and SR-BI by PAPP-A in THP-1 macrophage-derived foam cells.Abstract: Pregnancy-associated plasma protein-A (PAPP-A) has been involved in the atherosclerotic process through regulation of local expression of IGF-1 that mediates the activation of the phosphatidylinositol-3 (PI3-K) and Akt kinase (Akt) signaling cascades which lead to constitutive nitric oxide formation, with its attending vasodilator, antiplatelet and insulin-sensitizing actions. In addition, IGF-1 may decreased cholesterol efflux through reductions of expression in ABCA1 and SR-B1 by the PI3-K/Akt signaling pathway. In the current study, we examined whether PAPP-A was involved in LXRα regulation and in expression of ABCA1, ABCG1 or SR-B1 through the IGF-I-mediated signaling pathway (IGF/PI3-K/Akt). Results showed that PAPP-A significantly decreased expression of ABCA1, ABCG1 and SR-BI at both transcriptional and translational levels in a dose-dependent and time-dependent manner. Cellular cholesterol content was increased while cholesterol efflux was decreased by PAPP-A treatment. Moreover, LXRα which can regulate the expression of ABCA1, ABCG1 and SR-B1, was also down-regulated by PAPP-A treatment. LXRα-specific activation by LXRα agonist almost rescued the down-regulation of ABCA1, ABCG1 and SR-B1 expression by PAPP-A. In addition, PAPP-A can induce the IGF-1/PI3-K/Akt pathway in macrophages. Furthermore, our results indicate that the decreased levels observed in LXRα, ABCA1, ABCG1 and SR-B1 mRNA and protein levels upon treating cells with PAPP-A were strongly impaired with the PI3-K inhibitors or IGF-1R siRNA while the MAPK cascade inhibitor did not execute this effect, indicating that the process of ABCA1, ABCG1 and SR-BI degradation by PAPP-A involves the IGF-1/PI3-K/Akt pathway. In conclusion, PAPP-A may first down-regulate expression of LXRα through the IGF-1/PI3-K/Akt signaling pathway and then decrease expression of ABCA1, ABCG1, SR-B1 and cholesterol efflux in THP-1 macrophage-derived foam cells. Therefore, our study provided one of the mechanisms for understanding the critical effect of PAPP-A in pathogenesis of atherosclerosis.

Statins synergize dexamethasone-induced adipocyte fatty acid binding protein expression in macrophages - Corrected Proof

2012-04-16

Highlights: ► Statins inhibit macrophage FABP4 expression by reducing cellular cholesterol levels. ► Dexamethasone induces macrophage FABP4 expression by activating pGRE in the gene. ► However, statins synergize dexamethasone-induced FABP4 expression. ► The synergistic induction is completed by enhancing GR translocation and inhibiting nGRE.Abstract: Objective: Macrophage adipocyte fatty acid binding protein (FABP4) plays an important role in the development of atherosclerosis. We previously reported that dexamethasone induces macrophage FABP4 mRNA expression. Statins inhibit FABP4 expression. However, it remains unknown that if statins can antagonise dexamethasone-induced macrophage FABP4 expression.Methods and results: We determined the effect of co-treatment of statins and dexamethasone on macrophage FABP4 expression. Unexpectedly, statins did not block the induction of macrophage FABP4 expression by dexamethasone. In contrast, statins synergized dexamethasone-induced FABP4 expression. In vivo, pitavastatin synergized dexamethasone-induced FABP4 expression in both peritoneal macrophages and adipose tissues. Cholesterol and mevalonate, but not farnesylation and geranylgeranylation, inhibited the synergistic induction. Promoter assay disclosed a putative negative glucocorticoid regulatory element (nGRE) in FABP4 gene. Pitavastatin had little effect on expression of glucocorticoid receptor (GR). However, pitavastatin enhanced dexamethasone-mediated GR nuclear translocation but inhibited the binding of GR with nGRE.Conclusion: Our study defines an important mechanism involved in the regulation of macrophage FABP4 expression by a glucocorticoid and statins.

Statin therapy prevents expansive remodeling in venous bypass grafts - Corrected Proof

2012-04-16

Highlights: ► Atorvastatin significantly reduced expansive remodeling. ► Intimal luminal cross-sectional area by histomorphometry was significantly reduced by atorvastatin. ► macrophage infiltration, MMP-2 activity and metalloelastase activity were reduced in the atorvastatin treated group.Abstract: Background: Venous grafts (VG) have high failure rates by 10 years in aortocoronary bypass surgery. We have previously shown that expansive remodeling followed by increased LDL retention are early atherosclerotic changes in experimental VG placed in the arterial circulation. The objective of this study was to determine whether statin therapy prevents these expansive remodeling changes.Methods and results: Reversed jugular vein-to-common carotid artery interposition graft was constructed in 27 cholesterol-fed (0.5%) rabbits. Rabbits were randomized either to control or atorvastatin (5mg/kg/day) groups, starting two weeks prior to vein graft implantation and continuing until sacrifice at 1 or 12 weeks post-surgery. Ultrasound measurements of arterial luminal cross-sectional area (CSA) were done at day 3 and at 4, 8 and 12 weeks post-surgery. Histomorphometric measurements were performed following sacrifice at 12 weeks. Atorvastatin treatment significantly decreased total plasma cholesterol levels at 4, 8 and 12 weeks (12 weeks: 6.7±4.2mmol/L versus control 38.7±10.6mmol/L, p<0.0002). Atorvastatin significantly reduced expansive remodeling at 4, 8 and 12 weeks (lumen CSA: 44.6±6.6mm2 versus control 77.6±10.7mm2, p<0.0001). Intimal CSA by histomorphometry was also significantly reduced by atorvastatin at 12 weeks (5.59±2.19mm2 versus control 9.57±2.43mm2, p<0.01). VG macrophage infiltration, MMP-2 activity and metalloelastase activity were reduced in the atorvastatin treated group.Conclusion: Atorvastatin inhibits both expansive remodeling and intimal hyperplasia in arterialized VG, likely through inhibition of macrophage infiltration and reduction of tissue proteolytic activity. The mechanism proposed above may be important for preventing VG atherosclerosis and late VG failure.

Evaluation of cytochrome P450-derived eicosanoids in humans with stable atherosclerotic cardiovascular disease - Corrected Proof

2012-04-16

Highlights: ► We evaluated clinical factors that influence CYP-derived eicosanoids in CAD patients. ► Obesity and advancing age were associated with lower EET levels. ► Obesity and cigarette smoking were associated with lower epoxide:diol ratios. ► Renin–angiotensin system inhibitor use was associated with lower 20-HETE levels. ► CAD patients exhibited lower sEH metabolic activity relative to healthy individuals.Abstract: Objective: Preclinical and genetic epidemiologic studies suggest that modulating cytochrome P450 (CYP)-mediated arachidonic acid metabolism may have therapeutic utility in the management of coronary artery disease (CAD). However, predictors of inter-individual variation in CYP-derived eicosanoid metabolites in CAD patients have not been evaluated to date. Therefore, the primary objective was to identify clinical factors that influence CYP epoxygenase, soluble epoxide hydrolase (sEH), and CYP ω-hydroxylase metabolism in patients with established CAD.Methods: Plasma levels of epoxyeicosatrienoic acids (EETs), dihydroxyeicosatrienoic acids (DHETs), and 20-hydroxyeicosatetraenoic acid (20-HETE) were quantified by HPLC–MS/MS in a population of patients with stable, angiographically confirmed CAD (N=82) and healthy volunteers from the local community (N=36). Predictors of CYP epoxygenase, sEH, and CYP ω-hydroxylase metabolic function were evaluated by regression.Results: Obesity was significantly associated with low plasma EET levels and 14,15-EET:14,15-DHET ratios. Age, diabetes, and cigarette smoking also were significantly associated with CYP epoxygenase and sEH metabolic activity, while only renin-angiotensin system inhibitor use was associated with CYP ω-hydroxylase metabolic activity. Compared to healthy volunteers, both obese and non-obese CAD patients had significantly higher plasma EETs (P<0.01) and epoxide:diol ratios (P<0.01), whereas no difference in 20-HETE levels was observed (P=NS).Conclusions: Collectively, these findings suggest that CYP-mediated eicosanoid metabolism is dysregulated in certain subsets of CAD patients, and demonstrate that biomarkers of CYP epoxygenase and sEH, but not CYP ω-hydroxylase, metabolism are altered in stable CAD patients relative to healthy individuals. Future studies are necessary to determine the therapeutic utility of modulating these pathways in patients with CAD.

Follow-up association study of linkage regions reveals multiple candidate genes for carotid plaque in Dominicans - Corrected Proof

2012-04-16

Highlights: ► We investigate genetic associations in four linkage regions for carotid plaque among Dominican families. ► Associations with carotid plaque are marked for NAV2, EFCAB11, AGBL1, PTPN9, LINGO1 genes. ► Replications are found for EFCAB11, NAV2, AGBL1 and other several genes.Abstract: Objective: Carotid plaque is a marker of subclinical atherosclerosis with a genetic component. The aim of this follow-up fine mapping study was to identify candidate genes for carotid plaque within four linkage regions.Methods: We successfully genotyped 3712 single nucleotide polymorphisms (SNPs) under the four linkage regions that were previously identified in 100 extended Dominican families. Family-based association tests were performed to investigate their associations with carotid plaque. Promising SNPs were evaluated in an independent population-based subcohort (N=941, 384 Dominicans) from the Northern Manhattan Study (NOMAS).Results: In the family study, evidence for association (p<0.0005) was found regarding several genes (NAV2, EFCAB11/TDP1, AGBL1, PTPN9, LINGO1 and LOC730118), with the strongest association at rs4143999 near EFCAB11/TDP1 (p=0.00001 for carotid presence and 0.00003 for plaque area, multiple testing corrected p≤0.02). The association in AGBL1 and PTPN9 was mainly driven by the families with linkage evidence (p=0.00008–0.00001 and 0.76–0.32, respectively, in the families with and without linkage evidence). However, these associations explained only a small portion of the observed linkage. In NOMAS, replication (p<0.05 in the whole NOMAS subcohort and p<0.10 in the smaller Dominican subcohort) was found for SNPs within/near EFCAB11, NAV2, AGBL1 and other genes.Conclusion: This follow-up study has identified multiple candidate genes for carotid plaque in the Dominican population. Many of these genes have been implicated in neurodegenerative and cardiovascular diseases. Further studies with in-depth re-sequencing are needed to uncover both rare and common functional variants that contribute to the susceptibility to atherosclerosis.

Evaluation of possible subclinical atherosclerosis in adolescents with a family history of premature atherosclerosis - Corrected Proof

2012-04-16

Highlights: ► We aimed to evaluate the adolescents who are at high risk of premature atherosclerosis. ► Thirty-three subjects who have a father with a history of obstructive coronary artery disease before the age 40 and 30 controls were enrolled to the study. ► Carotid intima-media thickness and pulse wave velocity values were higher in the risky group, but these differences were not statistically significant. ► Present methods failed to show expected subclinical atherosclerosis in risky adolescents.Abstract: Objective: To evaluate possible subclinical atherosclerosis using biomarkers and ultrasound-guided methods in a group of adolescents having fathers with premature atherosclerosis.Methods: Thirty-three subjects whose fathers had a history of premature coronary artery disease and 30 counterparts whose fathers had no history of coronary artery disease were included in the study.Results: The homocysteine levels, high-sensitivity C-reactive protein levels, and cardiac chamber sizes and functions did not differ between the two groups. The carotid stiffness index β (CSI), the intima-media thickness (CIMT) and aortic pulse wave velocity (PWV) values were higher in the group with a family history of coronary artery disease, but only the difference in the CSI was statistically significant (CSI 3.07±1.33 vs 3.88±1.25, P=0.015; CIMT 0.53±0.09mm vs 0.57±0.08mm, P=0.068; PWV 3.49±0.53m/s vs 3.78±0.63m/s, P=0.053).Conclusion: Among several markers of subclinical atherosclerosis, the CSI was significantly higher in adolescents who had a family history of premature atherosclerosis. The small sample size, the multifactorial nature of atherosclerosis or the insufficient power of these methods may explain these results.

Visceral abdominal fat accumulation predicts the progression of noncalcified coronary plaque - Corrected Proof

2012-04-13

Highlights: ► Excess visceral abdominal tissue (VAT) is more strongly associated with risk factors of coronary artery disease (CAD) than other measures of obesity. ► This prospective cohort study comprised 553 CAD patients with coronary plaque with ≤50% coronary stenosis on CT angiography. ► VAT accumulation was more strongly associated with progression of coronary noncalcified plaque than BMI or waist circumference. ► These results suggest combining VAT measurements and CAD risk factors may provide better risk assessments of noncalcified plaque progression.Abstract: Background: Excess visceral abdominal tissue (VAT) is more strongly associated with risk factors of coronary artery disease (CAD) than body mass index (BMI) or waist circumference. However, whether adding VAT measurements to CAD risk factors provides better risk assessment for CAD progression has not been fully evaluated.Methods and results: This prospective cohort study comprised 553 CAD patients with coronary plaque with ≤50% coronary stenosis as assessed by computed tomography (CT) angiography. Quantification of VAT area was performed together with CT angiography using abdominal CT scanning. After a mean 38±8months follow-up, 320 patients underwent repeated CT scans for worsening angina symptoms without findings of positive ischemia. Increased segments of noncalcified plaque were seen in 152 (48%) and an increased calcium score was seen in 261 (82%) patients. The risk for progression of noncalcified plaque increased steadily with higher VAT quartiles, independent of CAD risk factors. Patients in the higher quartiles were at increased risk of progression of noncalcified plaque (quartiles IV OR 4.7; 95% CI 2.3–9.4, p-value<0.001). In contrast, increases above the median calcium score showed no independent correlation to VAT. Compared to VAT, progression of noncalcified plaque showed no phased increase with higher waist circumference and weaker increase with higher BMI quartiles.Conclusion: VAT accumulation was positively associated with progression of coronary noncalcified plaque, but not of calcified plaque. This suggests that risk assessment of progression of noncalcified plaque can be improved by combining VAT measurements and CAD risk factors.

Platelets enter atherosclerotic plaque via intraplaque microvascular leakage and intraplaque hemorrhage: A histopathological study in carotid plaques - Corrected Proof

2012-04-12

Highlights: ► We studied the presence of platelets inside carotid plaques using immunohistochemistry (CD42b). ► Platelets were observed in intraplaque hemorrhages, around plaque microvessels, mostly without leakage of erythrocytes and in mural thrombi. ► Platelet positive staining was associated with a higher plaque microvesseldensity, and elevated plaque-levels of interleukin-8.Abstract: Background: Platelets foster an inflammatory environment that influences atherosclerotic lesion progression and facilitates plaque rupture, in addition to their role in acute thrombus formation. The route of entry of platelets into the atherosclerotic plaque and their exact location inside the plaque are however not completely understood.Methods and results: 188 carotid plaques were examined for the presence of platelets using immunohistochemistry (CD42b), and 76/188 (40.4%) were platelet positive. Platelets were observed in intraplaque hemorrhages, around plaque microvessels, mostly without leakage of erythrocytes; and in mural thrombi. Platelet positive staining was associated with a higher plaque microvessel density, and elevated plaque-levels of interleukin-8.Conclusion: Due to their short life span, platelets reflect recent bleeding. It can be hypothesized that platelets might serve as a marker for leaky microvessels inside atherosclerotic plaques that are at risk for development, or progression of plaque hemorrhage.

Inhibition of hepatic scavenger receptor-class B type I by RNA interference decreases atherosclerosis in rabbits - Corrected Proof

2012-04-11

Highlights: ► Small hairpin RNA specific for SR-BI nucleotides 214-232 reduces SR-BI expression in vitro. ► SiRNA treatment reduces liver expression of SR-BI in New Zealand White rabbits. ► Liver specific inhibition of SR-BI leads to a 50% reduction of atherosclerosis in cholesterol fed rabbits. ► The role of SR-BI in rabbits may be different from the one found in rodents.Abstract: Objective: Scavenger receptor-class B type I (SR-BI), the receptor for HDL-cholesterol, plays a key role in HDL metabolism, whole body cholesterol homeostasis, and reverse cholesterol transport. We investigated the in vivo impact of hepatic SR-BI inhibition on lipoprotein metabolism and the development of atherosclerosis employing RNA interference.Methods: Small hairpin RNA plasmid specific for rabbit SR-BI was complexed with galactosylated poly-l-lysine, allowing an organ-selective, receptor-mediated gene transfer. Rabbits were fed a cholesterol-rich diet, and were injected with plasmid-complexes once a week.Results: After 2 weeks of treatment hepatic SR-BI mRNA levels were reduced by 80% accompanied by reduced SR-BI protein levels and a modulation of the lipoprotein profile. Rabbits treated with SR-BI-specific plasmid-complexes displayed higher cholesteryl ester transfer from HDL to apoB-containing lipoproteins, lower HDL-cholesterol, and higher VLDL-cholesterol levels, when compared to controls. In a long-term study, this gene therapeutic intervention led to a similar modulation of the lipoprotein profile, to lower total cholesterol levels, and most importantly to a 50% reduction of the relative atherosclerotic lesion area.Conclusion: Our results are another indication that the role of SR-BI in lipoprotein metabolism and atherogenesis in rabbits – a CETP-expressing animal model displaying a manlike lipoprotein profile may be different from the one found in rodents.

Human group X secreted phospholipase A2 induces dendritic cell maturation through lipoprotein-dependent and -independent mechanisms - Corrected Proof

2012-04-11

Highlights: ► Phospholipolyzed LDL induces DC maturation resulting in pro-Th1 cells. ► hGX-sPLA2 can directly activate DC through membrane phospholipids hydrolysis. ► LPC and FFA modulation of DC maturation depends on their fatty acid content.Abstract: Objective: Increased secreted phospholipase A2 (sPLA2) activity has been documented in several inflammatory disorders. Among sPLA2s, the human group X (hGX)-sPLA2 has the highest catalytic activity towards phosphatidylcholine (PC), the major phospholipid of cell membranes and blood lipoproteins. hGX-sPLA2 has been detected in human atherosclerotic lesions, indicating that sPLA2s are an important link between lipids and inflammation, both involved in atherosclerosis. The presence of dendritic cells (DC), the most potent antigen presenting cells, in atherosclerotic lesions has raised the question about their role in disease progression.Methods and results: In this study, we show that hGX-sPLA2-treated LDL induces human monocyte-derived DC maturation, resulting in a characteristic mature DC phenotype and enhanced DC ability to activate IFNγ secretion from T cells. hGX-sPLA2 phospholipolysis of LDL produces high levels of lipid mediators, such as lysophosphatidylcholine (LPC) and free fatty acids (FFAs), which also modulate DC maturation. The major molecular species of LPC containing a palmitic or stearic acid esterified in the sn-1 position induce DC maturation, whereas the FFAs can positively or negatively modulate DC maturation depending on their nature. hGX-sPLA2 added alone can also activate DC in vitro through the hydrolysis of the DC membrane phospholipids leading, however, to a different cytokine profile secretion pattern than the one observed with hGX-sPLA2-phospholipolysed LDL.Conclusion: hGX-sPLA2 secreted in inflamed tissues can contribute to local DC maturation, resulting in pro-Th1 cells, through the production of various lipid mediators from hydrolysis of either LDL and/or cell plasma membrane.

Advances in cell-based therapy for peripheral vascular disease - Corrected Proof

Arnon Blum, Wayne Balkan, Joshua M. Hare — 2012-04-11

Abstract: Evidence is accumulating to support cell-based therapies as a new approach for chronic diseases. Perhaps the area of greatest impact, in terms of patient numbers, is cardiovascular disorders. This review considers cell transplantation as a potential treatment for peripheral vascular disease, including ischemic stroke and erectile dysfunction. Bone marrow derived cells are required for endogenous repair in adult individuals affected by angiopathies. Clinical trials using progenitor cells generated from monocytic or non-monocytic cells indicate that both are effective, suggesting that angiogenesis is the result of cross talk between different cells and pathways. Currently, there are 14 registered clinical trials (ClinicalTrials.gov) examining different approaches to stem cell therapy to cure peripheral artery disease, of which 6 have completed enrollment. Here, we will review published clinical studies that used cell transplantation for peripheral vascular ischemic disorders.

Benefits from intracoronary as compared to intravenous abciximab administration for STEMI patients undergoing primary angioplasty: A meta-analysis of 8 randomized trials - Corrected Proof

Giuseppe De Luca, Monica Verdoia, Harry Suryapranata — 2012-04-09

Highlights: ► Abciximab has been shown to improve survival in STEMI patients undergoing primary angioplasty. ► Intracoronary administration, by providing of larger amount of drug at the occlusion, seems to be more effective as compared to intravenous administration. ► Contrasting data have been reported in small randomized trials. ► This is meta-analysis of 8 randomized trials shows that intracoronary abciximab is safe. Despite benefits in myocardial perfusion, it does not improve survival.Abstract: Background: Adjunctive abciximab administration has been demonstrated to reduce mortality and reinfarction in patients with ST-elevation myocardial infarction (STEMI) referred to invasive management. Standard abciximab regimen consists of an intravenous (IV) bolus followed by a 12-h IV infusion. Experimental studies and small clinical trials suggest the superiority of intracoronary (IC) injection of abciximab over IV route. Therefore, the aim of the current study was to perform a meta-analysis of randomized trials (RCTs) to assess the clinical efficacy and safety of IC vs IV abciximab administration in STEMI patients undergoing primary angioplasty.Methods: We obtained results from all RCTs enrolling STEMI patients undergoing primary percutaneous coronary intervention (PCI). The primary endpoint was mortality, while recurrent myocardial infarction, postprocedural epicardial (TIMI 3) and myocardial (MBG 2–3) perfusion were identified as secondary endpoints. The safety endpoint was the risk of major bleeding complications.Results: A total of 8 randomized trials were finally included in the meta-analysis, enrolling a total of 3259 patients. As compared to IV route, IC abciximab was associated with a significant improvement in myocardial perfusion (OR [95% CI]=1.76 [1.28–2.42], p<0.001), without significant benefits in terms of mortality (OR [95% CI]=0.85 [0.59–1.23], p=0.39), reinfarction (OR [95% CI]=0.79 [0.46–1.33], p=0.37), or major bleeding complications (OR [95% CI]=1.19 [0.76–1.87], p=0.44). However, we observed a significant relationship between patient's risk profile and mortality benefits from IC abciximab administration (p=0.011).Conclusions: The present updated meta-analysis showed that IC administration of abciximab is associated with significant benefits in myocardial perfusion, but not in clinical outcome at short-term follow-up as compared to IV abciximab administration, without any excess of major bleedings in STEMI patients undergoing primary PCI. However, a significant relationship was observed between patient's risk profile and mortality benefits from IC abciximab administration. Therefore, waiting for long-term follow-up results and additional randomized trials, IC abciximab administration cannot be routinely recommended, but may be considered in high-risk patients.

ApoA-1 infusion reduces arterial cholesterol and myocardial lesions in a rat model of cardiac dysfunction and insulin resistance - Corrected Proof

2012-04-09

Highlights: ► Assessed the ability of rHDL to improve vascular complications in the JCR:LA-cp rat. ► Short-term infusion of rHDL temporarily improves LV-dysfunction. ► rHDL acutely reduced arterial cholesterol deposition. ► rHDL reduced the development of early-stage myocardial lesions.Abstract: Objective: Low plasma high-density lipoprotein cholesterol (HDL-C) concentration is associated with the metabolic syndrome (MetS) and increased prevalence of cardiovascular disease (CVD). Animal and human studies report infusion of apolipoprotein A-1 (apoA-1) can reduce endothelial dysfunction, and/or induce regression of atherosclerosis. However, the direct mechanisms underlying the vascular benefits of either apoA-1 or HDL-C remain unclear. In this study, we assessed the ability of reconstituted HDL (rHDL) to improve vascular complications of MetS, including left ventricular (LV)-hypertrophy, arterial cholesterol deposition and myocardial lesion development.Methods and results: Obese insulin resistant (IR) JCR:LA-cp rats were infused with rHDL (0.4mg/kg) over 3 days before assessing cardiac function (Echocardiography) at days 7 and 50 post-infusion, as well as haematoxylin and eosin staining of myocardial lesions at day 50. Acute ex vivo arterial cholesterol deposition was assessed with acute infusion of rHDL ex-vivo. Infusion of rHDL partially corrected abnormal diastolic compliance (18%; *p<0.05) and improved parameters of cardiac function in IR rats. Further, acute rHDL infusion in carotid vessels reduced remnant lipoprotein associated-cholesterol deposition (30–86%; **p<0.01) ex vivo in IR and male Wistar rats and reduced (41%; *p<0.05) the frequency of early-stage myocardial lesions in IR rats.Conclusion: Short-term infusion of rHDL may beneficially reduce chronic vascular sequelae of MetS, including temporary improvement in LV-dysfunction, acute reduction of acute arterial cholesterol deposition and the development of early-stage myocardial lesions in the JCR:LA-cp rat.

Asymmetric dimethylarginine in adults with cystathionine β-synthase deficiency - Corrected Proof

2012-04-09

Highlights: ► Hyperhomocysteinemia is an independent risk factor for cardiovascular diseases, but the underlying mechanisms remain unknown. ► Increased ADMA levels, an endogenous nitric oxide synthase inhibitor, may contribute to the homocysteine-induced endothelial dysfunction. ► Adult CBS deficient patients have strongly elevated levels of homocysteine in plasma, but ADMA levels are within the normal range. ► The negative vascular effects of homocysteine appear to be ADMA-independent.Abstract: In hyperhomocysteinemia (HHcy), an independent risk factor for cardiovascular diseases, endothelial dysfunction due to reduced bioavailability of nitric oxide is a consistent finding. However, the underlying mechanisms remain unknown.Increased levels of the nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA) have been associated with HHcy, and may contribute, at least in part, for the homocysteine-induced endothelial dysfunction, but whether cystathionine β-synthase (CBS) deficiency is associated with increased ADMA has hardly been investigated.To address this question, we measured total homocysteine (tHcy), ADMA and symmetric dimethylarginine (SDMA) in plasma of 22 adult CBS deficient patients, using established HPLC techniques. Results showed that in CBS deficient patients with elevated levels of tHcy (median (total range): 33 (14–237)μmol/L), both ADMA and SDMA levels were normal. Moreover, tHcy and ADMA concentrations were not correlated (rs=0.017, p=0.94). Our results favor the hypothesis that the negative vascular effects of HHcy have an ADMA-independent etiology.

Pharmacological LXR activation reduces presence of SR-B1 in liver membranes contributing to LXR-mediated induction of HDL-cholesterol - Corrected Proof

2012-04-05

Highlights: ► In mice, induction of fecal neutral sterol secretion by the pharmacological liver X receptor (LXR) ligand T0901317 critically depends on LXRα but not on LXRβ. ► Pharmacological LXR activation results in a time-dependent raise in plasma HDL cholesterol-concentrations with the appearance of more and larger HDL particles. ► LXR activation reduces SR-B1 in hepatic membranes of mice, probably contributing to elevated HDL cholesterol. ► SR-B1 is not required to enhance fecal neutral sterol secretion upon pharmacological LXR activation in mice.Abstract: Objective: Pharmacological LXR activation has anti-atherosclerotic actions in animal models. Part of these beneficial effects may be explained by accelerated reverse cholesterol transport since both plasma high density lipoprotein (HDL) cholesterol and fecal neutral sterol secretion are higher upon LXR activation. Mechanisms underlying these LXR-mediated effects have not been fully elucidated.Methods: We investigated the roles of the isoforms LXRα and LXRβ and the HDL cholesterol uptake receptor SR-B1 in modulation of cholesterol metabolism upon treatment of mice with the LXR ligand T0901317.Results: HDL cholesterol was maximally 60% increased in a time-dependent fashion due to appearance of more and larger HDL particles. Fecal neutral sterol secretion was maximally induced after 1week treatment. T0901317 treatment induced fecal neutral sterol secretion by ∼300% in wild-type but not in Lxrα deficient mice. Surprisingly, LXR activation reduced SR-B1 protein amount in hepatic membranes, suggesting that this might contribute to elevated HDL cholesterol. However, T0901317 still elevated plasma HDL cholesterol in Sr-b1 deficient mice, suggesting that SR-B1 is not the only step involved in LXR-mediated induction of plasma HDL cholesterol. In addition, SR-B1 is not essential for LXR-induced cholesterol removal from the body.Conclusion: Induction of fecal neutral sterol secretion by T0901317 critically depends on LXRα but not on LXRβ. LXR activation reduces SR-B1 in hepatic membranes, probably partly contributing to elevated HDL cholesterol. SR-B1 is not required to enhance fecal neutral sterol secretion.

Elevated level of C-reactive protein is associated with risk of prediabetes in Indians - Corrected Proof

2012-04-05

Highlights: ► We investigated association of hsCRP levels with Impaired Fasting Glucose (IFG) and Impaired Glucose Tolerance (IGT) in 1,726 Indians. ► We determined whether relationship is influenced by risk factors like obesity, lipid profile, hypertension, family history of diabetes. ► Elevated hsCRP level is found to be associated with the risk of IFG and IGT in Indians. ► The association of hsCRP with prediabetes is independent of the effect of traditional risk factors of hyperglycemia. ► Our study supports the role of inflammation in the pathogenesis of hyperglycemia.Abstract: Background: Relationship of high sensitivity C-reactive protein (hsCRP) with prediabetes has not been explored extensively in Indians. Here we sought to investigate the association of hsCRP levels with prediabetes, as represented by impaired fasting glucose (IFG) and impaired glucose tolerance (IGT), and the influence of risk factors like obesity, decreased HDL cholesterol, hypertension, family history of diabetes and current smoking habit on the relationship.Methods: A cross-sectional study on 1726 Indians, comprising of 1276 individuals with normal glucose tolerance (NGT), 250 IFG and 200 IGT individuals. Subjects were defined according to WHO criteria based on fasting plasma and 2h glucose levels.Results: Median levels of hsCRP were significantly higher in IFG (2.20mg/l) and IGT (2.32mg/l) compared to NGT (1.64mg/l) subjects. Individuals with high risk hsCRP levels (>3mg/l) had an odds ratio (OR) (95% confidence interval (CI)) of 2.60 (1.56–5.34) [P=1.3×10−4] for IGT after adjusting the effect of age, sex, medication, body mass index (BMI), waist circumference (WC) and risk factors like decreased high-density lipoprotein cholesterol (HDL-cholesterol), hypertension, family history of diabetes and current smoking. Significant increase in risk of IGT was found with a unit increase in natural log transformed hsCRP levels after adjustment for covariates [OR (95%CI)=1.57 (1.27–1.94), P=3.0×10−5]. When subjects were stratified on the basis of risk factors, we found stronger association of elevated hsCRP levels with risk of IFG and IGT in subjects having HDL-cholesterol ≤50mg/dl and with hypertension.Conclusions: Our study demonstrates that elevated hsCRP levels are independently associated with risk of IFG and IGT in Indians.

ω-6 lipids regulate PPAR turnover via reciprocal switch between PGC-1 alpha and ubiquitination - Corrected Proof

Jia Fei, Carla Cook, Nalini Santanam — 2012-04-05

Highlights: ► Dietary ω-6 lipids exhibit both pro- and anti-atherogenic effects on vascular cells. ► ω-6 lipids are biological ligands peroxisome proliferators-activated receptors (PPAR). ► ω-6 lipids modulates PPAR turnover and transactivity in a ligand and time dependent mechanism. ► ω-6 lipids modulates reciprocal interaction of PPARs between PGC-1 and ubiquitination.Abstract: Objective: Dietary ω-6 lipids such as linoleic acid and its oxidized forms (13-HPODE OxLA) interact with peroxisome proliferator-activated receptors (PPARs) and elicit pro and anti-atherogenic effects in vascular cells. Ligand-dependent PPAR protein turnover is promoted by ubiquitination, but attenuated by binding to its co-activator, peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1α). The objective of our study was to investigate if the dual atherogenic effects of ω-6 lipids are due to its regulation of PPAR turnover.Methods and results: In rat aortic smooth muscle cells (RASMCs), oxidized linoleic acid (OxLA) at 10–50μM induced and stabilized PPARα protein at earlier time points (0–4h) but suppressed it at 12h. Conversely, it activated PPARγ protein turnover at a later time point (12h). Pre-treatment with the proteasome inhibitor (MG132) prevented OxLA mediated loss of PPAR stability and transactivity. Co-immunoprecipitation studies indicated a ligand mediated time-dependent reciprocal exchange of PPAR interaction between ubiquitination and PGC-1α. This ω-6 lipid mediated time-dependent switch between PPAR degradation versus stability helped modulate the pro and anti-atherogenic effects of these dietary lipids.Conclusion: Our findings provide insights into the dual pro and anti-atherogenic effects of dietary ω-6 lipids on vascular cells by the regulation of PPAR turnover.

Apolipoprotein E gene mutations in subjects with mixed hyperlipidemia and a clinical diagnosis of familial combined hyperlipidemia - Corrected Proof

2012-04-05

Highlights: ► Rare APOE mutations are responsible for approximately 3.5% of FCHL cases in our population. ► APOE R136S mutation induces autosomal dominant familial dysbetalipoproteinemia. ► APOE p.Leu149del induces a phenotype indistinguishable from familial combined hyperlipidemia.Abstract: Objective: Rare mutations in the APOE gene, undetectable with the usual genotyping technique, are responsible for dominant familial dysbetalipoproteinemia (FD) and therefore could be easily misclassified as familial combined hyperlipidemia (FCHL). We aimed to identify APOE mutations associated with dominant combined hyperlipoproteinemia and to establish their frequency in subjects with a clinical diagnosis of FCHL.Methods and results: In 279 unrelated subjects with FCHL in whom a functional LDLR mutation was excluded, sequencing of the entire APOE gene detected 9 carriers of a rare mutation: 5 subjects (1.8%) with the R136S mutation (arginine at residue 136 changed to serine) and 4 subjects (1.4%) with the p.Leu149del mutation, a 3-bp inframe deletion that results in the loss of leucine at position 149. Both genetic defects were detected with similar frequency (2.5% and 1.3%, respectively) in an independent group of 160 FCHL subjects from other locations in Spain. Family studies demonstrated cosegregation of these APOE mutations with hyperlipoproteinemia. R136S carriers showed dysbetalipoproteinemia, while the lipid phenotype of p.Leu149del carriers was IIa or IIb.Conclusions: Rare APOE mutations are responsible for approximately 3.5% of FCHL cases in our population. APOE R136S and p.Leu149del induce autosomal dominant FD and a phenotype indistinguishable from FCHL, respectively.

Reconsidering metabolic diseases: The impacts of persistent organic pollutants - Corrected Proof

Jérôme Ruzzin, Duk-Hee Lee, David O. Carpenter, David R. Jacobs — 2012-04-03

Taking effective action against type 2 diabetes and obesity has remained an enormous challenge. Latest estimates indicate that type 2 diabetes is globally affecting at least 366 million people, and that around 500 million adults are obese . Alarmingly, over 20% of US children aged 2–5years are overweight or obese . Better understanding of the causes of these diseases is essential for both prevention and treatment .

Herpes simplex virus type 2 (HSV-2) as a coronary atherosclerosis risk factor in HIV-infected men: Multicenter AIDS Cohort Study - Corrected Proof

2012-04-03

Highlights: ► We assessed associations between herpesviruses and subclinical coronary atherosclerosis among HIV-infected men. ► Coronary artery calcium (CAC) was measured by non-contrast coronary CT imaging to evaluate coronary atherosclerosis. ► This is the first report suggesting herpes simplex virus type 2 (HSV-2) may be a risk factor for coronary atherosclerosis in HIV-infected men. ► Infection with multiple herpesviruses may contribute to the increased burden of atherosclerosis.Abstract: We assessed associations of herpes simplex virus types 1 and 2 (HSV-1 and -2), cytomegalovirus (CMV), and human herpesvirus 8 (HHV-8) infection with subclinical coronary atherosclerosis in 291 HIV-infected men in the Multicenter AIDS Cohort Study. Coronary artery calcium (CAC) was measured by non-contrast coronary CT imaging. Markers for herpesviruses infection were measured in frozen specimens collected 10–12 years prior to case identification. Multivariable logistic regression models and ordinal logistic regression models were performed. HSV-2 seropositivity was associated with coronary atherosclerosis (adjusted odds ratio [AOR]=4.12, 95% confidence interval [CI]=1.58–10.85) after adjustment for age, race/ethnicity, cardiovascular risk factors, and HIV infection related factors. Infection with a greater number of herpesviruses was associated with elevated CAC levels (AOR=1.58, 95% CI=1.06–2.36). Our findings suggest HSV-2 may be a risk factor for subclinical coronary atherosclerosis in HIV-infected men. Infection with multiple herpesviruses may contribute to the increased burden of atherosclerosis.

Non-linear association between ankle-brachial pressure index and prevalence of silent cerebral infarction in Japanese patients with type 2 diabetes - Corrected Proof

2012-03-29

Highlights: ► We examined the association between ABI abnormality and SCI in diabetic patients. ► Low and high ABI had greater risks of prevalent SCI as compared with normal ABI. ► The association was unchanged even after adjusting for cardiovascular risk factors. ► Low and high ABI have predictive values of prevalent SCI in diabetic patients.Abstract: Objective: Patients with peripheral artery disease (PAD), defined as having low ankle-brachial pressure index (ABI), have increased risk for incident stroke compared with those without PAD. We aimed to reveal whether ABI abnormality, especially high ABI is associated with prevalent silent cerebral infarction (SCI) in type 2 diabetic patients.Methods: We studied 538 Japanese type 2 diabetic patients, 227 women and 311 men, with a mean [±SD] age of 64±11 years. All patients underwent cranial magnetic resonance imaging (MRI). Values of ABI were classified as low (<0.9), normal (0.9≤ and <1.3), and high (1.3≤). Logistic regression model was used to calculate odds ratio and 95% confidence interval (95% CI) for prevalent SCI.Results: The mean ABI among the overall 538 patients was 1.09±0.16. Low and high ABI values were found in 52 (9.7%) and 33 (6.1%) patients, respectively. SCI was detected in 297 (55.2%) patients. The prevalence in patients with low, normal, and high ABI values were 88.5%, 49.7%, and 78.8 (p<0.001), respectively. In the multivariate logistic regression analysis, both patients with high and low ABI were significantly increased risk of prevalent SCI (odds ratio 4.53, 95% CI 1.67–12.34, p=0.003 and odds ratio 3.50, 95% CI 1.50–10.29, p=0.005), independently of other traditional cardiovascular risk factors, than those with normal ABI.Conclusions: Both high and low ABI may be strongly associated with prevalent SCI in Japanese patients with type 2 diabetes.

Different prognostic value of white blood cell subtypes in patients with acute cerebral infarction - Corrected Proof

2012-03-29

Highlights: ► Neutrophil and lymphocyte counts, which were measured at admission in patients with acute cerebral infarction, had different predictive values. ► Higher total WBC and neutrophil counts were associated with more severe stroke at admission. ► Lower lymphocyte counts were associated with poor neurologic improvement in early phase and worse long-term functional outcome.Abstract: Objective: We aimed to investigate the relationship of each white blood cells (WBC) subtype with neurologic severity and outcome in acute stroke.Methods: We included 779 patients with first-ever acute cerebral infarction within 72h after symptom onset. We investigated the association between counts for WBC subtypes in peripheral blood at admission and (1) initial stroke severity; (2) early change in stroke severity within one week; and (3) functional outcome at three months.Results: Higher total WBC and neutrophil counts were associated with more severe stroke at admission (p<0.001). In contrast, lower lymphocyte counts were associated with a lesser improvement during the first week after admission (p<0.05) and with poor functional outcome at three months (OR=0.706 per 1000 lymphocyte counts/mm3, p=0.020).Conclusions: Our study merits further investigation on the role of each WBC subtype in ischemic injury and different prognostic value of WBC subtypes measured at admission in acute stroke.

Detection of subclinical atherosclerosis in familial hypercholesterolemia using non-invasive imaging modalities - Corrected Proof

2012-03-29

Highlights: ► Despite treatment FH patients have higher atherosclerotic burden than controls. ► Aortic MRI allows to characterize atherosclerotic plaques in FH patients. ► In FH patients there is a correlation between aortic wall volume and cIMT. ► Lipid plaques in FH were associated with coronary artery disease in relatives.Abstract: Objectives: To investigate the extent of subclinical atherosclerosis in asymptomatic familial hypercholesterolemia (FH) patients using non-invasive images techniques.Patients, methods and results: The atherosclerotic burden of 36 molecularly defined FH patients (18 males, 45.7±10.9 years) without evidence of cardiovascular disease receiving lipid-lowering treatment and 19 (47.8±11.3 years) controls was investigated. Descending thoracic aorta magnetic resonance imaging (MRI) was performed in a 1.5T equipment with T1 and T2 sequences to characterize atherosclerotic plaques and to measure aortic wall volumen. Carotid intima-media thickness (cIMT) and presence of plaques were measured using B-mode carotid ultrasound.Mean aortic wall volumen, cIMT and atherosclerotic plaques in aorta were significantly higher in FH cases (P<0.001). A significant correlation between aortic wall volume and cIMT was observed (P<0.01). Aortic MRI detected plaques in 94% and carotid ultrasound in 14% of cases. Lipid-rich plaques were observed only in FH cases (33%) and were associated with family history of premature coronary artery disease (P<0.05).Conclusions: Asymptomatic middle-aged FH patients have significantly higher atherosclerotic burden than controls. cIMT has shown a significant correlation with aortic wall volume and MRI allowed the detection of lipid-rich plaques in FH subjects that were associated with family history of premature coronary artery disease.

The role of endoglin in atherosclerosis - Corrected Proof

Petr Nachtigal, Lenka Zemankova (Vecerova), Jana Rathouska, Zbynek Strasky — 2012-03-29

Abstract: Endoglin (CD 105, TGF-β receptor III) is a homodimeric transmembrane glycoprotein that plays a regulatory role in TGF-β signaling. Its functional role in the context of atherosclerosis has yet to be defined and should be stated here. Therefore, we focused on the role of endoglin in atherosclerosis in both humans and experimental animals. Endoglin expression was demonstrated in atherosclerotic vessels predominantly in endothelial cells and smooth muscle cells in various types of blood vessels in mice and humans, suggesting its participation in atherogenesis. Endoglin expression was also related to the expression of eNOS in endothelium, repair of the vessel wall, plaque neoangiogenesis, production of collagen and stabilization of atherosclerotic lesions. In addition, increased levels of soluble endoglin were associated with hypercholesterolemia, atherosclerosis, acute myocardial infarction and were related to inhibition of TGF-β signaling in the vessel wall. Moreover, soluble endoglin levels were significantly lowered after a series of extracorporeal eliminations in patients with familial hypercholesterolemia. Additionally, statin treatment decreased levels of soluble endoglin and increased its expression in aorta, which was related to reduced atherosclerosis in mice. In conclusion, we propose that measurement of soluble endoglin might give information about progression of the atherosclerotic process or the efficacy of therapeutic interventions, which is the task that must be answered in clinical trials.

Chronic inflammation, albuminuria, and functional disability in older adults with cardiovascular disease: The National Health and Nutrition Examination Survey, 1999–2008 - Corrected Proof

Hsu-Ko Kuo, Soham Al Snih, Yong-Fang Kuo, Mukaila A. Raji — 2012-03-29

Highlights: ► Inflammation and albuminuria independently correlate with disability in older adults with cardiovascular disease (CVD). ► There is a joint effect of inflammation and albuminuria on multiple domains of disability and metabolic risks. ► Albuminuria may amplify the effect of inflammation on disability in older adults with CVD.Abstract: Objective: Although C-reactive protein (CRP) and albuminuria are well-documented cardiovascular risk markers, the functional implications of these biomarkers and their combination on functional disability and metabolic risks in patients with cardiovascular disease (CVD) are unknown.Methods: Data were from 1403 adults (≥60 years, mean 73.2 years) with CVD, ascertained by self-reported diagnosis of angina, coronary heart disease, congestive heart failure, myocardial infarction or stroke, in the National Health and Nutrition Examination Survey 1999–2008. Disability in activities of daily living (ADL), instrumental activities of daily living (IADL), leisure and social activities (LSA), general physical activities (GPA), and lower-extremity mobility (LEM) were obtained from self-reports. The urinary albumin-to-creatinine ratio (UACR) was calculated by dividing the urinary albumin value by the urinary creatinine concentration. CRP levels were quantified by latex-enhanced nephelometry.Results: Inflammation and albuminuria were associated with disability. In the full-adjusted models, odds ratios (ORs) (95% confidence intervals [CIs]) of disability in ADL, LSA, and LEM were 1.60 (1.13–2.28), 1.76 (1.22–2.55) and 2.31 (1.62–3.31), respectively, comparing participants in the highest CRP quartile to the lowest (p values for trend across CRP quartiles<0.01). The corresponding ORs (95% CI) for disability in ADL, IADL, LSA, and LEM were 1.71 (1.20–2.45), 1.72 (1.21–2.45), 1.46 (1.01–2.12) and 2.50 (1.73–3.62), respectively, comparing participants in the highest UACR quartile to the lowest. We found combined association of inflammation and albuminuria with disability and with metabolic risks. Based on medians of both UACR and CRP, subjects with both higher levels of both markers had higher odds of disability and a more unfavorable metabolic profile than those with lower levels.Conclusions: Elevated levels of CRP and UACR independently correlate with disability among older adults with CVD. There is a combined association of inflammation and albuminuria on multiple domains of disability and metabolic risks, suggesting the presence of elevated UACR may amplify the association of inflammation with disability and with metabolic risk in older adults living with CVD.

Inflammation modulates human HDL composition and function in vivo - Corrected Proof

2012-03-28

Highlights: ► Endotoxin challenge in humans results in marked alterations in HDL particle composition with reduced phospholipids and increased serum amyloid A but without significant change in cholesterol of apo-AI. ► Endotoxin induced selective remodeling of HDL particles with induction of specific HDL lipases and reductions in CETP mass and LCAT activity. ► HDL efflux function was reduced after endotoxin challenge with reduced capacity of particles to mediate efflux via ABCA1 and SR-BI cholesterol transporter pathways; reduced efflux correlated with alterations in HDL composition and reduction in specific HDL sub-populations.► Overall, these data support the concept that atherogenic HDL dysfunction and impaired RCT occur in human inflammatory syndromes, independent of significant change in plasma HDL-C levels.Abstract: Objectives: Inflammation may directly impair HDL functions, in particular reverse cholesterol transport (RCT), but limited data support this concept in humans.Methods and results: We employed low-dose human endotoxemia to assess the effects of inflammation on HDL and RCT-related parameters in vivo. Endotoxemia induced remodelling of HDL with depletion of pre-β1a HDL particles determined by 2-D gel electrophoresis (−32.2±9.3% at 24h, p<0.05) as well as small (−23.0±5.1%, p<0.01, at 24h) and medium (−57.6±8.0% at 16h, p<0.001) HDL estimated by nuclear magnetic resonance (NMR). This was associated with induction of class II secretory phospholipase A2 (∼36 fold increase) and suppression of lecithin:cholesterol acyltransferase activity (−20.8±3.4% at 24h, p<0.01) and cholesterol ester transfer protein mass (−22.2±6.8% at 24h, p<0.001). The HDL fraction, isolated following endotoxemia, had reduced capacity to efflux cholesterol in vitro from SR-BI and ABCA1, but not ABCG1 transporter cell models.Conclusions: These data support the concept that “atherogenic-HDL dysfunction” and impaired RCT occur in human inflammatory syndromes, largely independent of changes in plasma HDL-C and ApoA-I levels.

Effect of everolimus on pre-existing atherosclerosis in LDL-receptor deficient mice - Corrected Proof

2012-03-27

Highlights: ► Animal study investigating mTOR-inhibition for treatment of advanced atherosclerosis. ► Everolimus reduced inflammatory cell content locally but pre-existing atherosclerosis progressed obstructively. ► Anti-atherosclerotic efficiency of pharmacotherapeutic mTOR-inhibition is suggested stage-dependent. ► Higher potential might be obscured by concomitant hypercholesterolemia.Abstract: Objective: Proliferation signal inhibitors/mTOR-inhibitors have been shown to reduce de novo development of hypercholesterolemic atherosclerosis in animal models. However, their effect on pre-existing atherosclerosis has not yet been studied.Methods and results: Feeding LDL-R-KO mice a high cholesterol diet for 12 weeks resulted in formation of moderate fibroatheroma (induction phase). Sixty mice received either everolimus (1 or 5mg/kg) or no everolimus for further 12 weeks (treatment phase). Everolimus significantly enhanced hypercholesterolemia (plasma cholesterol +45%, p<0.001). Atherosclerosis progressed obstructively in treated and non-treated mice. Everolimus (5mg/kg) tended to reduced progression in aortic root lesions (0.28±0.02 vs. 0.33±0.03mm2, p=ns) and brachiocephalic lesions (0.044±0.006 vs. 0.066±0.012mm2, p=ns) but without significance. Everolimus (5mg/kg) resulted in an arrest of CD68 positive plaque area (p=0.03) and nearly halved CD68 fraction (p=0.05) in aortic root lesions but not in brachiocephalic lesions. Taken together, despite a trend to reduced progression and inflammatory cell content there was less conclusive net effect of everolimus treatment than expected.Conclusion: A higher potential of everolimus in the treatment of atherosclerosis might be obscured by its concomitant hypercholesterolemia. Considering stronger effects in previous studies we suggest that everolimus might exert more potent anti-atherogenic properties in earlier stages of atherogenesis than in advanced atherosclerosis.

Lipid-altering efficacy of ezetimibe plus statin and statin monotherapy and identification of factors associated with treatment response: A pooled analysis of over 21,000 subjects from 27 clinical trials - Corrected Proof

2012-03-14

Abstract: Objective: Patients with dyslipoproteinemia constitute the largest risk group for development of atherosclerosis and cardiovascular disease (CVD). Despite extensive statin use, many patients with CVD risk do not achieve guideline-recommended low-density lipoprotein cholesterol (LDL-C) targets. This pooled analysis of 27 previously published clinical trials conducted between 1999 and 2008 evaluated the lipid-altering efficacy and factors related to treatment response of ezetimibe combined with statin and statin monotherapy.Methods: Patient-level data were combined from double-blind, placebo-controlled or active comparator studies randomizing adult subjects to ezetimibe 10mg plus statin (n=11,714) versus statin alone (n=10,517) for 6–24 weeks (mean=9 weeks). Association of factors with treatment response, percent change from baseline LDL-C and other lipids, and attainment of guideline-recommended lipid and lipoprotein targets were evaluated.Results: Higher baseline LDL-C, diabetes mellitus, Black race, greater age, and male gender were associated with small but significantly greater percent reductions in LDL-C regardless of treatment. Treatment influenced efficacy, with ezetimibe plus statin producing significantly greater reductions in LDL-C, total-cholesterol, non-HDL-C, ApoB, triglycerides, lipid ratios, hs-CRP; significantly larger increases in HDL-C and ApoA1; and significantly higher achievement of LDL-C (<70mg/dl, <100mg/dl), non-HDL-C (<100mg/dl, <130mg/dl), and ApoB (<80mg/dl, <90mg/dl) targets than statin monotherapy at statin potencies compared (p<0.0001 for all). Differential treatment effects were seen with first-/second-line therapy and statin potency.Conclusion: These results suggest that patient characteristics have a limited influence on response to lipid-lowering therapy and demonstrate the consistent treatment effect of ezetimibe combined with statin and statin monotherapy across a diverse patient population.