Faculty Bibliography

Ultrasound assessment of vascular biomarkers has been implemented for screening, prevention and improvement of cardiovascular risk stratification beyond classical risk factors including smoking, diabetes, hypercholesterolemia and hypertension. Thus, the presence of vascular damage at the sub-clinical, asymptomatic stages can identify a "vulnerable" patient, and aid in implementing cardiovascular prevention strategies. Increased intima-media thickness of the common carotid artery is a well-known marker of early atherosclerosis, which significantly correlates with the development of coronary or cerebro-vascular disease. More recently, guidelines for cardiovascular prevention in hypertension also introduced other vascular parameters evaluating both mechanical and functional arterial properties of peripheral arteries. Increased arterial stiffness, which can be detected by ultrasound at the common carotid, has been shown to predict future cardiovascular events and it is already considered a subclinical target organ of hypertensive patients. Even earlier vascular abnormalities such as endothelial dysfunction in the peripheral arteries, detected as reduced flow-mediated dilation of the brachial artery by ultrasound, have also been mentioned for their possible clinical use in the future. This manuscript reviews clinical evidence supporting the use of these different vascular markers for cardiovascular risk stratification, focusing on the need for an accurate, robust and reliable methodology for the assessment of vascular markers, which could improve their predictive value and increase their use in routine clinical practice.

OBJECTIVE:To evaluate local carotid stiffness (CS) and intima-medial thickness (C-IMT) in hypertensive patients with different cardiovascular risk profile, using a new user-friendly ultrasound-based system, previously validated vs. RF-based echotracking device. METHODS:We investigated a population with different cardiovascular risk: 45 healthy normotensives (NT), 90 non-diabetic hypertensives (HT), and 48 patients with hypertension and type-2 diabetes (DM). Framingham risk factor score (FRS) was calculated. PWV was assessed by applanation tonometry. The relative stroke change in diameter (ΔD) and C-IMT were measured on carotid scans. Distensibility coefficient (DC) was calculated as ΔA/(A*ΔP), where A = diastolic lumen area, ΔA = stroke change in lumen area, and ΔP = carotid pulse pressure. CS (m/s) was calculated as (ρ*DC) - 1/2 (ρ = blood density). RESULTS:CS, C-IMT, PWV were significantly increased in HT and DM vs. NT. C-IMT and PWV were significantly higher in DM than HT. ΔD and DC were significantly lower in HT and DM vs. NT. FRS ≥10% group showed increased carotid diameter, C-IMT and CS than the FRS <10%. FRS was (p < 0.001) correlated with CS (r = 0.35); ΔD (r = -0.36), DC (r = 0.35), C-IMT (r = 0.48), PWV (r = 0.38). CS correlated (p < 0.05) with PWV in the entire population (r = 0.37), in the NT (r = 0.35), in the HT and DM (r = 0.20). PWV (r = 0.50) and CS (r = 0.33) were correlated with age. Determinants of aortic and carotid stiffness were identified by multivariate stepwise analysis. CONCLUSIONS:The proposed B-mode ultrasound-based system is a reliable and user-friendly method that could serve to investigate the predictive value of CS for cardiovascular events in future large clinical studies.

OBJECTIVE:The aim of this study was to compare the effects of HDL(Milano) and HDL(wild-type), on regression and stabilization of atherosclerosis. METHODS:Atherosclerotic New Zealand White rabbits received 2 infusions, 4 days apart, of HDL(Milano) (75mg/kg of apoA-I(Milano)), HDL(wild-type) (75mg/kg apoA-I(wild-type)) or placebo. Pre- and post-treatment plaque volume was assessed by MRI. Markers of plaque vulnerability and inflammation were evaluated. Liver and aortic cholesterol content, aortic ABCA-1 and liver SR-BI were quantified. The effect of apoA-I Milano and wild-type proteins on MCP-1 and COX-2 expression by macrophages was evaluated in vitro. RESULTS:Both forms of HDL induced aortic plaque regression (-4.1% and -2.6% vs. pre-treatment in HDL(Milano) and HDL(wild-type) respectively, p<0.001 and p=0.009). A similar reduction in cholesterol content of aorta and liver was observed with both treatments vs. placebo. The expression of aortic ABCA-1 and hepatic SR-BI was significantly higher in both treated groups vs. placebo. A significantly reduced plaque macrophage density was observed in the HDL(Milano) vs. both HDL(wild-type) and placebo groups. Plaque levels of COX-2, MCP-1, Caspase-3 antigen and MMP-2 activity were significantly reduced in the HDL(Milano) vs. both HDL(wild-type) and placebo groups. In vitro studies showed that apoA-I(Milano) protein significantly reduced expression of COX-2 and MCP-1 in oxLDL loaded macrophages vs. apoA-I(wild-type). CONCLUSIONS:Despite a similar effect on acute plaque regression, the infusion of HDL(Milano) exerts superior anti-inflammatory and plaque stabilizing effects than HDL(wild-type) in the short term.

AIMS/OBJECTIVE:The aim of this study was to investigate the effects of liver X receptors (LXRs)-β preferential activation by LXR-623 (WAY-252623), a novel LXRs agonist, on plaque progression/regression in a rabbit model of advanced atherosclerosis. METHODS AND RESULTS/RESULTS:Advanced atherosclerosis was induced in New Zealand White Rabbits (n= 41). At the end of atherosclerosis induction, animals underwent a baseline magnetic resonance imaging (MRI) and were randomized to receive LXR-623 (1.5, 5, or 15 mg/kg/day), simvastatin (5 mg/kg/day), or placebo. The combination of LXR-1.5/simvastatin was also tested. After a final MRI, animals were euthanized and their aortas processed for further analysis. Simvastatin significantly reduced lesion progression (-25%; P< 0.01) in comparison with the placebo group. A similar effect was observed in the LXR-1.5 and -5 groups. A significant regression (16.5%; P< 0.01) of existing atherosclerosis was observed in the LXR-1.5/simvastatin group. Histological and molecular analysis showed plaque stabilization in the animals treated with the LXR-1.5 and -5, and LXR-1.5/simvastatin. The effects of LXR-623 were observed in the presence of a non-significant effect on total-cholesterol, low-density lipoproteins-cholesterol, and triglyceride levels. CONCLUSION/CONCLUSIONS:The results of the present study show that LXR-623 significantly reduces the progression of atherosclerosis and induces plaque regression in combination with simvastatin. These observations could drive future development of novel anti-atherosclerotic therapeutic approaches.

BACKGROUND:The structural secuelae of acute myocardial infarction (AMI) is mostly dictated by left ventricular (LV) remodelling, leading to heart failure. Monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play a critical role in LV remodelling. β-blockers are first line therapy for AMI and heart failure; however, the mechanisms responsible for their benefits remain poorly understood. Different β-blocker agents have been shown to exert beneficial activities both in AMI and heart failure, however, their role in early remodelling after ischemia/reperfusion is to be fully elucidated. We sought to compare the effect of 2 of the most prescribed β-blocker agents in early markers of LV remodelling after AMI. METHODS:A reperfused AMI was induced in Yorshire pigs, being randomized to early intravenous carvedilol, metoprolol or placebo. Twenty-four hours after reperfusion markers of early remodelling were addressed in the LV. RESULTS:The early administration of both β-blockers is able to significantly reduce macrophage infiltration as well as the expression and activity of MCP-1 and MMP-2 compared to placebo. The effects of carvedilol were much stronger than those of metoprolol. Conversely, carvedilol upregulated the expression TIMP-2 to a greater extent than metoprolol. CONCLUSIONS:In an AMI model closely mimicking human pathophysiology, the early administration of carvedilol reduced the expression of markers associated with early LV remodelling to greater extent than metoprolol. These findings may explain the superior clinical benefits exerted by carvedilol in heart failure.

Cardiovascular disease (CVD) is currently the second most frequent cause of death (after cancer) among HIV-positive subjects. The clinical use of highly active antiretroviral therapy (HAART) has dramatically reduced mortality and morbidity in HIV-positive population, leading to prolonged and improved quality of life. However, as mortality and morbidity from AIDS-related conditions improve, CVD assumes increasing magnitude. It is estimated that by 2015 more than 50% of HIV-positive patients will be older than 50 years. Since age is a major unmodifiable cardiovascular risk factor, the risk for CVD in this population will significantly and progressively increase in the near future. A large part of the risk for cardiovascular events appears to be a result of lipid abnormalities characterizing HIV-positive persons. This review focuses on HIV-associated lipid abnormalities and CVD. Lipid abnormalities may be related to either viral infection, HAART or both. Dyslipidemia characterizing HIV-infected patients has become a therapeutic target to reduce cardiovascular risk of HIV-treated patients. HAART-treated patients show an atherogenic lipid profile comprised of low HDL-cholesterol levels, hypertrigliceridemia and increased levels of small-LDL particles. Current guidelines for the treatment of dyslipidemia and reducing cardiovascular risk in HIV-positive patients suggest that when lifestyle modifications (i.e., diet and exercise) and switching antiretroviral therapy are not enough, statins should be the first-line therapy for dyslipidemia. HDL raising interventions (niacin and fibrates) should be considered to raise HDL levels and lower triglyceride in HIV-infected patients. Implications of lipid-related interventions in HIV-treated patients to avoid drug interactions and their adverse effects are also discussed.

Reduction of low-density lipoprotein cholesterol by statin therapy has only modestly decreased coronary heart disease (CHD)-associated mortality in developed countries, which has prompted the search for alternative therapeutic strategies for CHD. Epidemiologic and interventional studies have clearly established an inverse association between plasma levels of high-density lipoprotein (HDL) cholesterol and incidence of atherosclerosis. The atheroprotective benefits of HDL are not only dependent on HDL concentrations (quantity), but also on HDL function (quality). Therefore, several techniques have been recently developed to assess the different properties of HDL. Because reverse cholesterol transport (RCT) is considered a key player in the beneficial action of HDL, this review focuses on the different methods used to evaluate cholesterol efflux. Measuring the in vivo function of HDL could be of significant importance for both the clinical evaluation of an individual patient and to evaluate the effectiveness of different RCT-enhancing therapeutic approaches.

Apolipoprotein A-I (ApoA-I)/high-density lipoprotein (HDL)-raising treatments are effective antiatherosclerotic strategies. We have compared the antiatherogenic effects of human ApoA-I (hApoA-I) overexpression by intraportal and intramuscular gene transfer in atherosclerotic ApoE-knockout mice. Atherosclerotic lesions were induced by atherogenic diet. After atherosclerosis induction, a group of animals was killed and served as atherosclerosis baseline-control group. The remaining animals were randomized into the following groups: (1) atherosclerosis-progression-control, (2) intraportal/vector administration, and (3) intramuscular/vector administration. Aortas and hearts were processed for atherosclerotic quantification by en face Sudan IV and Oil Red-O, respectively. Liver and muscle specimens were processed for protein/gene expression analysis. A sustained increase in hApoA-I/HDL plasma levels was observed in both transduced groups. hApoA-I overexpression abolished plaque progression versus progression-control group. hApoA-I overexpression significantly reduced lesion macrophage, feature indicative of plaque stabilization. Scavenger receptor class-B type I (SR-BI), but not ATP-binding cassette, sub-family A (ABCA), member 1 (ABCA-1), was significantly upregulated in treated groups versus progression-controls. The results of this study show a similar effect of hApoA-I/HDL overexpression on plaque progression/stabilization by 2 different routes of administration. Our results showing similar effects using either intramuscular administration and intraportal route of administration may have significant clinical implications, given the reduced medical risk to patient and cost of intramuscular injections.

OBJECTIVES/OBJECTIVE:The aims of this study were to investigate the feasibility of contrast-enhanced ultrasound (CEU) imaging for in vivo visualization of intraplaque neovascularization and to correlate the in vivo observations with histological assessment of neovessel density and plaque composition in an experimental animal model of advanced atherosclerosis. BACKGROUND:Recent evidence has linked plaque angiogenesis with enhanced atherosclerotic plaque progression and vulnerability. Increased neovascularization has been detected in ruptured human lesions and is associated with clinical manifestations of plaque rupture. METHODS:Advanced aortic atherosclerosis was induced in New Zealand white rabbits (n = 21; high cholesterol-rich diet/double-balloon aortic denudation). Animals underwent standard and CEU imaging at the end of the atherosclerosis induction period. Six age-matched animals served as control subjects. Within 24 h, animals were euthanized and aortas processed for histopathological evaluation of plaque composition and neovascularization. Imaged plaques were classified as contrast enhanced (CE) positive or CE negative, according to their contrast enhancement on CEU imaging. The lesions were also classified as class III (predominantly echogenic) or class II (predominantly echolucent), according to their echogenicity on non-CEU images. RESULTS:No contrast enhancement was observed in control animals. In atherosclerotic animals, class III lesions showed an increased contrast enhancement compared with class II lesions and CE-positive lesions showed greater neovascularization than CE-negative plaques. Macrophage density, but not smooth muscle cell density, was significantly higher in CE-positive than CE-negative lesions. As expected, class III lesions showed increased macrophage density compared with class II plaques. Intraplaque neovessel density at histology was significantly higher in CE-positive than in CE-negative lesions. Class III plaques showed a significantly higher neovessel density compared with class II lesions. A strong correlation between intraplaque neovessels and contrast enhancement was found. CONCLUSIONS:CEU imaging is a feasible noninvasive imaging modality to evaluate intraplaque neovascularization. A noninvasive imaging modality to assess lesion neovascularization could be of great importance to identify vascularized, "high-risk" lesions before rupture.