Established Atherosclerosis Research Articles

Coronary atherosclerosis burden by coronary computed tomography in type II diabetes with preclinical non-obstructive carotid atherosclerosis and without inducible myocardial ischemia.

To quantify cardiovascular risk in persons with type-2 diabetes (DM) and established carotid atherosclerosis, without inducible myocardial ischemia by stress imaging. We selected clinically healthy DM and subjects without DM (nonDM, controls) with non-obstructive carotid atherosclerosis and without significant coronary artery disease (CAD) by stress echocardiography. Coronary flow velocity reserve (CFR) was assessed during stress echocardiography. In those with negative stress imaging, coronary artery calcium (CAC) assessment by computed tomography (CT) was proposed. Outcome at 180-day follow-up was performed by visits or telephone interview. Total CAC was greater in DM (n=19) than in nonDM (n=19, CAC: 225±158 vs 78±65Agatston units, 156±161 vs 53±51 in the left anterior descendent artery, both p<0.05). Coronary stenosis burden was greater in DM than in nonDM, albeit <75% in all cases; CFR was lower in DM than nonDM (p<0.05). Mean age was slightly higher (69±6 vs 64±12, p>0.2), and male gender tended to be less frequent (63 vs 79%) with DM than with nonDM; all subjects showed dyslipidemia, and almost all had arterial hypertension. Based on CAC, a 10-year cardiovascular risk was 10-20% in 78% of the DM and in 28% of the non-DM, and was >20% in 11% of the DM but in none of the nonDM. A single non-fatal myocardial infarction was recorded within 180days from test, among DM. Based on CAC, in the presence of non-obstructive carotid atherosclerosis, asymptomatic DM may show significantly higher CAD burden than nonDM even in the absence of inducible myocardial ischemia.

Early atheroprotection by SYK inhibition fails in established disease when local macrophage proliferation dominates lesion progression

Macrophages in the arterial intima sustain chronic inflammation during atherogenesis. Under hypercholesterolemic conditions murine Ly6Chigh monocytes surge in the blood and spleen, infiltrate nascent atherosclerotic plaques, and differentiate into macrophages that proliferate locally as disease progresses. Spleen tyrosine kinase (SYK) may participate in downstream signaling of various receptors that mediate these processes. We tested the effect of the SYK inhibitor fostamatinib on hypercholesterolemia-associated myelopoiesis and plaque formation in Apoe−/− mice during early and established atherosclerosis. Mice consuming a high cholesterol diet supplemented with fostamatinib for 8 weeks developed less atherosclerosis. Histologic and flow cytometric analysis of aortic tissue showed that fostamatinib reduced the content of Ly6Chigh monocytes and macrophages. SYK inhibition limited Ly6Chigh monocytosis through interference with GM-CSF/IL-3 stimulated myelopoiesis, attenuated cell adhesion to the intimal surface, and blocked M-CSF stimulated monocyte to macrophage differentiation. In Apoe−/− mice with established atherosclerosis, however, fostamatinib treatment did not limit macrophage accumulation or lesion progression despite a significant reduction in blood monocyte counts, as lesional macrophages continued to proliferate. Thus, inhibition of hypercholesterolemia-associated monocytosis, monocyte infiltration, and differentiation by SYK antagonism attenuates early atherogenesis but not established disease when local macrophage proliferation dominates lesion progression.

Covering the Cover

Covering the Cover

Established atherosclerosis might be a prerequisite for chicory and its constituent protocatechuic acid to promote endothelium-dependent vasodilation in mice.

Chicory (Cichorium intybus L. var. foliosum, Belgian endive), a typical Mediterranean vegetable, and its constituent protocatechuic acid (PCA) can inhibit established atherosclerosis progression. We thus investigated whether chicory can improve vascular relaxation, a critical pathway for combating atherosclerosis, and whether PCA is a contributor to a chicory-induced effect. Apolipoprotein E-deficient (ApoE-/- ) mice with established atherosclerosis and C57BL/6J mice without atherosclerosis were fed an AIN-93G diet, or AIN-93G plus 0.5% freeze-dried chicory or 0.003% PCA for 1 wk. In ApoE-/- mice, both chicory and PCA consumption increased endothelium-dependent vasodilation and endothelial nitric oxide synthase (eNOS) activity independent of eNOS and phospho-eNOS Ser1177 and Thr495 protein expression. Chicory- or PCA-induced eNOS activities were associated with increased vascular tetrahydrobiopterin (BH4 ) levels that result from reduced BH4 oxidation partially through preventing eNOS uncoupling. In C57BL/6J mice, neither chicory nor PCA consumption affected endothelium-dependent vasodilation and eNOS activity. Notably, in vitro studies showed that PCA increases eNOS activity in mouse aortic endothelial cells in co-culture with macrophage foam cells, but not in aortic endothelial cells alone. Chicory improves eNOS-mediated endothelium-dependent vasodilation by increasing BH4 levels in mice with established atherosclerosis, which might be partially ascribed to its constituent PCA.

Evaluation of endothelial dysfunction in patients with type 2 diabetes mellitus – Analysis of 100 cases

Background: Atherosclerosis which is a precursor for macro-vascular disease involves both functional as well as structural changes in the vasculature. Functional changes involve abnormalities in endothelium, vascular smooth muscle cells and platelet functions. Endothelial dysfunction was reported in type 2 DM cases. Ultrasound assessment of brachial artery FMD (Flow Mediated Dilatation) provides important information about vascular function in health and disease. Low FMD values predict independently an established atherosclerosis. Materials and methods: A total of 150 individuals included in this study; 100 were diabetes mellitus patients and 50 were age matched persons without diabetes, hypertension or vascular disease. Of 100 diabetes patients 50 each were grouped in two groups with good (HbA1c 7.5%) control of hyperglycemia. Brachial artery flow mediated vasodilatation (FMD) was studied in all patient after overnight fasting (8 hours) using 7.5 MHz phased array transducer. The medial epicondyle was used as anatomical landmark for brachial artery. Observations : In this study it is observed that, the mean age for FMD 4.5% is 56.538.37. Age is higher in diabetics with FMD % 4.5%. Smoking in diabetics with FMD 4.5 % was 15%. Hypertension was equally found 40%. Family history of diabetes with FMD % 4.5% were none. Summary and conclusions : The data suggest that endothelial function is a useful prognostic marker in coronary artery disease patients. In these patients, it has been reported that endothelial dysfunction is an independent prognostic factor and may predict future events, irrespective of the angiographic severity of the disease. Moreover, blunted endothelial function may reflect early atherosclerosis, and should lead to a detailed evaluation.

Drug-Eluting Stents Are the Default Strategy for Superficial Femoral Artery Intervention Now

Drug-eluting stents (DES) are the default strategy for superficial femoral artery (SFA) intervention in 2015 because they have been evaluated in a large number of patients over a long follow-up period with outcomes superior to other treatment options. No other therapy can make that claim. Response by Garcia on p 329 The SFA and the contiguous popliteal artery constitute the femoropopliteal (FP) segment, which is among the human body’s most hostile vascular environments. It is extremely long, exposed to external compression, and undergoes marked conformational changes every time the leg is flexed. Because it courses posterior to the knee joint, there is obligatory longitudinal compressive force with knee flexion (Figure 1).1 Magnetic resonance angiography studies demonstrated 13% shortening of the SFA between supine and fetal positions.2 There are 60° of SFA torsion produced by simultaneous knee and hip flexion.2 Given these anatomic and dynamic challenges, it is hardly surprising that treatment strategies for established FP atherosclerosis suffer from marked limitations. Figure 1. A , The femoropopliteal arterial segment (represented by the solid black line) is not coaxial with the femur and tibia. B , With knee flexion this relationship creates an excess of arterial length, partially compensated by vessel tortuosity ( C ). Reproduced from Wensing et al1 with permission from the publisher. Copyright © 1995, John Wiley & Sons Ltd. When a disease process lacks 1 definitive cure, a wide variety of marginally effective solutions frequently surface. Such is the case with FP occlusive disease. The spectrum of treatment options is broad, ranging from exercise therapy3 as the least intrusive option, to bypass surgery as the most intrusive.4 In between these extremes lies a seemingly endless variety of endovascular options. ### Less Is Less Perhaps the most universally available and cheapest treatment strategy for symptomatic lower extremity vascular disease is …

Atheroprotective effects of pure tocotrienol supplementation in the treatment of rabbits with experimentally induced early and established atherosclerosis

BackgroundAtherosclerosis is the main cause of coronary artery disease -related deaths worldwide. The atheroprotective properties of pure tocotrienols (T3) in the absence of alpha-tocopherol (α-TCP) in vitamin E has not been extensively examined.AimTo determine the atheroprotective properties of T3 in early and established atherosclerosis rabbits.MethodsThirty New Zealand white rabbits were divided into two groups, B1 and B2 which represent early [fed 1% high cholesterol diet (HCD) for 2 weeks] and established (fed 1% HCD for 8 weeks) atherosclerosis. Each group was subdivided into three intervention arms: 1) T3–4 mg/kg, 2) T3–15 mg/kg and 3) vehicle without T3 (T3 negative) for 8 weeks. Serial fasting blood samples were obtained for lipid profile, and whole lengths of aorta were used to determine tissue markers of endothelial activation, inflammation and plaque stability.ResultsIn B1, atherosclerotic lesion in T3–4 mg/kg group was significantly reduced (p=0.008), while aortic tissue expression of vascular cellular adhesion molecule 1 (VCAM-1), interleukin-6 (IL-6) and matrix metalloproteinase (MMP-12) was reduced in T3–4 mg/kg compared to T3-negative rabbits group (0.2±0.1 vs. 28.5±3.1%; 3.0±1.6 vs. 14.0±1.7%; and 5.2±2.2 vs. 27.7±0.8%, respectively, p<0.05). T3–15 mg/kg group showed reduction in VCAM-1, E-selectin, IL-6 and MMP-12 (3.9±1.9 vs. 28.5±3.1%; 10.3±0.5 vs. 59.8±8.5%; 2.6±1.7 vs. 14.0±1.7%; and 16.2±3.2 vs. 27.7 0.8%, respectively, p<0.05). In B2, T3–4 mg/kg group reduced aortic tissue expression of intercellular adhesion molecule 1 (ICAM-1), E-selectin, IL-6, MMP-12 and MMP-9 compared to T3-negative rabbits group (29.9±2.4 vs. 55.3±1.3%; 26.7±1.5 vs. 60.5±7.6%; 15.7±0.7 vs. 27.7±4.8%; 34.8±2.7 vs. 46.5±3.4%; and 25.89±3.9 vs. 45.9±1.7%, respectively, p<0.05). T3–15 mg/kg group showed reduced VCAM-1, ICAM-1, E-selectin, IL-6, MMP-12 and MMP-9 (20.5±3.3 vs. 35.6±2.5%; 24.9±1.3 vs. 55.3±1.3%; 29.9±6.7 vs. 60.5±7.6; 11.3±2.2 vs. 27.7±4.8%; 23.0±1.7 vs. 46.5±3.4%; and 17.6±1.9 vs. 45.9±1.7%, respectively, p<0.05.ConclusionThese findings suggest the possible atheroprotective role T3 plays as an adjunct supplementation to standard treatment in the prevention of CAD.

Voluntary Exercise Stabilizes Established Angiotensin II-Dependent Atherosclerosis in Mice through Systemic Anti-Inflammatory Effects.

We have previously demonstrated that exercise training prevents the development of Angiotensin (Ang) II-induced atherosclerosis and vulnerable plaques in Apolipoprotein E-deficient (ApoE-/-) mice. In this report, we investigated whether exercise attenuates progression and promotes stability in pre-established vulnerable lesions. To this end, ApoE-/- mice with already established Ang II-mediated advanced and vulnerable lesions (2-kidney, 1-clip [2K1C] renovascular hypertension model), were subjected to sedentary (SED) or voluntary wheel running training (EXE) regimens for 4 weeks. Mean blood pressure and plasma renin activity did not significantly differ between the two groups, while total plasma cholesterol significantly decreased in 2K1C EXE mice. Aortic plaque size was significantly reduced by 63% in 2K1C EXE compared to SED mice. Plaque stability score was significantly higher in 2K1C EXE mice than in SED ones. Aortic ICAM-1 mRNA expression was significantly down-regulated following EXE. Moreover, EXE significantly down-regulated splenic pro-inflammatory cytokines IL-18, and IL-1β mRNA expression while increasing that of anti-inflammatory cytokine IL-4. Reduction in plasma IL-18 levels was also observed in response to EXE. There was no significant difference in aortic and splenic Th1/Th2 and M1/M2 polarization markers mRNA expression between the two groups. Our results indicate that voluntary EXE is effective in slowing progression and promoting stabilization of pre-existing Ang II-dependent vulnerable lesions by ameliorating systemic inflammatory state. Our findings support a therapeutic role for voluntary EXE in patients with established atherosclerosis.

A CD1d-dependent lipid antagonist to NKT cells ameliorates atherosclerosis in ApoE-/- mice by reducing lesion necrosis and inflammation.

Atherosclerosis-related deaths from heart attacks and strokes remain leading causes of global mortality, despite the use of lipid-lowering statins. Thus, there is an urgent need to develop additional therapies. Reports that NKT cells promote atherosclerosis and an NKT cell CD1d-dependent lipid antagonist (DPPE-PEG350, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N[methoxy(polyethyleneglycol)-350]) reduces allergen-induced inflammation led us to investigate its therapeutic potential in preventing the development and progression of experimental atherosclerosis. DPPE-PEG350 was administered to hyperlipidaemic ApoE(-/-) mice with/without established atherosclerosis. Atherosclerosis and immune cells were assessed in the aortic sinus lesions. Lesion expression of monocyte chemoattractant protein-1 (MCP-1) and vascular cell adhesion protein-1 (VCAM-1) responsible for inflammatory immune cell recruitment as well as mRNA expression of IFNγ and its plasma levels were investigated. Necrotic cores and lesion smooth muscle and collagen contents important in plaque stability were determined as were plasma lipid levels. DPPE-PEG350 reduced atherosclerosis development and delayed progression of established atherosclerosis without affecting plasma lipids. CD4 and CD8 T cells and B cells in atherosclerotic lesions were decreased in DPPE-PEG350-treated mice. Lesion MCP-1 and VCAM-1 protein expression and necrotic core size were reduced without affecting lesion smooth muscle and collagen content. IFNγ and lymphocytes were unaffected by the treatment. The attenuation of progression of established atherosclerosis together with reduced development of atherosclerosis in hyperlipidaemic mice by the NKT antagonist, without affecting NKT cell or other lymphocyte numbers, suggests that targeting lesion inflammation via CD1d-dependent activation of NKT cells using DPPE-PEG350 has a therapeutic potential in treating atherosclerosis.

Chicory, a typical vegetable in Mediterranean diet, exerts a therapeutic role in established atherosclerosis in apolipoprotein E-deficient mice.

Since protocatechuic acid exerts an atheroprotective role, we investigated how chicory (Cichorium intybus L. var. foliosum, Belgian endive) rich in protocatechuic acid, a typical vegetable in Mediterranean diet, affects preestablished atherosclerosis progression. Apolipoprotein E-deficient mice fed AIN diets containing 0.5% freeze-dried chicory for 10 weeks displayed a reduction in lesion size with a concomitant improvement in lesion stability indicated by fewer macrophages and more collagen content. Chicory consumption suppressed aortic cholesterol accumulation and intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and monocyte chemoattractant protein-1 expression, whereas it increased aortic ATP-binding cassette transporter A1 (ABCA1) and G1 (ABCG1) expression. Furthermore, chicory consumption improved peritoneal macrophage phenotype with less cellular cholesterol associated with an enhancement of cholesterol efflux capacity through upregulation of ABCA1 and ABCG1, less cellular oxidative stress associated with an inhibition of nicotinamide adenine dinucleotide phosphate oxidase activity, and weaker inflammatory responses associated with an inhibition of nuclear factor-κB activation. Interestingly, ABCA1 and ABCG1 silencing tended to completely block beneficial effects of chicory in peritoneal macrophages. Chicory exerts an atheroprotective role in mice possibly by regulating lesional macrophage content and phenotype, suggesting that chicory is one underrated contributor to Mediterranean Diet-induced atheroprotection.

PET imaging of atherosclerosis.

Atherosclerosis is a chronic, progressive, multifocal disease of the arterial wall, which is mainly fuelled by local and systemic inflammation, often resulting in acute ischemic events following plaque rupture and vessel occlusion. When assessing the cardiovascular risk of an individual patient, we must consider both global measures of disease activity and local features of plaque vulnerability, in addition to anatomical distribution and degree of established atherosclerosis. These parameters cannot be measured with conventional anatomical imaging techniques alone, which are designed primarily to identify the presence of organic intraluminal obstruction in symptomatic patients. However, molecular imaging with PET, using specifically targeted radiolabeled probes to track active in vivo atherosclerotic mechanisms noninvasively, may potentially provide a method that is better suited for this purpose. Vascular PET imaging can help us to further understand aspects of plaque biology, and current evidence supports a future role as an emerging clinical tool for the quantification of cardiovascular risk in order to guide and monitor responses to antiatherosclerosis treatments and to distinguish high-risk plaques.

Abstract 16514: Magnetic Resonance Imaging-Detected Carotid Plaque Characteristics Predict Systemic Cardiovascular Outcomes in Patients With Clinically Established Atherosclerosis in the AIH-HIGH Study

Background: Although vulnerable plaques are considered the pathological substrate for acute cardiovascular events, the pursuit of imaging surrogate markers for systemic atherothrombotic risk is currently focused on plaque burden. The prognostic value of vulnerable plaque characteristics as novel markers for systemic cardiovascular outcomes remains elusive. Advances in magnetic resonance imaging (MRI) have enabled characterization of the vulnerable carotid plaque. In a prospective study, we tested whether MRI-detected carotid plaque characteristics predict subsequent cardiovascular events. Methods: As part of an event-driven clinical trial, subjects with clinically established atherosclerotic disease were recruited. A multi-sequence protocol was used to measure the volumes of calcification and necrotic core (NC), and to identify the presence of intraplaque hemorrhage (IPH) and thin/ruptured fibrous cap (FC) using published criteria. The primary endpoint included fatal and non-fatal myocardial infarction or ischemic stroke, hospitalization for acute coronary syndrome, and symptom-driven revascularization. Cox regression analysis was used to present results as hazard ratio (HR) with 95% confidence interval (CI). Results: Of 232 subjects recruited, 214 (92.2%) with diagnostic image quality constituted the study population (mean age: 61±9 years; male: 82%; statin use: 94%). Calcification, NC, IPH, and thin/ruptured FC were detected in 48%, 52%, 8% and 14% of subjects, respectively. During a median follow-up of 35.1 months, 18 (8.4%) subjects reached the primary endpoint. MRI-detected plaque characteristics associated with the primary endpoint included larger NC (HR per 1 standard deviation increase in volume [1-SD]: 1.43 [1.16, 1.75], p&lt;0.001), thin/ruptured FC (HR: 4.31 [1.67, 11.12], p=0.003), and marginally IPH (HR: 3.00 [0.99, 9.13], p=0.053). Although calcium deposition is traditionally pursued as a marker for atherosclerosis, subjects with larger calcification (HR per 1-SD: 0.60 [0.26, 1.41], p=0.20) did not show increased risk. Conclusions: Vulnerable plaque characteristics are prevalent in carotid arteries of patients with established atherosclerosis and are predictive of systemic cardiovascular outcomes.

Voluntary exercise slows the progression of established angiotensin II-dependent advanced atherosclerotic lesions

Objectives: We have previously demonstrated that exercise training prevents the development of Angiotensin (Ang) II-induced atherosclerosis and vulnerable plaques in ApoE-/- mice. In the present study, we investigated whether exercise attenuates the progression of established Ang II-dependent atherosclerosis.

The relationship of novel adipokines, RBP4 and omentin-1, with carotid atherosclerosis severity and vulnerability

ObjectiveWe investigated the relationship of circulating novel adipokines, retinol-binding protein 4 (RBP4) and omentin-1, with advanced carotid atherosclerosis and ultrasound indexes of severity (total plaque area-TPA) and plaque echogenicity and vulnerability (Gray-Scale median – GSM score). MethodsWe enrolled 225 patients with high-grade carotid stenosis (HGCS) who underwent carotid revascularization (73 Symptomatic patients, 152 asymptomatic patients) and 75 age- and sex-matched, asymptomatic individuals with low-grade (<50%) carotid stenosis (LGCS). Seventy-three individuals without current manifestations of atherosclerotic disease served as control group (COG). All participants underwent carotid ultrasound with TPA and GSM score assessment. Moreover, clinical parameters, metabolic profile, and circulating levels of hsCRP and adipokines were assessed. ResultsRBP4 was significantly elevated in HGCS (51.44 ± 16.23 mg/L) compared to LGCS (38.39 ± 8.85 mg/L), independent of symptoms existence, whereas RBP4 levels in COG were even lower (25.74 ± 10.72 mg/L, p < 0.001 compared to either HGCS or LGCS). Inversely, serum omentin-1 levels were significantly lower across HGCS (490.41 ± 172 ng/ml) and LGCS (603.20 ± 202.43 ng/ml) than COG (815.3 ± 185.32, p < 0.001). Moreover, the considerable difference between HGCS and LGCS (p < 0.001) was exclusively attributed to the excessive suppression of omentin-1 concentrations in symptomatic versus asymptomatic (p = 0.004) patients. HGCS and LGCS did not differ in the rest of clinical and biochemical parameters. In multiple regression analysis, RBP4 (beta = 0.232, p = 0.025) and hsCRP (beta = 0.300, p = 0.004) emerged as independent determinants of TPA in patients with carotid atherosclerosis. Low serum levels of omentin-1 correlated with GSM score and symptoms but that association was lost in multivariate analysis.. ConclusionRBP4 serum levels were significantly elevated in patients with established carotid atherosclerosis and were positively associated with atherosclerosis severity. The association of low serum omentin-1 with carotid plaque echolucency requires further investigation..ClinicalTrials.gov Identifier: NCT00636766.

Quantitative contrast-enhanced ultrasound of intraplaque neovascularization in patients with carotid atherosclerosis.

Intraplaque neovascularization (IPN) is an increasingly studied marker of the vulnerable atherosclerotic plaque, and contrast-enhanced ultrasound (CEUS) is an in vivo imaging technique for the assessment of IPN. The purpose of this study was to test novel quantification methods for the detection of carotid IPN using CEUS. 25 patients with established carotid atherosclerosis underwent bilateral carotid CEUS using a Philips iU-22 ultrasound system with an L9 - 3 transducer. Visual scoring of IPN was performed using a 3-point score. Quantification of IPN was performed using novel custom developed software. In short, regions of interest were drawn over the atherosclerotic plaques. After motion compensation, several IPN features were calculated. Statistical analysis was performed using Spearman's rho. Reproducibility of the quantification features was calculated using intra-class correlation coefficients and mean differences between calculations. 45 carotid arteries were available for the quantification of IPN. The quantification of IPN was feasible in all 45 carotid plaques. The IPN area, IPN area ratio and neovessel count had a good correlation with the visual IPN score (respectively ρ = 0.719, ρ = 0.538, ρ = 0.474 all p < 0.01). The intra-observer and inter-observer agreement was good to excellent (p < 0.01). The intra-observer and inter-observer variability was low. The quantification of carotid IPN on CEUS is feasible and provides multiple features on carotid IPN. Accurate quantitative assessment of IPN may be important to recognize and to monitor changes during therapy in vulnerable atherosclerotic plaques.

Abstract 478: Single-Cell Isolation and Analysis of Viable Proliferating Macrophages From Atherosclerotic Plaques

INTRODUCTION: We have recently shown that the proliferation of macrophages within the atherosclerotic plaque, rather than the recruitment of new monocytes from the blood is the key driver of plaque growth in established atherosclerosis. The study of proliferating macrophages has been hampered by the lack of a technique for identifying proliferating macrophages and isolating them from the atherosclerotic plaque in a viable state. OBJECTIVE: Develop and validate a process for identifying proliferating macrophages in atherosclerotic lesions, and isolating them as single, viable cells, then perform a molecular characterization of these cells. METHODS: ApoE-/- and LDLR-/- mice were fed a diet high in fat and cholesterol. Atherosclerotic aortas were collected, and cells were isolated by mechanical and enzymatic dissociation. Cells were stained using fluorescent markers and DNA-binding dyes, then analyzed and isolated by fluorescence-activated cell sorting. RESULTS: Mincing and enzymatic digestion in collagenase I, collagenase XI, DNAse, and hyaluronidase optimally isolates macrophages from aortic explants. Cell-surface staining with fluorescent antibodies against B220, F4/80, Ly6c, MHC II, and CD11b allows identification of lesional macrophages. DNA staining with Vybrant DyeCycle Violet allows identification of proliferating cells. Combined, these processes allow for the FACS-based sorting of viable proliferating macrophages from atherosclerotic plaque, and subsequent RNA or protein analysis, or culture. CONCLUSIONS: The process described here allows the first-ever viable isolation of proliferating macrophages from atherosclerotic plaques. This will permit the investigation of molecular targets to interfere with the pathogenesis of atherosclerosis.

Abstract 26: Anti-TIM-1 Monoclonal Antibody Therapy Expands Atheroprotective B1a B Cells in vivo and Attenuates Development and Progression of Atherosclerosis

Background: B1a B cells attenuate atherosclerosis by secreting natural IgM, but their therapeutic application is limited by lack of availability. Regulatory B cells identified by Tim-1 expression and expanded through Tim-1 ligation by anti-TIM-1 low affinity monoclonal antibody (RMT1-10 mAb) induced tolerance. Here, we examined the capacity of this mAb to expand B1a B cells to inhibit atherosclerosis development and progression of established atherosclerosis. Methods and Results: Six-week old male ApoE-deficient mice were treated with RMT1-10 mAb and fed a high-fat diet (HFD) for 8 weeks. B1a TIM-1+IgM+ B cells and B1a TIM-1+IgM+IL-10+ B cells were selectively expanded. These effects reduced lesion size, markedly increased plasma and lesion IgM and decreased lesion oxidatively modified LDL. Lesion CD4+ and CD8+ T cells, macrophages and MCP-1, VCAM-1, proinflammatory cytokine expression, apoptotic cell numbers and necrotic cores were reduced. Splenectomy indicated that these effects were B1a B cell-dependent. B1a B cell stimulation in vitro with RMT1-10 mAb promoted dose-response B1a B cell proliferation and B1a-derived IgM production. To determine whether treatment attenuated developed atherosclerosis progression, 6 week-old male ApoE-deficient mice were fed a HFD for 6 weeks, and treated with anti-TIM-1 mAb for another 6 weeks while continuing the HFD. Treatment also increased B1a TIM-1+IgM+ B cells, B1a TIM-1+IgM+IL-10+ B cells and IgM levels and greatly attenuated atherosclerosis progression. Conclusions: Anti-TIM-1 treatment attenuates atherosclerosis development and progression by selectively expanding atheroprotective B1a B cells and modulating its immunoinflammatory component. TIM-1 mAb therapy could be an attractive approach for treating atherosclerosis.

PW198 Physical Activity Motivation: Impact of educational program in Primary care

Introduction: Cardiovascular disease has been recognised to be a major cause of morbidity and mortality in Chronic Kidney Disease (CKD). Lipid abnormalities were originally considered as complications of end stage renal disease; these changes can be present in early stages of CKD and may actively participate in the pathogenesis of serious complications such as atherosclerotic vascular disease. The dyslipidaemia can be influenced by several intrinsic (nephrotic range proteinuria, diabetes mellitus, hereditary disorders of lipid metabolism) or exogenous (erythropoietin administration, steroids, calcineurin inhibitors, etc.) factors. Objectives: Study of Carotid Intimal Medial thickness(CIMT) in different stages of CKD and its relation with lipid profile and comparing with similar control group. Methods: Inclusion criteria: Patients with CKD between the age group 18-85years, Exclusion criteria: patients with acute kidney injury, ischemic heart disease disease, stroke, on hypolipidaemic drugs. 70 cases of CKD were selected on basis of simple random sampling technique. 30 age and sex matched healthy control were taken. CKD patients were classified into different stages according to glomerular filtration rate. Results: Study showed 18 (25.7%) of the patients belonged to 4 and 5 decade. There was male preponderance 39 (55.7%). Most common etiology of CKD was Diabetic Nephrology 21 (30%). 16 (22.9%) patients were found to have Hypercholesterolemia, 42 (60%) had Triglyceride level 150 mg/dl. 23 (32.9%) patients had serum HDL-C 30 mg/dl. The mean CIMT in patients was 0.84 0.23mm and control was 0.46 0.19mm. There was statistically significant (p < 0.001) difference in CIMT between two groups. There was significant Univariate positive co-relation between CIMT and age (r1⁄40.605, p<0.001), BMI (r1⁄40.377, p1⁄40.001). Total cholesterol (r1⁄40.236, p<0.018) and serum Triglyceride (r1⁄40.387, p<0.001). Statistically significant correlation was found between CIMT and eGFR, VLDL and HDL-C. Conclusion: CIMT is significantly higher in CKD patients and is very well co-related with traditional cardiovascular risk factors like age, BMI, total Cholesterol and Triglyceride. Assessment of CIMT in different stages of CKD may act as a marker for premature or established atherosclerosis. Disclosure of Interest: None Declared

PW195 Lipid profile and Carotid Intimal Medial thickness in different stages of Chronic Kidney Disease

Introduction: Cardiovascular disease has been recognised to be a major cause of morbidity and mortality in Chronic Kidney Disease (CKD). Lipid abnormalities were originally considered as complications of end stage renal disease; these changes can be present in early stages of CKD and may actively participate in the pathogenesis of serious complications such as atherosclerotic vascular disease. The dyslipidaemia can be influenced by several intrinsic (nephrotic range proteinuria, diabetes mellitus, hereditary disorders of lipid metabolism) or exogenous (erythropoietin administration, steroids, calcineurin inhibitors, etc.) factors. Objectives: Study of Carotid Intimal Medial thickness(CIMT) in different stages of CKD and its relation with lipid profile and comparing with similar control group. Methods: Inclusion criteria: Patients with CKD between the age group 18-85years, Exclusion criteria: patients with acute kidney injury, ischemic heart disease disease, stroke, on hypolipidaemic drugs. 70 cases of CKD were selected on basis of simple random sampling technique. 30 age and sex matched healthy control were taken. CKD patients were classified into different stages according to glomerular filtration rate. Results: Study showed 18 (25.7%) of the patients belonged to 4 and 5 decade. There was male preponderance 39 (55.7%). Most common etiology of CKD was Diabetic Nephrology 21 (30%). 16 (22.9%) patients were found to have Hypercholesterolemia, 42 (60%) had Triglyceride level 150 mg/dl. 23 (32.9%) patients had serum HDL-C 30 mg/dl. The mean CIMT in patients was 0.84 0.23mm and control was 0.46 0.19mm. There was statistically significant (p < 0.001) difference in CIMT between two groups. There was significant Univariate positive co-relation between CIMT and age (r1⁄40.605, p<0.001), BMI (r1⁄40.377, p1⁄40.001). Total cholesterol (r1⁄40.236, p<0.018) and serum Triglyceride (r1⁄40.387, p<0.001). Statistically significant correlation was found between CIMT and eGFR, VLDL and HDL-C. Conclusion: CIMT is significantly higher in CKD patients and is very well co-related with traditional cardiovascular risk factors like age, BMI, total Cholesterol and Triglyceride. Assessment of CIMT in different stages of CKD may act as a marker for premature or established atherosclerosis. Disclosure of Interest: None Declared

Proliferating Macrophages Populate Established Atherosclerotic Lesions

A new study reveals that the majority of macrophages in established atherosclerotic lesions are derived from local proliferation, rather than from the influx of blood-borne monocytes. While the factors driving proliferation remain to be understood, the findings suggest that targeting macrophage proliferation could represent a new therapeutic opportunity in established atherosclerosis.