Circulating miRNA levels differ with respect to carotid plaque characteristics and symptom occurrence in patients with carotid artery stenosis and provide information on future cardiovascular events.

INTRODUCTION: Circulating microRNAs (miRNAs) levels are potentially important biomarkers and therapeutic targets of cerebral ischemic event (CIE) in patients with internal carotid artery stenosis (ICAS). AIM: This prospective study investigated associations between circulating miRNAs and symptomatic and asymptomatic ICAS, carotid plaque morphology and future cardiovascular events. MATERIAL AND METHODS: Circulating miRNAs (miR-1-3p, miR-16-5p, miR-34a-5p, miR-124-3p, miR-133a-3p, miR-133b, miR-134-5p, miR-208b-3p, miR-375 and miR-499-5p) were analyzed in 92 consecutive patients with significant ICAS referred for revascularization. Group I comprised 65 subjects (41 males, age 69.3 ±9.7 years) with a recent CIE. Group II comprised 27 patients (15 males, age 68.2 ±8.4 years) with asymptomatic ICAS. The ICAS degree and plaque morphology was assessed by ultrasonography. The incidences of cardiovascular death (CVD), myocardial infarction (MI) and recurrent CIE (CVD/MI/CIE) were recorded prospectively (mean: 38.7 ±3.8 months). RESULTS: Groups II and I differed significantly in levels of miR-124-3p (p = 0.036), miR-133a-3p (p = 0.043) and miR-134-5p (p = 0.02). Hypoechogenic, as compared to echogenic, plaques differed in levels of miR-124-3p (p = 0.038), miR-34a-5p (p = 0.006), miR-133b (p = 0.048), miR-134-5p (p = 0.045), and miR-375 (p = 0.016), while calcified plaques differed in miR-16-5p (p = 0.023). Ulcerated plaques showed higher levels of miR-1-3p (p = 0.04) and miR-16-5p (p = 0.003), while thrombotic plaques showed lower levels of miR-1-3p (p = 0.032). CVD/MI/CIE occurred in 14 (15.5%) out of 90 follow-up patients. Multivariate Cox and ROC analysis showed associations between miR-1-3p and CVD (AUC = 0.634; HR = 4.84; 95% CI: 1.62-14.5; p = 0.005), MI (AUC = 0.743; HR = 7.8; 95% CI: 2.01-30.0; p = 0.003), and CVD/MI/CIE (AUC = 0.560; HR = 4.6; 95% CI: 1.61-13.1; p = 0.004), while miR-133b was associated with recurrent CIE (AUC = 0.581; HR = 2.25; 95% CI: 1.01-5.02; p = 0.047). CONCLUSIONS: A significant difference in levels of selected miRNAs is observed in symptomatic vs. asymptomatic ICAS. Plaque morphology and structure is associated with change of miRNA levels. The expression of miR-1-3p may be a potential prognostic factor for future cardiovascular events.

Diagnostic and prognostic micro-RNAs in ischaemic stroke due to carotid artery stenosis and in acute coronary syndrome: a four-year prospective study.

BACKGROUND: Circulating microRNAs (miRs) levels are potentially important diagnostic and prognostic biomarkers in acute coronary syndrome (ACS) or cerebral ischaemic events (CIE) resulting from internal carotid artery stenosis (ICAS). AIM: This four-year prospective study aimed to compare the levels of circulating miRs in ACS vs. CIE patients, and investigate miRs potentially associated with risk of recurrent cardiovascular events. METHODS: The circulating miRs levels (miR-1-3p, miR-16-5p, miR-34a-5p, mir-122-5p, miR-124-3p, miR-133a-3p, miR-133b, miR-134-5p, miR-208b-3p, miR-375, and miR-499-5p) were compared in 43 (34 men, 57.6 ± 10.1 years) patients with ACS, and in 71 (47 men, 69.5 ± 9.6 years) with CIE due to ICAS. A four-year prospective evaluation of miRs associated with risk of cardiovascular death (CVD), myocardial infarction (MI), CIE, or all (CVD/MI/CIE) was performed. RESULTS: In ACS vs. CIE patients, the levels of miR-124-3p (p < 0.001), miR-134-5p (p = 0.012), miR-208b-3p (p < 0.001), miR-34a-5p (p < 0.001), and miR-499-5p (p < 0.001) were higher, while levels of miR-16-5p (p < 0.001) and miR-122-5p (p < 0.001) were lower. Levels of miR-1-3p (p = 0.195), miR-133a-3p (p = 0.333), miR-133b (p = 0.056), and miR-375 (p = 0.055) were non-statistically different. During follow-up (median 57 months, Q1-Q3: 54-60), CVD/MI/CIE occurred in 23 subjects, including eight CVDs, five non-fatal CIEs, and 10 non-fatal MIs. The multivariate Cox proportional hazard analysis (relative risk [RR]; 95% confidence interval [CI]) revealed that miR-208b-3p (1.225; 1.092-1.375), miR-34a-5p (0.963; 0.935-0.992), and miR-499-5p (0.077; 0.025-0.239) were independently associated with risk of CVD/MI/CIE, as well as risk of each event. Furthermore, miR-133b (1.009; 1.003-1.015) was associated with risk of CVD. CONCLUSIONS: This study shows that although most investigated miRs levels differ significantly between patients with ACS and CIE, similar levels of circulating miR-1-3p, miR-133a-3p, miR-133b, and miR-375 were observed; furthermore, we identified several common miRs as possible risk factors for recurrent cardiovascular events.

Multiterritory Atherosclerosis and Carotid Intima-Media Thickness as Cardiovascular Risk Predictors After Percutaneous Angioplasty of Symptomatic Subclavian Artery Stenosis.

OBJECTIVES: To identify independent predictors of cardiovascular events among patients with subclavian artery stenosis. METHODS: Two hundred eighteen consecutive patients with subclavian artery stenosis referred to angioplasty were examined for coexistent coronary, renal, or lower extremity artery stenosis of 50% or greater. Initial carotid intima-media thickness and internal carotid artery (ICA) stenosis were assessed. Intima-media thickness was reassessed in 108 randomly chosen patients to evaluate the change over time. The incidence of cardiovascular death, myocardial infarction (MI), ischemic stroke, and symptomatic lesion progression was recorded. RESULTS: The patients included 116 men and 102 women (mean age ± SD, 62.1 ± 8.4 years). Isolated subclavian artery stenosis and involvement of 1, 2, and 3 or 4 other territories with stenosis of 50% or greater were found in 46 (21.1%), 83 (38.1%), 55 (25.2%), and 34 (15.6%) patients, respectively. Internal carotid artery stenosis of 50% or greater (relative risk [RR], 1.54; 95% confidence interval [CI], 1.39-1.70; P < .001) and initial intima-media thickness (RR, 1.16; 95% CI, 1.05-1.28; P = .005) were identified as independent markers of multiterritory atherosclerosis. The optimal intima-media thickness cutoff for atherosclerosis extent was 1.3 mm (sensitivity, 75.6%; specificity, 76.1%). During follow-up of 57 ± 35 months, cardiovascular death, MI, and ischemic stroke occurred in 29 patients (13.3%). Those patients had significantly higher intima-media thickness progression (+0.199 ± 0.57 versus +0.008 ± 0.26 mm; P = .039) and more widespread initial atherosclerosis (mean territories, 1.8 ± 1.1 versus 1.3 ± 1.1; P = .042). Independent predictors of cardiovascular death, MI, ischemic stroke, and lesion progression were coronary artery disease (RR, 1.32; 95% CI, 1.10-1.58; P = .003) and intima-media thickness progression (RR, 1.22; 95% CI, 1.02-1.46; P = .033; sensitivity, 75.0%; specificity, 61.8%). CONCLUSIONS: In patients with symptomatic subclavian artery stenosis, baseline carotid intima-media thickness and ICA stenosis of 50% or greater are associated with multiterritory atherosclerosis, whereas intima-media thickness progression is associated with the risk of cardiovascular events.

Decision-making microRNAs (miR-124, -133a/b, -34a and -134) in patients with occluded target vessel in acute coronary syndrome.

BACKGROUND: Coronary artery occlusion does not always manifest with ST-elevation, and some patients can have patent coronary vessel. AIM: We evaluated circulating microRNA (miRNA) profiles to discriminate subjects with infarct-related artery (IRA) occlusion. METHODS AND RESULTS: Patients (n = 43) with uncomplicated acute coronary syndrome and positive troponins were classified with respect to patent vs. occluded IRA or ST-elevation vs. non-ST elevation MI (STEMI vs. NSTEMI). Expression levels of serum miRNAs (miR-1, -16, -34a, -122, -124, -208b, -133a/b, -375, and -499) were analysed. Out of 16 STEMI and 27 NSTEMI patients, IRA occlusion was noted in 12 and 15 patients, respectively. The remaining four STEMI and 12 NSTEMI patients had patent IRA. STEMI patients had higher troponin T levels and a 3.83-fold higher miR-134 expression (p < 0.025). Patients with the occluded vs. patent IRA had higher levels of miR-133a (fold change: 7.00), miR-133b (4.57), miR-34a (5.50), miR-124 (2.55), and miR-134 (3.45) but no difference in troponin T levels. Receiver operator characteristic analysis identified decision-making miRNAs in occluded vessels: miR-124 (AUC: 0.787, p < 0.001), miR-133b (AUC: 0.704, p = 0.006), and miR-134 (AUC: 0.686, p = 0.016). With respect to STEMI, only miR-134 showed a discriminating value (AUC: 0.725, p = 0.002). CONCLUSIONS: The degree of IRA occlusion determines circulating miRNA expression, and specific miRNAs may be useful in indicating patients requiring urgent coronary revascularisation.

Low prevalence of collateral cerebral circulation in the circle of Willis in patients with severe carotid artery stenosis and recent ischemic stroke.

INTRODUCTION: The circle of Willis is thought to play a key role in development of collateral flow in patients with internal carotid artery stenosis (ICAS). AIM: To assess flow in the circle of Willis in patients with recent ischemic stroke (IS). MATERIAL AND METHODS: The study included 371 patients, 102 symptomatic with severe ICAS and recent IS (within the last 3 months) (group I) and 269 asymptomatic with severe ICAS (group II). Flow in the middle (MCA), anterior (ACA) and posterior (PCA) cerebral arteries and pattern of the cross-flow through anterior (ACoA) and posterior (PCoA) communicating arteries were assessed with transcranial color-coded Doppler ultrasonography (TCCD). RESULTS: The ACoA or PCoA was less prevalent in group I than in group II (54% vs. 78%, p < 0.001 and 20% vs. 42%, p < 0.001, respectively), resulting in lower peak-systolic velocity (PSV) in the MCA in group I vs. group II (p = 0.015). Any collateral pathway was present in 67% of patients in group I, compared to 86% in group II (p < 0.001). Both PSV and end-diastolic (EDV) flow velocity in the ACA were lower in patients with recent IS, compared to asymptomatic subjects (71 ±24 cm/s vs. 86 ±34 cm/s, p < 0.001 and 32 ±12 cm/s vs. 37 ±17 cm/s, p = 0.038, respectively). Presence of ACoA or PCoA and higher PSV in the MCA and ACA were associated with significant risk reduction of IS (RR = 0.28 (95% CI = 0.16-0.49, p < 0.001), RR = 0.28 (95% CI = 0.15-0.52, p < 0.001), RR = 0.97 (95% CI = 0.96-0.99, p < 0.001), RR = 0.99 (95% CI = 0.98-0.99, p < 0.032), respectively). However, ROC curves failed to show reliable MCA or ACA PSV cut-offs for IS risk assessment. CONCLUSIONS: The ACoA and PCoA seem to play a key role in the evaluation of IS risk in subjects with severe ICAS.

Assessment of left ventricle function in aortic stenosis: mitral annular plane systolic excursion is not inferior to speckle tracking echocardiography derived global longitudinal peak strain.

BACKGROUND: Early detection of left ventricle (LV) systolic dysfunction is essential for management of patients with aortic stenosis (AS). Two- dimensional speckle tracking derived global longitudinal peak strain (GLPS) is more sensitive than ejection fraction (EF) but requires good image quality and is not easily accessible. The aim of the study was to compare GLPS with traditional echocardiographic parameter- mitral annular plane systolic excursion (MAPSE) in AS. MATERIAL AND METHODS: In consecutive patients with moderate to severe AS and LV ejection fraction ≥ 50% standard echocardiography and two-dimensional speckle tracking echocardiography were performed. Mitral annular plane systolic excursion and global longitudinal peak strain were obtained from apical echocardiographic views. RESULTS: A total of 82 patients were examined, median age was 68 (60-78), 56% of them were men. There was a positive correlation between aortic valve area index (AVAI) and: MAPSE (r = 0.334, p = 0.002), MAPSE indexed for body surface area- MAPSEI (r = 0.349, p = 0.001) and GLPS (r = 0.342, p = 0.002) but not EF (r = 0.031, p = 0.782). A positive correlation was found between GLPS and MAPSE (r = 0.558, p < 0.001) and between GLPS and MAPSEI (r = 0.543, p < 0.001). All above parameters were significantly lower in symptomatic patients compared to asymptomatic subjects (GLPS: -13.82 ± 3.56 vs. -16.39 ± 3.16%, p = 0.002, MAPSE: 10.49 ± 1.91 vs. 11.95 ± 1.82 mm, p = 0.001 and MAPSEI: 5.66 (4.83-6.6) vs. 6.46 ± 0.97 mm/m2, p = 0.005). CONCLUSION: Despite the development of the modern echocardiographic techniques, mitral annular plane systolic excursion can still be used as a sensitive tool to detect early longitudinal LV systolic dysfunction.

The effect of physical activity on serum levels of selected biomarkers of atherosclerosis.

BACKGROUND: Physical activity is associated with a lower risk of cardiovascular disease but the mechanism underlying this association is unclear. These benefits of physical activity might result from its effects on inflammation and endothelial function. AIM: We investigated whether cardiorespiratory fitness and the level of physical activity are associated with biomarkers of atherosclerosis in athletes and nonathletes. METHODS: Forty six athletes and 46 age- and sex-matched subjects who did not exercise regularly were studied. All subjects underwent anthropometric measurements and maximal treadmill cardiopulmonary exercise tests. Physical activity level was assessed using the International Physical Activity Questionnaire-Short Form (IPAQ-SF). Blood samples were taken before and immediately after exercise. Serum interleukin (IL)-6, soluble CD40 ligand (sCD40L) and soluble intercellular adhesion molecule- 1 (sICAM-1) levels were determined using the ELISA method. RESULTS: In all participants, IL-6 level was significantly increased after exercise as compared to baseline (1.35 ± 2.6 vs. 1.46 ± ± 2.1 pg/mL, p = 0.01). Resting IL-6 and sCD40L levels were lower in athletes as compared to nonathletes (0.7 ± 0.92 vs. 1.8 ± 3.52 pg/mL, p = 0.003, and 888.8 ± 892.9 vs. 2367.7 ± 8743.4 pg/mL, p = 0.005, respectively), while sICAM-1 levels did not differ between the two groups. IL-6 level correlated negatively with peak oxygen consumption (r = -0.25, p = 0.035) and the IPAQ-SF score (r = -0.26, p = 0.02), and sCD40L level correlated negatively with the IPAQ-SF score (r = -0.4, p = 0.004). CONCLUSIONS: Intensive exercise training and high exercise capacity are associated with lower serum IL-6 and sCD40L levels. This may constitute an important factor limiting progression of atherosclerosis.

Assessment of left ventricle function in patients with symptomatic and asymptomatic aortic stenosis by 2-dimensional speckle-tracking imaging.

BACKGROUND: Global longitudinal peak strain (GLPS) quantifies left ventricle (LV) long-axis contractility. Early detection of LV systolic dysfunction is pivotal in diagnosis and treatment of patients with aortic stenosis (AS). This study was performed to assess LV longitudinal systolic function by GLPS derived from 2-dimensional speckle tracking imaging (2D-STI) in AS patients in comparison to standard echocardiographic parameters. MATERIAL/METHODS: Laboratory tests, standard echocardiography, tissue Doppler imaging (TDI) and 2D-STI examinations with GLPS calculation were performed in 49 consecutive patients with moderate to severe AS with LV ejection fraction ≥50% and 18 controls. RESULTS: While LVEF do not differentiate AS patients from controls, GLPS was significantly decreased in the AS group (-15.30 ± 3.25% vs. -19.60 ± 2.46% in controls, p<0.001). GLPS was significantly reduced in symptomatic AS patients as compared to the asymptomatic AS group [-15.5 (11.8-16.8) vs. -17.5 (14.7-18.9)%, p=0.02]. CONCLUSIONS: In aortic stenosis patients, despite normal left ventricle ejection fraction, long-axis left ventricular function is impaired, which manifests in global longitudinal peak strain reduction. GLPS reveals that LV function impairment is more pronounced in symptomatic as compared to asymptomatic AS patients. Further studies are needed to determine the prognostic significance of early LV function impairment in aortic stenosis patients showed by GLPS.