Healthy Arteries Research Articles

Image-based quantification of 3D morphology for bifurcations in the left coronary artery: Application to stent design.

Improved strategies for stent-based treatment of coronary artery disease at bifurcations require a greater understanding of artery morphology. We developed a workflow to quantify morphology in the left main coronary (LMCA), left anterior descending (LAD), and left circumflex (LCX) artery bifurcations. Computational models of each bifurcation were created for 55 patients using computed tomography images in 3D segmentation software. Metrics including cross-sectional area, length, eccentricity, taper, curvature, planarity, branching law parameters, and bifurcation angles were assessed using open-sources software and custom applications. Geometric characterization was performed by comparison of means, correlation, and linear discriminant analysis (LDA). Differences between metrics suggest dedicated or multistent approaches should be tailored for each bifurcation. For example, the side branch of the LCX (i.e., obtuse marginal; OM) was longer than that of the LMCA (i.e., LCXprox) and LAD (i.e., first diagonal; D1). Bifurcation metrics for some locations (e.g., LMCA Finet ratio) provide results and confidence intervals agreeing with prior findings, while revised metric values are presented for others (e.g., LAD and LCX). LDA revealed several metrics that differentiate between artery locations (e.g., LMCA vs. D1, LMCA vs. OM, LADprox vs. D1, and LCXprox vs. D1). These results provide a foundation for elucidating common parameters from healthy coronary arteries and could be leveraged in the future for treating diseased arteries. Collectively the current results may ultimately be used for design iterations that improve outcomes following implantation of future dedicated bifurcation stents. © 2015 Wiley Periodicals, Inc.

Nanopartículas calcificantes como factor etiológico del desarrollo de hiperplasia y calcificación vascular

ObjectiveCalcifying nanoparticles (NP) have been detected recently in calcified human arterial specimens, and are involved in the process of calcification. This study was designed to test the hypothesis that human-derived NP could worsen the response to arterial endothelial injury and induce vascular calcification. MethodsThe right carotid artery of 24 New Zealand rabbits was injured with an angioplasty balloon. Animals were perfused intravenously with saline (100mL) during the experiment and divided into 3 groups: group A, control; group B, exposed to NP (2mL) obtained from calcified aortic valves; and group C, exposed to NP (2mL) and treated post-operatively with atorvastatin (2.5mg/kg/24h). At 30 days, both carotid arteries were removed and examined histologically. Blood measurements were monitored during the study. ResultsThe intimal hyperplasia area was significantly larger in the injured right carotid artery compared with the left un-operated carotid artery in all groups. There was no significant variation in medial area between groups. Morphometrically, the intima/media ratio (IMR) was significantly higher in damaged carotids compared with controls. A significant increase in the IMR was found in group B (1.81±0.41) compared with group A (0.38±0.59; P=.004) or group C (0.89±0.79; P=.035). Differences between groups C and A were not significant (P=.064). Calcifications were observed in 6 animals, all of which had been exposed to NP (4 in group B, and 2 in group C, P=.027). Plasma levels of cholesterol and triglycerides remained stable. ConclusionsThis study confirms the ability of systemic inoculation of human-derived NP to accelerate hyperplasia and stimulate calcification in localised areas of arteries previously submitted to endothelial damage, while it was harmless in healthy arteries. Atorvastatin was demonstrated to slow down this process.

Abstract 18004: The Next Generation of Stretchable Sensors for Intravascular Electrochemical Impedance Spectroscopy of Varying Levels of Lipid Burden and Atherosclerosis

Introduction: Atherosclerosis, a chronic inflammatory arterial disease, persists as the leading cause of acute coronary events or stroke in developed countries. Developing a reliable, reproducible, safe and simple strategy to detect lipid-rich, metabolically active, and mechanically unstable plaques remains an unmet bioengineering challenge. It is recognized that oxidized low-density lipoprotein (oxLDL) and macrophages produce changes in the electrochemical properties of vessel walls that can be measured by electrochemical impedance spectroscopy (EIS). Hypothesis: We tested the hypothesis that a new, 2-point intravascular micro-electrode configuration enables flexible frequency-dependent EIS characterization of lipid-rich lesions. Methods: A simplified yet robust sensor design with higher differentiating capability, featuring individually addressable, flexible round electrodes (Æ = 240 μm), was developed and microfabricated for intravascular EIS investigation. We documented in vivo deployment of stretchable EIS sensors for endoluminal interrogation and impedimetric investigation at multiple levels of the aortic tree characterized by differing stages of atherosclerotic disease in a New Zealand White rabbit model, compared EIS values in vivo and ex vivo, confirmed endoluminal atherosclerotic lesions by intravascular ultrasound, histology and immunohistochemistry, and obtained EIS profiles in epicardial fat tissue. Results: The frequency sweep revealed overlapping tissue resistivity from 1-10 kHz, but distinct capacitive property from 10-300 kHz, thereby providing complementary differentiation in both impedance (Ω) and phase (f) between healthy arteries and varying levels of atherosclerotic lesions. The EIS measurements revealed a significant increase in impedance in the presence of oxLDL-rich lesions from 5219 Ω in control to 5949 Ω in mild, 6988 Ω in moderate and 8351 Ω in severe plaque at 300 kHz (P < 0.05, n = 5 controls, n = 3 atherosclerotic). The correlation between in vivo and ex vivo analyses using Spearman was r = 0.97 (P < 0.0001, n = 8 aortic segments). Conclusion: We demonstrate a new, paired-electrode based EIS sensor for intravascular interrogation of lipid-laden lesions to complement diagnostic angiography.

Abstract 18689: Lower Carotid Artery Compliance and Greater Carotid β-stiffness Index are Associated With Slower Processing Speed and Reduced Working Memory Performance in Older Healthy Adults

Introduction: Higher central elastic artery stiffness is associated with greater brain white matter hyperintensity volume and lower cognitive performance in older adults. However, it is unknown if higher central artery stiffness is related to regional decreases in brain white matter integrity (WMI) and reductions in working memory or processing speed. We hypothesized that greater large elastic artery stiffness would be associated with lower working memory and slower processing speed as well as lower regional WMI in older adults. Methods: In young (n=19; 23.8 ± 0.7 yrs) and older (n=22; 64.4 ± 4.2 yrs; range 60-71 yrs) healthy adults, aortic stiffness (carotid-femoral pulse wave velocity, cfPWV, via tonometry) and carotid artery compliance and β-stiffness index (carotid artery ultrasound and tonometry) were determined non-invasively. WMI was assessed by fractional anisotrophy (FA) (3T MRI) from diffusion tensor images. An N-Back task and letter and pattern comparison tests were performed as measures of working memory and processing speed, respectively. d’Prime, a measure of signal detection, was calculated on the N-Back task as a ratio of positive ‘hits’ to ‘false alarm’ responses. Results: cfPWV (8.4 ± 0.5 vs. 5.2 ± 0.2 m/sec, P<0.01) and carotid β-stiffness (11.3 ± 0.9 vs. 6.1 ± 0.7 U, P<0.01) were greater and carotid compliance (0.16 ± 0.02 vs. 0.07 ± 0.01 mm/mmHg, P<0.01) was lower in older vs. young adults. Carotid β, but not aortic, stiffness was associated with slower processing speed on letter (10.7 ± 0.5 vs. 13.5 ± 0.6 units/ 30 sec, p<0.01) and pattern (16.6 ± 0.6 vs. 22.0 ± 0.7 units/30 sec, p<0.01) comparison tasks and reduced working memory (1.8 ± 0.2 vs. 3.1± 0.2, P<0.01) in older vs. young adults. Lower carotid compliance was associated with slower performance on the letter (r=0.55, P<0.01) and pattern (r=0.54, P<0.01) tests and lower d’Prime (r=0.48, P<0.05). Greater β-stiffness was related to pattern comparison scores only (r=-0.54, P<0.01). Aortic stiffness, carotid compliance and β-stiffness were not related to WMI in any brain region. Conclusions: Our results suggest that lower carotid artery compliance and greater β-stiffness are associated with reduced working memory performance and slower processing speed but not regional WMI in older adults.

Histone acetylation and methylation significantly change with severity of atherosclerosis in human carotid plaques.

The aim of the study was to analyze histone acetylation, methylation, and the expression of their corresponding transferases in atherosclerotic plaques of patients with carotid artery stenosis. Atherosclerotic tissue from our biobank (n=80) was divided into various segments covering all plaque stages and classified according to the American Heart Association. The plaques were assigned to early (types I-III) or advanced (types V-VII) stage group of atherosclerosis. Ten healthy carotid arteries from transplant donors served as controls. The expression of histone acetyltransferases (GNAT group: GCN5L, P300/CBP group: P300, MYST group: MYST1 and MYST2) and histone methyltransferases (H3K4: MLL2/4, SET7/9, and hSET1A; H3K9: SUV39H1, SUV39H2, ESET/SETDB1, and EHMT1; H3K27: EZH2 and G9a) was analyzed by SYBR-green-based real-time polymerase chain reaction. Histone acetylation/methylation in the cells within atherosclerotic plaques was determined by immunohistochemistry. Increased histone acetylation was observed on H3K9 and H3K27 in smooth muscle cells (SMCs) in advanced atherosclerotic lesions compared to healthy vessels (P=.002 and .034). H3K9 acetylation in SMCs and macrophages was associated with plaque severity of atherosclerosis (P=.048 and <.001). Expression of GCN5L and MYST1 also correlated with the severity of atherosclerosis (P<.001). Methylation of H3K9 and H3K27 was significantly reduced in atherosclerotic plaques in SMCs and inflammatory cells (P<.001 and .026). Methylation on H3K4 was significantly associated with the severity of atherosclerosis. Expression of methyltransferase MLL2/4 was increased in advanced stages of atherosclerosis (P<.001). Histone acetylation and methylation seem to play a decisive role in atherosclerosis, showing significant differences between healthy vessels and vessels at different stages of atherosclerosis.

Effects of healthy aging and coronary artery disease on sympathetic axonal recruitment strategies during prolonged end-inspiratory apnea

Effects of healthy aging and coronary artery disease on sympathetic axonal recruitment strategies during prolonged end-inspiratory apnea

Quantitative analysis of human cutaneous vasomotor innervation

Background: A neural recruitment strategy exists within the sympathetic nervous system (SNS) of young individuals, whereby elevated sympathetic outflow during reflex-mediated stress is associated with recruitment of latent sub-populations of largeramplitude axons and acute modifications in synaptic delays. Aim: We investigated whether these recruitment patterns persist with healthy aging and coronary artery disease (CAD). Methods: Muscle sympathetic nerve activity (MSNA; microneurography) was measured in 14 young healthy individuals (YH; 25 ± 4 yrs), 14 older healthy individuals (OH; 59 ± 9 yrs), and 15 CAD patients (63 ± 11 yrs) at baseline and during a maximal end-inspiratory apnea. Action potential (AP) patterns were studied using a novel AP analysis technique. Results: Baseline burst frequency was greater in OH (35 ± 10 bursts/min) and CAD (39 ± 5 bursts/min), compared to YH (17 ± 7 bursts/min; all P b 0.001). APs/min and APs/burst increased during apnea (time effect; all P b 0.001). When APs were binned according to peak-to-peak amplitude (i.e., into ‘clusters’), total ‘clusters’ increased during apnea in YH (Δ6 ± 3; P b 0.001) and OH (Δ 2±1 ; P b 0.01), but was unchanged in CAD (Δ0 ± 1; P N 0.05). The number of active clusters/burst increased during apnea in YH (Δ 2±1 ; P b 0.001), OH (Δ1 ± 1; P b 0.01), and CAD (Δ1 ± 1; P = 0.01) groups, although greater change occurred in YH versus OH and CAD (all P b 0.01). In all groups, a pattern emerged where the cluster size-latency profile was shifted downwards for every corresponding cluster during apnea. Conclusion: The capacity of the SNS to recruit latent sub-populations of larger-amplitude axons is reduced with healthy aging and lost in CAD, while the ability to acutely modify synaptic delays appears unaltered.

Vascular compression syndromes.

Dealing with vascular compression syndromes is one of the most challenging tasks in Vascular Medicine practice. This heterogeneous group of disorders is characterised by external compression of primarily healthy arteries and/or veins as well as accompanying nerval structures, carrying the risk of subsequent structural vessel wall and nerve damage. Vascular compression syndromes may severely impair health-related quality of life in affected individuals who are typically young and otherwise healthy. The diagnostic approach has not been standardised for any of the vascular compression syndromes. Moreover, some degree of positional external compression of blood vessels such as the subclavian and popliteal vessels or the celiac trunk can be found in a significant proportion of healthy individuals. This implies important difficulties in differentiating physiological from pathological findings of clinical examination and diagnostic imaging with provocative manoeuvres. The level of evidence on which treatment decisions regarding surgical decompression with or without revascularisation can be relied on is generally poor, mostly coming from retrospective single centre studies. Proper patient selection is critical in order to avoid overtreatment in patients without a clear association between vascular compression and clinical symptoms. With a focus on the thoracic outlet-syndrome, the median arcuate ligament syndrome and the popliteal entrapment syndrome, the present article gives a selective literature review on compression syndromes from an interdisciplinary vascular point of view.

MPST but not CSE is the primary regulator of hydrogen sulfide production and function in the coronary artery.

Hydrogen sulfide (H2S) has emerged as an important gasotransmitter in the vasculature. In this study, we tested the hypothesis that H2S contributes to coronary vasoregulation and evaluated the physiological relevance of two sources of H2S, namely, cystathionine-γ-lyase (CSE) and 3-mercaptypyruvate sulfertransferase (MPST). MPST was detected in human coronary artery endothelial cells as well as rat and mouse coronary artery; CSE was not detected in the coronary vasculature. Rat coronary artery homogenates produced H2S through the MPST pathway but not the CSE pathway in vitro. In vivo coronary vasorelaxation response was similar in CSE knockout mice, wild-type mice (WT), and WT mice treated with the CSE inhibitor propargylglycine, suggesting that CSE-produced H2S does not have a significant role in coronary vasoregulation in vivo. Ex vivo, the MPST substrate 3-mercaptopyruvate (3-MP) and H2S donor sodium hydrosulfide (NaHS) elicited similar coronary vasoreactivity responses. Pyruvate did not have any effects on vasoreactivity. The vasoactive effect of H2S appeared to be nitric oxide (NO) dependent: H2S induced coronary vasoconstriction in the presence of NO and vasorelaxation in its absence. Maximal endothelial-dependent relaxation was intact after 3-MP and NaHS induced an increase in preconstriction tone, suggesting that endothelial NO synthase activity was not significantly inhibited. In vitro, H2S reacted with NO, which may, in part explain the vasoconstrictive effects of 3-MP and NaHS. Taken together, these data show that MPST rather than CSE generates H2S in coronary artery, mediating its effects through direct modulation of NO. This has important implications for H2S-based therapy in healthy and diseased coronary arteries.

Effect of Pulmonary Conduit Oversizing on Hemodynamics in Children

Implanting the largest valved conduit possible - oversizing - to reconstruct an absent connection from the right ventricle to the pulmonary artery in certain types of congenital heart defects has been suggested as a compensating measure for somatic outgrowth of the patient. However, one effect that has not been investigated yet is the hemodynamic consequence. For this purpose, virtual implantation and flow simulations were conducted in this study. To isolate the effects of conduit oversizing on the hemodynamics observed after conduit implantation and outgrowth, calculated wall shear stresses (WSS) of image-based computational fluid dynamic (CFD) simulations were used as indicators. Three different sizes of valved conduits (20 mm, 22 mm, and 24 mm), including the largest possible conduit size, virtually implanted in a child-sized healthy pulmonary artery and the corresponding adult-sized model were investigated. The child and adult models show a decrease of the mean WSS (approx. 26%) in the whole domain with an increase of the conduit size. When looking at the mean WSS at the anastomosis, for the child model the WSS is significantly increased (approx. 40%) when oversizing (Z-score +3.21). In contrast, the stresses are decreased for the adult model (34%) when using the largest conduit (Z-score +0.25). Based on the results of this study, it must be considered that choosing a prosthesis size that will lead to high WSS and an associated intimal reaction, possibly leading to stenosis, can defeat the benefit of having a nominally larger orifice area directly after implantation.

Glucose-Dependent Insulinotropic Polypeptide Stimulates Osteopontin Expression in the Vasculature via Endothelin-1 and CREB.

Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone with extrapancreatic effects beyond glycemic control. Here we demonstrate unexpected effects of GIP signaling in the vasculature. GIP induces the expression of the proatherogenic cytokine osteopontin (OPN) in mouse arteries via local release of endothelin-1 and activation of CREB. Infusion of GIP increases plasma OPN concentrations in healthy individuals. Plasma endothelin-1 and OPN concentrations are positively correlated in patients with critical limb ischemia. Fasting GIP concentrations are higher in individuals with a history of cardiovascular disease (myocardial infarction, stroke) when compared with control subjects. GIP receptor (GIPR) and OPN mRNA levels are higher in carotid endarterectomies from patients with symptoms (stroke, transient ischemic attacks, amaurosis fugax) than in asymptomatic patients, and expression associates with parameters that are characteristic of unstable and inflammatory plaques (increased lipid accumulation, macrophage infiltration, and reduced smooth muscle cell content). While GIPR expression is predominantly endothelial in healthy arteries from humans, mice, rats, and pigs, remarkable upregulation is observed in endothelial and smooth muscle cells upon culture conditions, yielding a "vascular disease-like" phenotype. Moreover, the common variant rs10423928 in the GIPR gene is associated with increased risk of stroke in patients with type 2 diabetes.

Expression of the NLRP3 Inflammasome in Carotid Atherosclerosis

The aim of the present study was to investigate NLRP3 inflammasome expression in human carotid atherosclerotic plaques and its relationship to plaque vulnerability. Carotid atherosclerotic plaques collected from 30 patients scheduled for carotid endarterectomy (CEA) were subjected to immunohistochemical, mRNA, and protein expression studies. Ten mesenteric arteries from intestinal cancer patients served as controls for the immunohistochemical studies. Twenty individuals who had no carotid stenosis or coronary artery stenosis served as controls for analyzing atherosclerotic risk factors and for enzyme-linked immunosorbent assay (ELISA) studies. Serum samples were collected from all patients to determine interleukin-1β (IL-1β) and IL-18 levels. The NLRP3 inflammasome signaling pathway components NLRP3, ASC, caspase-1, IL-1β, and IL-18 were strongly expressed in carotid atherosclerotic plaques, but not in healthy mesenteric arteries. Immunohistochemical, mRNA, and protein expression studies revealed higher expression levels of NLRP3, ASC, caspase-1, IL-1β, and IL-18 in unstable compared to stable plaques. The NLRP3 inflammasome was localized in the cytoplasm of macrophages and foam cells and was associated with cholesterol crystal clefts inside and outside of cells. ELISA showed that the serum levels of the cytokines IL-1β and IL-18 were higher in the CEA group compared to controls. These results demonstrated for the first time the close relationship between the expression of NLRP3 signaling pathway and human carotid atherosclerotic plaques. NLRP3, ASC, caspase-1, IL-1β, and IL-18 were associated with plaque vulnerability and atherogenesis. The serum levels of IL-1β and IL-18 may be useful predictors of atherosclerosis.

Mechanical Study of Nitinol Stents in Situation of Inflection on arterial Models

The tests which are presented to us to evaluate the nitinol stents used in the superficial femoral artery (SFA) are carried out without taking into account of their environment of deployment. The objectives of this work were (1) the creation of a model of healthy and pathological artery, (2) the creation of dynamometric tests reproducing the mechanical constraints of the SFA and (3) the study and comparison of different nitinol stents under these conditions.

Safety and Efficacy of New Sirolimus-eluting Stent Models in a Preclinical Study

Introduction and objectivesInitial preclinical studies are required during the process of improving polymers, platforms, and drug-eluting systems for new coronary stent designs. Our objective was to analyze the efficacy and safety of new drug-eluting stent models compared with a conventional stent and commercialized drug-eluting stents in an experimental model with healthy porcine coronary arteries. MethodsSixty stents (conventional stent, new sirolimus-eluting stents: drug-eluting stents 1, 2 and 3; Cypher® and Xience®) were randomly placed in the coronary arteries of 20 Large White domestic pigs. Angiographic and histomorphometric studies were done 28 days later. ResultsThe stents were implanted at a stent/artery ratio of 1.34±0.15, with no significant differences between groups. The new stents showed less late loss and angiographic restenosis than conventional stents (P=.006 and P<.001, respectively). Histologically, restenosis and neointimal area were lower with all the new platforms than with the conventional stents (P<.001 for each variable), and no differences were found vs the drug-eluting stents on the market. Safety data showed that endothelialization was lower with drug-eluting stents than with conventional stents, except for drug-eluting stent 3 (P=.084). Likewise, inflammation was lower with drug-eluting stent 3 than with other stents. ConclusionsThe new drug-eluting stent platforms studied are associated with less restenosis than conventional stents and showed no significant differences in safety or efficacy vs commercialized drug-eluting stents.

Seguridad y eficacia de nuevos modelos de stents liberadores de sirolimus en el modelo preclínico

Introduction and objectivesInitial preclinical studies are required during the process of improving polymers, platforms, and drug-eluting systems for new coronary stent designs. Our objective was to analyze the efficacy and safety of new drug-eluting stent models compared with a conventional stent and commercialized drug-eluting stents in an experimental model with healthy porcine coronary arteries. MethodsSixty stents (conventional stent, new sirolimus-eluting stents: drug-eluting stents 1, 2 and 3; Cypher® and Xience®) were randomly placed in the coronary arteries of 20 Large White domestic pigs. Angiographic and histomorphometric studies were done 28 days later. ResultsThe stents were implanted at a stent/artery ratio of 1.34±0.15, with no significant differences between groups. The new stents showed less late loss and angiographic restenosis than conventional stents (P=.006 and P<.001, respectively). Histologically, restenosis and neointimal area were lower with all the new platforms than with the conventional stents (P<.001 for each variable), and no differences were found vs the drug-eluting stents on the market. Safety data showed that endothelialization was lower with drug-eluting stents than with conventional stents, except for drug-eluting stent 3 (P=.084). Likewise, inflammation was lower with drug-eluting stent 3 than with other stents. ConclusionsThe new drug-eluting stent platforms studied are associated with less restenosis than conventional stents and showed no significant differences in safety or efficacy vs commercialized drug-eluting stents.Full English text available from:www.revespcardiol.org/en

Pharmacokinetic Study of Paclitaxel Concentration after Drug-Eluting Balloon Angioplasty in the Iliac Artery of Healthy and Atherosclerotic Rabbit Models

PurposeTo assess whether the presence of an atherosclerotic lesion may alter the deposition kinetics of paclitaxel on the arterial wall after drug-eluting balloon (DEB) angioplasty, as well as paclitaxel concentrations in serum and in the recovered balloons. Materials and MethodsThree New Zealand White rabbit models were created: an atheroma group (arterial mechanical injury and hyperlipidic diet; group A), a prelesional group (fat arterial infiltration, hyperlipidic diet; group B), and a control healthy group (group C). Forty-five animals underwent DEB angioplasty in the iliac artery. Arteries and serum samples were analyzed by liquid chromatography/tandem mass spectrometry at 1, 24, 48, 72, and 96 hours (arteries) and at 1, 6, 12, and 24 hours (serum). Recovered balloons were analyzed by UV chromatography. Histologic and statistical analyses were also performed. ResultsGroup A showed significantly higher arterial paclitaxel concentrations in the first hour after DEB angioplasty (632.05 ng/mg ± 125.75 in group A vs 179.55 ng/mg ± 45.64 and 168.54 ng/mg ± 83.48 in groups B and C, respectively; P < .05). Paclitaxel was undetectable in serum at 24 hours in all groups, but the amount was significantly higher (P < .05) in group B at 1, 6, and 12 hours. The paclitaxel amount in navigated balloons from group A was significantly lower than in other groups (P < .05). ConclusionsPaclitaxel concentration in an atherosclerotic lesion model immediately after DEB angioplasty is nearly fourfold higher than in a healthy artery. Paclitaxel remains in the bloodstream longer when a universal state of fat arterial infiltration is achieved. These findings could have clinical implications, as studies testing commercial drug-eluting devices on healthy animals may be underestimating paclitaxel arterial uptake.

Determinants of Aortic Root Dilatation and Reference Values Among Young Adults Over a 20-Year Period: Coronary Artery Risk Development in Young Adults Study.

Aortic size increases with age, but factors related to such dilatation in healthy young adult population have not been studied. We aim to evaluate changes in aortic dimensions and its principal correlates among young adults over a 20-year time period. Reference values for aortic dimensions in young adults by echocardiography are also provided. Healthy Coronary Artery Risk Development in Young Adults (CARDIA) study participants aged 23 to 35 years in 1990-1991 (n=3051) were included after excluding 18 individuals with significant valvular dysfunction. Aortic root diameter (ARD) by M-mode echocardiography at year-5 (43.7% men; age, 30.2 ± 3.6 years) and year-25 CARDIA exams was obtained. Univariable and multivariable analyses were performed to assess associations of ARD with clinical data at years-5 and -25. ARD from year-5 was used to establish reference values of ARD in healthy young adults. ARD at year-25 was greater in men (33.3 ± 3.7 versus 28.7 ± 3.4 mm; P<0.001) and in whites (30.9 ± 4.3 versus 30.5 ± 4.1 mm; P=0.006). On multivariable analysis, ARD at year-25 was positively correlated with male sex, white ethnicity, age, height, weight, 20-year gain in weight, active smoking at baseline, and 20-year increase in diastolic, systolic, and mean arterial pressure. A figure showing the estimated 95th percentile of ARD by age and body surface area stratified by race and sex is provided. This study demonstrates that smoking, blood pressure, and increase in body weight are the main modifiable correlates of aortic root dilation during young adulthood. Our study also provides reference values for ARD in young adults.

O82. Association between anti-angiogenic factor and signs of arterial aging in women with preeclampsia

Introduction Preeclampsia (PE) is a state of exaggerated inflammation and is associated with increased risk of cardiovascular disease later in life. Soluble fms-like tyrosine kinase-1 (s-Flt1) is an antiangiogenic factor and is increased, whereas placental growth factor (PlGF) is an angiogenic factor and is decreased in PE. Objectives Aim of this study was to assess s-Flt1, PlGF and s-Flt1/PlGF and its correlation with common carotid artery (CCA) intima, media and intima/media ratio (I/M) in women with and without PE. Methods We measured serum s-Flt1 and PlGF using commercially available enzyme-linked immunosorbent assay kits and individual CCA intima and media thicknesses were estimated by high-frequency (22 MHz) ultrasound in 55 women at PE diagnosis and in 64 women with normal pregnancies at a similar gestational age and about one year postpartum. A thick intima, thin media and high I/M are signs of a less healthy artery wall. Results During pregnancy, we found higher values of s-Flt1, lower values of PlGF and thicker intima, thinner media and higher I/M in women with PE than in women with normal pregnancy (all p 0.0001 ). Further, s-Flt1 and s-Flt1/PlGF showed a positive correlation with intima thickness and I/M but a negative correlation with media thickness (all p 0.0001 ). About one year postpartum, s-Flt1 and s-Flt1/PlGF values had decreased in both groups but there was still a significant group difference (all p 0.0001 ). No significant correlation with CCA wall layers (except for s-Flt1 and intima thickness, p = 0.007 ) was found at postpartum. Conclusions We found substantially higher values of s-Flt1 and s-Flt1/PlGF in women with PE and these higher values were associated with signs of arterial aging. Our findings support the findings of increased risk of cardiovascular disease in women with PE.

Membrane-bound Klotho is not expressed endogenously in healthy or uraemic human vascular tissue.

Cardiovascular disease (CVD) is the leading cause of death in patients with chronic kidney disease (CKD), a disease state that is strongly associated with loss of renal and systemic (alpha-)Klotho. Reversely, murine Klotho deficiency causes marked medial calcification. It is therefore thought that Klotho conveys a vasculoprotective effect. Klotho expression in the vessel wall, however, is disputed. We assessed Klotho expression in healthy human renal donor arteries (n = 9), CKD (renal graft recipient) arteries (n = 10), carotid endarterectomy specimens (n = 8), other elastic arteries (three groups of n = 3), and cultured human aortic smooth muscle cells (HASMCs) (three primary cell lines), using immunohistochemistry (IHC), immunofluorescence, quantitative reverse transcriptase-polymerase chain reaction, and western blotting (WB). We have extensively validated anti-Klotho antibody KM2076 by comparing staining patterns with other anti-Klotho antibodies (SC-22220, SC-22218, and AF1819), competition assays with recombinant Klotho, IHC on Klotho-deficient kl/kl mouse kidney, and WB with recombinant Klotho. Using KM2076, we could not detect full-length Klotho in vascular tissues or HASMCs. On the mRNA level, using primers against all four exon junctions, klotho expression could not be detected either. Fibroblast growth factor 23 (FGF23) injections in mice induced FGF23 signalling in kidneys but not in the aorta, indicating the absence of Klotho-dependent FGF23 signalling in the aorta. Using several independent and validated methods, we conclude that full-length, membrane-bound Klotho is not expressed in healthy or uraemic human vascular tissue.

Maldi-msi identifies the molecular anatomy of healthy and atherosclerotic arteries and reveals tmsb4x early accumulation in intima layer during atherosclerosis development

Maldi-msi identifies the molecular anatomy of healthy and atherosclerotic arteries and reveals tmsb4x early accumulation in intima layer during atherosclerosis development