Objective. Previous studies suggest a protracted course of recovery after mechanical endothelial injury; confounders may include degree of injury and concomitant endothelial dysfunction. We sought to define the time course of endothelial function recovery using flow-mediated dilation (FMD), after ischaemia-reperfusion (IR) and mechanical injury in patients and healthy volunteers. The contribution of circulating CD133+/CD34+/VEGFR2+ “endothelial progenitor” (EPC) or repair cells to endothelial repair was also examined. Methods. 28 healthy volunteers aged 18–35 years underwent transient forearm ischaemia induced by cuff inflation around the proximal biceps and radial artery mechanical injury induced by inserting a wire through a cannula. A more severe mechanical injury was induced using an arterial sheath and catheter inserted into the radial artery of 18 patients undergoing angiography. Results. IR and mechanical injury produced immediate impairment of FMD (from 6.5 ± 1.2% to 2.9 ± 2.2% and from 7.4 ± 2.3% to 1.5 ± 1.6% for IR and injury, resp., each P < 0.001) but recovered within 6 hours and 2 days, respectively. FMD took up to 4 months to recover in patients. Circulating EPC did not change significantly during the injury/recovery period in all subjects. Conclusions. Recovery of endothelial function after IR and mechanical injury is rapid and not associated with a change in circulating EPC.

BACKGROUND At the APOE gene, encoding apolipoprotein E, genotypes of the ε2/ε3/ε4 alleles associated with higher LDL-cholesterol (LDL-C) levels are also associated with higher coronary risk. However, the association of APOE genotype with other cardiovascular biomarkers and risk of ischaemic stroke is less clear. We evaluated the association of APOE genotype with risk of ischaemic stroke and assessed whether the observed effect was consistent with the effects of APOE genotype on LDL-C or other lipids and biomarkers of cardiovascular risk. METHODS We conducted a systematic review of published and unpublished studies reporting on APOE genotype and ischaemic stroke. We pooled 41 studies (with a total of 9027 cases and 61,730 controls) using a Bayesian meta-analysis to calculate the odds ratios (ORs) for ischaemic stroke with APOE genotype. To better evaluate potential mechanisms for any observed effect, we also conducted a pooled analysis of primary data using 16 studies (up to 60,883 individuals) of European ancestry. We evaluated the association of APOE genotype with lipids, other circulating biomarkers of cardiovascular risk and carotid intima-media thickness (C-IMT). RESULTS The ORs for association of APOE genotypes with ischaemic stroke were: 1.09 (95% credible intervals (CrI): 0.84-1.43) for ε2/ε2; 0.85 (95% CrI: 0.78-0.92) for ε2/ε3; 1.05 (95% CrI: 0.89-1.24) for ε2/ε4; 1.05 (95% CrI: 0.99-1.12) for ε3/ε4; and 1.12 (95% CrI: 0.94-1.33) for ε4/ε4 using the ε3/ε3 genotype as the reference group. A regression analysis that investigated the effect of LDL-C (using APOE as the instrument) on ischaemic stroke showed a positive dose-response association with an OR of 1.33 (95% CrI: 1.17, 1.52) per 1 mmol/l increase in LDL-C. In the separate pooled analysis, APOE genotype was linearly and positively associated with levels of LDL-C (P-trend: 2 × 10(-152)), apolipoprotein B (P-trend: 8.7 × 10(-06)) and C-IMT (P-trend: 0.001), and negatively and linearly associated with apolipoprotein E (P-trend: 6 × 10(-26)) and HDL-C (P-trend: 1.6 × 10(-12)). Associations with lipoprotein(a), C-reactive protein and triglycerides were non-linear. CONCLUSIONS In people of European ancestry, APOE genotype showed a positive dose-response association with LDL-C, C-IMT and ischaemic stroke. However, the association of APOE ε2/ε2 genotype with ischaemic stroke requires further investigation. This cross-domain concordance supports a causal role of LDL-C on ischaemic stroke.

Raised blood pressure (BP) is the world's leading mortality risk factor. Childhood BP substantially predicts adult levels, and although both prenatal and postnatal growth influence it, their relative importance is debated. In a longitudinal study (Avon Longitudinal Study of Parents and Children) of 12 962 healthy children, we aimed to assess the relative contribution of different growth periods and of standardized measures of height versus weight-for-height (an adiposity marker) to BP at age 10 years. Conditional growth modeling was used in the 3230 boys and 3346 girls with BP measurements. Systolic BP was inversely associated with birth weight and weight-for-height but not length (−0.33, −0.27, and −0.12 mm Hg · SD −1 ; P =0.003, 0.035, and 0.35, respectively). In infancy, weight, weight-for-height, and height gains were all positively associated with systolic BP (0.90, 0.41, and 0.82 mm Hg · SD −1 , respectively; all P <0.001). After infancy, all of the growth modalities were positively associated with systolic BP (weight, 1.91; weight-for-height, 1.56; height, 1.20 mm Hg · SD −1 ; all P <0.001). Similar but weaker associations were found with diastolic BP. Although BP at 10 years was associated with both prenatal and early postnatal growth, their influence was small compared with that of later growth. Because BP ranking relative to the population is substantially determined in the first decade of life, a focus on strategies to reduce the development of adiposity from infancy onward, rather than an emphasis on the nutrition and weight of mothers and infants, should bring greater reductions in population BP.

Vasoprotective effects of erythropoietin in animal models are mediated by endothelium-derived NO and/or mobilization of EPCs (endothelial progenitor cells) and may be enhanced by ischaemia: whether they are present in humans is unknown. We examined whether the erythropoietin analogue darbepoetin improves FMD (flow-mediated dilatation), a measure of endothelium-derived NO, and whether this is influenced by preceding I/R (ischaemia/reperfusion). A total of 36 patients (50-75 years) with stable coronary artery disease were randomized to receive a single dose of darbepoetin (300 μg) or saline placebo. FMD was measured at the brachial artery using high-resolution ultrasound. CD133⁺/CD34⁺/VEGFR2⁺ (vascular endothelial growth factor receptor 2) circulating EPCs were enumerated by flow cytometry. Measurements were made immediately before darbepoetin/placebo and at 24 h, 72 h and 7 days. At 24 h, FMD was repeated after 20 min of I/R of the upper limb. A further group of 11 patients was studied according to the same protocol, all receiving darbepoetin, with omission of forearm I/R at 24 h. Immunoreactive erythropoietin peaked at 24 h and remained elevated at approximately 50-fold of baseline at 72 h. FMD did not differ significantly between groups at 24 h (before I/R). At 72 h (48 h after I/R), FMD was greater (by 2.3±0.5% in the darbepoetin compared with the placebo group, a 66% increase over baseline; P<0.001) and greater than FMD at the same time point without preceding I/R (P<0.01). Increases in CD133⁺/CD34⁺/VEGFR2⁺ cells after darbepoetin did not differ according to the presence or absence of preceding I/R. Preceding I/R is required for darbepoetin to enhance endothelial function, possibly by increasing expression of the erythropoietin receptor and by a mechanism likely to involve Akt/NO rather than circulating EPCs.

Introduction During healthy pregnancy there is marked and progressive vasodilatation leading to increased blood flow to maternal organs, in particular the utero-placental circulation. While using a measure of flow-mediated dilatation (FMD) to assess endothelial function in pregnancy, we noticed vasoconstriction of the brachial artery during low-flow, induced by distal cuff occlusion. Technological improvements in measuring and analysing vascular responses to shear stress allowed us to quantify these novel changes that are so far, unique to pregnancy. Methods Endothelial function was prospectively assessed using FMD in 40 healthy, non-smoking pregnant women from 10 weeks gestation, at 4–6 weekly intervals throughout pregnancy and at 15 weeks postpartum. The distal brachial artery was occluded for 5 min and brachial artery diameter was measured continuously for 1 min prior to occlusion, during vessel occlusion and for 5 min after cuff release. Maternal heart rate was measured during this time, and blood flow and reactive hyperaemia responses were calculated. Results A novel and unexpected observation was vasoconstriction of the brachial artery at low-flow, proximal to cuff occlusion. As peripheral blood flow increased, this vasoconstriction became more marked with advancing gestation (ANOVA p<0.001) and fell postpartum. As previously observed, maternal heart rate increased and reactive hyperaemia decreased as pregnancy advanced (ANOVA p<0.001 in both cases). FMD was unchanged during pregnancy but fell postpartum. Conclusions Reducing peripheral blood flow in pregnancy leads to proximal artery vasoconstriction. This observation supports the view that high flow shear stress mediates at least part of the progressive vasodilatation of healthy pregnancy.

Rationale Circulating endothelial progenitor cells (EPC) increase after cardiovascular (CV) events. Whether this results from ischaemia or endothelial injury is unknown. We investigated effects of limb ischaemia-reperfusion (IR) and endothelial injury (EI) on endothelial function and EPC. Methodology Healthy subjects (n=42, 18–35 yr) underwent either limb IR generated by cuff inflation, or EI, induced by wire rotation through a cannula. Endothelial function was assessed by flow mediated dilation (FMD) and circulating CD34+/CD133+/VEGFR2+ EPC were enumerated by flow cytometry at baseline, 10 min, 1 h, 6 h, 2 days and 7 days. FMD and cytometry were also performed in patients (n=23, 40–75 yr) after EI from radial sheath insertion for coronary angiography, at baseline, 2 days, 7 days, 1 month and 4 months. Results Endothelial dysfunction occurred immediately after IR (50% reduction from baseline at 10 min, p<0.001), recovering to baseline after 6 h. After EI FMD fell by 80% (p<0.001), recovering by 2 days. In patients, a sustained fall in FMD after EI remained significant at 1 month. After IR CD133+/CD34+ cells increased after 7 days compared to 1 h (82.1±13/100 000 vs 62.2±8.8/100 000 cells, p<0.05). After EI CD133+/CD34+ cells increased after 7 days compared to baseline (105.3±15.8/100 000 vs 77.9±10.3/100 000 cells, p<0.05). In patients CD133+/VEGFR2+ cells fell at 2 days (by 27.6±11.8/100 000 cells, p<0.05) compared with baseline. There was no significant change in CD133+/CD34+ cells. Conclusions IR and EI cause significant endothelial dysfunction, proportional to the insult. More severe EI reduces CD133+/VEGFR2+ cells, consistent with consumption during repair. In health, CD133+/CD34+ cells expand after arterial insult. No expansion occurs in patients with CV risk factors, possibly reflecting impaired EPC mobilisation.

Nitroglycerin (NTG) selectively vasodilates muscular conduit arteries. Whether other nitric oxide (NO) donors and natriuretic peptides have similar effects is unknown. The aim of this study was to compare effects of NTG, sodium nitroprusside (NP) and brain natriuretic peptide (BNP) on the radial artery (a muscular conduit artery) and forearm resistance vasculature. Phentolamine (PHT), a vasodilator with minimal vasodilator effects on conduit arteries was used as a control. Healthy normotensive men aged 19–45 years were studied. The right brachial artery was cannulated using a 27 gauge needle and an intra-arterial infusion of each vasodilator (PHT, 10, 30 and 100μg/min, n = 9; NTG 0.03, 0.1, 0.3, 1.0, 3.0 μg/min, n = 8. NP, 0.3, 1, 3 μg/min, n = 11; BNP 0.03, 0.1, 0.3, 1, 3 μg/min n = 8) given on separate occasions or after washout. Forearm blood flow (FBF) was measured by venous occlusion plethysmography and change in radial artery diameter by ultrasound. The percentage change in diameter for different drugs was compared at doses producing the same change in FBF (DFBF). The efficacy of dilation of the radial artery was NTG>NP >BNP>PHT. Radial artery dilation by NTG and NP but not BNP was greater than that by PHT (P <0.05) and radial dilation by NTG greater than that by BNP (P<0.05). These results demonstrate that drugs acting on the guanylyl cyclase – cGMP pathway have differential actions on muscular conduit arteries.

s e351 PP.22.198 DIFFERENTIAL EFFECTS OF VASODILATORS ON HUMAN FOREARM MUSCULAR CONDUIT ARTERIES AND RESISTANCE VESSELS H. Fok, B. Jiang, A. Donald, P. Chowienczyk. King’s College London, London-United Kingdom Actions of vasodilator drugs on human forearm resistance vessels are well characterised. By contrast, little is known about their actions on muscular conduit arteries; these may be important since pulse pressure is influenced by the vascular tone of muscular arteries. Vasodilators that have preferential action on larger arteries may be useful in subjects with isolated systolic hypertension. The aim of this study was to compare effects of the alpha-adrenergic antagonist phentolamine (PHT), the nitrovasodilators nitroglycerin (NTG) and nitroprusside (NP), the calcium channel blocker verapamil (VER), brain natriuretic peptide (BNP) and hydralazine (HDZ) on the radial artery (muscular conduit artery) and forearm resistance vasculature. Healthy normotensive men aged 19-45 years were studied. The right brachial artery was cannulated using a 27 gauge needle and an intra-arterial infusion of each vasodilator (PHT, 10, 30 and 100 μg/min, n = 9; NTG 0.03, 0.1, 0.3, 1.0, 3.0 μg/min, n = 8. NP, 0.3, 1, 3 μg/min, n = 11; VER 3, 10, 30, 60 μg/min, n = 8; BNP 0.03, 0.1, 0.3, 1, 3 μg/min n = 8. HDZ, 10, 30 and 100 μg/min, n = 8) was given on separate occasions or after washout. Forearm blood flow (FBF, as a measure of resistance vessel vasodilation) was determined by venous occlusion plethysmography and change in radial artery diameter (muscular conduit artery) by high resolution ultrasound. The study was approved by the local research ethics committee and all subjects gave written informed consent. Changes in FBF were expressed as change from baseline (ΔFBF) during infusion of saline vehicle. Percent change in diameter was compared for different drugs at doses producing the same ΔFBF. Despite producing similar ΔFBF, PHT produced little increase in diameter. By contrast, there was an increase in diameter produced by NTG, NP, HDZ, BNP and VER with efficacy NTG > NP > HDZ > BNP > VER. The actions of NTG, NP and HDZ on the radial artery were significantly greater than PHT (P < 0.05). Thus, within the human forearm, nitrovasodilators are relatively selective dilators of muscular arteries compared to PHT. The activation of soluble guanylyl cyclase appeared to be a more potent pathway for radial artery dilatation than particulate guanylyl cyclase activation, as evident by a smaller vasodilatory effect of BNP on radial artery diameter compared to NTG (P < 0.05). Relative lack of effect of PHT on muscular arteries may relate to the density of sympathetic innervation of muscular and resistance vasculature. However, other mechanisms are required to explain the differential actions of NTG, NP, HDZ, BNP and VER. Classification of the relative specificity of vasodilators using this methodology may help target therapy in relation to the pulsatile and steady components of blood pressure. PP.22.199 PERIPHERAL ARTERIAL DISEASE IN HYPERTENSIVE PATIENTS IN THE VALENCIAN COMMUNITY V. Pallares-Carratala, V. Gil-Guillen, D. Orozco-Beltran, C. De La Sen, F. Valls-Roca, J. Redon, Jl Martinez, J. Navarro-Perez, C. Fluixá, C. Carratala, A. Fernandez, Jc Andres. Union De Mutuas, Burriana-Spain, Universidad Miguel Hernandez, San Juan-Spain, Cs San Gabriel, AlicanteSpain, Cs Beniganim, BeniganimSpain, Hospital Clinico Universitario, Valencia-Spain, Dirección Médica Atención Primaria Departamento De Salud Valencia-Clinico-Malvarrosa, Valencia-Spain, Cs Benimaclet, Valencia-Spain Background and Purpose: The screening of peripheral arterial disease (PAD) by using anklebrachial index (ABI) is very important as PAD has been considered a predictor of cardiovascular morbidity and mortality. The aim of the study was to estimate the prevalence of PAD in hypertensive patients, and to assess severity, degree of under diagnosis, percentage of symptomatic patients and associated factors. Material and Methods: A cross-sectional, observational, multicenter study. Setting: 20 primary care physicians from 10 health centers in Valencia (Spain). ABI determination is made by eco-doppler, the clinical situation is assessed by Fontaine Stages and their interpretation according to Rutherford classification. The sample size was 647 hypertensive patients. Multivariate statistical analysis was performed with binomial logistic regression taking as dependent variable altered or ABI < 0.9. Results: The prevalence of altered ABI was 14.5% (95% CI 11.8 to 17.2), 13.1% in mild-moderate category and 1.4% in severe category, no case more critical. 34% were known and 14.9% were newly diagnosed despite of being symptomatic, and 51.1% were newly diagnosed and asymptomatic. The multivariate model was highly significant (p = 0.000) explaining 58.1% ischemic ABI variability. Associated factors were: age (0.043), BMI (0.010), snuff (0.001), diabetes (0.016), total cholesterol (0.002), HDL cholesterol (0.003), pain in calf (0.000), (0.043), secondary prevention CV (0.000), creatinine (0.042). PP.22.196 THE IMPORTANCE OF AORTIC PULSE WAVE VELOCITY FOR CARDIOVASCULAR RISK STRATIFICATION J. Strizova, J. Filipovsky, J. Seidlerova, M. Dolejsova, O. Mayer, Jr., R. Cifkova. University Hospital Pilsen, Czech Republic, Pilsen-Czech Republic, Department of Preventive Cardiology, Thomayer University Hospital Prague, Czech Republic-Czech Republic Objective: Increased aortic PWV has been shown to predict cardiovascular, and in some cases all-cause, mortality in individuals with hypertension, diabetes mellitus, end stage renal failure. We aimed to study the distribution of aortic PWV values depending on blood pressure (BP) and to establish whether aortic PWV brings additional information about CV risk to the assessment based on conventional risk system. Design and Method: In total 1007 subjects, aged 25-74, were examined in 2008/2009 in the Post-MONICA study. BP was measured three times in the sitting position on the right arm using standard mercury sphygmomanometer. Large artery properties were measured using the Sphygmocor device, with the patient in supine position in the aorta, i.e. between carotid and femoral arteries (aortic PWV). The cardiovascular risk by SCORE system was used. High PWV, defined by the 4th quartile, was done. Results: : In the group aged up to 55 years with optimal BP high PWV was measured in 9% of subjects. In the group with normal and high normal BP high PWV was found in 27,4%. In the group above 55 years with optimal BP high PWV was measured in 15,4%. In the group with normal and high normal BP high PWV was found in 26,3%. After dividing our sample into two groups according to cardiovascular risk SCORE system we found high PWV in 18,9% subjects in group with low risk, aged up to 55 years. In the group above 55 years we found high PWV in 36,8% subjects with low SCORE risk. Conclusions: Pathologic values of aortic PWV in relevant part of our sample, i.e. subjects with normal BP or low SCORE risk, were found. We found that aortic PWV measurement can improve the evaluation of cardiovascular disease risk, better than existing SCORE system. PP.22.197 AGE-RELATED PROGRESSION OF CENTRAL BLOOD PRESSURE COMPONENTS AND THE IMPACT OF NITROVASODILATION: A PROSPECTIVE STUDY IN FEMALE TWINS M. Cecelja, B. Jiang, T Spector, P Chowienczyk. King’s College London, London-United Kingdom Background: Arterial pulse pressure, particularly central arterial pulse pressure (cPP) is closely linked to cardiovascular events. cPP can be partitioned into the height of the first systolic shoulder (P1), generated by a forward pressure and related to arterial stiffness, and augmentation pressure (AP) thought to result from reflected pressure waves and dependent on the calibre of muscular arteries. Objectives: Using a prospective study design, the objective was to examine the contributions of P1 and AP to age-related increases in cPP and the degree to which this may be reversed by nitrovasodilators. Methods: Subjects were 411(49 monozygotic, 162 dizygotic pairs) female twins, that had cPP, P1, and AP estimated using the SphygmoCor system from transformed radial waveforms at 2 time-points. First between 1996 2001 (mean ± SD age 47.6 ± 9.4 years) and between 2006 – 2010, as part of the Twins UK programme of research. Progression in cPP was analysed for the total cohort and separately for subjects < 50 and ≥50 years of age. In 42 women, measurements of cPP, arterial diameters (by ultrasonography) and aortic pulse wave velocity (PWV) were measured before and after sublingual nitroglycerin (NTG, 400 μg). Results: Average duration of follow-up was 10.8 ± 1.2 years. cPP increased more in comparison to pPP (increases of 10.3 and 9.2 mmHg, respectively, P < 0.0001) in the total cohort and in women < 50 years of age, but not in women ≥50 years. In women < 50 years, AP was a greater contributor to progression in cPP in comparison to P1. After age 50, progression of AP and P1 were comparable and accounted for the observed plateau in augmentation index (AIx) with age. NTG had a moderate effect on diastolic blood pressure (reduction from 74.1 ± 7.6 to 70.3 ± 7.1 mmHg, P < 0.0001) but reduced cPP from 43.1 ± 10.0 to 33.1 ± 8.3 mmHg (P < 0.0001). This decrease was entirely explained by a decrease in AP (15.2 ± 6.3 to 5.9 ± 4.3 mmHg, P < 0.0001) with no significant change in P1, or in PWV but an increase in large artery diameters of 5-18% (each P < 0.0001). Conclusion: These results suggest that AP is a major determinant of agerelated widening of cPP in women but can be effectively reduced by selective dilatation of muscular arteries, independently of any effect on PWV. e352 Journal of Hypertension Vol 29, e-Supplement A, June 2011 Parallel degradation of elastin and increase in collagen content and medial elastocalcinosis were observed. Conclusion: In LPK rats, ACE inhibition by perindopril significantly reversed MCSA, prese

AIMS:The endothelium plays a role in regulating vascular tone. Acute and dynamic changes in low-flow-mediated constriction (L-FMC) and how it changes with regard to traditional flow-mediated dilatation (FMD) have not been described. We aimed to investigate the changes in brachial artery L-FMC following percutaneous coronary intervention (PCI) and during recovery from non-ST-segment elevation myocardial infarction (NSTEMI).METHODS AND RESULTS:FMD was performed in accordance with a previously described technique in patients before and after PCI and in the recovery phase of NSTEMI, but in addition, L-FMC data were acquired from the last 30 s of cuff inflation. About 135 scans were performed in 96 participants (10 healthy volunteers and 86 patients). Measurement of brachial L-FMC was reproducible over hours. L-FMC was greater among patients with unstable vs. stable coronary atherosclerosis (-1.33 ±1.09% vs. -0.03 ± 1.26%, P < 0.01). Following PCI, FMD reduced (4.43 ± 2.93% vs. 1.66 ± 2.16%, P < 0.01) and L-FMC increased (-0.33 ± 0.76% vs. -1.63 ± 1.15%, P = 0.02). Furthermore, during convalescence from NSTEMI, L-FMC reduced (-1.37 ± 1.19% vs. 0.01 ± 0.82%, P = 0.02) in parallel with improvements in FMD (2.54 ± 2.19% vs. 5.15 ± 3.07%, P < 0.01).CONCLUSION:Brachial L-FMC can be measured reliably. Differences were observed between patients with stable and unstable coronary disease. L-FMC was acutely increased following PCI associated with reduced FMD and, in the recovery from NSTEMI, L-FMC reduced associated with increased FMD. These novel findings characterize acute and subacute variations in brachial L-FMC. The pathophysiological and clinical implications of these observations require further study.

AIMSTo assess the feasibility and reproducibility of non-invasive vascular assessment in a childhood population setting and identify the determinants of vascular phenotype in early life.METHODS AND RESULTSWe studied 7557 children (age 9.8-12.3 years) participating in the Avon Longitudinal Study of Parents and Children (ALSPAC). Six research technicians underwent a 5-month training protocol to enable study of brachial artery endothelial function by flow-mediated dilatation (FMD) and arterial stiffness by carotid to radial pulse wave velocity (PWV) and brachial distensibility [distensibility coefficient (DC)]. Reproducibility studies were performed at the beginning, the middle, and the end of the study. A blinded repeat evaluation of a random selection of 3% of the cohort was also undertaken throughout the study. The effect of anthropometric and environmental factors on each measure was examined. Successful measures were obtained in 88, 95, and 87% of the studied children for FMD, PWV, and DC, respectively. The coefficients of variation between technicians for FMD, PWV, and DC were 10.5, 4.6, and 6.6% at the beginning of the study and reached 7.7, 4.1, and 10% at the end. Baseline vessel diameter and gender were important determinants of all the vascular measures, with a small effect of room and skin temperatures on FMD and PWV. Boys consistently had lower FMD and DC and higher PWV measures (P < 0.01 for all).CONCLUSIONReproducible, high-quality assessments of vascular structure and function in children can be made on a large scale in field studies by suitably trained non-specialist operators. This study provides an invaluable resource for assessing the impact of early influences, genetic, and environmental factors on arterial phenotype.

Background— Preeclampsia is a life-threatening pregnancy syndrome of uncertain origin. To elucidate the pathogenesis, we evaluated the temporal relationships between changes in vascular function and circulating biomarkers of angiogenic activity before and after the onset of preeclampsia and gestational hypertension. Methods and Results— Maternal mean arterial pressure, uterine artery pulsatility index, brachial artery flow-mediated dilatation, and serum concentrations of placental growth factor (PlGF), soluble fms-like tyrosine kinase 1 (sFlt-1), and soluble endoglin were prospectively measured in 159 women from 10 weeks gestation until 12 weeks postpartum. At 10 to 17 weeks, women who developed preterm preeclampsia had lower serum PlGF (P=0.003), higher soluble endoglin (P=0.006), and higher sFlt-1:PlGF ratio (P=0.005) compared with women who later developed term preeclampsia, gestational hypertension, or normotensive pregnancy. At 10 to 17 weeks, mean arterial pressure inversely correlated with serum PlGF (r=−0.19, P=0.02); at 18 to 25 weeks, with soluble endoglin (r=0.18, P=0.02); and at 26 to 33 weeks, with sFlt-1 (r=0.28, P<0.001). At 23 to 25 weeks, uterine artery pulsatility index correlated with serum soluble endoglin (r=0.19, P=0.02) and sFlt-1 levels (r=0.17, P=0.03). Flow-mediated dilatation was higher during a pregnancy with gestational hypertension compared with preeclampsia (P=0.001). Twelve weeks postpartum, serum PlGF was higher in women who had a hypertensive pregnancy compared with a normotensive pregnancy (P<0.001). Conclusions— These observations support a role for placenta-derived angiogenic biomarkers in the control of maternal vascular resistance of preeclampsia. Gestational hypertension develops differently, with a hyperdynamic circulation and angiogenic biomarker profile similar to normotensive pregnancy. Larger studies of unselected women are needed to ascertain whether measures of these angiogenic biomarkers assist with the prediction and prognosis of preeclampsia and whether postpartum measures of serum PlGF have a role in predicting future cardiovascular disease.

Objective: To assess the influence of mannose-binding lectin (MBL) genotype on endothelial function in the presence and absence of infection in childhood.Methods: We studied 2176 children aged 10 years drawn from the Avon Longitudinal Study of Parents and Children. Endothelial function was assessed by flow mediated dilatation (FMD). Exon 1 and promoter polymorphisms in the MBL gene were determined by heteroduplexing procedures. Children were classified as AA (wild type) AO (heterozygotes) and OO (homozygotes).Results: During the vascular assessment, 544 children presented with current or recent (<2 weeks) infection (INF). FMD was reduced in the INF group compared to controls (10% reduction in FMD, p < 0.001). MBL genotype was not associated with FMD in controls, although a relationship with the degree of impairment during INF was observed (8.0%, 7.6% and 26.6% lower FMD compared to controls for groups AA, AO, OO respectively, p < 0.05). After multivariate analysis, OO was associated with reduced FMD in the INF group (odds ratio 2.95 [1.33, 6.52], p < 0.001).Conclusion: Homozygosity for MBL variant alleles is associated with greater impairment in FMD during infection in childhood. This suggests a gene-environment interaction operating in early life that may have relevance for the initiation and progression of atherosclerosis. (c) 2009 Elsevier Ireland Ltd. All rights reserved.

Objectives: Aortic pulse wave velocity (PWV) is usually estimated by sequential (ECG referenced) carotid-femoral tonometry using the SphygmoCor system (Atcor, Australia). This can be technically challenging and operator dependant. Here we evaluate a simple, operator independent method of estimating central PWV based upon simultaneous recording from upper arm and thigh cuffs (Vicorder, Skidmore Medical, UK). Methods: PWV was measured using the Vicorder and SphygmoCor systems (each measurement in triplicate) in 133 adults (mean age 53, range 21–70 years). SphymoCor PWV was calculated using the suprasternal notch (sn) to femoral distance. Two distances were used to calculate PWV from the Vicorder: cuff to cuff measured with arm at the side (cc) and sn to thigh cuff minus sn to arm cuff (notch to cuff difference, ncd). Reproducibility of the Vicorder was further assessed by repeat measures in 9 subjects. Results: Mean values of PWV obtained by SphygmoCor, Vicorder (cc) and Vicorder (ncd) were 9.0±1.6, 12.0±2.8 and 8.7±1.9m/s respectively. Both Vicorder (cc) and Vicorder (ncd) were closely correlated with SphygmoCor PWV (each r=0.7). The mean difference between SphygmoCor and Vicorder (ncd) was 0.2±1.4m/s. The within subject standard deviation for repeated measures for Vicorder (ncd) was 0.54m/s. Conclusion: There is a high correlation between values obtained using the Vicorder and SphygmoCor and good reproducibility for Vicorder measurements. Differences between the methods are likely due to errors in the estimation of path length. Vicorder PWV is quick and easy to perform with minimal training and offers a simple alternative to applanation tonometry.

Background— Endothelial dysfunction develops early and has been shown to predict the development of clinical complications of atherosclerosis. However, the relationship between early endothelial dysfunction and the progression of arterial disease in the general population is unknown. We investigated endothelial dysfunction, risk factors, and progression of carotid intima-media thickness (cIMT) in late-middle-aged individuals at low to intermediate cardiovascular risk in a prospective study between 1997 and 2005. Methods and Results— Brachial artery flow-mediated dilatation and cIMT were measured in 213 nonsmoking British civil servants recruited from a prospective cohort (Whitehall II study). Participants (age, 45 to 66 years) were free of clinical cardiovascular disease and diabetes mellitus. Risk factors and Framingham Risk Score were determined at baseline. cIMT was repeated 6.2±0.4 years later. At baseline, age, blood pressure, low-density lipoprotein cholesterol, and Framingham Risk Score correlated with cIMT. However, only flow-mediated dilatation, not risk factors or Framingham Risk Score, was associated with average annual progression of cIMT. This relationship remained significant after adjustment for risk factors whether entered as separate variables or as Framingham Risk Score. Further adjustment for waist circumference, triglycerides, and employment grade had no significant effect. Conclusions— Systemic endothelial function was associated with progression of preclinical carotid arterial disease over a 6-year period and was more closely related to cIMT changes than conventional risk factors. Thus, the relationship between endothelial dysfunction and adverse outcome is likely to be due not only to destabilization of established disease in high-risk populations but also to its impact on the evolution of the atherosclerotic substrate. Flow-mediated dilatation testing provides an integrated vascular measure that may aid the prediction of structural disease evolution and represents a potential short- to intermediate-term outcome measure for evaluation of preventive treatment strategies.

INTRODUCTION Many patients with Peyronie's disease (PD) have one or more risk factors (RFs) for atherosclerosis and endothelial dysfunction. It is well recognized that such RFs commonly lead to the development of systemic vascular abnormalities. While not necessarily so, this may implicate vascular dysfunction in its pathogenesis. The cause of PD remains obscure despite intense research over the years and investigating the role of vascular dysfunction in the pathogenesis of PD is a novel approach worth undertaking. AIM To test our hypothesis that PD is associated with systemic vascular changes even in the absence of RFs for atherosclerosis and endothelial dysfunction. METHODS Vascular function was assessed using high-resolution brachial artery ultrasound in 23 PD patients (aged 30-65 years) without RFs for endothelial dysfunction and atherosclerosis, and 23 age-matched healthy controls. Endothelium-dependent, flow-mediated brachial artery dilation was measured in response to increased shear stress (reactive hyperemia induced by 5 minutes of forearm ischemia). This response was contrasted with that of 400 microg sublingual glyceryl trinitrate, an endothelium-independent vasodilator. Anthropometric characteristics, blood pressure, fasting lipids, and glucose were also measured. MAIN OUTCOME MEASURE Endothelium-dependent, flow-mediated brachial artery dilation and glyceryl trinitrate-induced endothelium-independent vasodilation. RESULTS Endothelium-dependent flow-mediated dilation (FMD) was impaired in PD patients compared to controls (5.62 +/- 0.58% vs. 7.46 +/- 0.56%, P = 0.03). In contrast, responses to glyceryl trinitrate were similar in PD patients and controls as were blood pressure, lipid, and glucose values. FMD remained impaired after multivariable adjustment for potential confounders. CONCLUSION Patients with Peyronie's disease have evidence of systemic vascular changes in the way of systemic conduit artery endothelial impairment even in the absence of RFs for atherosclerosis and endothelial dysfunction. These wider vascular abnormalities in PD are likely to be of clinical relevance and require further study.

BACKGROUND:C-reactive protein (CRP), a marker of systemic inflammation, is associated with risk of coronary events and sub-clinical measures of atherosclerosis. Evidence in support of this link being causal would include an association robust to adjustments for confounders (multivariable standard regression analysis) and the association of CRP gene polymorphisms with atherosclerosis (Mendelian randomization analysis). METHODOLOGY/PRINCIPAL FINDINGS:We genotyped 3 tag single nucleotide polymorphisms (SNPs) [+1444T>C (rs1130864); +2303G>A (rs1205) and +4899T>G (rs 3093077)] in the CRP gene and assessed CRP and carotid intima-media thickness (CIMT), a structural marker of atherosclerosis, in 4941 men and women aged 50-74 (mean 61) years (the Whitehall II Study). The 4 major haplotypes from the SNPs were consistently associated with CRP level, but not with other risk factors that might confound the association between CRP and CIMT. CRP, assessed both at mean age 49 and at mean age 61, was associated both with CIMT in age and sex adjusted standard regression analyses and with potential confounding factors. However, the association of CRP with CIMT attenuated to the null with adjustment for confounding factors in both prospective and cross-sectional analyses. When examined using genetic variants as the instrument for serum CRP, there was no inferred association between CRP and CIMT. CONCLUSIONS/SIGNIFICANCE:Both multivariable standard regression analysis and Mendelian randomization analysis suggest that the association of CRP with carotid atheroma indexed by CIMT may not be causal.

Objective There is growing evidence that the presence of the cell stress protein heat shock protein (HSP) 60 in the circulation is associated with risk of coronary heart disease. In this study, we measured the association between plasma HSP60 and carotid arterial stiffness in middle-aged men and women. Methods Six hundred and forty-seven men and women aged 50–72 years and free of cardiovascular disease and medication were tested. Carotid artery distensibility coefficient was assessed ultrasonically as a measure of arterial stiffness, and plasma HSP60 was assessed using a sensitive immunoassay. Results We found a significant, independent association between high plasma levels of HSP60 and increased carotid stiffness. Carotid distensibility coefficient was also related to diabetes, adiposity, blood pressure, lipids, plasma interleukin-6 and C-reactive protein. After adjusting for these factors, the odds of HSP60 concentration of at least 1000 ng/ml were 1.79 (95% confidence intervals 1.06–3.04) for participants in the lowest compared with the highest tertile of the distensibility coefficient. Conclusion HSP60 is a potent activator of vascular endothelial cells and smooth muscle cells. Thus, it is possible that long-term stimulation of these cell populations by blood-borne HSP60 acts to drive blood vessel changes resulting in decreased arterial elasticity.

Objectives Our aim was to determine reproducibility of the flow-mediated dilation (FMD) response profile, and discriminatory ability of the components.Background Brachial FMD is widely used to study conduit artery endothelial function. Automated B-mode image edge detection (B-ED) provides a full response profile. Reproducibility and biological relevance of these additional components have not been fully explored.Methods Forty-two healthy adults underwent FMD using B-ED repeated at fixed time intervals up to 3 months. The FMD profile was assessed for diameter changes, area under the curve, and time course. Measures were compared in 25 adults with hypercholesterolemia, 25 subjects with diabetes, and 50 matched control subjects.Results The maximum change in FMD was the most reproducible (coefficient of variation = 9.8%, 10.6%, 6.6%, and 9.2% at 4 to 6 h, 1 week, 1 month, and 3 months, respectively). Most of the variability occurred between subjects rather than within. All FMD measures except time course were significantly reduced in hypercholesterolemia and diabetes. Power curves were generated to indicate the appropriate number of subjects for parallel and crossover study designs.Conclusions Maximum FMD percentage change from baseline is the most reproducible of the response curve measures and best identifies those with risk factors. Flow-mediated dilation measured by B-ED is robust and practical to assess the effect of interventions on endothelial function in clinical trials.