tbuckley/cabot-search huggingface dataset:
Abstract 5489: Provisional (bail-out) Use versus Routine Use of Glycoprotein 2b/3a Blockers in Stemi: A Sub-analysis From the On-time 2 Trial
Authors: Rik Hermanides, Ton Heestermans, Jurrien ten Berg, Marcel Gosselink, Jan Paul Ottervanger, Gert van Houwelingen, Evelien Kolkman, Pieter Stella, Thorsten Dill, Christian Hamm, Arnoud W van ‘t Hof
Journal: Circulation (2009)

Background: In daily practice, in patients (pts) with acute ST-elevation myocardial infarction, glycoprotein 2b/3a blockers use is often limited to the provisional use in case of complications after PCI. However, it is unknown whether this strategy is of any value as compared to the routine use of these agents before PCI. Methods: The On-TIME 2 trial compared routine pre-hospital initiation of high dose Tirofiban (HDT) versus provisional use of HDT in a randomized placebo controlled design. Thrombotic bail-out use of study medication was pre-defined and part of the combined clinical end-point. Within the pts who received blinded bail-out treatment, we compared pts who were pre-treated with placebo and received bail-out HDT (Tirofiban bail-out) versus those who were pre-treated with HDT and received bail-out placebo (Placebo bail-out). Results: Blinded bail-out use of study medication was used in 24% (237/980) of pts, with a significant higher use in pts pre-treated with placebo (29% (140/492) versus 20% (97/488), p=0.002). The indication for bail-out use of study medication was mostly distal embolisation or no-reflow after PCI and did not differ between the groups (73.6% vs 75.3% p=NS). As expected, bail-out use of study medication was associated with more residual ST deviation (5.5±7.2 vs 3.7±4.8mm), larger infarct size (median (IQR): 1750 (674–3117) vs 948 (379–2278) IU), and worse outcome (MACE 12.2% vs 5.6%, p&lt;0.01 for all comparisons) as compared to no-bail-out treatment, table . Conclusion: Routine pre-hospital treatment with HDT significantly reduced the use of blinded bail-out study medication. Bail-out study medication was associated with larger infarct size and poor outcome, especially in those pts pre-treated with placebo. Therefore, routine pre-treatment with HDT is preferred over provisional use of HDT in pts with STEMI. It is highly questioned whether provisional use of glycoprotein 2b3a blockers in case of complications after PCI is effective at all. Table
Abstract 5596: Differential Paracrine Dysfunction and Hypoxic Desensitization in Diabetic CD34+ and CD14+ Bone Marrow Derived Cells (BMDC)
Authors: Yagna P Jarajapu, Prajitha Thampi, Sergio Caballero, Michael E Boulton, Maria B Grant
Journal: Circulation (2009)

Purpose: To test the hypothesis that BMDC of diabetic (D) origin have impaired responsiveness to the hypoxic microenvironment and ineffective paracrine signaling. Methods: FACS sorted CD34+ and CD14+ cells from D and non diabetic (n=10) (ND) individuals (n=10) were exposed to normoxia (pO2=40 mmHg) or hypoxia (pO2=5 mmHg). Levels of HIF1- α and surface expression of VEGFR1, VEGFR2 and CXCR4 were evaluated. CD34+ and CD14+ cells were placed in culture (24 hr) and conditioned-medium (CM) was concentrated and analyzed for cytokines and growth factors. The effects of CM were assessed on CD34+ cell and CD14+ cell migration, and proliferation. Streptozotocin-induced diabetic mice of 11 months duration or age matched controls received intravitreal injection of fluorescently labeled ND or D CD34+ cells or CD14+ cells. Cells were localized 48 hrs post-injection and the degree of vascular homing to acellular capillaries was evaluated immunohistologically and quantitated. Results: With hypoxia (4hr), expression of VEGFR2 (not VEGFR1), CXCR4 and HIF1- α were significantly increased in ND CD34+ cells but this response was markedly blunted in D CD34+ cells. In contrast, both ND and D CD14+ cells showed decreases in surface expression for all 3 receptors following hypoxia. TNF α , MCP1 and TGF β were 10-fold higher and SCF and HGF were 3-fold lower in CM from D CD34+ cells compared to ND CD34+ cells. CM from D CD34+ cells increased migration of ND CD14+ cells but not their proliferation (P&lt;0.01). D CD14+ cells did not secrete SCF and Flt-3L and did not support proliferation. ND CD14+ secreted higher levels of IL6, IL1 β , IL8 and TGF β 1 compared to D CD34+. ND CD34+ and both ND and D CD14+cells, but not DM CD34+ cells migrated through the vitreous and homed to areas of injured vasculature in diabetic mice, but not control mice (P&lt;0.05). Conclusions: D CD34+ are unresponsive to hypoxia and cannot home to areas of retinal injury, whereas D CD14+ remain hypoxia responsive, can home as efficiently as ND CD14+ and ND CD34+ and can support migration but not proliferation. Our studies show complex paracrine interactions between BMDC and distinct desensitization of CD34+ cells to hypoxia which supports their observed in vivo dysfunction.
Abstract 5525: Caveolin-1 Downregulation Reduces NADPH Oxidase Assembly and Reverses eNOS Uncoupling
Authors: Irina Lobysheva, Geraldine Rath, Belaid Sekkali, Olivier Feron, Chantal Dessy, Bernard Gallez, Jean-Luc Balligand
Journal: Circulation (2009)

Activation of the renin-angiotensin system generates endothelial dysfunction through increased oxidant stress. We examined the spatial co-localization of eNOS and NADPH oxidase in endothelial caveolae and the role of caveolin-1 (cav1) on eNOS activation and uncoupling in response to Angiotensin II (AII) in vitro and in vivo. NO and O2- production at the membrane of BAECs and HUVECs was measured by EPR spin-trapping (Fe(II)-[DETC]2 and DMPO, respectively). AII increased NO to 158±12% and O2- to 209±5% of control (all from min. 3 exp; P&lt;0.05). AII-stimulated NO production was sensitive to inhibition of NADPH oxidase assembly (with apocynin, to 64±11% of ctl; P&lt;0.05), as well as of the redox-sensitive PI3 kinase (with LY294002), cSrc (with PP2) and cAbl kinases (by siRNA targeting; all P&lt;0.05). Reciprocally, the NOS inhibitor, L-NAME inhibited AII-stimulated O2- (by 47±11%;P&lt;0.05), indicating eNOS uncoupling, as confirmed by increased eNOS monomer/dimer ratio. To resolve the role of eNOS and NADPH compartmentation for uncoupling, endothelial cell extracts were separated by isopycnic ultracentrifugation. Upon AII stimulation, p47phox, a NADPH subunit, was redistributed with eNOS to cav1-enriched fractions. siRNA downregulation of cav1 by ~50% while preserving eNOS confinement, inhibited AII-stimulated p47phox translocation, O2 - production (by 2.5-fold) in cav-1-enriched fractions and abrogated the L-NAME-sensitive O2 - production, indicating reversal of eNOS uncoupling. To test similar effects of moderate cav1 depletion in vivo, heterozygote cav-1+/− mice (and WT) were treated with AII infusion (osmotic minipump), their blood pressure recorded by telemetry and plasma Hb-NO measured by ESR. While AII infusion produced hypertension and decreased Hb-NO in WT (to 55±15% of ctl; p&lt;0.05), cav-1+/− mice were fully preserved. We conclude that, in endothelial cells, co-localization of NADPH oxidase and eNOS in caveolae-enriched fractions activates eNOS through ROS-sensitive PI3K and cSrc/cAbl kinases but produces eNOS uncoupling in response to AII. Moderate downregulation of cav-1 prevents NADPH assembly and eNOS uncoupling, while preserving NO production, underlining the interest for cav-1 modulation to treat endothelial dysfunction.
Abstract 5504: Transient Decrease in Circulating Dendritic Cell Precursors After Acute Stroke - Potential Recruitment Into the Brain
Authors: Atilla Yilmaz, Tanja Fuchs, Barbara Dietel, Regina Altendorf, Iwona Cicha, Christian Stumpf, Peter Schellinger, Ingmar Blümcke, Stefan Schwab, Werner G. Daniel, Christoph D. Garlichs, Rainer Kollmar
Journal: Circulation (2009)

Background: The role of dendritic cells (DC) as potent mediators of inflammation has not been sufficiently investigated in stroke. Methods: Circulating myeloid (mDCP), plasmacytoid (pDCP), and total DCP (tDCP) were flow cytometrically analyzed in healthy controls (n=29), patients with asymptomatic A. carotis interna stenosis (ACI-S, n=46), transient ischemic attack (TIA, n=39), acute ischemic stroke (AIS, n=73), and acute hemorrhagic stroke (AHS, n=31). The National Institutes of Health Stroke Scale (NIHSS) and the infarction size in CT scan were evaluated after stroke. In a patient subgroup, postmortem immunohistochemical brain analyses were performed to detect mDC (CD209), pDC (CD123), T cells (CD3), and HLA-DR. Results: In AIS and AHS, circulating mDCP, pDCP, and tDCP (each p&lt;0.005) were significantly reduced. A significant inverse correlation was found between the NIHSS and circulating DCP (p&lt;0.02), as well as between hsCRP and circulating DCP (p&lt;0.001). Patients with large stroke size in CT scan had significantly lower mDCP, pDCP, and tDCP (each p&lt;0.01) than those with smaller stroke. Follow-up analysis showed a significant recovery of circulating DCP in the first days after stroke. In the infarcted brain, a dense infiltration of mDC colocalized with T cells, single pDC, and high HLA-DR expression were observed. Conclusions: Acute stroke leads to a decrease in circulating DCP. Potentially, circulating DCP are recruited from the blood into the infarcted brain, and probably trigger cerebral immune reactions there.
Abstract 5594: Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences Undergo Cardiogenesis in Defined Conditions
Authors: Jianhua Zhang, Andrew G Soerens, Gisela F Wilson, Junying Yu, James A Thomson, Timothy J Kamp
Journal: Circulation (2009)

Human induced pluripotent stem (iPS) cells hold great promise for cardiovascular research and therapies, but the lentiviral generated iPS cells contain transgene and vector sequences inserted into the genome that could result in insertional mutagenesis. The recent generation of iPS cells using oriP/EBNA1(Epstein-Barr nuclear antigen-1)-based episomal vectors avoids genomic integrations and brings iPS cells closer to clinical applications. However, the ability of vector-free iPS cells to generate cardiomyocytes (CMs) is unknown. The aim of this study was to test the ability of vector-free iPS cells to undergo cardiac differentiation in embryoid bodies (EBs) as well using directed differentiation in defined medium. Four iPS clones derived from foreskin fibroblasts (iPS-DF6 –9 –9T, iPS-DF6 –9 –12T, iPS-DF19 –9 –7T, iPS-DF19 –9 –11T) were maintained either on mouse embryonic fibroblasts for EB formation, or in feeder-free TeSR medium for directed differentiation. EBs formed contracting outgrowths with variable efficiency among 4 clones (6.1±3.0%, 10.4±3.0%, 0.4±0.2% and 1.6±0.6%, respectively). Likewise, directed differentiation using sequential treatment with Activin A (100 ng/ml) and BMP4 (10 ng/ml) resulted in the formation of contracting CMs, but flow cytometry for cTnT-positive cells demonstrated 10 –15 fold more CMs were obtained using directed differentiation than with the EB method. RT-PCR comparing iPS cell-derived CMs with undifferentiated iPS cells demonstrated strong upregulation of cardiac genes (NKX2–5, TNNT2, MYH6, ACTN2, HPPA, PLN, MYL2 and MYL7) and concomitant downregulation of pluripotency genes (OCT4 and NANOG). Immunolabeling of cTnT, alpha-actinin, MLC2a and MLC2v showed organized sarcomeres in single iPS cell-derived CMs. Sharp microelectrode recordings revealed spontaneous action potentials comparable to those observed in embryonic stem cell-derived CMs (n = 34 cells of 5 microdissected EB outgrowths). Our findings demonstrate that vector-free iPS cells readily undergo cardiogenesis in EBs and that directed differentiation under defined conditions greatly increases cardiogenesis efficiency. Thus, vector-free iPS cells have growing appeal for cardiovascular research and cardiac repair applications.
Abstract 5532: Apoptosis, Interstitial Collagen and Diastolic Function in Essential Hypertension
Authors: Miguel Rivera, Raquel Cortes, Pedro Morillas, Esther Rosello, Lilian Grigorian, Luis Martinez, Catheline Lauwers, Fernando Garcia de Burgos, Federico Soria, Jose Ramon Gonzalez Juanatey, Rafael Paya, Teresa Lozano, Placido Orosa, Vicente Miro, Isabel Antorrena, Beatriz de Rivas, Luis Mainar, Jose Leandro Perez Bosca, Gonzalo Baron, Vicente Climent, Jose Ramon Calabuig, Alejandro Jordan, Antonio Salvador, Manuel Portoles, Vicente Bertomeu
Journal: Circulation (2009)

Introduction: Sustained hypertension (HT) leads to left ventricular hypertrophy (LVH) and left ventricular dilatation, processes associated with loss of cardiac myocytes through apoptosis. Nevertheless, the relationship of apoptosis, myocardial fibrosis and diastolic function remains unknown. Therefore, the aims of the present study were to analyze the relationship between aminoterminal propeptide levels of type III procollagen (PIIINP), sFas, soluble TNF receptor 1 (sTNF-R1) and diastolic function in essential HT. Hypothesis: We assessed the hypothesis that increased cardiac collagen turnover would be associated with LVH, diastolic dysfunction and apoptosis even in asymptomatic HT. Methods: The asymptomatic hypertensive group consisted of 253 Caucasian patients (mean age 60±13 years, 139 males) from 11 hospitals. A routine physical examination, laboratory analyses, and echo-Doppler study were performed. Results: Serum concentrations of PIIINP were higher in hypertensive patients compared to control group [4.18 (3.55 – 5.04 μ g/L) vs. 3.54 (2.98 – 4.54 μ g/L), p=0.006] and also plasma levels of sFas and sTNF-R1 [1.40 (1.13 – 1.69 ng/mL) vs. 1.00 (0.84 – 1.27 ng/mL), p&lt;0.0001; 385 (291 – 545 pg/mL) vs. 236 (194 – 302 pg/mL), p&lt;0.0001]. Log-transformed concentrations of anti-apoptotic cytokines were correlated between them (r=0.404, p&lt;0.0001). Moreover, serum PIIINP concentrations were associated with log-transformed sFas (r=0.386, p&lt;0.0001) and sTNF-R1 (r=0.298, p&lt;0.001). Then, multivariate analyses included sFas (p&lt;0.0001) and sTNF-R1 (p&lt;0.0001) as independent factors of serum procollagen levels. Finally, marker concentrations were significantly correlated with diastolic parameters. Conclusions: This study showed increased circulating levels of PIIINP, sFas and sTNF-R1 in our group of asymptomatic hypertensive patients. Furthermore, sFas and sTNF-R1 are independent factors of serum type III procollagen, and diastolic parameters showed a highly significant relationship with myocardial fibrosis and anti-apoptotic cytokines.
Abstract 5629: Time Course Analysis of Bone-marrow Derived Progenitor Cell Transdifferentiation During Neointima Formation
Authors: Jan-Marcus Daniel, Harald Tillmanns, Daniel G Sedding
Journal: Circulation (2009)

Background: Transdifferentiation of bone marrow derived progenitor cells (BMPC) into smooth muscle cells (SMC) was suggested to contribute to vascular remodeling processes. However, the ability of these cells to account for highly differentiated contractile SMC remains elusive. In this study, we examined the time points of accumulation, differentiation and the long term contribution of BMPC compared to locally derived SMC in the process of neointima formation. Methods and Results: C57BL/6 mice were lethally irradiated and subsequently rescued by bone-marrow transplantation (BMTx) of eGFP+-BM. Arterial remodeling was induced by wire injury of the femoral artery, and vessels were harvested after 3 days, 1, 2, 3, 4, 6 and 16 weeks (n=10 animals per time point). Neointima/media ratio and the number of eGFP+-cells in the vessel wall were quantified. Following a careful analysis using deconvolution analysis of z-image stacks and confocal microscopy, the expression of α SMA in eGFP+-cells occurred to be a very rare event. This was also observed for the co-expression of CD31 in BM-derived cells. Moreover, some cells positive for eGFP and α SMA also expressed monocytic lineage markers like CD68 or CD11b. The number of BM derived cells in the neointima constantly declined at later timepoints, and most of the eGFP+-cells were indeed identified as monocytes/macrophages. At 16 weeks after dilatation, highly differentiated SMC were exclusively derived from GFP negative and thus pre-existing resident cells. In addition to these data, selective prevention of resident vascular cell proliferation by irradiation of mice before BMTx inhibited neointima formation in a dose dependent manner. Conclusions: The contribution of eGFP+-cells to the cellular compartment of the neointimal lesion is limited to a temporary time period of the inflammatory response to the vascular injury. Hence, BM derived cells contribute to arterial remodeling, but these cells do not transdifferentiate into definitive and highly differentiated SMC.
Abstract 5597: Transient Blockade of TGF- <i>β</i> in Hematopoietic Stem Cells and CD34+ Endothelial Precursor Cells Enhances Survival, Proliferation and Reparative Functions in Response to Ischemic Vascular Injury
Authors: Sergio Li Calzi, Ashay D Bhatwadekar, Aqeela Afzal, Sergio Caballero, Stephen Bartelmez, Carl J Pepine, Maria B Grant
Journal: Circulation (2009)

Endothelial precursor cells participate in vascular repair; however in diabetes these cells can become dysfunctional. Previously, we showed that transient blockade of endogenous transforming growth factor-beta1 (TGF-beta1) by antisense phosphorodiamidate morpholino oligomers (PMOs) accelerated homing and engraftment of hematopoietic stem cells (HSC) in experimental models of bone marrow transplant. We asked whether transient blockade of TGF-beta1 could enhance mouse HCS and human CD34 + -endothelial progenitor cell (EPC) recruitment and re-endothelialization to areas of vascular injury and correct vascular reparative defects of diabetic cells. Cells were treated with either control-PMO, anti-TGF-beta-PMO, or saline. Cell proliferation in response to the above treatments was studied in the colony forming assay for 1 week. CXCR-4 expression was examined by immunostaining and migration in response to stromal derived factor-1 (SDF-1) was tested using Boyden chamber assay. Mice were injected systemically with modified HSCs and then underwent either laser rupture of Bruch’s membrane to initiate choroidal neovascularization (CNV) or modified CD34 + cells of diabetic and nondiabetic origin and then received the retinal ischemia reperfusion (I/R) injury. Neutralizing endogenous TGF-beta1 in cells sustained their survival in the absence of growth factors with no loss of proliferative potential. Pre-treatment with anti-TGF-beta-PMO increased expression of CXCR-4 in both diabetic and nondiabetic CD34 + cells and increased their migration to SDF-1. HSC treated with anti-TGF-betaPMO demonstrated increased homing to CNV lesions and increased reendothelialization of acellular capillaries in the I/R model by both diabetic and nondiabetic cells. Blocking endogenous TGF-beta1 production by PMO induced increased CXCR-4 expression in cells and increased nitric oxide generation facilitating cell migration and enhanced vascular repair. Blocking TGF-beta1 in HSC/CD34 + EPC may represent a novel therapeutic strategy in enhancing vascular repair in diabetes. This research has received full or partial funding support from the American Heart Association, Greater Southeast Affiliate (Alabama, Florida, Georgia, Louisiana, Mississippi, Puerto Rico &amp; Tennessee).
Abstract 5571: Induction of Macrophage Activation and Foam Cell Formation by High Glucose is Mediated by Rho-associated Coiled-coil Forming Kinase
Authors: Cheng-I Cheng, Shu-Meng Cheng, Yuxin Li, Qian Zhou, James K Liao
Journal: Circulation (2009)

Background: Vascular complications are major causes of morbidity and mortality in patients with diabetes mellitus. Although the precise mechanism by which diabetes leads to vascular disease remains unknown, hyperglycemia has been shown to accelerate atherogenesis, a process involving the uptake of modified low-density lipoproteins (LDL) by macrophages and formation of macrophage-derived foam cells. We have recently shown that deletion of Rho-associated coiled-coil forming kinase (ROCK)-1 in macrophages leads to decreased LDL uptake, foam cell formation, and atherosclerosis. Therefore, we hypothesize that high glucose may enhance atherogenesis by inducing uptake of LDL by macrophages and increasing foam cell formation through activation of ROCK. Methods &amp; Results: To investigate the effects of high glucose on ROCK and macrophage function, we isolated bone marrow-derived macrophage (BMDM). Treatment with high glucose but not mannitol increased ROCK activity in BMDM in a dose-dependent manner, with initial increase observed after 8 hrs. Downstream metabolites of glucose such as sorbitol and L-glutamine, as well as intracellular oxidative stress and protein kinase C all induced ROCK activation in BMDM. Indeed, co-treatment with the ROCK inhibitor, fasudil (30 uM), decreased glucose-induced oxidative stress. High glucose increased BMDM adhesion and migration, which was inhibited by co-treatment with fasudil. Proliferation and phagocytic activity of BMDM as determined by [ 3 H]-tritium incorporation and zymosan uptake, respectively, were increased in high glucose culture medium, but these effects of high glucose were not affected by fasudil. High glucose (4.5 g/L) increased the size of BMDM compared to low glucose (1.0 g/L), and led to more foam cell formation in the presence of 50 ug/ml of cholesterol. In addition, the expression of TNFalpha in BMDM was increased by 2.7-fold in high glucose medium, which was suppressed by treatment with fasudil. Conclusion: These findings indicate that high glucose induces macrophage activation and foam cell formation is mediated by increased ROCK activity. These results suggest that targeting ROCK may have therapeutic benefits in ameliorating atherosclerosis in diabetic patients.