Identification of optimal culture conditions for the <i>ex-vivo</i> expansion of ACS PB-derived EPC/ECFC.

<p> Primary EPC/ECFC colonies were generated by plating patient PBMC in M⁵¹⁰⁰ medium, as detailed in the Method section. In <b>A</b>, after the colony identification (at day 5 after plating), medium was change (arrow) and replaced either with fresh M⁵¹⁰⁰, or M^EGM or M¹⁹⁹ and the development of the colonies was monitored over the time. The growth kinetics of a representative experiment out of five independent experiments is shown. At each indicated time point, the mean cell number/ECFC was determined by two independent operators; standard deviations were below 10% and are not shown. Asterisk, p<0.05. In <b>B</b>, immunocytochemical analysis of <i>in vitro</i> expanded EPC/ECFC documenting positivity for CD105 antigen (original magnification: 20X) and for the specific endothelial marker Factor VIII (original magnification: 40X). In <b>C</b>, FISH analysis performed on <i>in vitro</i> expanded EPC/ECFC by using the centromeric enumeration probe CEP9 (white arrows) documenting a normal diploid chromosomal pattern (original magnification: 40X).</p

Phalaena sp.

<div><h3>Background</h3><p>The current understanding of the functional characteristics of circulating endothelial progenitor cells (EPC) is limited, especially in patients affected by cardiovascular diseases. In this study, we have analyzed the <em>in vitro</em> clonogenic capacity of circulating EPC, also known as endothelial colony-forming cells (ECFC), in patients with acute coronary syndrome (ACS), in comparison to the colony forming unit-endothelial-like cells (CFU-EC) of hematopoietic/monocytic origin.</p> <h3>Methodology/Principal Findings</h3><p>By culturing peripheral blood mononuclear cells (PBMC) of patients with ACS (n = 70), CFU-EC were frequently isolated (from 77% of ACS patients), while EPC/ECFC were obtained only in a small subset (13%) of PBMC samples, all harvested between 7–14 days after the acute cardiovascular event. Notably, <em>ex-vivo</em> generation of EPC/ECFC was correlated to a higher <em>in vitro</em> release of PDGF-AA by the corresponding ACS patient PBMC. By using specific endothelial culture media, EPC/ECFC displayed <em>in vitro</em> expansion capacity, allowing the phenotypic and functional characterization of the cells. Indeed, after expansion, EPC/ECFC exhibited a normal diploid chromosomal setting by FISH analysis and an immunophenotype characterized by: <em>i</em>) uniform positivity for the expression of CD105, CD31, CD146 and Factor VIII, <em>i</em>) variable expression of the CD34, CD106 and CD184 markers, and <em>iii</em>) negativity for CD45, CD90, CD117 and CD133. Of interest, in single-cell replanting assays EPC/ECFC exhibited clonogenic expansion capacity, forming secondary colonies characterized by variable proliferation capacities.</p> <h3>Conclusion/Significance</h3><p>Our data indicate that a careful characterization of true EPC is needed in order to design future studies in the clinical autologous setting of patients with ACS.</p> </div

Subcloning potential of EPC/ECFC generated from the PBMC of ACS patients.

<p>After <i>ex-vivo</i> expansion, primary EPC/ECFC colonies were trypsinized and assessed for clonogenic potential capacity by single cells replating assay. In <b>A</b>, single cells derived from EPC/ECPF colonies were seeded in collagen I coated wells and monitored day by day (<b>a</b>: day 1; <b>b</b>: day 2; <b>c</b>: day 3; <b>e–f</b>: day 4; <b>a–e</b>: original magnification 25X; <b>f</b>: original magnification 40X). One representative experiment is shown. In <b>B</b>, secondary clones were classified on the basis of their proliferation properties. Data are mean±SD derived from six independent experiments.</p

Supplemental Material - Impact of Contrast-Associated Acute Kidney Injury on One-Year Outcomes in Very Elderly STEMI Patients: Insights From a Multicenter Registry in Northern Italy

Supplemental Material for Impact of Contrast-Associated Acute Kidney Injury on One-Year Outcomes in Very Elderly STEMI Patients: Insights From a Multicenter Registry in Northern Italy by Alessandro Ruzzarin, Simone Muraglia, Enrico Fabris, Giorgio Caretta, Filippo Zilio, Andrea Pezzato, Gianluca Campo, Matthias Unterhuber, and Luca Donazzan in Angiology.</p

A counseling program on nuisance bleeding improves quality of life in patients on dual antiplatelet therapy: A randomized controlled trial

<div><p>Background</p><p>Nuisance bleeding is a major determinant of quality of life and drug discontinuation in patients on dual antiplatelet therapy (DAPT). However, no randomized trial has been focused on the impact of nuisance bleeding on quality of life.</p><p>Methods</p><p>BATMAN is an investigator-driven, randomized, controlled, single-center, open trial (NCT02554006). Four hundred and forty-eight consecutive patients with indication to at least 6 months of DAPT were randomized to: i) multimodal counseling program focused on nuisance bleedings (interventional arm); ii) usual discharge process (control arm). The primary endpoint was the one-month health-related quality of life assessed by the EuroQol-5 Dimension (EQ-5D) visual analog scale (VAS) score. Secondary endpoints were EQ-5D at 1 and 6 months, EQ-5D VAS at 6 months, DAPT withdrawal, need of information regarding DAPT and/or nuisance bleedings, 6-month ischemic and bleeding adverse events.</p><p>Results</p><p>The EQ5D-VAS was significantly higher in the interventional arm compared to the control arm at 1 and 6 months (81[74–88] vs. 73[64–80], p < 0.001 at 1 month; 82[76–88] vs. 74[65–81], p < 0.001 at 6 months). Patients in the interventional arm had also significantly lower pain/discomfort and anxiety/depression at the EQ-5D both at 1 and 6 months. Patients in the control arm withdrew DAPT significantly more (7 (3%) vs. 1 (0.4%), p = 0.03) and looked for information regarding DAPT and/or about nuisance bleeding more frequently than those in the interventional arm (178 (79%) vs.19 (8%), p < 0.001).</p><p>Conclusions</p><p>The systematic utilization of a multimodal counseling program improved quality of life and reduced the DAPT withdrawal rate in patients on DAPT.</p></div

CONSORT study flow chart.

<p>* were included in the primary endpoint analysis. ** were included in the secondary endpoints 6-month analysis. PCI: percutaneous coronary intervention. DES: drug eluting stent.</p

EQ-5D results at one and six months.

<p>EQ-5D results at one and six months.</p

Additional file 1: of Physical activity intervention for elderly patients with reduced physical performance after acute coronary syndrome (HULK study): rationale and design of a randomized clinical trial

Brochure for patients at hospital discharge. (DOC 499 kb