Soluble CD40 ligand levels (sCD40L) in patients without and with the primary and secondary endpoints in the subgroup of patients with stent implantation (n = 205).

<p>Continuous data are given as mean ± SEM.</p><p>Soluble CD40 ligand levels (sCD40L) in patients without and with the primary and secondary endpoints in the subgroup of patients with stent implantation (n = 205).</p

Soluble CD40 ligand levels (sCD40L) in patients without and with the primary and secondary endpoints in the overall study population (n = 562).

<p>Continuous data are given as mean ± SEM.</p><p>Soluble CD40 ligand levels (sCD40L) in patients without and with the primary and secondary endpoints in the overall study population (n = 562).</p

<i>In Vivo</i> and protease-activated receptor-1-mediated platelet activation in patients presenting for cardiac catheterization

<p>Pathways of platelet activation that are not targeted by current antithrombotic therapy may be crucial for the development of ischemic events in patients undergoing coronary angiography. We therefore investigated whether <i>in vivo</i> and thrombin receptor activating peptide (TRAP)-stimulated platelet activation and monocyte-platelet aggregate (MPA) levels can serve as independent risk markers for adverse outcomes in aspirin-treated patients presenting for cardiac catheterization. <i>In vivo</i> and TRAP-stimulated platelet surface P-selectin, activated glycoprotein IIb/IIIa (GPIIb/IIIa) and MPA levels were determined in 682 consecutive patients undergoing cardiac catheterization and in 47 healthy controls. Two-year follow-up data were obtained from 562 patients. <i>In vivo</i> platelet surface P-selectin, activated GPIIb/IIIa and MPA levels were significantly higher in patients with angiographically-proven coronary artery disease than in healthy controls (all <i>p</i>≤0.02). Patients with an acute coronary syndrome (ACS; <i>n</i>=125) had significantly higher levels of <i>in vivo</i> MPA than patients without ACS (<i>n</i>=437; <i>p</i>=0.01). In the overall study population (<i>n</i>=562) the surface expression of P-selectin and activated GPIIb/IIIa, and the levels of MPA <i>in vivo</i> and in response to TRAP were similar in patients without and with subsequent ischemic events (all <i>p</i>>0.05). Similar results were obtained when only patients with angiographically-proven coronary artery disease (<i>n</i>=459), stent implantation (<i>n</i>=205) or ACS (<i>n</i>=125) were analyzed. Receiver-operating characteristic curve analyses did not reveal cut-off values for P-selectin, activated GPIIb/IIIa, and MPA levels for the prediction of ischemic events. In conclusion, <i>in vivo</i> and TRAP-stimulated platelet activation and MPA levels did not predict adverse ischemic outcomes in aspirin-treated patients presenting for cardiac catheterization.</p

Soluble CD40 ligand levels (sCD40L) in patients without and with the primary and secondary endpoints in the subgroup of patients with stent implantation (n = 205).

<p>Continuous data are given as mean ± SEM.</p><p>Soluble CD40 ligand levels (sCD40L) in patients without and with the primary and secondary endpoints in the subgroup of patients with stent implantation (n = 205).</p

Patient characteristics of the overall study population, and of patients with aspirin monotherapy (ASA) vs. dual antiplatelet therapy (ASA+Clo).

<p>Continuous data are shown as mean ± SEM. Dichotomous data are shown as %.</p><p>Abbreviations: ACE, angiotensin converting enzyme; BMI, body mass index; CRP, C-reactive protein; MI, myocardial infarction.</p><p>Patient characteristics of the overall study population, and of patients with aspirin monotherapy (ASA) vs. dual antiplatelet therapy (ASA+Clo).</p

Stability and metabolism of the four diastereomers of compound 1 in human plasma at 37 °C.

<p>t_1/2, half-life of first order elimination. For other abbreviations, see Figure 1.</p

³¹P NMR Spectra (¹H decoupled) of the diastereomers of compound 1 and their racemic mixture.

<p>In the left panel are the P¹,P⁴ regions, and in the right panel are the P²,P³ regions of the spectra. The traces are, from bottom to top: racemic mixture (RM), diastereomer <b>1.1</b>, <b>1.2</b>, <b>1.3</b>, and <b>1.4</b>.</p

Platelet-related properties of the four diastereomers of compound <b>1</b>.

a<p>VASP, vasodilator-stimulated phosphoprotein.</p>b<p>All comparisons <i>vs.</i> diastereomer <b>1.1</b>.</p>c<p>p<0.05.</p>d<p>p<0.01.</p>e<p>p<0.001.</p

Effects of compound 1 diastereomers on platelet aggregation and platelet ADP receptor signaling.

<p>Inhibition by the diastereomers of compound <b>1</b> of: A, 3 µM ADP-stimulated platelet aggregation; B, P2Y₁₂ mediated ADP-induced decrease of VASP phosphorylation; and C, P2Y₁-mediated ADP-induced intraplatelet Ca²⁺ level increase. VASP, vasodilator-stimulated phosphoprotein. For other abbreviation, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094780#pone-0094780-g001" target="_blank">Figure 1</a>.</p

Reversed-phase HPLC separation of the diastereomers of compound 1.

<p>Panel A: chromatogram of the racemic mix of the diastereomers of compound <b>1</b> analyzed by reversed-phase HPLC using a XBridge C18 column, 3.5 µm, 4.6×150 mm (Waters Inc., Waltham, Mass. USA), linear gradient from 1 to 7% methanol in 20 mM potassium phosphate buffer, pH 7, 1 ml/min, detection by UV at 260 nm; Panel B: chromatograms of isolated individual diastereomers (<b>1.1</b>–<b>1.4</b>) of compound <b>1</b> analyzed using the same column and flow rate, but with isocratic elution with 7% methanol in 20 mM potassium phosphate buffer, pH 7.</p