1. Intracoronary shear-related up-regulation of platelet P-selectin and platelet-monocyte aggregation despite the use of aspirin and clopidogrel
by Andy S. C. Yong, Gabrielle J. Pennings, Michael Chang, Afiqah Hamzah, Tommy Chung, Miao Qi, David Brieger, Masud Behnia, Steven A. Krilis, Martin K.C. Ng, Harry C. Lowe, and Leonard Kritharides
Blood
Volume 117(1):11-20
January 6, 2011
©2011 by American Society of Hematology

2. Representative patient.
Representative patient. (A) Coronary angiogram showing a lesion in the mid left anterior descending artery. (B) Two-dimensional quantitative coronary angiography showing 52% DS (arrow). (C) Three-dimensional quantitative coronary angiography showing cross-sectional area stenosis (CS) of 62% and lesion length of mm. (D) Shear stress map generated from flow field calculated by computational fluid dynamics analysis, showing peak wall shear stress of 33.1 Pa. (E-G) Flow cytometry density plot of platelet CD62P expression in the coronary artery proximal (E) and distal (F) to the lesion, and in the coronary sinus (G). (H) Histogram overlay of platelet CD62 expression in the 3 sites. MFI indicates mean fluorescence intensity.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

3. Comparison between coronary sinus samples taken before and after sampling in the coronary arteries.
Comparison between coronary sinus samples taken before and after sampling in the coronary arteries. (A) Platelet CD62P, (B) platelet PAC-1, (C) platelet-monocyte aggregates (PM-Agg), (D) platelet-granulocyte aggregates (PG-Agg), and (E) soluble CD62P. MFI indicates mean fluorescence intensity. P = not significant for all comparisons shown. Solid bars and error bars represent mean ±SD.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

4. Comparison between femoral vein and coronary sinus.
Comparison between femoral vein and coronary sinus. (A) Platelet CD62P, (B) platelet PAC-1, (C) platelet-monocyte aggregates (PM-Agg), (D) platelet-granulocyte aggregates (PG-Agg), and (E) soluble CD62P. MFI indicates mean fluorescence intensity; *P < .05. Solid bars and error bars represent mean ±SD.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

5. Comparison between proximal, distal, and coronary sinus.
Comparison between proximal, distal, and coronary sinus. (A) Platelet CD62P, (B) platelet PAC-1, (C) platelet-monocyte aggregates (PM-Agg), (D) platelet-granulocyte aggregates (PG-Agg), and (E) soluble CD62P. MFI indicates mean fluorescence intensity; *P < .05. Solid bars and error bars represent mean ±SD.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

6. Correlation between %DS and absolute increase from the proximal artery to the distal artery.
Correlation between %DS and absolute increase from the proximal artery to the distal artery. (A) Platelet CD62P, (B) platelet PAC-1, (C) percentage of platelet-monocyte aggregates (PM-Agg), (D) platelet-granulocyte aggregates (PG-Agg), and (E) soluble CD62P. MFI indicates mean fluorescence intensity; PA (proximal artery); DA (distal artery). Solid lines shown are derived from linear regression analyses; dotted lines intersect y-axis = 0; r and P values shown are derived from correlation analyses.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

7. Comparison between lesions with ≤ 50% stenosis and > 50% stenosis for absolute increase from the proximal artery to the distal artery.
Comparison between lesions with ≤ 50% stenosis and > 50% stenosis for absolute increase from the proximal artery to the distal artery. (A) Platelet CD62P and (B) platelet-monocyte aggregates (PM-Agg). MFI indicates mean fluorescence intensity; PA, proximal artery; DA, distal artery; *P < .05. Box and error bars represent mean ±SD. Dotted lines intersect y-axis = 0.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

8. Correlation between log peak shear stress and absolute increase from the proximal artery to the distal artery.
Correlation between log peak shear stress and absolute increase from the proximal artery to the distal artery. (A) Platelet CD62P, (B) percentage of platelet-monocyte aggregates (PM-Agg). PA indicates proximal artery; DA, distal artery. Solid lines shown are derived from linear regression analyses; r and P values shown are derived from correlation analyses. Dotted lines intersect y-axis = 0.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

9. Monocyte CD11b results.
Monocyte CD11b results. (A) Comparison between levels in the coronary artery proximal and distal to the lesion. (B) Correlation between %DS and absolute increase in mean fluorescence intensity (MFI) from the proximal artery (PA) to the distal artery (DA). (C) Comparison between lesions with ≤ 50% stenosis and > 50% stenosis for absolute increase from PA to DA. (D) Correlation between log peak shear stress and absolute MFI increase from PA to DA. DS indicates diameter stenosis. Solid bars or boxes with error bars represent mean ±SD. r and P values were obtained from correlation analysis; solid lines were derived from linear regression analysis; dotted lines intersect y-axis = 0. *P < .05.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

10. Platelet and leukocyte activation in the presence of heparin and high doses of aspirin and clopidogrel.
Platelet and leukocyte activation in the presence of heparin and high doses of aspirin and clopidogrel. Comparison of levels of platelet aggregation between healthy controls and subjects on antiplatelet agents for (A) arachidonic acid and (B) ADP-induced maximum percentage of platelet aggregation. (C) Thrombin generation induced by low and high concentrations of phospholipids in controls and patients on heparin. Patients on heparin had undetectable low phospholipid dose-induced thrombin generation. **P < .05 for 1-way analysis of variance between groups and post hoc Tukey test between patient and control groups and between patient groups subjected to low and high doses of phospholipid-induced thrombin generation. (D-F) Comparison between proximal and distal coronary site for levels of platelet CD62P, PM-Agg, and monocyte CD11b. *P < .05. Solid bars and error bars represent mean ±SD.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology

11. Shear-induced activation in the presence of CD62P-blocking antibody and soluble platelet agonist inhibition.
Shear-induced activation in the presence of CD62P-blocking antibody and soluble platelet agonist inhibition. (A) Platelet CD62P expression, (B) PM-Agg formation, and (C) monocyte CD11b expression. Flow cytometry was performed in 3 healthy donors under control conditions, after shear stress with and without CD62P blocking antibody or soluble platelet agonist inhibition. Solid bars and error bars represent mean ±SD. P < .05 for 1-way analysis of variance and *P < .05 for difference compared with shear group by post hoc Tukey test.
Andy S. C. Yong et al. Blood 2011;117:11-20
©2011 by American Society of Hematology