Online Appendix for the following JACC article
TITLE: Detection of Antecedent Myocardial Ischemia With Multiselectin Molecular Imaging
AUTHORS: Brian P. Davidson, MD, Beat A. Kaufmann, MD, J. Todd Belcik, BS, RCS, RDCS, Aris Xie, MS, Qi
Yue, MD, Jonathan R. Lindner, MD
APPENDIX
Supplemental Figures
Low MI
High MI (continuous or intermittent)
Signal Intensity
Continuous
High MI
Continuous
Low MI
B
Continuous
Low MI
A
Intermittent
High MI
C
D
imaging paused
Time
Supplemental Figure I. Method for Detecting Signal from Retained Microbubbles at Low
and High MI. After bolus injection and observation of first pass through the myocardial
circulation using low-MI imaging, imaging was paused for 8 min. Imaging was resumed using
low MI imaging and several end-systolic frames were obtained and averaged (A) representing a
combination of retained and freely circulating microbubbles. End-systolic triggered imaging was
then initiated and the MI was increased to 1.4 while shielding the myocardium. The first two
frames obtained were then obtained and averaged (B). Continuous high-MI imaging was then
used to destroy microbubbles and, subsequently, several end-systolic frames at a pulsing interval
of 10 cardiac cycles were obtained and averaged (C) to measure high-MI signal attributable to
freely circulating microbubbles. The MI was then returned to low MI settings and several endsystolic images were obtained at least 5 seconds after reducing the MI to measure low MI signal
attributable to freely circulating microbubbles. The signal attributable to retained microbubbles
alone was determined by digital subtraction of averaged frames D from A for low MI imaging,
and C from B for high MI imaging. Relative signal intensities are not necessarily to scale.
Supplemental Figure II. Intravascular Residence Time for Targeted Microbubble Agents.
Data demonstrate LV cavity signal from each agent after intravenous injection of 1×105
microbubbles.
MBctr
MBAb
MBYSPSL
Supplemental Figure III. Size Distribution of Microbubble Agents. Data represent diameter
vs number histograms obtained by Coulter Multisizer analysis.
IR
PO
Supplemental Figure IV. Examples Illustrating Lack of Myocardial Infarction on TTC
Staining. Post-mortem TTC staining of short-axis slices obtained at 1 mm increments through
the ventricles from a mouse undergoing ischemia-reperfusion protocol (IR) illustrating lack of
myocardial infarction. The bottom set of images illustrates a positive control using permanent
occlusion (PO) of a more distal portion of the LAD for 90 min where infarction is illustrated by
the region lacking TTC staining (arrows).
A
B
Low-Power Imaging
0.5
Low-Power Imaging
0.5
0.4
MBYSPSL
0.3
Signal Intensity (VIU)
Signal Intensity (VIU)
0.4
MBAb
0.2
0.1
0.0
1.5 hr
3 hr
6 hr
18 hr
1.5 hr
3 hr
6 hr
0.2
P-sel -/- Mice
0.0
18 hr
Remote Region
Ischemic Region
Control
3 hr
6 hr
Sham
D
3 hr
6 hr
Ischemic Region
3 hr
6 hr
Remote Region
High-Power Imaging
High-Power Imaging
0.5
0.5
0.4
0.4
MBYSPSL
0.3
Signal Intensity (VIU)
Signal Intensity (VIU)
MBAb
0.3
0.1
C
MBAb
0.2
MBYSPSL
MBAb
0.3
0.2
P-sel -/- Mice
0.1
0.0
MBYSPSL
0.1
1.5 hr
3 hr
6 hr
Ischemic Region
18 hr
1.5 hr
3 hr
6 hr
Remote Region
18 hr
0.0
Control
3 hr
Sham
6 hr
3 hr
6 hr
Ischemic Region
Supplemental Figure V. Myocardial Molecular Imaging Data After Log-Linear
Conversion.
3 hr
6 hr
Remote Region