p=0.0001
19
7±
29
.1
Po
st
hr
s
24
24
hr
s
27
0±
28
.2
29
6±
29
.8
Pr
ePC
I
35
0±
29
.4
4
Po
st
I
-P
C
Aggregation Units (AU)
1000
500
p>0.05
1000
500
24
68
1±
50
.8
Po
st
81
4±
52
.6
hr
s
Po
st
-P
C
I
hr
s
Po
st
93
4±
45
.5
0
93
9±
65
.7
0
Po
st
-P
C
I
Pr
ePC
I1
00
0±
51
.6
Po
st
-P
C
I
86
9±
42
.1
24
hr
s
Po
st
79
2±
41
.0
0
p=0.01
1500
p<0.05
11
48
±6
6.
1
500
TRAP induced Impedence aggregometry
Patients without HTPR
p=0.01
1500
Aggregation Units (AU)
1000
Po
st
TRAP induced Impedence aggregometry
Patients with HTPR
p=0.0002
p<0.05
56
3±
49
.
72
9±
42
.3
Pr
ePC
I
Pr
ePC
I4
99
±4
0.
3
Po
st
-P
C
I4
07
±3
7.
7
24
hr
s
Po
st
26
9±
24
.6
Aggregation Units (AU)
1500
200
0
0
TRAP induced Impedence aggregometry
All patients
fig2
200
400
I
0
400
600
88
5±
65
.2
200
600
Pr
ePC
I
400
p<0.05
800
Aggregation Units (AU)
Aggregation Units (AU)
600
p<0.05
p>0.05
800
Po
st
-P
C
p>0.05
Pr
ePC
I
Aggregation Units (AU)
800
ADP induced Impedence aggregometry
Patients without HTPR
ADP induced Impedence aggregometry
Patients with HTPR
p=0.0002
24
ADP induced Impedence aggregometry
All patients
fig1
Figure 1: ADP (adenosine diphosphate) and Figure 2: TRAP (thrombin receptor agonist
peptide) induced aggregation as measured by MEA (multi electrode aggregometry). Each
agonist response in ALL patients (left panel), those with HTPR (centre panel), and those
without HTPR (right panel). Aggregation units calculated as area under the curve after 6
minutes (impedence:time). Abbreviations: HTPR: High on treatment platelet reactivity, PCI:
percutaneous coronary intervention
40
30
30
30
hr
s
15
10
5
0
-5
I
15
10
5
0
-5
55
2
2.
2.
1
2
IQ
R
IQ
R
1.
2.
3.
1
2.
6
I
Po
st
-P
C
st
hr
s
Po
Po
hr
s
24
st
-P
C
I
st
I
3.
2
2.
8
IQ
R
IQ
R
IQ
R
7.
6
5
5.
5.
8
Po
Pr
ePC
I
st
Po
24
hr
s
3.
1
IQ
R
2.
6
5
2.
I
st
-P
C
IQ
R
3.
5
5
1.
IQ
R
IQ
R
Po
ns
20
-10
-10
3.
6
-10
% CD62P Positive
-5
ns
25
ns
20
Pr
ePC
0
24
24
% CD62P Positive
5
% platelets expressing surface p-selectin
Patients without HTPR
ns
25
10
2.
7
31
.9
±2
.2
Pr
ePC
I
Po
st
I
Po
st
-P
C
24
hr
s
31
.2
±2
.4
34
.5
±3
.4
Pr
ePC
I
Po
st
31
.8
±1
.9
Po
st
-P
CI
Pr
ePC
I
ns
15
Pr
ePC
I
% platelets expressing surface p-selectin
Patients with HTPR
ns
20
32
.5
±3
.2
0
36
.1
±3
.9
0
36
.1
±3
.0
10
0
32
.1
±1
.7
10
25
% CD62P Positive
20
10
Po
st
20
ns
31
.6
±2
.9
ng/ml
40
ng/ml
40
20
ns
50
ns
hr
s
ns
50
% platelets expressing surface p-selectin
All patients
fig4
Serum concentration Soluble p-selectin
Patients without HTPR
24
p=0.023
ns
50
ng/ml
Serum concentration Soluble p-selectin
Patients with HTPR
Po
st
-P
C
Serum concentration Soluble p-selectin
All patients
fig3
Figure 3 (top panel): Soluble p-selectin concentration pre, post and 24hrs post-PCI showing
ongoing p-selectin secretion throughout the 24hr period, mean±SEM.
Figure 4 (bottom panel): Platelet bound p-selectin expressed as percentage of platelets that
were CD62P positive median (IQR) (bottom row).
All patients (left column), those with HTPR (middle column) and those without HTPR.
Abbreviations: HTPR: High on treatment platelet reactivity, PCI: percutaneous coronary
intervention, CD62P: p-selectin protein present on platelet membrane
Serum concentration 12-HETE
All patients
Serum concentration 12-HETE
Patients with HTPR
p=0.017
4
3
3
3
2
2
2
0
-1
-1
-1
06
-0
.1
0.
09
0.
15
t
Pr
ePC
I
st
Po
s
hr
24
(0
.
4)
-1
.4
0.
9
2
0.
13
-P
C
I
Pr
e
st
Po
s
hr
24
(0
.1
1
8)
-0
.3
(0
.0
7
0.
1
1
0.
15
-P
C
I
(0
.0
90.
24
)
0
(0
.0
80.
86
)
0
4)
1
Po
s
1
p=0.009
hr
s
1
ng/ml
4
ng/ml
4
(0
.0
90.
36
)
ng/ml
p=0.63
Pr
e
Serum concentration 12-HETE
Patients without HTPR
24
fig5
Fig 5: Serum 12-HETE concentration in All patients (left panel), HTPR patients (middle panel)
and patients without HTPR (right panel). Data points are pre-PCI and 24 hrs post-PCI. Data
expressed as median (IQR), ng/ml. Wilcoxin test used throughout. Abbreviations: 12-HETE:
12- Hydroxyeicosatetraenoic acid, HTPR: High on treatment platelet reactivity, PCI:
percutaneous coronary intervention
24 hrs post-PCI ADP induced Aggregation
vs
Soluble P-selectin at 24 hours
Pre-PCI ADP induced Aggregation
vs
Soluble P-selectin at 24 hours
Pre-PCI ADP AU
1250
Pearson r 0.391
CI 0.071 to 0.64
R2 0.152
p=0.0185
1000
24 hrs Post-PCI ADP AU
fig6
750
500
250
0
0
20
40
60
80
100
24 hrs Post-PCI Sol p-selectin ng/ml
1250
Pearson r 0.155
CI -0.19 to 0.47
R2 0.024
p=0.389
1000
750
500
250
0
0
20
40
60
80
100
24 hrs Post-PCI Sol p-selectin ng/ml
Figure 6:
Correlation between pre-PCI ADP induced aggregation and p-selectin measured at 24hrs
post-PCI (left panel), and 24hr post-PCI ADP induced aggregation and p-selectin measured at
24hrs (right panel). Pre-PCI ADP correlates with degree of platelet activation as indicated by
soluble p-selectin concentration at 24hrs (Pearson r 0.391, R 2 0.152, p=0.0185)
Table 1: Baseline characteristics of patient population including selected paired
characteristics at 24 hrs post-PCI
Pre-PCI
24 hrs post-PCI
p value
n=42
Age
65.9 ± 12.6
Male
31 (73.8)
Haemaglobin g/L
145.7 ± 11.2
139 ±13.6
0.11
Platelets x 109/L
258.4 ± 71.4
258.7 ± 58.5
0.98
CRP mg/L
4.4 (3.0 – 4.0)
Creatinine μmol/L
95.3 ± 28.2
Symptoms to PCI days
5 (5 – 7)
NSTEMI
42 (100)
Left ventricular function (EF)
52.7 ± 3.5
Hypertension
25 (59.5)
Diabetes
14 (33.3)
Hypercholesterolaemia
31 (73.8)
Smoking history
20 (47.6)
Beta-blocker
32 (76.2)
Calcium channel blocker
6 (14.3)
ACE Inhibitor
35 (83.3)
Statin
36 (85.7)
Clopidogrel
42 (100)
Baseline clinical and laboratory characteristics of study population. Both pre-PCI and 24 hr
post-PCI haemoglobin and total platelet count are included with no significant difference seen
between either of these values. Abbreviations: PCI - percutaneous coronary intervention, CRP
– C reactive protein, NSTEMI – non-ST-elevavation myocardial infarction, EF - ejection
fraction, ACE - angiotensin converting enzyme inhibitor
Table 2: Summary table of key findings for aggregometry, soluble and
platelet bound p-selectin (ALL patients)
n=42
Pre PCI
(SEM)
Post PCI
(SEM)
24hrs post
PCI (SEM)
P ANOVA
95% CI of
diff
a/ Pre vs Post PCI
b/ Pre vs 24 hr PCI
c/ Post vs 24 hr PCI
ADP aggregation
(AUC)
499.1 ±
46.3
407.6 ±
37.7
269.1 ±
24.6
a/ p>0.05
b/ p=0.0002*
c/ p<0.05*
a/ -34.1 to 217
b/ 104.5 to
355.6
c/ 13 to 264.1
TRAP
aggregation
(AUC)
1000 ±
51.6
869.2 ±
42.1
792.0 ±
41.0
a/ p<0.05*
b/ p=0.0002*
c/ p>0.05
Soluble pselectin (ng/ml)
32.1 ± 1.7
31.8 ± 1.9
36.1 ± 3.0
a/ p>0.05
b/ p=0.0225
c/ p>0.05
a/ 0.74 vs
261.4
b/ 114.1 to
302.6
c/ -29.5 to
184.1
a/ -3.7 to 3.7
b/ 0.82 to
10.0
c/ -0.9 to 12.0
% p-selectin +ve
plts
n=16
2.7 (1.45.0)
2.5 (2.13.6)
2.6 (2.05.5)
a/ ns
b/ ns
c/ ns
Row 1 and 2 ADP and TRAP induced aggregation (AU), Row 3 soluble p-selectin (ng), row 4
percentage of platelets expressing p-selectin. Mean ± SD or median (IQR). PANOVA values with
Tukey’s multiple comparison (a= Pre-PCI vs Post-PCI, b=Pre-PCI vs Post-PCI). Except were
not normally distributed or missing values prevent (Soluble p-selectin).