Introduction
Experimental procedures
Patients
Design
Patient management
Blood sample management and biochemistry analysis
Animals
Subjects
Experimental groups
ES model: surgical procedures
pMCAO model: surgical procedures
Physiological monitoring: mortality
Functional evaluation
Evaluation of infarct volume by hematoxylin-eosin (H&E)
Cell death
Quantification of biochemical markers
Data analysis
Results
Patients
Stroke | Controls |
P
| |
---|---|---|---|
(n = 51) | (n = 17) | ||
Baseline, demographic data and risk factors
| |||
Men, n (%) | 24 (47) | 8 (47) | 1 |
Age, mean (SD) | 73.5 (10.3) | 70.4 (15) | 0.353 |
Symptoms onset to first blood sample, h; mean (SD) | 4.8 (3.3) | 5 (3.3) | 0.795 |
Symptoms Onset to second blood sample, h; mean (SD) | 71.2 (4.9) | 73.5 (4.1) | 0.394 |
Systolic blood pressure on admission, mean (SD) | 154.3 (19) | 149.2 (32.3) | 0.467 |
Blood glucose, mmol/L; median (IQR) | 123 (38) | 133 (106) | 0.100 |
Hypertension, n (%) | 34 (66.7) | 12 (70.6) | 0.765 |
Diabetes Mellitus, n (%) | 10 (19.6) | 6 (35.3) | 0.187 |
Hyperlipidemia, n (%) | 17 (33.3) | 7 (41.2) | 0.558 |
Atrial Fibrillation, n (%) | 22 (43.1) | 0 (0) | 0.001 |
Current smoking, n (%) | 13 (25.5) | 5 (29.4) | 0.751 |
Alcohol abuse, n (%) | 2 (3.9) | 2 (11.8) | 1 |
Coronary heart disease, n (%) | 8 (15.7) | 2 (11.8) | 1 |
Peripheral arterial disease, n (%) | 1 (2) | 2 (5.9) | 0.440 |
Disease severity
| |||
NIHSS score on admission, mean (SD) | 11.3 (6) | - | - |
APACHE II score on admission, mean (SD) | 7.4 (2.6) | 7 (2.5) | 0.555 |
90-days outcome
| |||
mRS 0-2, n (%) | 27 (53) | 10 (58.8) | 0.723 |
Barthel index ≥ 90, n (%) | 22 (43.1) | 6 (35.3) | 0.569 |
Mortality, n (%) | 7 (13.7) | 0 (0) | 0.175 |
Brain infarction size, ml; median (IQR)
| 27.8 (78) | - | |
First and second tertile, ml, range
| 3 - 40 | - | |
Third tertile, ml, range
| 50 - 405 | - |
Biochemical markers
Correlations between biochemical markers, stroke severity, volume of infarct and stroke outcome
Values of spearman rho correlations: | IL-6 | TNF-α | Glutamate | ||||
---|---|---|---|---|---|---|---|
IL-6 | IL-6 | TNF-α | TNF-α | Glu | Glu | ||
<12 h | 72 h | <12 h | 72 h | <12 h | 72 h | ||
Clinical severity (APACHE II) | All patients | 0.261 |
0.326**
| 0,130 | 0,155 | −0.076 | −0.029 |
−0.061 |
0.484**
| −0,085 | 0,068 |
0.421*
| −0.193 | ||
Large infarcts |
0.630***
| 0.433 | 0.244 | 0.347 | −0.052 | 0.149 | |
−0.369 | 0.197 | −0,821 | −0,462 | −0.350 |
−0.727**
| ||
Medium infarcts | −0.066 | 0.241 | 0.190 | 0.044 | −0.042 | −0.007 | |
0.261 |
0.759**
| - | 0.400 | 0.374 | 0.410 | ||
Stroke severity (NIHSS) | All patients | 0.068 |
0.497***
| 0.312 | 0.247 | 0.193 |
0.261*
|
- | - | −0.488 | −0.488 | - | - | ||
Large infarcts | 0.247 | 0.020 | 0.388 | 0.433 | 0.165 | −0.013 | |
- | - | −0.866 | −0.866 | - | - | ||
Medium infarcts | −0.223 |
0.505***
| −0.023 | −0.108 | 0.217 |
0.371**
| |
- | - | −0.866 | −0.866 | - | - | ||
Volume of infarct§
| All patients | 0.207 |
0.290**
|
0.476**
| 0.275 | 0.120 | 0.151 |
Large infarcts | 0.398 | 0.299 |
0.592*
| 0364 | 0.325 | 0.324 | |
Medium infarcts | 0.069 | −0.113 | 0.097 | 0.178 | 0.001 | 0.001 | |
Clinical outcome: 3-months mRS | All patients | 0.230 |
0.444***
|
0.352*
| 0.150 | 0.101 | 0.006 |
-
| - | 0.546 | −0.095 | - | - | ||
Large infarcts |
0.453*
| 0.455 | 0.370 | 0.283 | 0.235 | 0.295 | |
- | - | 0.105 | 0.316 | - | - | ||
Medium infarcts | 0.042 | 0.273 | 0.338 | 0.120 | −0.023 | −0.080 | |
-
| - | 0.316 | −0.738 | - | - | ||
Clinical outcome: 3-months Barthel index | All patients | −0.121 | −0.245 | −0.152 | −0.109 | −0.164 | −0.017 |
−0.492*
| −0.303 | −0.536 | −0.009 | 0.092 |
0.433*
| ||
Large infarcts | −0.366 | −0.366 | 0.119 | −0.444 | −0.268 | 0.000 | |
−0.449 | −0.338 | −0.200 | −0.300 | −0.570 |
0.591*
| ||
Medium infarcts | −0.053 | −0.078 | −0.262 | 0.168 | −0.042 | 0.006 | |
−0.702*
| 0.025 | −0.316 | 0.738 | 0.368 | −0.495 |
Animal models
ES (n = 6) | pMCAO (n = 6) |
P
| |
---|---|---|---|
Volume of infarct in percent of the contralateral hemisphere, median (IQR)
| 58.6 (9.6) | 29 (5.1) | <0.001 |
Cell death as number of TUNEL positive cells
| |||
Cortex, median (IQR) | 103 (20.1) | 133.3 (18.1) | 0.022 |
Striatum, median (IQR)* | 59.1 (10.4) | - | - |
Functional evaluation**
| |||
At 24h, median (IQR) | 4 (1) | 3 (0) | 0.009 |
At 48 h, median (IQR) | 4 (1) | 3 (0) | 0.009 |
At 72 h, median (IQR) | 4 (1) | 3 (1) | 0.041 |
Percent post-operative weight loss, mean (SD)
| 34.8 (11.5) | 42.8 (12) | 0.286 |
Biochemical markers
Correlations between biochemical markers, functional evaluation, volume of infarct and cell death
Correlations: values of spearman R | IL-6 | TNF-α | Glutamate | |||
---|---|---|---|---|---|---|
IL-6 3h | IL-6 72h | TNF-α 3h | TNF-α 72h | Glu 3h | Glu 72 h | |
Clinical score pre-euthanasia (72 h)
| 0.171 | −0.600 | 0.348 | −0.522 | 0.429 | 0.388 |
0.507 | 0.169 | 0.676 | 0.338 | −0.169 | 0.169 | |
Volume of infarct as percentage of contralateral hemisphere
| 0.348 | 0.319 | −0.088 | −0.794 | 0.116 |
0.943***
|
0.058 | −0.087 | 0.522 | 0.000 | −0.232 | 0.232 | |
Cell death TUNEL in cortex
|
0.928***
| −0.058 |
0.765*
| −0.353 |
0.783*
| 0.486 |
TUNEL in striatum
†
| −0.714 | −0.257 | −0.714 |
−0.886**
| 0.086 | −0.257 |
−0.580 | 0.290 | −0.294 | −0.177 | −0.348 | 0.429 | |
- | - | - | - | - | - |