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Table 1.

Baseline characteristics of control and stroke groups at admission, including haematological parameters, vascular risk factors and admission details.

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Fig 1.

Comparison of GPVI expression between control and stroke cohorts and GPVI-dimer as a function of total GPVI in control and stroke cohorts.

A) Total GPVI and B) GPVI-dimer are both significantly higher in the stroke population at admission compared to the control cohort (P <0.0001). P-values were calculated using an unpaired t-test and the error bars represent the mean MFI of each of the cohorts ± SD. C) Correlation between platelet surface total GPVI-dimer and GPVI-dimer. These data were calculated using Pearson correlation coefficient and the regression line is presented for each set of data. The control platelets show a narrow range of GPVI-dimer levels, which is negatively correlated to total GPVI (P <0.0001). In contrast, the surface GPVI-dimer of stroke patients’ platelets demonstrate strong positive correlation to the amount of total GPVI (P <0.0001) and in general have higher GPVI-dimer levels than the controls. MFI = mean fluorescence intensity.

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Fig 2.

Comparison of P-selectin exposure between control and stroke cohorts and correlation between GPVI-dimer and P-selectin exposure.

A) Comparison of P-selectin surface exposure of the stroke patients with the controls show that the stroke patients have more activated “resting” platelets (i.e., before stimulation by exogenous agonist) and after CRP-XL (4 μg/mL) addition compared to the control group (P<0.0001). P-selectin exposure in response to ADP (0.5 μM) was not significantly different compared to controls. The error bars represent the median (Q1–Q3) %PP of each of the cohorts and P-values were calculated by the Mann-Whitney U-test for non-parametric data. B) GPVI-dimer expression and resting P-selectin exposure in both control and stroke patients are strongly correlated at admission (P <0.0001 for both). C) GPVI-dimer expression is correlated with P-selectin exposure in response to CRP-XL in the stroke patients at admission (P = 0.004). Correlations were calculated using Spearman’s rank correlation coefficient and regression line is presented for each data set. %PP = percentage of platelets positive, ns = not significant.

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Fig 3.

Comparison of GPVI expression and P-selectin exposure in stroke patients at day-0 and at day-90 (n = 51).

A) Total GPVI expression is not different at day-90 compared to day-0 (P = 0.52). B) GPVI-dimer is significantly higher at day-90 compared to day-0 (P <0.0001). The error bars represent the mean MFI of each of the cohorts ± SD and P-values were calculated by a paired t-test. C) Resting P-selectin exposure (P <0.0001), P-selectin exposure induced by 0.5 μM ADP (P <0.0001), and P-selectin exposure induced by 4 μg/mL CRP-XL (P = 0.001) are all significantly lower at day-90. The error bars represent the median (Q1–Q3) %PP of each of the cohorts and P-values were calculated by the Wilcoxon signed-rank test for non-parametric data. ns = not significant. D) Comparison of P-selectin expressed on the platelet surface of stroke patients at day-0 and at day-90 after stroke and P-selectin of resting normal platelets. The P-selectin exposure on stroke patients at day-0 are strongly correlated to GPVI-dimer expression (P <0.0001) and higher than those of resting control platelets. By day-90, the stroke patients’ P-selectin exposure, albeit still correlated with GPVI-dimer level (P <0.0001), has decreased. Correlations were calculated using Spearman’s rank correlation coefficient and regression line is presented for each data set. MFI = mean fluorescent intensity, %PP = percentage of platelets positive.

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Fig 4.

GPVI expression between control and stroke subtypes.

A) Total GPVI expression (P <0.0001) and B) GPVI-dimer expression are higher in all ischemic stroke etiologies as well as hemorrhagic strokes (P <0.0001). P-values were calculated using one-way ANOVA with Dunnett’s multiple comparisons testing and the error bars represent the mean MFI of each of the cohorts ± SD. C) Resting P-selectin expression is higher in ischemic and hemorrhagic stroke (P <0.0001). P-values were calculated by a Kruskal-Wallis test with Dunn’s multiple comparisons testing for non-parametric data and the error bars represent the median (Q1–Q3) %PP of each of the cohorts. Numbers in parentheses show the number of participants within each group. MFI = mean fluorescent intensity, %PP = percentage of platelets positive.

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Fig 4 Expand

Table 2.

Simple linear regression to identify associations between total GPVI or GPVI-dimer expression and a single predictor variable in the stroke population.

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