El CAD.HMGB1 and Atherosclerotic Plaque CompositionFigure 1. Weak correlations were observed Fruquintinib between calcium scoring with hs-CRP, hs-TnT and HMGB1 (a, c and f). A weak correlation was also noted between hs-CRP and A 196 chemical information non-calcified plaque burden (b), while stronger correlations were observed between the latter with hs-TnT and HMGB1 (d and f). doi:10.1371/journal.pone.0052081.g(p = 0.09) (Figure 2a). HsTnT and HMGB1 values significantly increased with increasing plaque presence and complexity. The highest values were observed in subjects with remodeled plaque (Figure 2). A correlation was observed between hs-TnT and HMGB1 (r = 0.26; p,0.01). No significant associations were noted between hs-TnT and hs-CRP or between hs-CRP and HMGB1 (data not shown).Prediction of Non-calcified Plaque Burden and of Plaque Composition by Biochemical MarkersUsing univariate analysis the total number of atherogenic risk factors, hsCRP, hsTnT and HMGB1 were associated with noncalcified plaque. By multivariate logistic regression analysis HMGB1 and hsTnT were independent predictors for the presence of non-calcified plaque burden, whereas risk factors and hs-CRP were no longer significant (Table 2).HMGB1 and Atherosclerotic Plaque CompositionFigure 2. Classifying patients by plaque composition, a trend was observed for higher hs-CRP values in patients with non-calcified plaque without however, reaching statistical significance (a). HsTnT and HMGB1 values on the other hand, increased with increasing plaque presence and complexity, yielding higher values in patients with non-calcified plaque versus purely calcified or no plaques and the highest values in subjects with remodeled non-calcified plaque (b and c). doi:10.1371/journal.pone.0052081.gPlaque characteristics by patient tertiles based on their hsTnT and HMBG1 values are presented in Table 3. Patients in the upper tertiles for both hsTnT and HMBG1 showed highercalcium scoring and non-calcified plaque burden versus those in the mid and lower tertiles. Furthermore, by combining both biomarkers, a very high negative predictive value for the presenceHMGB1 and Atherosclerotic Plaque CompositionTable 2. Uni- and multivariable logistic regression analysis for the prediction plaque composition (no plaque and only calcified versus non-calcified plaque with or without vascular remodeling).VariablesCoefficient Univariate analysisOdds Ratio95 Confidence Interval (CI)p-valueAge(yrs.) Male gender Arterial hypertension Hyperlipidemia Diabetes mellitus Positive family history Cigarette smoking Number of risk factors Hs-CRP 
Hs-TnT Hmbg0.03 0.30 0.73 0.58 0.44 0.47 0.24 0.41 0.17 0.14 1.23 Multivariable analysis1.03 1.35 2.07 1.79 1.55 1.60 1.27 1.51 1.18 1.16 3.0.99 to 1.07 0.70 to 2.59 0.89 to 4.79 0.93 to 3.46 0.51 to 4.69 0.84 to 3.05 0.66 to 2.43 1.11 to 2.03 1.02 to 1.37 1.07 to 1.24 2.29 to 5.0.06 NS 0.09 0.08 NS NS NS 0.008 0.03 0.0001 ,0.Age(yrs.) Number of risk factors Hs-CRP Hs-TnT Hmbg0.008 0.48 12926553 0.08 0.19 1.1.0 1.6 1.1 1.2 4.0.95 to 1.06 0.98 to 2.65 0.96 to 
1,21 1.07 to 1.37 2.43 to 7.NS 0.06 NS ,0.01 ,0.HR indicates risk ratios and CI the corresponding 95 confidence intervals. doi:10.1371/journal.pone.0052081.tof non-calcified and remodeled plaque (95 and 100 respectively) was noted in patients within the lower tertiles, which surpassed the negative predictive value of each biomarker separately. Similarly, patients in the upper tertiles for both biomarkers yielded high positive predictive values for non-calcifie.El CAD.HMGB1 and Atherosclerotic Plaque CompositionFigure 1. Weak correlations were observed between calcium scoring with hs-CRP, hs-TnT and HMGB1 (a, c and f). A weak correlation was also noted between hs-CRP and non-calcified plaque burden (b), while stronger correlations were observed between the latter with hs-TnT and HMGB1 (d and f). doi:10.1371/journal.pone.0052081.g(p = 0.09) (Figure 2a). HsTnT and HMGB1 values significantly increased with increasing plaque presence and complexity. The highest values were observed in subjects with remodeled plaque (Figure 2). A correlation was observed between hs-TnT and HMGB1 (r = 0.26; p,0.01). No significant associations were noted between hs-TnT and hs-CRP or between hs-CRP and HMGB1 (data not shown).Prediction of Non-calcified Plaque Burden and of Plaque Composition by Biochemical MarkersUsing univariate analysis the total number of atherogenic risk factors, hsCRP, hsTnT and HMGB1 were associated with noncalcified plaque. By multivariate logistic regression analysis HMGB1 and hsTnT were independent predictors for the presence of non-calcified plaque burden, whereas risk factors and hs-CRP were no longer significant (Table 2).HMGB1 and Atherosclerotic Plaque CompositionFigure 2. Classifying patients by plaque composition, a trend was observed for higher hs-CRP values in patients with non-calcified plaque without however, reaching statistical significance (a). HsTnT and HMGB1 values on the other hand, increased with increasing plaque presence and complexity, yielding higher values in patients with non-calcified plaque versus purely calcified or no plaques and the highest values in subjects with remodeled non-calcified plaque (b and c). doi:10.1371/journal.pone.0052081.gPlaque characteristics by patient tertiles based on their hsTnT and HMBG1 values are presented in Table 3. Patients in the upper tertiles for both hsTnT and HMBG1 showed highercalcium scoring and non-calcified plaque burden versus those in the mid and lower tertiles. Furthermore, by combining both biomarkers, a very high negative predictive value for the presenceHMGB1 and Atherosclerotic Plaque CompositionTable 2. Uni- and multivariable logistic regression analysis for the prediction plaque composition (no plaque and only calcified versus non-calcified plaque with or without vascular remodeling).VariablesCoefficient Univariate analysisOdds Ratio95 Confidence Interval (CI)p-valueAge(yrs.) Male gender Arterial hypertension Hyperlipidemia Diabetes mellitus Positive family history Cigarette smoking Number of risk factors Hs-CRP Hs-TnT Hmbg0.03 0.30 0.73 0.58 0.44 0.47 0.24 0.41 0.17 0.14 1.23 Multivariable analysis1.03 1.35 2.07 1.79 1.55 1.60 1.27 1.51 1.18 1.16 3.0.99 to 1.07 0.70 to 2.59 0.89 to 4.79 0.93 to 3.46 0.51 to 4.69 0.84 to 3.05 0.66 to 2.43 1.11 to 2.03 1.02 to 1.37 1.07 to 1.24 2.29 to 5.0.06 NS 0.09 0.08 NS NS NS 0.008 0.03 0.0001 ,0.Age(yrs.) Number of risk factors Hs-CRP Hs-TnT Hmbg0.008 0.48 12926553 0.08 0.19 1.1.0 1.6 1.1 1.2 4.0.95 to 1.06 0.98 to 2.65 0.96 to 1,21 1.07 to 1.37 2.43 to 7.NS 0.06 NS ,0.01 ,0.HR indicates risk ratios and CI the corresponding 95 confidence intervals. doi:10.1371/journal.pone.0052081.tof non-calcified and remodeled plaque (95 and 100 respectively) was noted in patients within the lower tertiles, which surpassed the negative predictive value of each biomarker separately. Similarly, patients in the upper tertiles for both biomarkers yielded high positive predictive values for non-calcifie.
