Thrombin (FIIa; A) and APC (B) generation, and TM (C, D), EPCR (E, F), and Protein S (G, H) expression, on the endocardial surface of wildtype, all HF (HF), early HF, acute HF, and chronic HF mice.

<p>Thrombin generation is increased in acute HF compared to wildtype mice (A). APC generation is decreased in all HF mice compared to wildtype mice (B). TM (C) and EPCR (E) <u>mRNA</u> expression is attenuated in all HF mice groups compared to wildtype mice. Protein S (G) transcripts are also decreased in early and chronic HF mice. <u>Protein</u> expression of TM (D) and Protein S (H) is decreased in chronic HF compared to wildtype mice. EPCR (F) protein expression is attenuated in early and acute HF mice relative to wildtype and chronic HF mice. *p<0.05, **p<0.01, ***p<0.005.</p

vWF (red; A,D,G,J,M), fibrin (green; B,E,H,K,N), and co-localized (C,F,I,L,O) photomicrographs of the LV.

<p>Images are from wildtype (A-C), early HF (D-I), acute HF (J-L), and chronic HF (M-O) mice. vWF staining is present within endocardial cells (within presumptive Weibel-Palade bodies) in all groups and very prominently in early HF (D, F, inset, arrows). Fibrin staining is limited on the endocardial surface of wildtype mice (B) but increased in HF mice (E, H, K, N). Small thrombi positive for both fibrin and vWF are present on the endocardial surface in both acute and chronic HF. vWF staining is limited on the endocardial surface in chronic HF mice (M-O). Note the heterogeneity in muscle fiber size in HF mice (I, O). Scale bar in O is 145 μm for all panels, except inset (84 μm).</p

CREB_A133 mice phenotype (A), and hematoxylin and eosin (B, D) and Masson’s trichrome (C, E) stained cross-sections through the ventricles of wildtype (B), early HF (C), acute HF (D), and chronic HF (E) mice.

<p>An acute HF mouse (A, right) is next to a wildtype littermate (A, left); note the significant anasarca in acute HF. In histological section, chambers are dilated and thinner with progression of HF. Insets in C show margins of an intracardiac thrombus in two different sectional planes. In the bottom inset panel, a fibrous band (blue) attaches the thrombus (150 μm x 100 μm) to the endocardium. Endocardial thrombi are present in early and acute HF mice with limited incidence in chronic HF. In E, post-surgical hemorrhage is seen within ventricles (orange/brown-staining with trichrome), without endocardial attachment. Scale bar in E is 1 mm for all panels (0.5 mm for insets).</p

Hemodynamic measurements from wildtype (WT) and CREB_A133 mice during cardiac echocardiography.

<p>Fractional area change (FAC); fractional shortening (FS); left ventricular ejection fraction (LVEF); stroke volume (SV); cardiac output (CO); LV diastolic (d) and systolic (s) internal diameter (LVID).</p><p>*p<0.05 vs WT,</p><p>**p<0.01 vs WT,</p><p>***p<0.001 vs WT,</p><p>^#p<0.05 vs Acute HF,</p><p>^##p<0.01 vs Acute HF,</p><p>^###p<0.001 vs Acute HF.</p><p>Hemodynamic measurements from wildtype (WT) and CREB_A133 mice during cardiac echocardiography.</p

Tail clot assay (A) and platelet aggregometry (B-D) in wildtype, all HF (HF), early HF, acute HF, and chronic HF mice.

<p>The bleeding index, reflecting time to clot and total blood loss, is decreased in early and acute HF mice compared to wildtype and chronic HF mice (A; p<0.005). Platelet aggregation with ADP is similar across all groups (B). In contrast, platelet agglutination with ristocetin is attenuated overall in HF mice (p<0.05; representative trace in C, data in D), particularly in acute HF mice.</p

Quantitative determination of vWF: in plasma (A; immunoblot), by immunostaining in sectional endocardium (B; IHC), on the endocardial surface (C; ELISA), and following thrombin (IIa)-promoted vWF secretion from endocardium (D; ELISA).

<p>Circulating vWF expression is higher in all groups of HF compared to wildtype mice (A). Sectional percentage of vWF staining is higher in early HF compared to both wildtype and chronic HF mice (B). Endocardial vWF protein levels are higher in acute HF compared to both wildtype and chronic HF mice (C). Thrombin-promoted vWF extrusion is higher overall than unstimulated endocardium (D). Thrombin-promotion of vWF extrusion is prominent in wildtype mice and in early HF mice (D). *p<0.05, **p<0.01.</p

Representative M-mode (A-D) and short-axis views (E-H, at end diastole) following echocardiography of wildtype (A, E), early HF (B, F), acute HF (C, G), and chronic HF (D, H) mice.

<p>M-mode images were obtained from a parasternal long-axis view to assess septal (top) and posterior (bottom) wall thickening. A mural echolucent mass on the posterior wall consistent with mural thrombus is seen (G, white arrow) as well as papillary muscle (G, grey arrow). Speckle within LV cavities represents spontaneous echo contrast. Scale bar is on the right axis (mm).</p