p66shc mediates CRIF1 deficiency-induced VCAM-1 expression in <i>Crif1</i> silenced cells.

<p>(A) and (B) CRIF1 deficiency increased VCAM-1 protein and mRNA expression levels, respectively. MS-1 cells were transfected with <i>Crif1</i> siRNA for 48 h. Protein expression was measured by Western blotting. VCAM-1 protein expression levels were quantified by densitometric analysis (A, lower panel). (C) and (D) p66shc mediates CRIF1 deficiency-induced VCAM-1 protein and mRNA expression levels, respectively. MS-1 cells were infected with Adp66shcRNAi or AdLacZ for 24 h, followed by transfection with <i>Crif1</i> siRNA for 48 h. VCAM-1 protein expression levels were quantified by densitometric analysis (C, right panel). All Western blots are representative of three independent experiments and data are presented as means ± SEM of three independent experiments, **P<0.01, *** P<0.001 compared with the control.</p

p66shc mediates CRIF1 deficiency-induced adhesion of monocytes to endothelial cells (HUVECs).

<p>(A) CRIF1 deficiency induced monocyte adhesion to HUVECs. HUVECs were transfected with <i>Crif1</i> siRNA for 48 h. Representative photomicrographs of U937 cells adherent to HUVECs (A, upper panel) and quantification of adherent cells (A, lower panel) are shown. (B) p66shc mediates CRIF1 deficiency-induced adhesion of monocytes to HUVECs. HUVECs infected with Adp66shcRNAi or AdLacZ were transfected with <i>Crif1</i> siRNA for 48 h and incubated with U937 cells for 30 min. Representative photomicrographs of U937 cells adherent to HUVECs (B, upper panel) and quantification of adherent cells (B, lower panel) are shown. Data represent means ± SEM of three independent experiments, *P<0.05 compared with the control.</p

Loss of OXPHOS complexes and mitochondrial dysfunction in <i>Crif1</i>-silenced cells.

<p>(A) CRIF1 deficiency decreased OXPHOS complex subunit I, III and IV OXPHOS complex subunits, as determined by Western blotting using the appropriate antibodies. β-actin is shown as a loading control. OXPHOS complexes protein expression levels were quantified by densitometric analysis (right panel). Western blots are representative of three independent experiments. (B) CRIF1 deficiency increased mitochondrial ROS. Relative fluorescence of MitoSOX red was used as a measure of mitochondrial ROS levels (C) CRIF1 deficiency increased the mitochondrial membrane potential. Relative fluorescence of TMRE was used as a measure of mitochondrial membrane potential. (D) Oxygen consumption rates (OCR) were measured using a Seahorse XF-24 flux analyzer. Oligo: Oligomycin, CCCP: Carbonyl cyanide <i>m</i>-chloro phenyl hydrazine, Rote: Rotenone. Data represent means ± SEM of three independent experiments, *P<0.05, ** P<0.01, compared with the control.</p

p66shc mediates CRIF1 deficiency-induced mitochondrial ROS and cytosolic ROS production in <i>Crif1</i>-silenced cells.

<p>MS-1 cells were transfected with <i>Crif1</i> siRNA for 48 h. (A) CRIF1 deficiency increased cytosolic ROS, measured using the Amplex Red H₂O₂ assay kit. Relative fluorescence of Amplex Red was used as a measure of the H₂O₂ released from the cells. (B) CRIF1 deficiency increased phosphorylation of p66shc (p-p66shc) on ser36. p-p66shc levels were measured by Western blotting, which are representative of three independent experiments, and quantified by densitometric analysis (B, lower panel). (C) and (D) p66shc mediated CRIF1 deficiency-induced ROS production. MS-1 cells were infected with Adp66shcRNAi or AdLacZ for 24 h, followed by transfection with <i>Crif1</i> siRNA for 48 h. Data represent means ± SEM of three independent experiments, *P<0.05, ** P<0.01 compared with the control.</p

Phosphorylation of eIF2α (p-eIF2α) and CHOP protein expression were up-regulated in <i>Crif1</i>-silenced cells.

<p>CRIF1 deficiency induced p-eIF2α and CHOP protein expression. p-eIF2α and CHOP protein expression levels were measured by Western blotting. p-eIF2α and CHOP protein expression levels were quantified by densitometric analysis (left panels). All Western blots are representative of three independent experiments, and data are presented as means ± SEM of three independent experiments, *P<0.05 compared with the control.</p

Brachial-ankle PWV for predicting clinical outcomes in patients with acute stroke

<p><b>Background:</b> Although brachial-ankle pulse wave velocity (baPWV) is well-known for predicting the cardiovascular mortality and morbidity, its anticipated value is not demonstrated well concerning acute stroke.</p> <p><b>Methods:</b> Total 1557 patients with acute stroke who performed baPWV were enrolled. We evaluated the prognostic value of baPWV predicting all-cause death and vascular death in patients with acute stroke</p> <p><b>Results:</b> Highest quartile of baPWV was ≥23.64 m/s. All-caused deaths (including vascular death; 71) were 109 patients during follow-up periods (median 905 days). Multivariate Cox regression analysis revealed that patients with the highest quartile of baPWV had higher risk for vascular death when they are compared with patients with all other three quartiles of baPWV (Hazard ratio with 95% confidence interval [CI] 1.879 [1.022–3.456], <i>p</i> = .042 for vascular death).</p> <p><b>Conclusion:</b> High baPWV was a strong prognostic value of vascular death in patients with acute stroke</p