Journal: Thrombosis and haemostasis

Article Title: Plasma Phospholipid Transfer Protein Promotes Platelet Aggregation

doi: 10.1055/s-0038-1675228

Figure Lengend Snippet: PS externalization was assessed by annexin V–FITC labeling followed by flow cytometry. (A) and (B) flow cytometry analysis and quantification of the effect of PLTP overexpression (Tg) on PS externalization on platelets. (C) Effect of PLTP deficiency on PS externalization. (D) The effect of rPLTP on PLTP deficient platelet PS externalization. (E) Effect of PLTP overexpression on shear-induced (SIPA) induced platelet aggregation. Flow cytometric plots for the measurement of platelet aggregation. SIPA was estimated as increase in platelet population outside the single platelet gate compared to static control mouse platelet rich plasma. (F) Flow cytometric plots for the measurement of platelet aggregation after gated with PE-CD61 for platelet population. Values represent the mean ± SD, n = 5–8, *P < 0.05, **P < 0.01.

Article Snippet: Reagents Fluorescein isothiocyanate (FITC)-labeled mouse fibrinogen was purchased from Fisher Scientific (Silver Spring, MD, USA), and apyrase, prostaglandin E1, human fibrinogen, ADP, bovine serum albumin (BSA), and HEPES were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Labeling, Flow Cytometry, Over Expression, Shear, Control, Clinical Proteomics

Journal: Journal of medicinal chemistry

Article Title: Discovery of Allosteric Modulators of Factor XIa by Targeting Hydrophobic Domains Adjacent to its Heparin-Binding Site

doi: 10.1021/jm301757v

Figure Lengend Snippet: A) Representative profiles of direct inhibition of human factor XIa (FXIa) by sulfated quinazolin-4(3H)-ones (QAO). The inhibition of FXIa was measured spectrophotometrically through a S2366 hydrolysis assay at pH 7.4 and 37 °C. Solid lines represent sigmoidal fits to the data to obtain IC50, YM, and YO using equation 1, as described in the experimental section. B) Proteolytic activity of human thrombin (Thr), factor Xa (FXa), trypsin (Tryp) and chymotrypsin (ChTryp) by 500 μM sulfated QAO (16S, 15S, and 13S) using chromogenic substrate assay. The assays were performed using substrates appropriate for the enzyme being studied under conditions closest to the physiological condition. The ratio of the proteolytic activity of an enzyme in the presence of the sulfated QAO to that in its absence was used to determine percent activity (%).

Article Snippet: Chromogenic substrates, Spectrozyme TH (H- D -cyclohexylalanyl-Ala-Arg- p -nitroanilide), Spectrozyme factor Xa (Methoxycarbonyl- D- cyclohexylglycyl- Gly-Arg- p -nitroanilide), and Spectrozyme CTY were obtained from American Diagnostica (Greenwich, CT).

Techniques: Inhibition, Hydrolysis Assay, Activity Assay