Journal: PLoS Pathogens

Article Title: Endothelial Cell Permeability during Hantavirus Infection Involves Factor XII-Dependent Increased Activation of the Kallikrein-Kinin System

doi: 10.1371/journal.ppat.1003470

Figure Lengend Snippet: Kallikrein-kinin system and BK inhibitors. A schematic illustration of three inhibitors that impact BK-mediated increase in endothelial cell permeability. CTI (corn trypsin inhibitor) inhibits FXIIa; PKSI-527 (plasma kallikrein specific inhibitor) inhibits KAL activity; Icatibant (HOE 140) is a peptidomimetic drug consisting of ten amino acids, which is a selective and specific antagonist of BKB2 receptors and is used in humans under orphan drug status for symptomatic treatment of hereditary angioedema.

Article Snippet: PKSI-527 is a specific inhibitor of KAL activity (Enzo Life Sciences) and corn trypsin inhibitor (CTI) inhibits FXIIa (Haematologic Technologies Inc.).

Techniques: Permeability, Activity Assay

Journal: PLoS Pathogens

Article Title: Endothelial Cell Permeability during Hantavirus Infection Involves Factor XII-Dependent Increased Activation of the Kallikrein-Kinin System

doi: 10.1371/journal.ppat.1003470

Figure Lengend Snippet: Kallikrein-kinin system activation and changes in permeability of hantavirus-infected HUVEC. Mock-, HTNV-, or ANDV-infected HUVEC were trypsinized, seeded onto ECIS chamberslides, and cultured until confluent. Media were removed and replaced with phenol red free EBM containing Zn 2+ . After an equilibration, 0.2 ml of phenol red free EBM containing Zn 2+ , FXII, PK, and HK (100 nM each) were added to cells (time zero) (black lines) (A). To measure inhibition of activation, some samples were treated with CTI (1 µM) (blue line), PKSI-527 (5 µM) (red line), or HOE 140 (1 µM) (green line) simultaneously with factors (B, C, D). Real time measurements frequency measurements were taken throughout the assay.

Article Snippet: PKSI-527 is a specific inhibitor of KAL activity (Enzo Life Sciences) and corn trypsin inhibitor (CTI) inhibits FXIIa (Haematologic Technologies Inc.).

Techniques: Activation Assay, Permeability, Infection, Electric Cell-substrate Impedance Sensing, Cell Culture, Inhibition

Journal: Thrombosis and haemostasis

Article Title: Nucleic Acids as Cofactors for Factor XI and Prekallikrein Activation: Different roles for high molecular weight kininogen

doi: 10.1160/TH16-09-0691

Figure Lengend Snippet: Effect of DNA on fXI-dependent thrombin generation in plasma (A) Thrombin generation measured in normal plasma after addition of 25 μg/ml DNA without (left hand panel) or with (right hand panel) 0.5 pM TF. Assays were run in the presence of vehicle (solid line), the anti-fXI IgG O1A6 (dotted line) or the fXIIa inhibitor CTI (dashed line). (B) Images of neutrophils stimulated with vehicle (control) or 100 ng/ml phorbol myristate acetate (PMA) with or without 100 U/mL DNAse. Cell permeable Hoechst stain (top row) indicates DNA within intact cell nuclei. Sytox Green staining (bottom row) demonstrates extracellular DNA or DNA within dying cells that have lost the ability to exclude the dye. PMA treated cells form neutrophil extracellular traps (NETs, middle column), which are not observed in the presence of DNase (right column). Images were made on an Inverted Fluorescence Microscope (89404-464) using a V10MP camera (VWR), and Motic Images Plus 2.0 software. (C) Thrombin generation in normal plasma supplemented with neutrophils treated with vehicle (gray lines, –PMA) or PMA (black lines, + PMA), without (solid lines) or with (dashed lines) DNAse in the absence of an inhibitor (No inhibitor), or in the presence of O1A6 or CTI. For reactions containing O1A6, no thrombin generation was detected with or without DNAse treatment. For reactions with CTI, some activity was detected in samples not treated with DNAse.

Article Snippet: Corn trypsin inhibitor (CTI), PK, fXIIa, HK, and HKa; Enzyme Research Lab.

Techniques: Staining, Fluorescence, Microscopy, Software, Activity Assay