Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Modelization of Blood-Borne Hypercoagulability in Myeloma: A Tissue-Factor-Bearing Microparticle-Driven Process

doi: 10.1055/s-0039-1700885

Figure Lengend Snippet: Effect of MPCs (1,000 cells/μL) or MPC-dMPs on thrombin generation in normal PPP and in PPP depleted of FVII or FXII

Article Snippet: Samples of fresh frozen normal platelet poor plasma (PPP; Ref 00539) and immunodepleted lyophilized plasma deficient of clotting factor VII (FVII) or FXII were purchased from Stago (Gennevilliers, France).

Techniques:

Journal: The Journal of Experimental Medicine

Article Title: Targeting coagulation factor XII provides protection from pathological thrombosis in cerebral ischemia without interfering with hemostasis

doi: 10.1084/jem.20052458

Figure Lengend Snippet: Infarct volumes and functional outcomes 24 h after focal cerebral ischemia in WT and FXII −/− mice, and in FXII −/− mice infused with human FXII. (A) Representative images of three corresponding coronal sections of WT (left), FXII −/− (middle), and FXII −/− mice reconstituted with human FXII (huFXII, 2 μg/g body weight i.v. 10 min before the MCAO; right) stained with TCC. (B) Brain infarct volumes in WT ( n = 18), FXII −/− ( n = 18), and FXII −/− mice reconstituted with huFXII ( n = 8); **P < 0.01. (C) Neurological Bederson score assessed at day 1 after tMACO for WT ( n = 18), FXII −/− ( n = 18), and huFXII-treated FXII −/− animals ( n = 8); **P < 0.01. n.s., not significant.

Article Snippet: In some experiments, human FXII (huFXII; American Diagnostics) was injected intravenously immediately before the experiment.

Techniques: Functional Assay, Staining

Journal: Scientific Reports

Article Title: Penicillin causes non-allergic anaphylaxis by activating the contact system

doi: 10.1038/s41598-020-71083-x

Figure Lengend Snippet: Penicillin activates the contact system in an FXII-dependent manner. ( A ) Penicillin-induced CSA in standard or FXII-deficient human plasma. 100 μL of plasma was pretreated with 100 μL of penicillin at various concentrations (diluted by Tris buffer: 50 mM Tris–HCl, 0.117 M NaCl, pH 7.8) at 37 °C. Ten minutes later, 100 μL of the chromogenic substrate S-2302 (1.5 mg/mL) was added and further incubated at 37 °C for 30 min. The reaction mix was centrifuged at 3,000 × g for 5 min. Supernatant absorbance was monitored at 405 nm. Kaolin was used as a positive control of the contact system. Buffer alone was included as the negative control. * P < 0.05 and ** P < 0.01 vs. negative control. ( B ) Plasma prototypical FXII level decreased after penicillin treatment. Standard human plasma was incubated with or without 4 KU/mL penicillin at 37 °C for 30 min and analyzed for FXII determination by western blotting. Transferrin was used as the internal reference. Full-length blots and the detailed information of the used antibodies were presented in Supplementary file . ( C ) Penicillin induced BK release in PI-HUVEC. HUVEC were incubated with 10% standard human plasma in the presence of 20 μM Zn 2+ at 37 °C for 1 h. The plasma was removed and the cells were washed twice. PI-HUVEC and non-PI-HUVEC were further incubated with penicillin at 37 °C for 30 min. Supernatant BK was determined by ELISA. ** P < 0.01.

Article Snippet: Antibodies for human FXII and transferrin were from GeneTex Inc. (San Antonio, TX, USA).

Techniques: Clinical Proteomics, Incubation, Positive Control, Negative Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Clinical Laboratory Analysis

Article Title: Evaluation of the Atellica COAG 360 coagulation analyzer in a specialized coagulation laboratory

doi: 10.1002/jcla.24276

Figure Lengend Snippet: Reagents used on the Atellica COAG 360 analyzer

Article Snippet: FXI and FXII OSA , Actin FS (Siemens) , SHP (Siemens) , Immunodepleted (Siemens).

Techniques: Activity Assay

Journal: The Journal of Experimental Medicine

Article Title: Targeting coagulation factor XII provides protection from pathological thrombosis in cerebral ischemia without interfering with hemostasis

doi: 10.1084/jem.20052458

Figure Lengend Snippet: Inhibition of FXII activity inhibits clotting in vitro and thrombus formation in vivo. (A) Normal human plasma (open symbols) and WT mouse plasma (filled symbols) were incubated with increasing concentrations of PCK (1–200 μg/ml final concentration), an inhibitor that blocks FXIIa activity and activation. Clotting was initiated by adding kaolin and CaCl 2 (triangles) or TF (squares) to determine the aPTT and the PT, respectively. (B and C) PCK (8 μg/g of body weight) was infused intravenously into WT mice ( n = 8) before tMCAO. 24 h after stroke, treated animals were analyzed and compared with untreated controls (ctrl, n = 18 per group). (B) Infarct volumes determined from TTC-stained sequential coronal sections (*P < 0.05) and (C) the neurological function assessed by the Bederson Score for PCK-treated and untreated mice (***P < 0.0001).

Article Snippet: The peptide-based FXII inhibitor PCK was obtained from Bachem and administered intravenously.

Techniques: Inhibition, Activity Assay, Coagulation, In Vitro, In Vivo, Incubation, Concentration Assay, Activation Assay, Staining

Journal: The Journal of Experimental Medicine

Article Title: Targeting coagulation factor XII provides protection from pathological thrombosis in cerebral ischemia without interfering with hemostasis

doi: 10.1084/jem.20052458

Figure Lengend Snippet: Inhibition of FXII activity does not affect normal hemostasis. (A) Tail bleeding times for PCK-treated (8 μg/g of body weight) and untreated control mice (ctrl; n = 12 per group; ***P < 0.0001). Heparin-infused (hep) mice are shown for comparison. (B) Serial coronal T2-weighted MRI brain images from untreated (ctrl), PCK-treated (8 μg/g of body weight), and FXII −/− mice at days 1, 3, and 7 after tMCAO ( n = 5 per group). The asterisk indicates hydrocephalus of the left lateral ventricle as an indicator of infarct-related swelling.

Article Snippet: The peptide-based FXII inhibitor PCK was obtained from Bachem and administered intravenously.

Techniques: Inhibition, Activity Assay