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  1. Article ; Online: Platelet Functional Testing Via High-Throughput Microtiter Plate-Based Assays.

    Tamang, Hem Kumar / Stringham, Emily N / Tourdot, Benjamin E

    Current protocols

    2023  Volume 3, Issue 2, Page(s) e668

    Abstract: Platelets play a critical role in hemostasis and thrombosis; therefore, in vitro assays that measure platelet reactivity are fundamental tools to gain insight into these physiologic processes, to diagnose platelet disorders, and to develop antithrombotic ...

    Abstract Platelets play a critical role in hemostasis and thrombosis; therefore, in vitro assays that measure platelet reactivity are fundamental tools to gain insight into these physiologic processes, to diagnose platelet disorders, and to develop antithrombotic therapies. However, conventional platelet assays such as aggregometry, the clinical gold standard for assessing platelet function, are low throughput and require specialized equipment. Since platelets have a finite life span ex vivo, processes to miniaturize and multiplex assays allow a much broader overview of platelet function in significantly less time than conventional assays. Several groups have developed simplified, high-throughput approaches to quantify platelet activation with standard laboratory equipment to lower the barrier of entry to study platelet biology. This article describes a panel of optimized and validated high-throughput microplate assays to comprehensively assess platelet functionality, independently or in combination, to increase throughput and reduce costs. Specifically, following stimulation of platelets, a plate reader can be used to measure light transmission aggregation via absorbance; dense-granule secretion based on ATP-dependent luminescence generation; and cytosolic calcium levels with a cell-permeant, fluorescent Ca
    MeSH term(s) Platelet Aggregation ; Platelet Function Tests/methods ; Calcium/metabolism ; Blood Platelets/metabolism ; Platelet Activation
    Chemical Substances Calcium (SY7Q814VUP)
    Language English
    Publishing date 2023-02-13
    Publishing country United States
    Document type Journal Article
    ISSN 2691-1299
    ISSN (online) 2691-1299
    DOI 10.1002/cpz1.668
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Anchoring IgG-degrading enzymes to the surface of platelets selectively neutralizes antiplatelet antibodies.

    Lynch, Donald R / Stringham, Emily N / Zhang, Boya / Balbin-Cuesta, Ginette / Curtis, Brian R / Palumbo, Joseph S / Greineder, Colin F / Tourdot, Benjamin E

    Blood advances

    2022  Volume 6, Issue 15, Page(s) 4645–4656

    Abstract: Immune thrombocytopenia (ITP) is an acquired bleeding disorder characterized by immunoglobulin G (IgG)-mediated platelet destruction. Current therapies primarily focus on reducing antiplatelet antibodies using immunosuppression or increasing platelet ... ...

    Abstract Immune thrombocytopenia (ITP) is an acquired bleeding disorder characterized by immunoglobulin G (IgG)-mediated platelet destruction. Current therapies primarily focus on reducing antiplatelet antibodies using immunosuppression or increasing platelet production with thrombopoietin mimetics. However, there are no universally safe and effective treatments for patients presenting with severe life-threatening bleeding. The IgG-degrading enzyme of Streptococcus pyogenes (IdeS), a protease with strict specificity for IgG, prevents IgG-driven immune disorders in murine models, including ITP. In clinical trials, IdeS prevented IgG-mediated kidney transplant rejection; however, the concentration of IdeS used to remove pathogenic antibodies causes profound hypogammaglobulinemia, and IdeS is immunogenic, which limits its use. Therefore, this study sought to determine whether targeting IdeS to FcγRIIA, a low-affinity IgG receptor on the surface of platelets, neutrophils, and monocytes, would be a viable strategy to decrease the pathogenesis of antiplatelet IgG and reduce treatment-related complications of nontargeted IdeS. We generated a recombinant protein conjugate by site-specifically linking the C-terminus of a single-chain variable fragment from an FcγRIIA antibody, clone IV.3, to the N-terminus of IdeS (scIV.3-IdeS). Platelets treated with scIV.3-IdeS had reduced binding of antiplatelet IgG from patients with ITP and decreased platelet phagocytosis in vitro, with no decrease in normal IgG. Treatment of mice expressing human FcγRIIA with scIV.3-IdeS reduced thrombocytopenia in a model of ITP and significantly improved the half-life of transfused platelets expressing human FcγRIIA. Together, these data suggest that scIV.3-IdeS can selectively remove pathogenic antiplatelet IgG and may be a potential treatment for patients with ITP and severe bleeding.
    MeSH term(s) Animals ; Bacterial Proteins/metabolism ; Bacterial Proteins/therapeutic use ; Blood Platelets/metabolism ; Humans ; Immunoglobulin G ; Mice ; Purpura, Thrombocytopenic, Idiopathic/drug therapy ; Streptococcus pyogenes/metabolism ; Thrombocytopenia/drug therapy
    Chemical Substances Bacterial Proteins ; Immunoglobulin G
    Language English
    Publishing date 2022-06-22
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 2915908-8
    ISSN 2473-9537 ; 2473-9529
    ISSN (online) 2473-9537
    ISSN 2473-9529
    DOI 10.1182/bloodadvances.2022007195
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 7153: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: GPR56/ADGRG1 is a platelet collagen-responsive GPCR and hemostatic sensor of shear force.

    Yeung, Jennifer / Adili, Reheman / Stringham, Emily N / Luo, Rong / Vizurraga, Alexander / Rosselli-Murai, Luciana K / Stoveken, Hannah M / Yu, Maiya / Piao, Xianhua / Holinstat, Michael / Tall, Gregory G

    Proceedings of the National Academy of Sciences of the United States of America

    2020  Volume 117, Issue 45, Page(s) 28275–28286

    Abstract: Circulating platelets roll along exposed collagen at vessel injury sites and respond with filipodia protrusion, shape change, and surface area expansion to facilitate platelet adhesion and plug formation. Various glycoproteins were considered to be both ... ...

    Abstract Circulating platelets roll along exposed collagen at vessel injury sites and respond with filipodia protrusion, shape change, and surface area expansion to facilitate platelet adhesion and plug formation. Various glycoproteins were considered to be both collagen responders and mediators of platelet adhesion, yet the signaling kinetics emanating from these receptors do not fully account for the rapid platelet cytoskeletal changes that occur in blood flow. We found the free N-terminal fragment of the adhesion G protein-coupled receptor (GPCR) GPR56 in human plasma and report that GPR56 is the platelet receptor that transduces signals from collagen and blood flow-induced shear force to activate G protein 13 signaling for platelet shape change.
    MeSH term(s) Animals ; Blood Platelets/metabolism ; Collagen/metabolism ; Hemostasis ; Humans ; Integrins/metabolism ; Mice ; Mice, Knockout ; Platelet Adhesiveness ; Platelet Aggregation ; Pseudopodia/metabolism ; Receptors, G-Protein-Coupled/genetics ; Receptors, G-Protein-Coupled/metabolism ; Signal Transduction ; Thrombosis/metabolism ; Transcriptome
    Chemical Substances ADGRG1 protein, human ; GPR56 protein, mouse ; Integrins ; Receptors, G-Protein-Coupled ; Collagen (9007-34-5)
    Language English
    Publishing date 2020-10-23
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.2008921117
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1148: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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