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https://www.readbyqxmd.com/read/28445754/probing-the-dynamics-of-clot-bound-thrombin-at-venous-shear-rates
#1
Laura M Haynes, Thomas Orfeo, Kenneth G Mann, Stephen J Everse, Kathleen E Brummel-Ziedins
In closed system models of fibrin formation, exosite-mediated thrombin binding to fibrin contributes to clot stability and is resistant to inhibition by antithrombin/heparin while still susceptible to small, active-site inhibitors. Each molecule of fibrin can bind ∼1.6 thrombin molecules at low-affinity binding sites (Kd = 2.8 μM) and ∼0.3 molecules of thrombin at high-affinity binding sites (Kd = 0.15 μM). The goal of this study is to assess the stability of fibrin-bound thrombin under venous flow conditions and to determine both its accessibility and susceptibility to inhibition...
April 25, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28382370/impact-of-fibrinogen-carbamylation-on-fibrin-clot-formation-and-stability
#2
Veronika Binder, Brith Bergum, Stéphane Jaisson, Philippe Gillery, Carsten Scavenius, Endy Spriet, Anne Karin Nyhaug, Helen M Roberts, Iain L C Chapple, Annelie Hellvard, Nicolas Delaleu, Piotr Mydel
Carbamylation is a non-enzymatic post-translational modification induced upon exposure of free amino groups to urea-derived cyanate leading to irreversible changes of protein charge, structure and function. Levels of carbamylated proteins increase significantly in chronic kidney disease and carbamylated albumin is considered as an important biomarker indicating mortality risk. High plasma concentrations and long half-life make fibrinogen a prime target for carbamylation. As aggregation and cross-linking of fibrin monomers rely on lysine residues, it is likely that carbamylation impacts fibrinogen processing...
May 3, 2017: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/28132723/eb1-contributes-to-proper-front-to-back-polarity-in-neutrophil-like-hl-60-cells
#3
Matthias Samereier, Michael Schleicher, Heike Roth, Doris Brechtefeld, Barbara Walzog, Annette Müller-Taubenberger
Directed migration of leukocytes towards a chemotactic source is largely dependent on coordinated actin cytoskeleton functions that provide the driving forces at the cell front and enable contractility at the rear. In contrast to the force-generating properties of the actin cytoskeleton, the microtubule network assumes a regulatory function in balancing front-to-back polarity. In migrating neutrophils, microtubules are mostly concentrated at the cell rear, and previously published work suggested that microtubules are stabilized and kept in place by a mechanism involving Cdc42, WASP, CD11b, and the end-binding protein 1 (EB1)...
March 2017: European Journal of Cell Biology
https://www.readbyqxmd.com/read/28030952/calcium-stearate-as-an-effective-alternative-to-poly-vinyl-alcohol-in-poly-lactic-co-glycolic-acid-nanoparticles-synthesis
#4
Claudia Cella, Irini Gerges, Paolo Milani, Cristina Lenardi, Simona Argentiere
Poly(d,l-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) are among the most studied systems for drug and gene targeting. So far, the synthesis of stable and uniform PLGA NPs has involved the use of a large excess of polyvinyl surfactants such as poly(vinyl alcohol) (PVA) and polyvinylpyrrolidone (PVP), whose removal requires multistep purification procedures of high ecological and economic impact. Hence the development of environment-friendly and cost-effective synthetic procedures for the synthesis of PLGA NPs would effectively boost their use in clinics...
January 18, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/27942000/free-fatty-acids-modulate-thrombin-mediated-fibrin-generation-resulting-in-less-stable-clots
#5
Anna Tanka-Salamon, Erzsébet Komorowicz, László Szabó, Kiril Tenekedjiev, Krasimir Kolev
Upon platelet activation, free fatty acids are released at the stage of thrombus formation, but their effects on fibrin formation are largely unexplored. Our objective was to characterize the kinetic effects of fatty acids on thrombin activity, as well as the structural and mechanical properties of the resultant fibrin clots. Thrombin activity on fibrinogen was followed by turbidimetry and detailed kinetic characterization was performed using a fluorogenic short peptide substrate. The viscoelastic properties of fibrin were measured with rotatory oscillation rheometer, whereas its structure was analyzed with scanning electron microscopy (SEM)...
2016: PloS One
https://www.readbyqxmd.com/read/27506588/in-vitro-effects-of-different-sources-of-fibrinogen-supplementation-on-clot-initiation-and-stability-in-a-model-of-dilutional-coagulopathy
#6
N Schäfer, A Driessen, U Bauerfeind, M Fröhlich, J Ofir, E K Stürmer, M Maegele
OBJECTIVES: To analyse which fibrinogen source may improve coagulation using an in vitro 33% dilutional coagulopathy model. BACKGROUND: Uncritical volume resuscitation in the context of trauma haemorrhage contributes to the iatrogenic arm of the acute trauma-induced coagulopathy through dilution and depletion of coagulation factors, with fibrinogen reaching critical levels first. MATERIALS AND METHODS: By using an experimental model of 33% dilutional coagulopathy, we have analysed which fibrinogen source may exert superior effects on improving haemocoagulative capacities and correcting depleted fibrinogen levels...
October 2016: Transfusion Medicine
https://www.readbyqxmd.com/read/27192476/hemostatic-therapy-using-tranexamic-acid-and-coagulation-factor-concentrates-in-a-model-of-traumatic-liver-injury
#7
Christian Zentai, Paola E J van der Meijden, Till Braunschweig, Nicolai Hueck, Markus Honickel, Henri M H Spronk, Rolf Rossaint, Oliver Grottke
BACKGROUND: The potential clinical benefits of targeted therapy with coagulation factor concentrates (e.g., fibrinogen) and antifibrinolytic agents (e.g., tranexamic acid [TXA]) for the treatment of trauma-induced coagulopathy are increasingly recognized. We hypothesized that human fibrinogen concentrate (FC) and prothrombin complex concentrate (PCC), administered as combined therapy with TXA, would provide additive effects for reducing blood loss in an animal trauma model. METHODS: Thirty-six pigs were subjected to 2 consecutive blunt liver injuries, resulting in severe hemorrhagic shock and coagulopathy...
July 2016: Anesthesia and Analgesia
https://www.readbyqxmd.com/read/26969525/significantly-reduced-adsorption-and-activation-of-blood-components-in-a-membrane-oxygenator-system-coated-with-crosslinkable-zwitterionic-copolymer
#8
Yan-Bing Wang, Ke-Hui Shi, Hong-Li Jiang, Yong-Kuan Gong
UNLABELLED: A crosslinkable zwitterionic copolymer PMBT was coated onto the surfaces of polypropylene hollow fiber membrane (PP-HFM) oxygenator and its connecting tubes. The PMBT copolymer coating on the oxygenator circuit formed a cell outer membrane mimetic surface with excellent stability. The hemocompatibility of the PMBT copolymer coated PP-HFM oxygenator circuit was evaluated by animal extracorporeal circulation. The concentrations of clotting components fibrinogen and platelet in the blood were almost unchanged during the circulation through the PMBT copolymer coated oxygenator circuits...
August 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/26840731/real-time-monitoring-of-platelet-activation-using-quartz-thickness-shear-mode-resonator-sensors
#9
Huiyan Wu, Guangyi Zhao, Hongfei Zu, James H-C Wang, Qing-Ming Wang
In this study, quartz thickness-shear mode (TSM) resonator sensors were adopted to monitor the process of platelet activation. Resting platelets adhering to fibrinogen-coated electrodes were activated by different concentrations of thrombin (1, 10, and 100 U/mL), and the corresponding electrical admittance spectra of TSM resonators during this process were recorded. Based on a bilayer-loading transmission line model of TSM resonators, the complex shear modulus (G' + jG″) and the average thickness (hPL) of the platelet monolayer at a series of time points were obtained...
February 2, 2016: Biophysical Journal
https://www.readbyqxmd.com/read/26689816/translation-of-cell-therapy-into-clinical-practice-validation-of-an-application-procedure-for-bone-marrow-progenitor-cells-and-platelet-rich-plasma
#10
REVIEW
Joerg Nowotny, Jana Farack, Corina Vater, Matthias Johnsen, Michael Gelinsky, Torsten Tonn, Philip Kasten
PURPOSE: Tissue regeneration can be improved by local application of autologous bone marrow derived progenitor cells (BMSC) and platelet rich plasma (PRP). However, there is a lack of standardized application procedures for clinical use. Therefore, a technique in accordance with the guidelines for advanced therapies medical products of the European Medicine Agency was developed and established. METHODS: In detail, a process for the isolation and formulation of autologous bone marrow cells (BMC) and PRP in a clinical setting was validated...
April 6, 2016: Journal of Applied Biomaterials & Functional Materials
https://www.readbyqxmd.com/read/26687734/haemocompatibility-and-cytotoxic-studies-of-non-metallic-composite-materials-modified-with-magnetic-nano-and-microparticles
#11
Maria Szymonowicz, Zbigniew Rybak, Aneta Fraczek-Szczypta, Danuta Paluch, Agnieszka Rusak, Katarzyna Nowicka, Marta Blazewicz
PURPOSE: Preventing the formation of blood clots on the surface of biomaterials and investigation of the reasons of their formation are the leading topics of the research and development of biomaterials for implants placed into the bloodstream. Biocompatibility and stability of a material in body fluids and direct effect on blood cell counts components are related both to the structure and physico-chemical state of an implant surface. The aim of this study was to determine haemocompatibility and cytotoxicity of polysulfone-based samples containing nano and micro particles of magnetite (Fe3O4)...
2015: Acta of Bioengineering and Biomechanics
https://www.readbyqxmd.com/read/26385924/calcium-regulates-the-activity-and-structural-stability-of-tpr-a-bacterial-calpain-like-peptidase
#12
Dominika Staniec, Miroslaw Ksiazek, Ida B Thøgersen, Jan J Enghild, Aneta Sroka, Danuta Bryzek, Matthew Bogyo, Magnus Abrahamson, Jan Potempa
Porphyromonas gingivalis is a peptide-fermenting asaccharolytic periodontal pathogen. Its genome contains several genes encoding cysteine peptidases other than gingipains. One of these genes (PG1055) encodes a protein called Tpr (thiol protease) that has sequence similarity to cysteine peptidases of the papain and calpain families. In this study we biochemically characterize Tpr. We found that the 55-kDa Tpr inactive zymogen proteolytically processes itself into active forms of 48, 37, and 33 kDa via sequential truncations at the N terminus...
November 6, 2015: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/26324704/factor-xiiia-dependent-retention-of-red-blood-cells-in-clots-is-mediated-by-fibrin-%C3%AE-chain-crosslinking
#13
James R Byrnes, Cédric Duval, Yiming Wang, Caroline E Hansen, Byungwook Ahn, Micah J Mooberry, Martha A Clark, Jill M Johnsen, Susan T Lord, Wilbur A Lam, Joost C M Meijers, Heyu Ni, Robert A S Ariëns, Alisa S Wolberg
Factor XIII(a) [FXIII(a)] stabilizes clots and increases resistance to fibrinolysis and mechanical disruption. FXIIIa also mediates red blood cell (RBC) retention in contracting clots and determines venous thrombus size, suggesting FXIII(a) is a potential target for reducing thrombosis. However, the mechanism by which FXIIIa retains RBCs in clots is unknown. We determined the effect of FXIII(a) on human and murine clot weight and composition. Real-time microscopy revealed extensive RBC loss from clots formed in the absence of FXIIIa activity, and RBCs exhibited transient deformation as they exited the clots...
October 15, 2015: Blood
https://www.readbyqxmd.com/read/26206586/determining-the-effect-of-storage-conditions-on-prothrombin-time-activated-partial-thromboplastin-time-and-fibrinogen-concentration-in-rat-plasma-samples
#14
Vinod Kumar Goyal, Somesh Kakade, Santosh Kumar Pandey, Anil Kalidas Gothi, Ramakrishna Nirogi
Coagulation parameters are usually included in clinical and preclinical safety studies to evaluate the effect of xenobiotics on the extrinsic or intrinsic pathways of coagulation. The analysis is generally performed at the time of terminal sacrifice where many activities are scheduled. Chances of delay in analysis are likely particularly when blood is collected for coagulation via the abdominal vena cava. This experiment was planned to assess the variations in coagulation parameters caused by delay in analysis as well as by storage conditions...
October 2015: Laboratory Animals
https://www.readbyqxmd.com/read/26178178/stability-of-coagulation-protein-activities-in-single-units-or-pools-of-cryoprecipitate-during-storage-at-20-24%C3%A2-c-for-up-to-24-h
#15
W P Sheffield, V Bhakta, C Jenkins
BACKGROUND AND OBJECTIVE: Cryoprecipitate is a concentrated source of fibrinogen and other plasma proteins. Cryoprecipitate must be transfused within 4-6 h of thawing and storage at 20-24°C. We compared plasma protein activities in single or pooled cryoprecipitate units stored at 20-24°C for 0, 4 or 24 h. MATERIALS AND METHODS: Individual cryoprecipitate units (n = 36) were thawed, diluted with sterile saline and sampled over time. Cryoprecipitate pools of eight individual units were assembled either by serial passage of diluent (Method A, n = 6 pools) or by separate dilution into a single collection bag (Method B, n = 6 pools)...
January 2016: Vox Sanguinis
https://www.readbyqxmd.com/read/26147092/transthyretin-amyloidosis-chaperone-concentration-changes-and-increased-proteolysis-in-the-pathway-to-disease
#16
Gonçalo da Costa, Cristina Ribeiro-Silva, Raquel Ribeiro, Samuel Gilberto, Ricardo A Gomes, António Ferreira, Élia Mateus, Eduardo Barroso, Ana V Coelho, Ana Ponces Freire, Carlos Cordeiro
Transthyretin amyloidosis is a conformational pathology characterized by the extracellular formation of amyloid deposits and the progressive impairment of the peripheral nervous system. Point mutations in this tetrameric plasma protein decrease its stability and are linked to disease onset and progression. Since non-mutated transthyretin also forms amyloid in systemic senile amyloidosis and some mutation bearers are asymptomatic throughout their lives, non-genetic factors must also be involved in transthyretin amyloidosis...
2015: PloS One
https://www.readbyqxmd.com/read/26010651/regulation-of-integrins-in-platelets
#17
REVIEW
Joel S Bennett
Blood platelets prevent bleeding after trauma by forming occlusive aggregates at sites of vascular injury. Platelet aggregation is mediated by the integrin heterodimer αIIbβ3 and occurs when platelet agonists generated at the injury site convert αIIbβ3 from its resting to its active conformation. Active αIIbβ3 is then able to bind macromolecular ligands such as fibrinogen that crosslink adjacent platelets into hemostatic aggregates. Platelets circulate in a plasma milieu containing high concentrations of the principal αIIbβ3 ligand fibrinogen...
July 2015: Biopolymers
https://www.readbyqxmd.com/read/25934730/characterization-of-thrombin-bound-dabigatran-effects-on-protease-activated-receptor-1-expression-and-signaling-in-vitro
#18
Buxin Chen, Antonio G Soto, Luisa J Coronel, Ashley Goss, Joanne van Ryn, JoAnn Trejo
Thrombin, the key effector protease of the coagulation cascade, drives fibrin deposition and activates human platelets through protease-activated receptor-1 (PAR1). These processes are critical to the progression of thrombotic diseases. Thrombin is the main target of anticoagulant therapy, and major efforts have led to the discovery of new oral direct inhibitors of thrombin. Dabigatran is the first oral anticoagulant licensed for the prevention of thromboembolisms associated with orthopedic surgery and stroke prevention in atrial fibrillation...
July 2015: Molecular Pharmacology
https://www.readbyqxmd.com/read/25909374/stability-of-hemostatic-proteins-in-canine-fresh-frozen-plasma-thawed-with-a-modified-commercial-microwave-warmer-or-warm-water-bath
#19
Medora B Pashmakova, James W Barr, Micah A Bishop
OBJECTIVE: To compare stability of hemostatic proteins in canine fresh-frozen plasma (FFP) thawed with a modified commercial microwave warmer (MCM) or warm water bath (37°C; WWB) or at room temperature (22°C). SAMPLE: Fresh-frozen plasma obtained from 8 canine donors of a commercial blood bank. PROCEDURES: A commercial microwave warmer was modified with a thermocouple to measure surface temperature of bags containing plasma. The MCM and a WWB were each used to concurrently thaw a 60-mL bag of plasma obtained from the same donor...
May 2015: American Journal of Veterinary Research
https://www.readbyqxmd.com/read/25900422/biochemical-characterization-of-a-new-glycosylated-protease-from-euphorbia-cf-lactea-latex
#20
Jaruwan Siritapetawee, Punchapat Sojikul, Sompong Klaynongsruang
A dimeric protease designated as EuP-82 was purified from Euphorbia cf. lactea latex. Since its proteolytic activity was inhibited by a serine protease specific inhibitor (PMSF), EuP-82 was classified as a serine protease. N-glycan deglycosylation tests revealed that EuP-82 was a glycosylated protein. MALDI-TOF MS showed that EuP-82 was a homodimer, which was its active form. The optimal conditions for fibrinogenolytic activity were at pH 11 and 35 °C. EuP-82 enzyme had broad range of pH stability from 4 to 12...
July 2015: Plant Physiology and Biochemistry: PPB
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