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Functional clotting assay

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https://www.readbyqxmd.com/read/28994570/clustering-of-cyclodextrin-functionalized-microbeads-by-an-amphiphilic-biopolymer-real-time-observation-of-structures-resembling-blood-clots
#1
Chandamany Arya, Camila A Saez Cabesas, Hubert Huang, Srinivasa R Raghavan
Colloidal particles can be induced to cluster by adding polymers in a process called bridging flocculation. For bridging to occur, the polymer must bind strongly to the surfaces of adjacent particles, such as via electrostatic interactions. Here, we introduce a new system where bridging occurs due to specific interactions between the side chains of an amphiphilic polymer and supramolecules on the particle surface. The polymer is a hydrophobically modified chitosan (hmC) while the particles are uniform polymeric microbeads (∼160 μm in diameter) made by a microfluidic technique and functionalized on their surface by α-cyclodextrins (CDs)...
October 10, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28971515/serum-proteomic-assessment-of-the-progression-of-fracture-healing
#2
Amira I Hussein, Christian Mancini, Kyle E Lybrand, Margaret E Cooke, Heather E Matheny, Brenna L Hogue, Paul Tornetta, Louis C Gerstenfeld
A targeted proteomic analysis of murine serum over a 35-day course of fracture healing was carried out to determine if serum proteomic changes could be used to monitor the biological progression of fracture healing. Transverse, closed femoral fractures where generated and stabilized with intramedullary fixation. A single stranded DNA aptamer-based multiplexed proteomic approach was used to assay 1310 proteins. The transcriptomic profiles for genes matching the 1310 proteins were obtained by microarray analysis of callus mRNA...
October 2, 2017: Journal of Orthopaedic Research: Official Publication of the Orthopaedic Research Society
https://www.readbyqxmd.com/read/28970801/polyphosphate-in-neonates-less-shedding-from-platelets-and-divergent-prothrombotic-capacity-due-to-lower-tfpi-levels
#3
Axel Schlagenhauf, Harald Haidl, Sina Pohl, Eva-Christine Weiss, Bettina Leschnik, Siegfried Gallistl, Wolfgang Muntean
Background: The neonatal hemostatic system exhibits a fragile balance featuring lower levels of clotting factors as well as inhibitors. Neonatal platelets show in-vitro hypoaggregability, but neonates exhibit well-functioning primary and secondary hemostasis despite this impairment. Recently, polyphosphate shed by activated platelets has been shown to induce a prothrombotic shift on the plasmatic coagulation system of adults. The impact of platelet derived polyphosphate might differ in neonates due to aforementioned peculiarities...
2017: Frontiers in Physiology
https://www.readbyqxmd.com/read/28964870/hemostasis-and-uterine-contraction-promoting-effect-of-the-extract-from-drugs-in-the-zi-yin-tiao-jing-granule-a-traditional-chinese-compound-preparation
#4
Qing Tang, Sheng Yang, Jie Tong, Xu Li, Junjie Wu, Senhong Wang, Jifen Zhang, Yi Chen, Xiaoyu Xu
ETHNOPHARMACOLOGICAL RELEVANCE: Zi-Yin-Tiao-Jing granule (ZG) is a traditional Chinese medicine compound preparation for perimenopausal dysfunctional uterine bleeding. It is made from 9 Chinese crude drugs based on a modified traditional Chinese prescription recorded in Fu Qingzhu Nvke as Guben Zhibeng Tang. AIM OF THE STUDY: This study aimed to investigate the hemostasis and uterine contraction promoting effect of quality controlled ZG extract on animals as a preclinical study...
September 28, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28942075/clinical-and-in-vitro-evidence-that-subclinical-hemolysis-contributes-to-lvad-thrombosis
#5
Carlo R Bartoli, David Zhang, Jooeun Kang, Samson Hennessy-Strahs, David Restle, Jessica Howard, Gretchen Redline, Christian Bermudez, Pavan Atluri, Michael A Acker
BACKGROUND: Recent data suggest that hemolysis contributes to left ventricular assist device (LVAD) thrombosis, but the mechanism is unknown. In a clinical study, we measured plasma free hemoglobin (pfHgb) and the incidence of LVAD thrombosis. In an in vitro study, we examined biophysical relationships between shear stress, pfHgb and von Willebrand factor (vWF) metabolism toward understanding mechanisms of LVAD thrombosis. METHODS: In the clinical study, blood samples were obtained from continuous-flow LVAD patients (n = 30)...
September 21, 2017: Annals of Thoracic Surgery
https://www.readbyqxmd.com/read/28911285/in-vitro-evaluation-of-the-effect-of-haemodilution-with-dextran-40-on-coagulation-profile-as-measured-by-thromboelastometry-and-multiple-electrode-aggregometry
#6
Pca Kam, Jpc Liou, Kxf Yang
We evaluated the effects of haemodilution with either dextran 40 or 0.9% normal saline on coagulation in vitro using rotational thromboelastometry (ROTEM®, Pentapharm Co., Munich, Germany) and multiple electrode aggregometry (Multiplate® Platelet Function Analyser, Dynabyte, Munich, Germany). Venous blood samples obtained from 20 healthy volunteers were diluted in vitro with dextran 40 or normal saline by 5%, 10% and 15%. Fibrinogen concentration, ROTEM-EXTEM® (screening test for the extrinsic coagulation pathway), FIBTEM® (an EXTEM-based assay of the fibrin component of clot) parameters including coagulation time, clot formation time, alpha angle, maximum clot firmness and lysis index were measured in the undiluted sample and at each level of haemodilution...
September 2017: Anaesthesia and Intensive Care
https://www.readbyqxmd.com/read/28899852/identification-of-extant-vertebrate-myxine-glutinosa-vwf-evolutionary-conservation-of-primary-hemostasis
#7
Marianne A Grant, David L Beeler, Katherine C Spokes, Junmei Chen, Harita Dharaneeswaran, Tracey E Sciuto, Ann M Dvorak, Gianluca Interlandi, José A Lopez, William C Aird
Hemostasis in vertebrates involves both a cellular and protein component. Previous studies in jawless vertebrates (cyclostomes) suggest that the protein response, which involves thrombin-catalyzed conversion of a soluble plasma protein, fibrinogen, into a polymeric fibrin clot, is conserved in all vertebrates. However, similar data are lacking for the cellular response, which in gnathostomes, is regulated by von Willebrand factor (VWF), a glycoprotein that mediates the adhesion of platelets to the subendothelial matrix of injured blood vessels...
September 12, 2017: Blood
https://www.readbyqxmd.com/read/28882875/class-iii-pi3k-positively-regulates-platelet-activation-and-thrombosis-via-pi-3-p-directed-function-of-nadph-oxidase
#8
Yangyang Liu, Mengjiao Hu, Dongjiao Luo, Ming Yue, Shuai Wang, Xiaoyan Chen, Yangfan Zhou, Yi Wang, Yanchun Cai, Xiaolan Hu, Yuehai Ke, Zhongzhou Yang, Hu Hu
OBJECTIVE: Class III phosphoinositide 3-kinase, also known as vacuolar protein sorting 34 (VPS34), is a highly conserved enzyme regulating important cellular functions such as NADPH oxidase (NOX) assembly, membrane trafficking, and autophagy. Although VPS34 is expressed in platelets, its involvement in platelet activation remains unclear. Herein, we investigated the role of VPS34 in platelet activation and thrombus formation using VPS34 knockout mice. APPROACH AND RESULTS: Platelet-specific VPS34-deficient mice were generated and characterized...
September 7, 2017: Arteriosclerosis, Thrombosis, and Vascular Biology
https://www.readbyqxmd.com/read/28880550/a-polymer-therapeutic-having-universal-heparin-reversal-activity-molecular-design-and-functional-mechanism
#9
Manu Thomas Kalathottukaren, Srinivas Abbina, Kai Yu, Rajesh A Shenoi, A Louise Creagh, Charles Haynes, Jayachandran N Kizhakkedathu
Heparins are widely used to prevent blood clotting during surgeries and for the treatment of thrombosis. However, bleeding associated with heparin therapy is a concern. Protamine, the only approved antidote for unfractionated heparin (UFH) could cause adverse cardiovascular events. Here, we describe a unique molecular design used in the development of a synthetic dendritic polycation named as universal heparin reversal agent (UHRA), an antidote for all clinically used heparin anticoagulants. We elucidate the mechanistic basis for the selectivity of UHRA to heparins and its nontoxic nature...
September 19, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28841240/preanalytic-processing-of-rat-plasma-influences-thrombin-generation-and-fibrinolysis-assays
#10
Marjory B Brooks, Alyssa P Stablein, Lynn Johnson, Albert Eric Schultze
BACKGROUND: Thrombin generation assays (TGA) have potential applications as measures of hemostatic balance in animal models. However, variations in plasma processing greatly influence human TGA, and may also impact on the translational value of TGA in animal studies. OBJECTIVES: The purpose of the study was to compare the performance characteristics of Sprague-Dawley rat plasma prepared by single vs double centrifugation protocols in TGA and fibrinolysis assays...
August 25, 2017: Veterinary Clinical Pathology
https://www.readbyqxmd.com/read/28837538/human-neutrophil-elastase-mediates-fibrinolysis-shutdown-through-competitive-degradation-of-plasminogen-and-generation-of-angiostatin
#11
Christopher D Barrett, Hunter B Moore, Anirban Banerjee, Christopher C Silliman, Ernest E Moore, Michael B Yaffe
BACKGROUND: A subset of trauma patients undergo fibrinolysis shutdown rather than pathologic hyperfibrinolysis, contributing to organ failure. The molecular basis for fibrinolysis shutdown in trauma is incompletely understood. Elastase released from primed/activated human neutrophils (HNE) has historically been described as fibrin(ogen)olytic. However, HNE can also degrade plasminogen (PLG) to angiostatin (ANG), retaining the Kringle domains but not the proteolytic function, and could thereby compete for generation of active plasmin by tPA...
August 23, 2017: Journal of Trauma and Acute Care Surgery
https://www.readbyqxmd.com/read/28836753/potency-and-cytotoxicity-of-a-novel-gallium-containing-mesoporous-bioactive-glass-chitosan-composite-scaffold-as-hemostatic-agents
#12
Sara Pourshahrestani, Ehsan Zeimaran, Nahrizul Adib Kadri, Nicola Gargiulo, Hassan Mahmood Jindal, Sangeetha Vasudevaraj Naveen, Shamala Devi Sekaran, Tunku Kamarul, Mark R Towler
Chitosan-based hemostats are promising candidates for immediate hemorrhage control. However, they have some disadvantages and require further improvement to achieve the desired hemostatic efficiency. Here, a series of 1% Ga2O3-containing mesoporous bioactive glass-chitosan composite scaffolds (Ga-MBG/CHT) were constructed by the lyophilization process and the effect of various concentrations of Ga-MBG (10, 30, and 50 wt %) on the hemostatic function of the CHT scaffold was assessed as compared to that of Celox Rapid gauze (CXR), a current commercially available chitosan-coated hemostatic gauze...
September 20, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28811641/joint-inflammation-related-citrullination-of-functional-arginines-in-extracellular-proteins
#13
Kalle H Sipilä, Vipin Ranga, Pekka Rappu, Markku Mali, Laura Pirilä, Ilona Heino, Johanna Jokinen, Jarmo Käpylä, Mark S Johnson, Jyrki Heino
We report the extent, specific sites and structural requirements of joint inflammation related citrullination in extracellular proteins. A total of 40 synovial fluid samples derived from chronically inflamed human joints were analysed by heparin-agarose fractionation and LC-MS/MS. Citrullination of 55 arginines in extracellular proteins was detected. Importantly, 20% of the sites have a characterized function related to the hallmarks of destructive joint inflammation. E.g. four arginine residues, shown here to be citrullinated, are also affected by mutations in inherited diseases causing haemolysis or blood clotting dysfunction...
August 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28809406/clot-entrapped-blood-cells-in-synergy-with-human-mesenchymal-stem-cells-create-a-pro-angiogenic-healing-response
#14
Melanie A Burkhardt, Isabel Gerber, Cameron Moshfegh, Miriam S Lucas, Jasmin Waser, Maximilian Y Emmert, Simon P Hoerstrup, Falko Schlottig, Viola Vogel
Blood clots stop bleeding and provide cell-instructive microenvironments. Still, in vitro models used to study implant performance typically neglect any possible interactions of recruited cells with surface-adhering blood clots. Here we study the interaction and synergies of bone marrow derived human mesenchymal stem cells (hMSCs) with surface-induced blood clots in an in vitro model by fluorescence microscopy, scanning and correlative light and electron microscopy, ELISA assays and zymography. The clinically used alkali-treated rough titanium (Ti) surfaces investigated here are known to enhance blood clotting compared to native Ti and to improve the healing response, but the underlying mechanisms remain elusive...
September 26, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/28804825/assessment-of-hereditary-thrombophilia-performance-of-protein-c-pc-testing
#15
Richard A Marlar, Jana N Gausman
Protein C (PC) is a plasma Vitamin K-dependent pro-enzyme protein that is synthesized in the liver. Upon activation, PC regulates the coagulation process by neutralizing the procoagulant activities of factors V and VIII in the presence of the cofactor Protein S. PC is a major regulator of the coagulation process. The clotting based Protein C assay, the protocol described in this chapter, quantitates the amount of functional PC present in the specimen in a proportional fashion based on the prolongation of the Activated Partial Thromboplastin Time (APTT)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28803708/anti-phospholipid-syndrome-current-opinion-on-mechanisms-involved-laboratory-characterization-and-diagnostic-aspects
#16
REVIEW
Jean Amiral, Marie Peyrafitte, Claire Dunois, Anne Marie Vissac, Jerard Seghatchian
Anti-phospholipid syndrome is a complex and severe clinical situation, associated with symptoms such as recurrent thrombosis, arterial or venous, at any site, pregnancy loss, and other related syndromes. These clinical burdens, are highly variable from patient to patient, and are associated with biological abnormalities, such as the presence of the Lupus Anticoagulant or phospholipid dependent antibodies, confirmed on two occasions at least 12 weeks apart. From the diagnosis standpoint, both, functional (clotting) or immunological assays, are difficult to standardize and to optimize, due to the absence of reference material, or a characteristic clinical group, and international reference preparations...
July 15, 2017: Transfusion and Apheresis Science
https://www.readbyqxmd.com/read/28758116/structure-and-function-of-trypsin-loaded-fibrinolytic-liposomes
#17
Anna Tanka-Salamon, Attila Bóta, András Wacha, Judith Mihály, Miklós Lovas, Krasimir Kolev
Protease encapsulation and its targeted release in thrombi may contribute to the reduction of haemorrhagic complications of thrombolysis. We aimed to prepare sterically stabilized trypsin-loaded liposomes (SSLT) and characterize their structure and fibrinolytic efficiency. Hydrogenated soybean phosphatidylcholine-based SSLT were prepared and their structure was studied by transmission electron microscopy combined with freeze fracture (FF-TEM), Fourier transform infrared spectroscopy (FT-IR), and small-angle X-ray scattering (SAXS)...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28738008/randomized-placebo-controlled-study-of-the-effects-of-yunnan-baiyao-on-hemostasis-in-horses
#18
SallyAnne L Ness, Amelia H Frye, Thomas J Divers, Mark Rishniw, Hollis N Erb, Marjory B Brooks
OBJECTIVE To determine effects of oral administration of Yunnan Baiyao on platelet activation, coagulation, and fibrinolysis in healthy horses. ANIMALS 12 healthy adult horses. PROCEDURES In a randomized blinded crossover study that included a 4-week washout period between treatments, horses were orally administered a paste containing Yunnan Baiyao (15 mg/kg) or placebo at 12-hour intervals for 3 days. Blood samples were collected before start of treatment (time 0) and at 24 and 72 hours for a CBC, measurement of fibrinogen concentration, coagulation screening tests, and a panel of assays to assess platelet activation (including ADP- and collagen-induced aggregation and closure times, flow-cytometric variables of platelet-leukocyte aggregates, platelet membrane P-selectin and phosphatidylserine expression, and microparticle release), von Willebrand factor (vWF) concentration, and cofactor activity...
August 2017: American Journal of Veterinary Research
https://www.readbyqxmd.com/read/28732864/blood-coagulation-evaluation-of-n-alkylated-chitosan
#19
Zihao Chen, Xinpei Yao, Lu Liu, Jing Guan, Mengyuan Liu, Zhihong Li, Jian Yang, Shujie Huang, Jimin Wu, Feng Tian, Miaolei Jing
N-Alkylated chitosan (NACS) may improve the haemostatic efficiency of chitosan (CS). To study its coagulation capability and function, a series of NACS with various carbon chain lengths and substitution degrees (SD) of alkyl groups were synthesized and characterized by FTIR, NMR, and elemental analysis. Haemolysis and toxicity assays revealed that NACS showed good biocompatibility. In vitro blood clotting tests indicated that NACS had better haemostatic activity than CS, of which N-octadecyl CS with 3.85% SD showed the best results...
October 1, 2017: Carbohydrate Polymers
https://www.readbyqxmd.com/read/28731369/experimental-evaluation-of-tranexamic-acid-loaded-porous-starch-as-a-hemostatic-powder
#20
Chaoyun Xi, Liguo Zhu, Yuan Zhuang, Shufang Wang, Guixiang Sun, Yaqian Liu, Deqing Wang
We evaluated the effectiveness of a novel hemostatic powder called Tranexamic Acid-loaded Porous Starch (TAPS) developed recently on blood clotting activity and hemostasis. The effectiveness of TAPS was evaluated by comparing hemostatic properties with those of Quick-acting Styptic Powder (QSP) and Compound Microporous Polysaccharide Haemostatic powder (CMPHP). The blood clotting activities of human blood were analyzed by thromboela-stogram (TEG) assays in vitro. The hemostatic effectiveness in vivo was evaluated using a rat model with hepatic traumatic hemorrhage...
January 1, 2017: Clinical and Applied Thrombosis/hemostasis
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