Meizothrombin Preparation and Its Role in Fibrin Formation and Platelet Aggregation

Korolova, Daria; Chernyshenko, Tamara; Gornytska, Olga V.; Chernyshenko, Volodymyr; Platonova, T. N.

doi:10.4236/abb.2014.57069

Cited by 7 publications

(4 citation statements)

References 23 publications

(20 reference statements)

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“…PRP was isolated by centrifugation of blood at 160 g for 20 min at 23°C. For the preparation of platelet poor blood plasma PRP was spinned-down at 1500 rpm during 30 min [ 14 ].…”

Section: Methodsmentioning

confidence: 99%

Silica Nanoparticles Effects on Blood Coagulation Proteins and Platelets

Gryshchuk

Галаган

2016

Biochemistry Research International

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Interaction of nanoparticles with the blood coagulation is important prior to their using as the drug carriers or therapeutic agents. The aim of present work was studying of the primary effects of silica nanoparticles (SiNPs) on haemostasis in vitro. We studied the effect of SiNPs on blood coagulation directly estimating the activation of prothrombin and factor X and to verify any possible effect of SiNPs on human platelets. It was shown that SiNPs shortened coagulation time in APTT and PT tests and increased the activation of factor X induced by RVV possibly due to the sorption of intrinsic pathway factors on their surface. SiNPs inhibited the aggregation of platelet rich plasma induced by ADP but in the same time partially activated platelets as it was shown using flow cytometry. The possibility of SiNPs usage in nanomedicine is strongly dependant on their final concentration in bloodstream and the size of the particles that are used. However SiNPs are extremely promising as the haemostatic agents for preventing the blood loss after damage.

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“…PRP was isolated by centrifugation of blood at 160 g for 20 min at 23°C. For the preparation of platelet poor blood plasma PRP was spinned-down at 1500 rpm during 30 min [ 14 ].…”

Section: Methodsmentioning

confidence: 99%

Silica Nanoparticles Effects on Blood Coagulation Proteins and Platelets

Gryshchuk

Галаган

2016

Biochemistry Research International

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“…This method is easy to use and does not require expensive devices. It was successfully used in our laboratory and allowed us to obtain data with high reproducibility [16,17].…”

Section: Methodsmentioning

confidence: 99%

Preparation of highly-concentrated autologous platelet-rich plasma for biomedical use

Chernyshenko¹,

Shteinberg²,

Lugovska³

et al. 2019

Ukr.Biochem.J

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Cell therapy with platelets is a widely accepted approach for wound healing and tissue regeneration in medicine. however, with most available methods poorly concentrated platelet suspensions (up to 0.3•10 6 1/ µl) or suspensions of mostly inactivated or lost platelets are obtained. In this study, we aimed to develop a simple and effective method for preparing a suspension of native and resting platelets with over 1•10 6 1/µl. Platelet-rich plasma (PRP) was obtained from fresh blood of healthy donors (n = 5) collected using different amounts of heparin as the anticoagulant. Samples of PrP were spun down and re-suspended in auto logous blood plasma. Count and vitality of platelets in each sample were determined by aggregation study on the Solar aP2110 aggregometer. Platelet shape and cytoplasmic granularity that indicate the nativity of platelets were monitored on the CoULter ePICS XL Flow Cytometer. this study of aggregation of platelets in PrP obtained using various amounts of heparin allowed us to reduce final concentrations to the amount that effectively prevented clotting and did not affect platelet reactivi ty (5 U/ml). PRP concentrated 5 times with a total concentration of cells of 1•10 6 1/µl was able to be activated by adenosine diphosphate (aDP) (aggregation rate 54 ± 7%). the amount of cells with altered shape and granularity in concentrated suspension was not higher than 20%. This finding means that the platelets would still be able to release a number of growth factors and other biologically active compounds after stimulation or injection into tissue during cell therapy. the decrease in heparin concentrations also minimizes haemorrhage in the injection site supporting biomedical use of the suspension. A simple and effective method for preparation of highly-concentrated PRP (1.2•10 6 1/ µl) for biomedical use was developed. Aggregometry and flow cytometry proved that obtained platelets were resting and able to be activated. Being autologous, the preparation can be widely used for cell therapy without additional precautions. k e y w o r d s: cell therapy, platelets, autologous, platelet rich plasma, preparation protocol.

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“…Ecamulin (prothrombin activator from Echis multisqumatis venom) is an enzyme that activates prothrombin, descarboxy-prothrombin and prethrombin 1. It makes it possible to determine the total prothrombin level [17]. Thromboplastin acts through the tissue factor pathway of coagulation and activates only functionally active carboxylated and uncleaved forms of prothrombin as it is in the "prothrombin index" test.…”

Section: Methodsmentioning

confidence: 99%

Blood coagulation parameters in rats with acute radiation syndrome receiving activated carbon as a preventive remedy

Chernyshenko¹,

Snezhkova²,

Mazur³

et al. 2019

Ukr.Biochem.J

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radiation-induced coagulopathy (rIC) is one of the major causes of death during acute radiation syndrome (arS). The aim of this study was to characterize the responses of the hemostasis system to arS of a moderate level on the 1 st and 9 th days after irradiation. We aimed to identify molecular markers of the blood coagulation system that are most affected by ARS and to estimate the enterosorption effect on the development of irradiation-induced changes. Platelet aggregation rate, activated partial thromboplastin time (aPTT) and fibrinogen concentration were determined by standard methods. Level of protein C (PC) was measured using chromogenic substrate S2366 (p-glu-Pro-arg-pNa) and agkistrodon halys halys snake venom activating enzyme. Functionally inactive forms of prothrombin (FIFPs) were determined using two activators in parallel-thromboplastin or prothrombin activator from echis multisqumatis venom. rats of both irradia ted groups had a higher risk of intravascular clotting in comparison to both control groups. Statistically significant shortening of clotting time in the APTT test (24 ± 4 s vs. 33 ± 5 s) and increased fibrinogen concentration (4.2 ± 0.6 mg/ml vs. 3.2 ± 0.3 mg/ml) were detected. Both parameters were normalized on the 9 th day after irradiation. however the platelet count was decreased (0.3•10 6 ± 0.05•10 6 1/μl vs. 0.145•10 6 ± 0.04•10 6 1/μl) due to the impaired megakaryocytic function. The level of PC was decreased after X-ray irradiation (70 ± 10%) and partly restored on the 9 th day after irradiation (87 ± 10%). administration of activated carbon (aC) inhibited the drop in the PC concentration after X-ray irradiation (86 ± 15%) and accelerated its restoration on the 9th day (103 ± 14%). The statistically significant accumulation of FIFPs was detected in blood plasma of irradia ted rats at the 1 st and 9 th days after irradiation. No FIFPs were found in any irradiated rat treated with aC. Characterization of the hemostasis system of rats that were exposed to a semilethal dose of X-rays allowed us to select parameters that can be used for monitoring of arS development. apart of from basic coagulation tests (APTT) and the measurement of platelet aggregation, fibrinogen and protein C level we can recommend the determination of FIFPs as a useful tool for estimation of the hemostasis response after irradiation with X-rays. This test indicates the intravascular thrombin generation and can help predict thrombotic complication or disseminated intravascular coagulation. determination of FIFPs in blood plasma of irradia ted rats allowed us to study the enterosorption effect on the development of irradiationinduced changes. It was shown that enterosorption with aC prevented accumulation of FIFPs which appears to be a newly discovered antithrombotic effect of therapy with AC. ARS influenced hemostasis by inducing thrombin generation (indicated by FIFPs generation), lowgrade inflammation (indicated by PC concentration decrease) and thrombocytopenia. Enterosorption with AC minimizes inflammation and pr...

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Meizothrombin Preparation and Its Role in Fibrin Formation and Platelet Aggregation

Cited by 7 publications

References 23 publications

Silica Nanoparticles Effects on Blood Coagulation Proteins and Platelets

Silica Nanoparticles Effects on Blood Coagulation Proteins and Platelets

Preparation of highly-concentrated autologous platelet-rich plasma for biomedical use

Blood coagulation parameters in rats with acute radiation syndrome receiving activated carbon as a preventive remedy

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