Tag Archives: thrombosis
Hemostatic system imbalance in the pre-eclampsia model in rats
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1SI “Institute of Pharmacology and Toxicology, National Academy
of Medical Sciences of Ukraine”, Kyiv;
2Palladin Institute of Biochemistry, National Academy
of Sciences of Ukraine, Kyiv;
3Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine.
*e-mail: anna_shayakhmetova@yahoo.com
Received: 03 June 2026; Revised: 17 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Preeclampsia (PE) is a multisystem disorder in pregnancy, that results from placental ischemia, oxidative stress-induced placental damage and placental factors dissemination that cause systemic endothelial dysfunction. Patients with preeclampsia often develop a severe prothrombotic state, the mechanisms of which are not fully elucidated. Objectives. The aim of this study was to assess the informativeness of hemostatic system parameters that characterize the degree of coagulation pathway activation and anticoagulant pathway capacity capacity in experimental preeclampsia model in rats for predicting the development of thrombotic complications in PE. Methods. Wistar females rats were used in the study. To induce a preeclampsia-like syndrome, N(ω)-nitro-L-arginine methyl ester (L-NAME), at a concentration of 0.3 g/l in the drinking water was administered ad libitum from the 8th to the 14th day of gestation. On gestational day 21 the rats were anesthetized and blood samples were collected. Urinary and serum concentrations of creatinine, urea, and uric acid were determined using a fully automated Chemistry Analyzer and commercially available kits. Fibrinogen, soluble fibrin monomeric complexes and protein C concentrations, prothrombin time and antithrombin III activity in the blood plasma were estimated with a standard spectrophotometric methods. Results. Increased plasma fibrinogen, significant shortening of plasma clotting time, decrease in both protein C and AT III and increase in SFMCs plasma levels were detected in rats with experimentally L-NAME induced preeclampsia, indicating not only activation of the coagulation system but also disruption of the balance between procoagulant and anticoagulant pathways. Conclusions. L-NAME-induced PE in rats resulted in a pronounced imbalance of the hemostatic system and reproduced key pathophysiological features of PE, including excessive procoagulant shift, intravascular thrombin generation and impairment of anticoagulant mechanisms, making this model a valuable tool for studying mechanisms of disease progression. Analysis of hemostasis parameters in this model, indicates that soluble fibrin, protein C and antithrombin III are the most informative markers for assessing the risk of intravascular coagulation during preeclampsia.
Markers of prothrombotic state of patients recovered after COVID-19
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1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Ivan Horbachevsky Ternopil Medical University, Ternopil, Ukraine;
3International Research and Innovation in Medicine Program, CSMC, Los-Angeles, CA, USA;
*e-mail: d.korolova@gmail.com
Received: 08 May 2026; Revised: 18 July 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. In recent period the attention of scientists and doctors was focused mainly on methods of COVID-19 treatment. However, clinical course of coronavirus disease strongly affects blood coagulation with the tendency for venous and arterial thrombosis, leading to long-term imbalance between coagulation and anticoagulation systems, which can cause serious danger for health of post-COVID-19 (PCD) patients. Objective. In our work we focused on the analysis of the molecular markers of hemostasis system activation in blood plasma of patients recovered after COVID-19. Methods. Blood plasma samples of 119 seropositive patients recovered from COVID-19 were analysed. The patients were divided into a high-risk group (HRG, n = 163) and a non-risk group (NRG, n = 28) according to WHO recommendations. Patients were examined in a recovery time scale of 1-30; 31-60 days and more than 2 months. Blood plasma samples of seronegative healthy volunteers were collected as the reference group (n = 19). The concentration of soluble fibrin (SF) and D-dimer was measured with sandwich ELISA, generation of activated protein C (PC) and prethrombin-1 (Pre-1)) level with assay using specific chromogenic substrates, and fibrinogen concentration with spectrophotometric method after initiation of coagulation by Ancistron-H. Results. Concentration of fibrinogen, SF and Pre-1 were increased in a blood plasma of patients of NRG and HRG groups in comparison to the reference group. Increasing of fibrinogen and Pre-1 concentrations correlated with COVID-19 stage. The fibrinogen and SF concentrations remained high more than 2 months after recovery in HRG group and elevated after 2 months after recovery in NRG group. D-dimer and protein C levels were elevated in 8% and 11% of PCD patients respectively. Conclusions. Long-term prethrombotic state was more expressed in HRG group of PCD patients. Monitoring of the main coagulation markers can indicate the risk of intravascular thrombus formation in PCD patients. SF, Pre-1 and elevated fibrinogen concentrations were selected as useful thrombosis markers in PCD patients.
Characterization of the blood coagulation system in morbidly obese patients
D. S. Korolova1, O. V. Hornytska1, A. S. Lavrik2,
N. M. Druzhyna1*, N. Prysyazhna3, T. M. Platonova1
1Palladin Institute of Biochemistry, National Academy of Science of Ukraine, Kyiv;
2Shalimov National Institute of Surgery and Transplantation,
National Academy of Medical Sciences of Ukraine, Kyiv;
3Shupyk National Healthcare University of Ukraine, Kyiv;
*e-mail: ndbiochem@gmail.com
Received: 18 April 2023; Revised: 05 May 2023;
Accepted: 7 September 2023; Available on-line: 12 September 2023
Obesity is a complex metabolic disorder that can be followed by blood coagulation disorders, atherosclerosis and atherothrombosis. In the present work, the levels of fibrinogen, soluble fibrin, D-dimer as well as protein C were measured in the blood plasma of 24 morbidly obese patients (the body mass index exceeds 40 kg/m2) to evaluate the risk of prothrombotic state. The study showed that near by 80% of patients had substantially increased fibrinogen concentration, 33% had increased concentration of soluble fibrin, 42% had increased level of D-dimer in blood plasma as compared to control. According to the results of individual analysis, the high level of fibrinogen and soluble fibrin while reduced protein C indicated the threat of thrombosis, which requires complex diagnostics to be identified. Therefore, simultaneous quantification of hemostatic system biomarkers in the blood plasma is the confident way to predict the risk of thrombotic complications in morbidly obese patients.
Validation of the diagnostics algorithm to monitor coagulation parameters in pregnant women
D. S. Korolova1, A. O. Pavlenko1, A. Altorjay2, S. I. Zhuk3,
I. V. Us3, Y. Tsaryk1, A. Suranyi2, V. O. Chernyshenko1*
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Albert Szent-Györgyi Medical School, University of Szeged, Hungary;
3P. L. Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine;
*e-mail: bio.cherv@gmail.com
Received: 19 May 2023; Revised: 05 June 2023;
Accepted: 07 June 2023; Available on-line: 11 July 2023
Thrombotic events are among the most dangerous complications of pregnancy. Therefore, selection of appropriate tests and standardization of techniques used for accurate diagnostics of blood coagulation system state is of great importance. In this present study, we monitored several molecular markers of the dangers of intravascular thrombus formation and estimated the platelet function in pregnant women during gestation. We performed independent measurements using the same methodology for different cohorts of patients recruited in Kyiv (Ukraine) and in Szeged (Hungary). D-dimer and soluble fibrin were measured using ELISA. Protein C (PC) level was estimated using chromogenic substrate assay. Fibrinogen concentration was measured by spectrophotometry using thrombin-like enzyme. Platelet function was estimated by aggregometry. Statistical data analysis was performed using the Kruskal-Wallis test. Statistically significant increases of fibrinogen concentration from first to third gestational trimester was shown for both studied cohorts of patients (5-6 mg/ml at third trimester on average). Applied methods allowed us to detect the same tendencies of decreases in PC level as well as the appearance of moderate amounts of D-dimer (up to 300 ng/ml) and SF (up to 10-15 ug/ml). Platelet function was increased on the first trimester of pregnancy and decreased during following trimesters slightly. Results indicated the changes in the blood coagulation system of pregnant women during gestation with the same effectiveness independently of the selected cohorts, time and place of measurements. The application of the proposed diagnostics algorithm may allow estimating the risk of thrombotic complications during pregnancy.
Thromboelastographic study of fibrin clot and molecular basis of maximum clot firmness
D. S. Korolova1*, Y. M. Stohnii1, V. I. Gryshchuk1, S. I. Zhuk2,
I. V. Us2, T. M. Chernyshenko1, O. P. Kostiuchenko1, K. P. Klymenko1,
O. M. Platonov1,3, O. I. Ivashchenko3, V. O. Chernyshenko1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine;
3ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: d.korolova@gmail.com
Received: 29 January 2021; Accepted: 23 April 2021
Maximum clot firmness (MCF) is the main parameter of thromboelastography (TEG) reflecting the stability of a clot. In this work, we looked for markers that can influence the enhancement of MCF and detected molecular markers and blood clotting parameters that can be involved in such mechanisms. Blood samples of pregnant women with placental disorders were collected in the Kyiv Perinatal Center. TEG was performed on whole blood in EXTEM and INTEM tests. APTT, INR, fibrinogen concentration and platelet aggregation were measured using traditional laboratory approaches. D-dimer was detected in sandwich ELISA using monoclonal antibodies III-3B and II-4D. The relative cross-linking activity of factor XIIIa was measured by the direct quantification of the cross-linked γ-chain of fibrin using Western-Blotting with monoclonal antibody II-4D. D-dimer and fibrinogen concentrations, clotting time in the APTT test, INR and rate of platelet aggregation did not correlate with the MCF. However, we found positive correlations of MCF with factor XIIIa activity: 0.51 and 0.87 for EXTEM and INTEM, respectively. These data indicate that for normal and slightly increased fibrinogen concentrations, fibrin clot firmness will depend mostly on the activity of factor XIIIa. Thus the direct determination of factor XIIIa activity in blood plasma of patients can be relevant for predicting the risk of intravascular coagulation. Evaluation of the content and activity of individual clotting factors or other components of the coagulation system can be useful additions to the TEG diagnostics and should not be neglected.
Inventive activity of the Departments of Chemistry and Biochemistry of Enzymes, and Protein Structure and Function of the Palladin Institute of Biochemistry of NAS of Ukraine. Part III. Diagnostic test-systems for analysis of fibrinolysis blood system and novel approaches to thrombosis treatment
V. M. Danilova, R. P. Vynogradova, I. Yu. Chernysh
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kiyv;
e-mail: valdan@biochem.kiev.ua
This article continues analysis of scientific achievements of the Institute of Biochemistry in the study of hemostasis system. Two previous articles were focused on the studies of blood coagulation proteins and development of the immune-enzyme test-systems for evaluation of the risk of thrombosis upon various pathologies. This article highlights the research on the blood fibrinolysis system and new approaches to thrombosis treatment, which were developed (and are under development) in the Palladin Institute of Biochemistry of the NAS of Ukraine, in particular, in the Department of Chemistry and Biochemistry of Enzymes headed previously by Dr.Sci.(Biol.) S. O. Kudinov and now by Dr.Sci.(Biol.) T .V. Grinenko, and also in the Department of Protein Structure and Function headed by Dr.Biol.Sci. E. M. Makogonenko. The fundamental knowledge of protein molecule functions and mechanisms of regulation of blood coagulation and fibrinolysis opens up new opportunities to diagnose hemostasis disorders and control the effectiveness of the cardiovascular disease treatment and also contributes to development of new techniques for isolation of new proteins – promising therapeutic agents.