Tag Archives: soluble fibrin

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.

Overall hemostasis potential of blood plasma and its connection to molecular markers of the hemostasis system in patients with stenosis of coronary artery

N. V. Storozhuk1, L. V. Pyrogova2, Т. М. Chernyshenko2,
O. P. Kostyuchenko2, T. M. Platonova2, O. B. Storozhuk1,
B. G. Storozhuk1, R. Yu. Marunich2,
G. K. Bereznytsky2, E. M. Makogonenko2*

1MI Pirogov Vinnytsia National Medical University, Vinnytsia, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: ymakogonenko@gmail.com

Received: 01 April 2021; Accepted: 22 September 2021

The correlation relationships between hemostatic potential parameters and concentrations of molecular markers of the hemostasis system: soluble fibrin (sf), D-dimer (DD), fibrinogen (Fg) and protein C (PC) in patients with stenosis of coronary artery 6 months after coronary angioplasty have been investigated. It was found  three directions of changes in the state of the patients hemostasis system: an increasing in fibrinolytic activity (C) ~18% of patients; an increasing in coagulative activity (B) ~31% of patients; and maintaining of the balance between coagulation and fibrinolysis (A) ~51% of patients. In patients with signs of stenosis without angina pectoris, a strong Pearson correlation was shown between the half-life of the clot and the overall hemostatic potential (OHP) (r = 0.75, P << 0.05), a moderate relationship between concentrations of sf and D-dimer (r = 0.67, P <0.05), almost complete connection between coagulation potential (CP) and OHP (r = 0.975, P << 0,05) and strong connection between CP and fibrinolytic potential (FP) (r = 0.80, P << 0.05). In patients with signs of stable angina pectoris, almost complete connection was found between the concentration of sf and D-dimer (r = 0.981, P << 0.05), CP and OHP (r = 0.979, P << 0.05) and a strong connection between CP and FP (r = 0.846, P << 0.05). Possible functional mechanisms of connection between these parameters are discussed.

Overall hemostasis potential of the blood plasma and its relation to some molecular markers of the hemostasis system in patients with chronic renal disease of stage VD

B. G. Storozhuk1, L. V. Pyrogova2, T. M. Chernyshenko2, O. P. Kostiuchenko2,
I. M. Kolesnikova2, T. M. Platonova2, O. B. Storozhuk1, L. O. Storozhuk1,
G. K. Bereznitsky2, P. Yu. Tsap2, O. O. Masenko2,
E. M. Makogonenko2, E. V. Lugovskoy2

1Pyrogov National Medical University of Vinnytsa, Ukraine;
2Palladin Instiute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: makogonenko@biochem.kiev.ua

The values of the coagulation, overall and fibrinolysis potentials were estimated by the method of the global potential of Blomback M., as well as the values of concentrations of molecular markers of the hemostasis system: soluble fibrin (sf), D-dimer, fibrinogen (Fg) and protein C (88 patients, 52 of them men, 36 women). It was shown that hemostasis system activity in women plasma is higher than that in men plasma. The division of patients into 3 groups, depending on the concentration of sf: less than normal – sf ≤ 3, about norm – 3 < sf < 4 and more than norm – sf > 4 μg/ml, allowed establishing the growth of the parameters of both the hemostatic potential and concentrations of molecular markers in accordance with concentration of sf in the groups of patients. Paerson’s correlation analysis of the relationship between the parameters of the hemostasis potential and concentrations of molecular markers revealed an increase in the correlation relationship to the strong and very strong between the parameters of coagulation, fibrinolysis and protein C systems with an increase in the concentration of soluble fibrin in plasma of patients.

Inventive activity of the Departments of Protein Structure and Function, and Molecular Immunology of the Palladin Institute of Biochemistry of NAS of Ukraine. Part II. National breakthrough in the study and diagnostics of human hemostasis system

N. E. Lugovska

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: nlugovsk@mail.ru

The scientists of Protein Structure and Function, and Molecular Immunology Departments of the Palladin Institute of Biochemistry (NAS of Ukraine) under the supervision of member of NASU and NAMSU, prof. S. V. Komisarenko  and corresponding member of NASU prof. E. V. Lugovskoy have made the real breakthrough in the field of research of the mechanisms of fibrin polymerization and formation of fibrin framework of thrombi.  The immunodiagnostic test-systems for the evaluation of the risk of thrombus formation  were developed for the first time. Researches have obtained the monoclonal antibodies to fibrinogen, fibrin, D-dimer and their fragments. These monoclonal antibodies were used as molecular probes for the localization of newly detected fibrin polymerization sites. Obtained antibodies with high affinity interact with fibrinogen, D-dimer and soluble fibrin – main markers of the risk of thrombus formation. They were used for the development of the immunodiagnostic test-systems to quantify these markers in human blood plasma for the evaluation of the state  of haemostasis system, detection of prethrombotic states, disseminated intravascular coagulation, detection of thrombosis and monitoring of antithrombotic and fibrinolytic therapy. The successful trial of developed test-systems was carried out in clinics of Ukraine, and the State registration was obtained for the implementation of them into the clinical practice. Presented works were awarded State prize of Ukraine in Science and technology.

Level of overall hemostasis potential in donor and patient plasma in pathology

L. V. Pyrogova, T. M. Chernyshenko, I. N. Kolesnikova, T. N. Platonova,
G. K. Bereznitsky, Y. M. Makogonenko, E. V. Lugovskoy

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: ymakogonenko@gmail.com

Coagulation potential (CP), overall hemostasis potential (OHP) and fibrinolysis potential (FP) in plasma of donors and patients with myocardial infarction (MI), stroke (St) and hip joint diseases (HJD) have been investigated using M. Blomback’s global hemostasis assay. Plasma samples of the patients were analyzed with APTT reagent in the presence or absence of t-PA. It was found that the ratio of values of СP, OHP and FP in plasma of patients to those of donors plasma were 78, 60 and 123% at MI; 157, 155 and 162% at St; 128, 131 and 124% at HJD. CP to FP ratio that indicated balance between coagulation and fibrinolytic systems activities were 4.13, 2.5, 4.0 and 4.26 in plasma of donors and MI, St and HJD patients, respectively. These results are evidence of increased fibrinolytic activity in plasma of MI patients. Lag-periods of plasma clotting of MI, St and HJD patients were prolonged by 2.3, 7.2 and 1.5-fold, respectively. Pearson’s correlation analysis between parameters, obtained in vitro studies using global hemostasis assay, and concentrations of the molecular markers (soluble fibrin and D-dimer), which formed in vivo in plasma of MI, St and HJD patients, did not reveal any relationship between them.