Tag Archives: protein C
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.
Reparative osteogenesis markers during bone defects substitution with germanium-doped ceramics under experimental osteoporosis
T. Todosiuk1*, V. Chemerovskiy1, А. Rublenko2,
N. Ulianchych3, V. Kolomiiets3, S. Firstov3
1Bila Tserkva National Agrarian University, Bila Tserkva, Ukraine;
2Vinnytsia Mykhailo Kotsiubynskyi State Pedagogical University, Vinnytsia, Ukraine;
3Frantsevich Institute for Problems of Materials Science,
National Academy of Sciences of Ukraine, Kyiv;
*e-mail: tatyana.todosyuk@gmail.com
Received: 04 June 2025; Revised: 05 August 2025;
Accepted: 30 January 2026; Available on-line: 23 February 2026
Osteoporosis, as a systemic skeletal disease, is characterized by the loss of bone mass, decreased mineral density, and microarchitecture changes. In cases of traumatic fractures and critical-size bone defects osteoporosis can lead to spontaneous fractures, impair regeneration and complication when using bone substituting materials. Ceramic implants, doped with germanium to impart osteoinductive properties, are among promising bone substituting materials. In this study we aimed to assess biochemical markers of reparative osteogenesis at bone defects substitution with germanium-doped ceramics in rabbits under osteoporosis. The study was conducted on California White rabbits with osteoporosis, induced by administration of 0.4% dexamethasone solution. The model defects were created in trabecular and cortical bones, following the exposure of the periosteum with drills of 3 and 4.2 mm diameters, respectively, in compliance with the anesthetic regimen and antiseptic rules. Calcium phosphate ceramic granules with a size of 700 μm, synthesized from hydroxyapatite and β-tricalcium phosphate and doped with 0.8 mass.% germanium (CPC-Ge) were used for healing. In the control group of animals (n = 9) bone defects were healed under a blood clot. In the experimental group (n = 9), the defects were replaced with CPC-Ge granules. Blood samples for biochemical studies were collected before modeling the bone defect and on the 7th, 14th, 30th, and 60th days of reparative osteogenesis. The activity of tartrate-resistant acid phosphatase, alkaline phosphatase and its bone isoenzyme, as well as circulating immune complexes, protein C and NO serum levels were determined. It was shown that substitution of both trabecular and cortical bones defects with CPC-Ge, as compared to healing under a blood clot, leads to reduced inflammatory and immune responses, prevented the depletion of protein C and promotes a more dynamic course of reparative osteogenesis in animals with glucocorticoid-induced osteoporosis.
C(60) fullerene effect on the functional activity of rat gastrocnemius muscle during its regeneration after the open injury
D. M. Nozdrenko1, O. O. Gonchar2, N. E. Nurishchenko1,
V. O. Stetska1, T. Yu. Matviienko1, Ya. V. Stepanyuk3,
K. I. Bogutska1, Yu. I. Prylutskyy1*
1ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
2Bogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, Kyiv;
3Medical Faculty, Lesya Ukrainka Volyn National University, Lutsk, Ukraine;
*e-mail: prylut@ukr.net
Received: 05 September 2024; Revised: 14 October 2024;
Accepted: 21 November 2024; Available on-line: 17 December 2024
Open injuries are one of the most common skeletal muscle traumas. The study aimed to estimate the effect of the oral administration of C60 fullerene aqueous solution (C60FAS) daily at a dose of 1 mg/kg on the restoration of rat skeletal muscle functional activity on the 5th, 10th and 15th day after the open trauma. Male Wistar rats were randomly divided into three groups of 12 animals in each: control, with muscle injury and with muscle injury+C60FAS. The isolated gastrocnemius muscle was subjected to open injury by transverse dissection with a depth of 1 mm. Stimulation of muscle efferents was carried out by electrical impulses generated using a strain gauge generator. The content of C-reactive protein, creatinine, lactate, reduced glutathione and the activity of catalase and superoxide dismutase in the rat blood were determined. According to the data obtained, application of C60FAS promotes the restoration of the functional activity of injured muscle, which was confirmed by a significant increase in gastrocnemius muscle force impulse, attenuation of the inflammatory and development of fatigue and normalization of pro- and antioxidant balance in the process of regeneration.
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.
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.
Recombinant single chain variable fragment antibodies (scFv) against Pro(144)-Leu(155) fragment of human protein C
O. S. Oliinyk, K. O. Palyvoda, N. E. Lugovskaya,
D. V. Kolibo, E. V. Lugovskoy, S. V. Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: lenaoliinyk@mail.ru
The aim of this work was to obtain the recombinant single chain variable fragments of antibodies (scFv) against human protein C, the key component of blood anticoagulation system. For this purpose a peptide that mimics a Pro144-Leu155 sequence of protein C was synthesized and the murine immune scFv library against this peptide was constructed. The protein C specific scFv 9E were selected from the constructed library by the phage-display method. The scFv 9E dissociation constant was found to be 2∙10-9 М. It was shown that scFv 9E were suitable for protein C detection by ELISA and Western blotting. Selected scFv could be further used for protein C investigation and for the development of quantitative methods for protein C detection in human blood.