A study of fibrinolytic system components in donor groups depending on various titers of circulating anti-SARS-CoV-2 IgG in the bloodstream

Rachkovska, A.; Krenytska, Daryna; Karbovskyy, Vitalii; Raksha, Nataliia; Halenova, Tetiana; Vovk, Tetiana; Savchuk, О.; Остапченко, Л. И.

doi:10.1097/mbc.0000000000001248

Cited by 3 publications

(3 citation statements)

References 28 publications

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“…Measurement of inhibitory potential of α-2-antiplasmin was performed by method described previously [20,26]. Briefly, plasma was diluted 1:3 with 0.05 M Tris-HCl buffer, pH 7.4.…”

Section: Methodsmentioning

confidence: 99%

Coagulation and Anticoagulation Parameters in Multiple Sclerosis Patients With and Without Covid-19

Halenova,

Raksha,

Vovk

et al. 2024

Clin. and prev. med.

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The aim. To investigate plasma levels of main coagulation and fibrinolytic factors in MS patients with and without COVID-19 history. Materials and methods. A total of 127 participants were enrolled in this study, including 97 MS patients and 30 healthy controls (HC). Patients with MS were divided into two groups: MS+Covid group (n=41) – patients with MS, who had a laboratory-verified diagnosis of COVID-19 in the past 3-6-month period and MS group (n=56) – patients with MS, who did not suffer from COVID-19 previously. Determination of plasma levels of prothrombin, plasminogen, tissue-type plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), protein C (PC), soluble thrombomodulin (TM) was performed by means of enzyme-linked immunosorbent assay. Spectrophotometric techniques were used to determine concentrations of fibrinogen, soluble fibrin monomeric complexes (SFMC) as well as plasminogen activity and inhibitory potential of α-2-antiplasmin. Results. The MS group was characterized by elevated levels of plasma prothrombin, fibrinogen, D-dimer, SFMC, soluble TM compared to HC, while PC concentration did not differ between MS and HC groups. Plasma plasminogen level as well as plasma level of the potential plasmin activity were significantly decreased in MS patients compared to HC group. The plasma tPA level was significantly reduced while plasma PAI-I level was significantly increased in MS patients compared to HC. Patients of MS group had an increased level of plasma α-2-antiplasmin activity compared with HC group. To note, most of studied parameters did not differ between two MS groups, except protein C, soluble thrombomodulin levels and plasma α-2-antiplasmin activity. Conclusions. The results of our study showed that MS patients have got altered hemostasis parameters; however, further study is necessary to find out the relationship between particular components of coagulation and fibrinolytic systems and pathophysiology of MS. Additionally, our findings demonstrated that a SARS-CoV-2 infection had a limited effect on hemostasis parameters in MS patients, causing changes in only a few parameters, including thrombomodulin and protein C levels as well as α-2-antiplasmin activity.

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“…Measurement of inhibitory potential of α-2-antiplasmin was performed by method described previously [20,26]. Briefly, plasma was diluted 1:3 with 0.05 M Tris-HCl buffer, pH 7.4.…”

Section: Methodsmentioning

confidence: 99%

Coagulation and Anticoagulation Parameters in Multiple Sclerosis Patients With and Without Covid-19

Halenova,

Raksha,

Vovk

et al. 2024

Clin. and prev. med.

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“…To test the technique we used blood plasmafrom donors, who had recovered from COVID-19. According to the available scientific literature (Rachkovska et al, 2023), the post-COVID period is characterized by hemostatic disorders, which can lead to an increase in proteolytic activity in the blood stream.…”

Section: Methodsmentioning

confidence: 99%

Optimization of an Enzyme Electrophoresis Methodfor Using Collagen as a Substrate

Kalasnikova,

Savchuk,

Karbovskiy

2023

BTSNUKSB

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Background. The optimized method of collagen-containing sodium dodecyl sulfate-polyacrylamide electrophoresis is introduced in this work. The main parameters that affect the efficiency and resolution of the methodare the degree of sample dilution, the concentration of separation gel,the concentration of collagen solution which was used as a substrate, time of electrophoretic separation elongation. In order to investigate all active and pro-active forms of plasmin, all samples were prepared in two variations: with supplementary activation by streptokinase (Sk) addition, and without. Methods. To obtain clearly visible and detectable lysis points, samples with Sk were diluted in a ratio of 1:16, and samples without Sk were diluted in a ratio of 1:8. To prevent substrate migration and the loss of proteins' electrophoretic mobility, the concentration of was 15 % separation gel, and the concentration of copolymerized collagen solution was 1 mg/ml. To obtain the most informative results, the time of electrophoresis elongation was 15 min. After electrophoresis gels were washed in 2,5 % Triton X-100 solution for 1 hour and stained according to a standard protocol. Results. As a result of the conducted research, the optimal conditions for carrying out this enzyme-electrophoresis modification were found, according to all analytical manipulations, and methodical approaches to the detection of latent proenzymatic forms of enzymes with collagenolytic activity were shown. Conclusions. This modified technique can be used for quantitative and qualitative analysis of the presence of collagenolytic activity in various samples and allows research of enzymes that possess this activity, both from a scientific point of view, and in the process of finding and developing technologies for obtaining collagenolytic enzymes for biotechnological purposes.

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“…Potential plasminogen activity was measured by streptokinase method based on plasminogen activation to plasmin by streptokinase; plasmin in turn cleaves a specific chromogenic substrate. The measured change in sample optical density is directly proportional to potential plasminogen activity ( 20 ). Potential plasminogen activity was determined using commercial reagents (RENAU, Kharkiv, Ukraine).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of proteolytic activity and serine proteases distribution in plasma from patients with bladder cancer

Synelnyk,

Vovk,

Halenova

et al. 2023

Front. Med.

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BackgroundBladder cancer (BC) is an aggressive disease with a poor prognosis. A bladder tumor, like other malignant neoplasms, is characterized by the presence of both cancer cells and stromal cells which secrete cytokines, chemokines, growth factors, and proteolytic enzymes. One such class of proteolytic enzymes are serine proteases, which take part in the tumor microenvironment formation via supporting and contributing to tumor progression. This study aims to evaluate the proteolytic activity and serine protease contribution in plasma from BC patients.MethodsThe research involved patients of Alexandrovsky city clinical hospital aged 52–76 with transitional cell carcinoma of the bladder. All examined patients were divided into five groups: the control group included conditionally healthy donors, while other patients were grouped according to their tumor stage (I, II, III and IV). Plasma plasminogen levels were determined by enzyme-linked immunosorbent assay, and the potential activity was measured by chromogenic plasminogen assay. Serine proteases fractions were obtained by the affinity chromatography method, and enzyme concentration in the selected fractions were determined by the Bradford method. Serine proteases distribution was investigated by electrophoresis in a polyacrylamide gel.ResultsIt was determined that the concentration, potential activity of plasminogen, and the total amount of serine proteases in plasma from BC patients were greater than the values of the corresponding indicators in healthy donors. This could be one of the factors contributing to increased proteolysis seen in the process of carcinogenesis. Plasminogen concentration in BC patients with stage IV disease; however, displayed a tendency to be reduced compared to earlier stages, and the potential activity of plasminogen was significantly lower in patients with stages III – IV BC. Futhermore, a tumor stage specific gradual decline in the serine protease plasma content was shown. The results of electrophoretic analysis established a significant diminishment in the percentage of high molecular weight components (under non-reducing conditions) and their complete disappearance (under reducing conditions) in plasma serine protease fractions from BC patients. A decline in the percentage of heavy and light plasmin chains in BC patients was also observed. Additionally, a rise in the degraded forms of plasminogen/plasmin content was seen in BC samples, as well as the presence of fractions corresponding to trypsin and NE (under non-reducing conditions) that were absent in the control samples.ConclusionThe results indicate significant changes in the proteolytic activity of plasma, from BC patients when compared to healthy controls, which is accompanied by alterations in serine protease distribution caused by tumor microenvironment pecularlities at the different stages of oncopathology.

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A study of fibrinolytic system components in donor groups depending on various titers of circulating anti-SARS-CoV-2 IgG in the bloodstream

Cited by 3 publications

References 28 publications

Coagulation and Anticoagulation Parameters in Multiple Sclerosis Patients With and Without Covid-19

Coagulation and Anticoagulation Parameters in Multiple Sclerosis Patients With and Without Covid-19

Optimization of an Enzyme Electrophoresis Methodfor Using Collagen as a Substrate

Evaluation of proteolytic activity and serine proteases distribution in plasma from patients with bladder cancer

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