COMPARISON OF PLASMA COAGULABILITY AFTER SHORT-TERM TREATMENT WITH ROSUVASTATIN VERSUS ATORVASTATIN IN UNSTABLE ANGINA PATIENTS

Statins are the integral medications for the management of patients with acute coronary syndrome including unstable angina (UA) with multiple pleiotropic effects. However, the influence of statins on the coagulation system is controversial. Our study aimed to explore the effects of atorvastatin and rosuvastatin in high doses on some coagulation parameters (prothrombin pool (PP) and soluble fibrin-monomer complexes (SFMC) concentration) after a 7-days follow-up period in patients with UA. We recruited 50 patients aged 55 to 70 years with progressive UA. Standard therapy according to ESC guidelines 2020 was recommended for all patients. Before treatment onset, they were divided into 2 groups: group A – 26 patients were prescribed atorvastatin, group R – 24 patients with rosuvastatin treatment. The blood samples to analyze the concentration of PP and SFMC were collected twice – before the treatment onset and 7 days after. We revealed significant decrease in PP concentration (p=0,02) and increase in SFMC concentration (p=0,01) in group A patients while there were no significant changes of investigated parameters (p=0,94, p=0,57 respectively) in group R. Additionally, we have noted significant negative correlation between baseline PP concentration and direction of PP changes (r=-0,803, p<0,001) as well as PP changes direction and SFMC concentration after treatment (r=-0,655, p<0,001). Thus, we may consider that atorvastatin and rosuvastatin are characterized by different influences on coagulation in patients with progressive UA with standard basic treatment. The rebound coagulation system activation after anticoagulant discontinuation is more pronounced in UA patients against a background of atorvastatin treatment in comparison with rosuvastatin.Statins are the integral medications for the management of patients with acute coronary syndrome including unstable angina (UA) with multiple pleiotropic effects. However, the influence of statins on the coagulation system is controversial. Our study aimed to explore the effects of atorvastatin and rosuvastatin in high doses on some coagulation parameters (prothrombin pool (PP) and soluble fibrin-monomer complexes (SFMC) concentration) after a 7-days follow-up period in patients with UA. We recruited 50 patients aged 55 to 70 years with progressive UA. Standard therapy according to ESC guidelines 2020 was recommended for all patients. Before treatment onset, they were divided into 2 groups: group A – 26 patients were prescribed atorvastatin, group R – 24 patients with rosuvastatin treatment. The blood samples to analyze the concentration of PP and SFMC were collected twice – before the treatment onset and 7 days after. We revealed significant decrease in PP concentration (p=0,02) and increase in SFMC concentration (p=0,01) in group A patients while there were no significant changes of investigated parameters (p=0,94, p=0,57 respectively) in group R. Additionally, we have noted significant negative correlation between baseline PP concentration and direction of PP changes (r=-0,803, p<0,001) as well as PP changes direction and SFMC concentration after treatment (r=-0,655, p<0,001). Thus, we may consider that atorvastatin and rosuvastatin are characterized by different influences on coagulation in patients with progressive UA with standard basic treatment. The rebound coagulation system activation after anticoagulant discontinuation is more pronounced in UA patients against a background of atorvastatin treatment in comparison with rosuvastatin

Blood Plasma Serotonin and von Willebrand Factor as Biomarkers of Unstable Angina Progression Toward Myocardial Infarction

Aim: To investigate the serotonin and von Willebrand factor (vWF) concentrations among unstable angina (UA) patients without and with progression toward myocardial infarction (outcome) and to assess the utility of both as prognostic markers of UA complications. Materials and methods: In observational cohort study, we recruited 103 patients with ischemic heart disease (the median age 65.0 (59.0-69.0) years, 45 females (43.7%)). After full set of investigations including high sensitive Troponin I test and 28-day follow-up period, we defined three groups: Group 1 - stable angina patients (n=22) as control, Group 2 - UA patients without outcome (n=71), Group 3 - UA patients with outcome (n=10). We analyzed the blood plasma serotonin content by the ion-exchange chromatography with measurement of serotonin on fluorescence spectrophotometer. VWF concentration was determined by ELISA. We compared the concentrations of observed parameters among the groups with the Kruskal-Wallis test (with post-hoc Mann-Whitney test with Bonferroni-Holm correction). We assessed binary logistic models, receiver operating characteristic curves, calculated sensitivity (Se), specificity (Sp), and positive likelihood ratio (LR+) for each indicator. Results: We registered elevation in serotonin concentration and decline in vWF concentration in Group 3 in comparison with Group 2 (22.670 (20.687-24.927) μg/ml vs 11.980 (8.120-15.000) μg/ml, p< 0.001, and 0.117 (0.109-0.120) rel.units/ml vs 0.134 (0.127-0.143) rel.units/ml, p < 0.001) and Group 1 (12.340 (10.052-13.619) μg/ml, p < 0.001, and 0.137 (0.127-0.156) rel.units/ml, p < 0.001), respectively. No significant differences in serotonin and vWF concentrations between Group 1 and Group 2 were detected (p=0.81 and p=0.36, respectively). The probability of outcome increased significantly (by 60.7% and 59.7%, LR+ 19.0 [6.0, 60.0] and 18.0 [3.9, 80.0]) if serotonin concentration was above 21.575 μg/ml (Se=80.0%, Sp=95.8%, AUC=0.975) and vWF concentration was below 0.114 rel.units/ml (Se=50.0%, Sp=97.2%, AUC=0.973), respectively. Conclusions: Serotonin and vWF as biomarkers are demonstrated promising results for rule-in the patients with risk of short-term UA progression toward myocardial infarction

Difference in coagulation markers in acute and one year post acute ischemic stroke

Background and purpose: Shifts of the balance between coagulation and fibrinolysis play a crucial role in pathogenesis of cerebral ischemia. However, its relevance to post-acute disease period requires further elucidation. This study aimed to characterize whether hypercoagulation markers differ for patients in acute and the same patients in the post acute phase of ischemic stroke. Materials and methods: Systemic generation of hemostasis markers was monitored and fraction composition was described during one year of disease development and treatment. Results: Increased concentration of the markers of hypercoagulation in blood plasma of patients in acute as well as one year past acute phase ischemic stroke were shown. Thus, not all markers of hypercoagulation become relevant to norm one year past ischemic stroke. It\u27s the first time characterization of the coagulation markers in acute and post acute period of the absolutely same group of patients was done. Our study demonstrates the difference in quantity and quality of the fractions of proteins with prothrombin origin and soluble fibrin monomer complexes which could be used as a potential target for the prevention of the stroke repetition. Conclusions: We assume that prothrombin plays the most important role in the development of pathology tested, hence it may provide future targets for therapeutic strategies.</p

Methodological approach to the isolation of functionally active proteins from the tissues of marine hydrobionts: an example of Adamussium colbecki

Enzymes from cold-adapted organisms have significant application potential. Because of their unique properties they have been found to be useful in various industries. Despite indisputable practical interest, cold active enzymes also represent a valuable model for fundamental research into protein folding and catalysis. Many investigators have focused their attention on marine hydrobionts, which are growing in importance as a promising source of enzymes. The nature of the source not only determines the availability and the cost of biomolecules of interest but also determines the choice of method for their extraction. A simple and convenient methodological approach of two-stage extraction of proteins has been tested on the Antarctic marine hydrobiont—Adamussium colbecki. This method extracts enough effective protein directly from primary raw materials, as well as when using leftover crude precipitates. The electrophoretic pattern of proteins showed the presence of molecules in a wide range of molecular weights in the samples of A. colbecki after the first and the second stage of extraction. The general proteolytic activity in the first and the second extracts were examined using a zymogram technique. Our experiments revealed that the second extract of A. colbecki contained thermo stable protease exhibiting a molecular weight of 95 kDa in a gelatin zymogram. Further biochemical assays, using different substrates, were conducted to partially identify the types of hydrolases present in the first and the second extracts. Our results revealed the presence of enzymes with collagenolytic and some amylolytic activities preserved in the second extracts. But no esterase or amidase trypsin-like activities were found in the second extract, in contrast to the first extract where this type of activity was significant

Fragmenti kolagena male molekularne mase poboljšavaju masene i upalne parametre masnog tkiva pretilih štakora

Research background. Despite clearly recognized links between increased body mass and increased risk for various pathological conditions, therapeutic options to treat obesity are still very limited. The aim of the present study is to explore the effect of low-molecular-mass collagen fragments obtained from the scales of Antarctic wild marine fish on rats\u27 visceral and subcutaneous white adipose tissue in a high-calorie diet-induced obesity model. Experimental approach. The study was conducted on outbred rats, which were divided into 3 experimental groups: (i) control, consuming standard food (3.81 kcal/g), (ii) obese group, consuming a high-calorie diet (5.35 kcal/g), and (iii) obese group, consuming a high-calorie diet (5.35 kcal/g) with intragastric administration of low-molecular-mass collagen fragments (at a dose 1 g/kg of body mass during 6 weeks). Low-molecular-mass collagen fragments were obtained by a procedure that included collagen extraction from fish scales and enzymatic hydrolysis with pepsin. Apart from hematoxylin and eosin staining, fibrosis level was assessed by histochemical Van Gieson’s trichrome picrofuchsin staining, and mast cells were analysed by toluidine blue O staining. Results and conclusions. Group treated with low-molecular-mass fragments of collagen exhibited decreased rate of mass gain, relative mass, area occupied by collagen fibre of both visceral and subcutaneous adipose tissue, and cross-sectional area of both visceral and subcutaneous adipocytes. Treatment with low-molecular-mass fragments of collagen reduced the infiltration of immune cells, number of mast cells and their redistribution back to the septa. This was also accompanied by a decreased number of the crown-like structures formed by the immune cells, which are markers of chronic inflammation that accompanies obesity. Novelty and scientific contribution. This is the first study that reports the anti-obesity effect of low-molecular-mass fragments produced as a result of controlled hydrolysis of collagen from the scales of Antarctic wild marine fish in the in vivo model. Another novelty of this work is the observation that the tested collagen fragments not only reduce the body mass, but also improve the morphological and inflammatory parameters (decrease in the number of crown-like structures, immune cell infiltration, fibrosis and mast cells). Altogether, our work suggests that low-molecular-mass collagen fragments are a promising candidate for amelioration of some comorbidities linked to obesity.Pozadina istraživanja. Usprkos jasno utvrđenoj povezanosti povećane tjelesne mase s većim rizikom razvoja različitih patoloških oboljenja, mogućnosti liječenja pretilosti su još uvijek vrlo ograničene. Svrha je ovoga rada bila ispitati učinak fragmenata kolagena male molekularne mase, dobivenih iz ljuski morskih riba s područja Antarktika, na visceralno i potkožno masno tkivo štakora s dijabetesom uzrokovanim visokokaloričnom prehranom. Eksperimentalni pristup. Istraživanje je provedeno na nesrođenim sojevima štakora, podijeljenim u tri eksperimentalne skupine: (i) kontrolna skupina, koja je primala standardnu prehranu (3,81 kcal/g), (ii) pretila skupina koja je primala visokokaloričnu prehranu (5,35 kcal/g), i (iii) pretila skupina koja je primala visokokaloričnu prehranu (5,35 kcal/g) i intragastrično fragmente kolagena male molekularne mase (1 g po kg tjelesne mase tijekom 6 tjedana). Fragmenti kolagena male molekularne mase dobiveni su ekstrakcijom kolagena iz ljusaka ribe i njegovom enzimskom hidrolizom pomoću pepsina. Stupanj fibroze masnog tkiva je, osim bojenjem hematoksilin eozinom, utvrđen Van Giesonovim bojenjem, a mastociti su ispitani bojenjem toluidinskim modrilom. Rezultati i zaključci. Skupina štakora koja je primala fragmente kolagena male molekularne mase sporije je dobivala na masi i relativnoj masi, a kolagenska vlakna su zauzimala manju površinu njihovog visceralnog i potkožnog masnog tkiva te presjeka visceralnih i potkožnih adipocita. Intragastričnom primjenom fragmenata kolagena male molekularne mase smanjili su se infiltracija imunoloških stanica, broj mastocita i njihova redistribucija u septe. Također se smanjio broj krunastih struktura koje tvore makrofagi, a koje su markeri kronične upale povezane s pretilošću. Novina i znanstveni doprinos. Ovo je prvo istraživanje in vivo o utjecaju fragmenata kolagena dobivenih kontroliranom hidrolizom ljusaka morskih riba iz područja Antarktika na smanjenje tjelesne mase. Još jedna novina ovoga rada je opažanje da ispitani fragmenti kolagena nisu samo smanjili tjelesnu masu, već i poboljšali morfološke i upalne parametre (smanjili su se broj krunastih struktura, infiltracija imunostanica, fibroza i broj mastocita). Zaključno, rad je pokazao da fragmenti kolagena male molekularne mase mogu ublažiti neke komorbiditete povezane s pretilošću

Fibrinolytic parameters under ischemic stroke with diabetes mellitus combination

Abstract Background: Fibrinolysis and thrombosis alterations include important parts of stroke pathophysiology. At the same time fibrinolytic system disorders are a common feature of patients with metabolic syndrome and diabetes. So it may increase the possibility of developing atherosclerotic lesions and occlusive intravascular thrombi. The present study investigated the influence of type 2 diabetes mellitus presence on the indicators of fibrinolytic parameters (plasminogen activator inhibitor 1 (PAI-1), tissue-type plasminogen activator (tPA) content, streptokinase-activated plasminogen and α2-antiplasmin activities, euglobulin clot lysis time (ECLT) and Hageman-factor-dependent fibrinolysis time) under ischemic stroke (IS). Materials and methods: Participants were 87 subjects with IS, 22 of them had diabetes mellitus. Blood samples besides for aforementioned parameters were analyzed for glucose and glycosylated haemoglobin content. Results: The research established increase of plasma PAI-1 and tPA levels, ECLT, Hageman-factor-dependent fibrinolysis time in IS and IS with diabetes mellitus patient groups in comparison with the control. PAI-1 concentration in plasma was positively correlated with both lysis time tests but tPA content was negative correlated with glucose level and PAI-1 for only IS patients. But there was a high negative correlation between tPA and ECLT as well as Hageman-factor-dependent fibrinolysis time for both investigated IS forms. Conclusions: The results showed important differences in the characteristics of the fibrinolytic mechanism in IS patients compared with healthy population. The major differences were elevated PAI-1 and t-PA contents and prolonged ECLT in IS patients but no significant differences in these parameters were observed between the patients with IS and IS with diabetes.</p

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

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

Proteolytic parameter changes in the plasma of patients with bladder cancer – depending on tumor stage

Bladder cancer (BC) is a worldwide common disease with a high mortality rate. Recognizing the dynamic changes in plasma that proteases and their inhibitors undergo might be valuable in understanding the carcinogenesis of invasive bladder cancer and in identifying BC patients with poor prognosis. This study aims to determine the activity of the proteolytic enzyme system and their inhibitors in patients with BC. In this paper, the total proteolytic activity, the activity of matrix metalloproteases (MMPs) and serine proteases was analyzed by the method of caseinolytic activity. For detection of activity of some inhibitors of proteolysis, we used the unified method for determining the activity of alpha-1-antitrypsin (α1A) and alpha-2-Macroglobulin (α2M) in human plasma. The level of medium-mass molecules (MMM) was assessed spectrophotometrically by applying the Nikolaichik method

Low-Molecular-Mass Fragments of Collagen Improve Parameters Related to Mass and Inflammation of the Adipose Tissue in the Obese Rat

Research background. Despite clearly recognized links between increased body mass and increased risk for various pathological conditions, therapeutic options to treat obesity are still very limited. The aim of the present study is to explore the effect of low-molecular-mass collagen fragments obtained from the scales of Antarctic wild marine fish on rats' visceral and subcutaneous white adipose tissue in a high-calorie diet-induced obesity model. Experimental approach. The study was conducted on outbred rats, which were divided into 3 experimental groups: (i) control, consuming standard food (3.81 kcal/g), (ii) obese group, consuming a high-calorie diet (5.35 kcal/g), and (iii) obese group, consuming a high-calorie diet (5.35 kcal/g) with intragastric administration of low-molecular-mass collagen fragments (at a dose 1 g/kg of body mass during 6 weeks). Low-molecular-mass collagen fragments were obtained by a procedure that included collagen extraction from fish scales and enzymatic hydrolysis with pepsin. Apart from hematoxylin and eosin staining, fibrosis level was assessed by histochemical Van Gieson’s trichrome picrofuchsin staining, and mast cells were analysed by toluidine blue O staining. Results and conclusions. Group treated with low-molecular-mass fragments of collagen exhibited decreased rate of mass gain, relative mass, area occupied by collagen fibre of both visceral and subcutaneous adipose tissue, and cross-sectional area of both visceral and subcutaneous adipocytes. Treatment with low-molecular-mass fragments of collagen reduced the infiltration of immune cells, number of mast cells and their redistribution back to the septa. This was also accompanied by a decreased number of the crown-like structures formed by the immune cells, which are markers of chronic inflammation that accompanies obesity. Novelty and scientific contribution. This is the first study that reports the anti-obesity effect of low-molecular-mass fragments produced as a result of controlled hydrolysis of collagen from the scales of Antarctic wild marine fish in the in vivo model. Another novelty of this work is the observation that the tested collagen fragments not only reduce the body mass, but also improve the morphological and inflammatory parameters (decrease in the number of crown-like structures, immune cell infiltration, fibrosis and mast cells). Altogether, our work suggests that low-molecular-mass collagen fragments are a promising candidate for amelioration of some comorbidities linked to obesity

Disturbances of extracellular protein metabolism in ceruleininduced pancreatitis

Chronic pancreatitis (CP) is still a serious clinical problem due to the significant difficulties in its diagnosis, especially in the initial stages of development. Among the mechanisms that mediate the pathogenesis of CP and lead to pancreatitis-related disorders is unregulated activation of proteolytic enzymes, namely, matrix metalloproteinases (MMPs). The aim of our study was to determine the disturbances of protein metabolism under the conditions of CP alone or in combination with diabetes type 1 (CP+DT1). Herein, CP was induced in the nonlinear male rats by intraperitoneal injection of cerulein (5 µg·kg−1 of body weight; five times during fives day). DT1 was modeled in the rats with CP by a single intraperitoneal injection of streptozotocin (65 mg·kg−1 of the body weight). The levels of MMP-2 and -9 were determined by enzyme-linked immune sorbent assay, and the level of low and middle molecular weight (LMMW) substance was measured spectrophotometrically, while the peptide fractions were analyzed by size exclusion chromatography. The present study revealed a significant increase of MMP-2 and MMP-9 levels in the serum, liver and pancreas of the rats with CP and CP+DT1. Elevated levels of MMPS may act as a factor for the initiation of subsequent cascade of events resulting in the development of pancreatitis-associated complications. Pathogenesis of chronic pancreatitis alone or in combination with diabetes type 1 has been accompanied by the formation and accumulation of LMMW substance, changes in peptide composition and level of individual peptides in the tissues of the rats. Such alterations are among key triggers of amplification of metabolic disorders under chronic pancreatitis