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

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

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

Вплив секретів шкірних залоз амфібій на показники хронометричних тестів

According to the extremely high mortality and disability rates associated with the abnormal functioning of the hemostasis system, the search for new approaches for the prevention and treatment of these conditions is one of the most acute problems of modern biochemistry. The active components of reptile poisons are actively used in the treatment of these diseases, but the study of the effects of amphibians’ skin glands secretion on the functioning of hemostasis system has not yet been carried out. So, the aim of this work was to assess the effects of the components of amphibian skin secretions on the functioning of the coagulation link of the hemostasis system. Methods. In this study the skin secretions of ten Ukrainian species of amphibians were collected: B. bombina, B. variegata, B. bufo, B. viridis, R. temporaria, P. ridibundus, P. esculentus, P. fuscus, S. salamandra and the hybrid of B. bombina and B. variegatа. The samples of crude skin secretions were prepared. The activated partial thromboplastin time (aPTT), thrombin time (TT) and prothrombin time (PT) tests were conducted in vitro using the coagulation analyzer (Rayto RT-2201C, China) and the standard set of reagents (RENAM, Russian Federation).Results. It was established that the components of crude skin secretions of B. bombina, B. variegata, their hybrid, R. temporaria and P. ridibundus prolonged the aPTT of clotting plug formation. The components of skin secretions of B. viridis, P. esculentus, P. fuscus and S. salamandra prolonged the TT of clotting plug formation.Conclusions. The fact that some amphibian species prolonged the aPTT and TT of clotting plug formation could be the indicator of the presence of inhibitors of certain factors of coagulation hemostasis or be the reason of the degradation of the components of coagulation hemostasis by active components of skin secretion. Such results prove that the amphibian crude skin secretions are a potential source of the compounds that can affect the hemostasis system. The identification of an active component and the elaboration of its mechanism of action are required in further investigationsУчитывая чрезвычайно высокие показатели распространенности, раннюю инвалидизацию и смертность от заболеваний, связанных с нарушением функционирования системы гемостаза, поиск новых подходов профилактики и лечения этих состояний является актуальным вопросом современной биохимии. Известно, что активные компоненты яда рептилий активно используются в терапии данных недугов, однако исследования влияния секрета кожных желез амфибий на систему гемостаза до сих пор не были проведены. Еще меньше научно обоснованной информации можно встретить относительно природы и механизмов действия биологически активных компонентов секрета кожных желез представителей отечественной фауны. Поэтому целью данной работы было оценить влияние секретов кожных желез представителей класса земноводных, распространенных на территории Украины, на функционирование коагуляционного звена системы гемостаза.Методы. Было получено секреты кожных желез десяти самых распространенных украинских видов амфибий: B. bombina, B. vriegata, B. bufo, B. viridis, R. temporaria, P. ridibundus, P. esculentus, P. fuscus, S. salamandra и гибрида B. bombina и B. variegatа. Время свертывания плазмы крови в тестах активированное частичное тромбопластиновое время (АЧТВ), протромбиновое время (ПВ), тромбиновое время (ТВ) под влиянием секретов кожных желез исследуемых видов амфибий определяли in vitro и фиксировали на коагулометричному анализаторе (Rayto RT-2201C, Китай), используя стандартные наборы реагентов (RENAM, Российская Федерация).Результаты. Было установлено, что компоненты секретов кожных желез амфибий видов B. bombina, B. variegata, их гибрида, R. temporaria и P. ridibundus удлиняли время образования фибринового сгустка в тесте АЧТВ. Компоненты секретов кожных желез амфибий видов B. viridis, P. esculentus, P. fuscus и S. salamandra удлиняли время свертывания плазмы в тесте ТТ.Выводы. Полученные результаты могут быть обусловлены наличием в секретах кожных желез исследуемых видов амфибий ингибиторов определенных факторов коагуляционного гемостаза, или могут быть связаны с усилением деградации компонентов коагуляционного гемостаза активными компонентами секретов. Такие результаты доказывают, что секреты земноводных – потенциальные источники для поиска соединений эффекторов системы гемостаза, что указывает на перспективность дальнейших исследований в направлении идентификации активных молекул и детализации механизмов их действия, с целью дальнейшего применения этих соединений в качестве основы для разработки новых, более эффективных фармацевтических агентов направленного действияЗважаючи на надзвичайно високі показники поширеності, ранню інвалідизацію та смертність від захворювань, пов’язаних з порушенням функціонування системи гемостазу, пошук нових підходів щодо профілактики та лікування цих станів є актуальним питанням сучасної біохімії. Відомо, що активні компоненти отрути рептилій активно використовуються у терапії даних недуг, однак дослідження впливу секрету шкірних залоз амфібій на систему гемостазу досі не були проведені. Ще менше науково обґрунтованої інформації можна зустріти щодо природи та механізмів дії біологічно-активних компонентів секрету шкірних залоз представників вітчизняної фауни. Тому метою даної роботи було оцінити вплив секретів шкірних залоз представників класу земноводних, поширених на території України, на функціонування коагуляційної ланки системи гемостазу.Методи. Було отримано секрети шкірних залоз десяти найпоширеніших українських видів амфібій: B. bombina, B. variegata, B. bufo, B. viridis, R. temporaria, P. ridibundus, P. esculentus, P. fuscus, S. salamandra та гібриду B. bombina та B. variegatа. Час зсідання плазми крові у тестах активований частковий тромбопластиновий час (АЧТЧ), протромбіновий час (ПЧ), тромбіновий час (ТЧ), за дії секретів шкірних залоз досліджуваних видів амфібій, визначали in vitro та фіксували на коагулометричному аналізаторі (Rayto RT-2201C, Китай), використовуючи стандартні набори реагентів (RENAM, Російська Федерація).Результати. Було встановлено, що компоненти секретів шкірних залоз амфібій видів B. bombina, B. variegata, їх гібриду, R. temporaria та P. ridibundus подовжували час утворення фібринового згустку у тесті АЧТЧ. Компоненти секретів шкірних залоз амфібій видів B. viridis, P. esculentus, P. fuscus та S. salamandra продовжували час зсідання плазми у тесті ТТ.Висновки. Отримані результати можуть бути обумовлені наявністю у секретах шкірних залоз досліджуваних видів амфібій інгібіторів певних факторів коагуляційного гемостазу, або можуть бути пов’язані з посиленням деградації компонентів коагуляційного гемостазу активними компонентами секретів. Такі результати доводять, що секрети земноводних є потенційним джерелом пошуку сполук-ефекторів системи гемостазу, що вказує на перспективність подальших досліджень у напрямку ідентифікації активних молекул та деталізації механізмів їх дії, з метою подальшого застосування цих сполук як основи для розробки нових, більш ефективних фармацевтичних агентів направленої ді

N-(9,10-Dioxo-9,10-dihydroanthracen-1(2)-yl)-2-(R-thio) Acetamides: Synthesis, Antioxidant and Antiplatelet Activity

The synthesis of new N-(9,10-dioxo-9,10-dihydroanthracen-1(2)-yl)-2-(R-thio) acetamides was carried out using reaction of 2-chloro-N-(9,10-dioxo-9,10-dihydroanthracene-1(2)-yl)acetamides with functionalized thiols in the presence of potassium carbonate in N,N-dimethylformamide (DMF) at room temperature. Evaluation of the synthesized compounds on such indicators of radical scavenging activity as lipid peroxidation (LP) and oxidative modification of proteins (OMP) in vitro in rat liver homogenate was carried out. It was determined that the compounds with a substituent in the first position of anthracedione core showed better antioxidant properties than their isomers with a substituent in the second position. The compounds 6 and 7 with the best indicators of radical-scavenging activity were determined. Antioxidant effect in OMP processes was also determined for compound 10. The antiplatelet activity study in vitro revealed compound 10 with the inhibited effect of ADP-induced aggregatio

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

The evaluation of lipid peroxidation and oxidative modification of proteins in blood serum under obesity development and the consumption of aqueous kidney beans Phaseolus vulgaris pods extract

Our interest has focused on the investigation of the anti-obese potential of kidney beans (P. vulgaris) pods extract. In the course of the study, obesity development in rats was induced with high-calorie diet. Control and obese rats then have consumed with aqueous kidney beans (P. vulgaris) pods extract during 6 weeks (200 mg/kg). Results show that the long-term consumption of P. vulgaris pods extract can lead to the reduction of hyperglycemia and insulin resistance development. Furthermore, we saw a normalization of lipid peroxidation parameters and oxidative modification of protein due to the consumption of the kidney beans (P. vulgaris) pods extract. Our experimental data demonstrate the ability of the kidney beans (P. vulgaris) pod extracts to mitigate obesity development but the details of this mechanism remains to be not fully understood

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

Hypoglycemic activity of Phaseolus vulgaris (L.) aqueous extract in type 1 diabetic rats

The aim of the present study was to evaluate the hypoglycemic activity of the aqueous extract from the fruit walls of Phaseolus vulgaris pods and to examine the potential mechanism underlying the improvement of the glycemic level. In the course of the study, diabetes mellitus was induced in rats with a single intraperitoneal injection of streptozotocin (45 mg·kg−1 b.w.). Diabetic and control rats were then orally administered with a single-dose or repeated-dose (28 day) of P. vulgaris extract (200 mg·kg−1). Results show that the extract was found to possess significant hypoglycemic activity, and the study of glucose utilization by isolated rat hemidiaphragm suggests that the aqueous extract may enhance the peripheral utilization of glucose. The subsequent experiments have revealed that the P. vulgaris extract could increase glucose transporter 4 (GLUT-4) content in skeletal muscle cells of control and diabetic rats. Our data also indicate that the P. vulgaris extract did not affect the content of the insulin receptor, but significantly reduced the total tyrosine kinase activity in skeletal muscle cells of both experimental groups of rats. The present results clearly indicated that P. vulgaris extract may be beneficial for reducing hyperglycemia through its potency in regulation of glucose utilization via GLUT-4, but the current mechanism remains to be unidentified