Total Fibrinolytic Activity Research Articles

FIBRINOLYTIC ACTIVITY IN HEART TISSUE IN HYPOTHYROID AND HYPERTHYROID RATS UNDER THE INFLUENCE OF EXOGENOUS MELATONIN AND BLINDING CONDITIONS (ENUCLEATION)

The aim of our scientific work was to study the effect of exogenous melatonin on the indicators of fibrinolytic processes in the heart tissues and to analyze the changes in fibrinolytic activity occurring in the heart tissues of enucleated hyper- and hypothyroid white rats. The experiments were carried out on white nonlinear male rats weighing 0.12-0.14 kg. 4 experimental groups of animals were formed. The control group consisted of 11 normothyroid rats, which were injected with a melatonin solution in appropriate volumes. The animals were euthanized under light ether anesthesia. Samples of the examined heart tissue were ground in a glass homogenizer with borate buffer (pH 9.0). The homogenate was used in biochemical analysis. Determination of the total, enzymatic and non-enzymatic fibrinolytic activity (TFA, EFA, NFA, respectively) in the heart tissues was carried out by the method of azofibrin lysis (LLC "Simko", Ukraine). Statistical processing of the results was carried out by the method of variation statistics using the Student's test. Experimental studies on nonlinear male white rats have shown that the introduction of exogenous melatonin, as well as modeling the conditions of endogenous melatonin overproduction by blinding (enucleation) cause an increase in the intensity of enzymatic and non-enzymatic fibrinolysis in heart tissue in normotyroid animals. At other hand, enucleation causes an increase in the intensity of fibrinolysis in the tissues of the heart in hypothyroid rats, but not in hyperthyroid.

Особливості фібринолітичної та протеолітичної активності плазми крові у пацієнтів з хронічним панкреатитом, поєднаним з гіпотиреозом

The purpose of the study is to analyze the state of proteolytic and fibrinolytic activities of blood plasma in patients with chronic pancreatitis combined with hypothyroidism. Materials and methods. 105 people participated in our study, of which group 1 consisted of patients with chronic pancreatitis (n = 27), group 2 – patients with hypothyroidism (n = 30), group 3 – patients with chronic pancreatitis combined with hypothyroidism (n = 28), group 4 – almost healthy individuals (n = 20). The state of fibrinolytic activity of blood plasma was studied by lysis of azofibrin, followed by determination of total fibrinolytic activity, non-enzymatic fibrinolytic activity and enzymatic fibrinolytic activity. Assessment of the state of the proteolysis system was studied by lysis of azoalbumin (breakdown of low molecular weight proteins), azocasein (breakdown of high molecular weight proteins) and azocol (breakdown of collagen). Results. When analyzing the results of the study, we observe a probable increase in lysis of azoalbumin by 1.89, 1.96 and 2.16 times (p <0.05) in groups 1, 2, 3 compared with the group of almost healthy individuals. In patients with chronic pancreatitis and hypothyroidism, the most pronounced degradation of low molecular weight proteins was observed, which was 13.86% and 9.75% (p <0.05) higher than in the first and second groups. Indicators of azocasein lysis by 52.48%, 56.35% and 95.03% (p <0.05) were found in groups 1, 2, 3 compared with almost healthy individuals. Azocasein lysis was higher by 27.89% and 24.73% (p <0.05) in patients with chronic pancreatitis combined with hypothyroidism than in patients in groups 1 and 2. Azocol lysis was significantly higher by 10.85%, 12.05%, 16.87% (p <0.05) in groups 1, 2, 3 compared with almost healthy individuals. In addition, in patients with comorbid pathology there was an increase in lysis of azocol by 5.3% and 4.3% (p <0.05) compared with the first and second groups. The total fibrinolytic activity of blood plasma was 8.3%, 6.7%, 16.26% (p <0.05) lower in patients of groups 1, 2, 3 compared with almost healthy individuals. Non-enzymatic fibrinolytic activity of blood plasma was 44.89%, 49.64%, 66.27% higher in groups 1, 2 and 3 than in almost healthy individuals. Enzymatic fibrinolytic activity of blood plasma was 44.28%, 42.25%, 90.57% (p <0.05) lower in group 1, 2, 3 compared with the group of almost healthy individuals (p <0,05). There was a decrease in the level of enzymatic fibrinolytic activity of blood plasma by 32.07% and 33.96% (p <0.05) in patients with chronic pancreatitis associated with hypothyroidism compared with participants in groups 1 and 2 without comorbid pathology. Conclusion. The most pronounced changes in proteolytic (increased lysis of azoalbumin, azocasein, azocol) and fibrinolytic (decrease in total, non-enzymatic and enzymatic) activities of blood plasma in patients with chronic pancreatitis associated with hypothyroidism were determined

Аналіз протеолітичної та фібринолітичної активності плазми крові у хворих на бронхіальну астму, поєднану з ожирінням

Мета дослідження — провести аналіз стану протеолітичної та фібринолітичної активностей плазми крові у хворих на бронхіальну астму (БА), поєднану з ожирінням. Матеріали та методи. Обстежено 40 хворих, які були розподілені на групи. Основну групу становили 20 хворих на БА в поєднанні з ожирінням (ІІ група), дві групи порівняння — 10 хворих на БА з нормальною масою тіла (І група) та 10 хворих з ожирінням і без патології бронхолегеневої системи (ІІІ група). Контрольну групу становили 10 практично здорових осіб. У хворих на БА досліджували показники протеолітичної та фібринолітичної активностІ плазми крові. Результати. Аналіз отриманих даних у хворих на БА показав зростання активності плазмових факторів згортання крові: cумарна фібринолітична активність зростала порівняно із контрольною групою та становила 1,46 ± 0,13 Е440/мл/год при показниках у практично здорових осіб 1,23 ± 0,16 Е440/мл/год (р < 0,05). Крім того, зростання відзначалося і в групі пацієнтів з ізольованим ожирінням (на 38,5 % порівняно з групою практично здорових осіб), що можна пояснити підвищенням активності запального процесу за рахунок імунних зрушень, пов’язаних із біологічною активністю жирової тканини, як додаткового джерела прозапальних цитокінів. Висновки. При поєднанні бронхіальної астми та ожиріння спостерігається активація фібринолітичної та протеолітичної систем крові, що сприяє виникненню мікроциркуляторних і гемостатичних розладів, посиленню запального процесу. При ожирінні як стані системної запальної відповіді відзначається активація фібринолітичної та протеолітичної систем крові більш виражено, ніж при бронхіальній астмі.

Патогенетичні особливості пептичної виразки шлунка та дванадцятипалої кишки в поєднанні з артеріальною гіпертензією та цукровим діабетом 2-го типу

Розвиток та перебіг пептичної виразки шлунка та дванадцятипалої кишки супроводжуються наявністю штамів H. pylori (СagA+ VacA+, СagA+ vacА–, CagA– VacA+, CagA– VacA–), що виявлено у хворих за наявності супутньої патології та коли її немає. У групі хворих на пептичну виразку шлунка та дванадцятипалої кишки в поєднані з артеріальною гіпертензією і цукровим діабетом 2-го типу виявлено штами СagA+ VacA+ у 14 осіб (45,16 %), СagA+ VacA– — у 4 (12,9 %), CagA– VacA+ — у 13 (41,43 %) та в групі хворих на пептичну виразку шлунка та дванадцятипалої кишки — відповідно в 4 (13,79 %), у 8 (27,59 %), у 16 (55,17 %) осіб. Вплив штамів H. pylori супроводжується змінами протеолітичної та фібринолітичної активності. З урахуванням супутньої патології та ролі штамів при розвитку основної патології виявлено вірогідне зниження рівня лізису альбуміну, лізису азоказеїну та підвищення лізису азоколу порівняно зі зниженням сумарної фібринолітичної активності, неферментативної фібринолітичної активності та підвищенням ферментативної фібринолітичної активності. Використання антихелікобактерної базисної терапії та пробіотиків (Lactobacterium, Bifidobacterium) дає позитивний результат.

EVALUATION OF PROTEOLYTIC AND FIBRINOLYTIC ACTIVITY OF BLOOD PLASMA IN COMORBID CHRONIC CORONARY SYNDROME, CHRONIC OBSTRUCTIVE PULMONARY DISEASE, AND OBESE

The aim - to assess the proteolytic and fibrinolytic activity of blood plasma in comorbidchronic coronary syndrome (CCS), chronic obstructive pulmonary disease (COPD), andobese.Material and methods. The study involved 110 patients, who were divided into 5 clinicalgroups: G1 – 22 CCS and obese patients, G2 – 22 COPD and normal body weightpatients, G3 – 22 CCS, COPD and normal body mass index (BMI) subjects, G4 – 22CCS, COPD and overweight patients and G5 – 22 CCS, COPD and obese I, II degreepatients. The control group consisted of 20 healthy individuals. Proteolytic activityof blood plasma was assessed by the lysis of azoalbumin, azocasein, and azocol, andfibrinolytic – by the lysis of azofibrin.Results. There was a significant increase 50,5%, 43,3%, 43,8% (p<0,05) azoalbuminlysis in the fifth participant’s group compared with the 1st, 2nd, and 4th groups. An increasein azocasein lysis 38,08% was observed (p<0,05) in CCS, COPD, and obese patientscompared with CCS, and obese patients. Azocol lysis was significantly higher 41,7%,13,8%, and 22,06% (p<0,05) in CCS, COP, and obese patients compared with the first,second, and fourth groups. Total fibrinolytic activity (TFA) of blood plasma was 17,7%,14,89%, 21,27% (p<0,05) lower in the 3rd, 4th, and 5th patients’ groups compared withthe 1st one. Enzymatic fibrinolytic activity (EFA) of blood plasma was 28,29%, 22,19%,35,85% (p<0,05) lower in the 3rd, 4th, and 5th patients groups compared with the 1st one.EFA level decrease 23,77% (p<0,05) was observed in the fifth group compared with thesecond one.Conclusions. The most pronounced changes in proteolytic (increased azoalbumin,azocasein, and azocol lysis) and fibrinolytic (decreased total and enzymatic) activitiesof the blood plasma in comorbid chronic сoronary syndrome, chronic obstructivepulmonary disease, and obese patients.

INVESTIGATION OF EXTERNAL RESPIRATORY FUNCTION AND HEMOSTASIS SYSTEM’S STATUS IN PATIENTS WITH CHRONIC OBSTRUCTIVE LUNG DISEASE WITH CONCOMITANT CHRONIC PANCREATITIS

INTRODUCTION: It can be assumed that the comorbidity course of сhronic obstructive pulmonary disease (COPD) and chronic pancreatitis (CP) can enhance the clinical symptoms of both diseases and lead to frequent relapses of the pathological process due to changes in the proteolysis intensity of high and low molecular weight plasma proteins and the state of the hemocoagulation.
 OBJECTIVE: To establish the features of hemocoagulation and proteolytic hemostasis at COPD with concomitant CP.
 METHODS: 120 patients with COPD and CP were examined. The function of external respiration, total coagulation potential of blood plasma, the state of enzymatic and non-enzymatic fibrinolysis, total fibrinolytic activity were investigated.
 RESULTS: CP contributes to the development of broncho-obstructive syndrome, and the maximum indicators of reduction of FEV1 relative to the proper values are observed in patients with a comorbid course of COPD and CP. Reducing the intensity of collagenolysis in patients of groups 1-2 contributed to the development of diffuse pulmonary fibrosis in response to chronic inflammation. The imbalanced increase in the intensity of proteolysis due to reduced expression of its inhibitors in COPD patients with CP led to progressive destruction of the cell membranes of alveolocytes, acinar epithelium of pancreas and epithelium of the bronchial mucosa, acceleration of their apoptosis and development of desquamation, atrophic changes, metaplasia, and the like. The above factors are active as inducers of inflammation, and the formation of pulmonary fibrosis and fibrosis of the pancreas.
 CONCLUSIONS: In COPD patients with accompanying CP are an increase in the lysis rate of low and high molecular weight proteins and a decrease in blood collagenolytic activity on the background of a significant imbalance in the activity of plasma proteinase inhibitors. Defined suppression of the activity of the anti-coagulation system factors and enzymatic, Hageman-factor-dependent fibrinolysis indicates the formation of hypercoagulation syndrome in these patients.

FIBRINOLYTIC AND PROTEOLYTIC ACTIVITY OF BLOOD PLASMA IN PEPTIC ULCER OF THE STOMACH, TAKING INTO ACCOUNT THE PATHOGENIC STRAINS OF HELICOBACTER PYLORI

The purpose of the study is to investigate changes in fibrinolytic and proteolytic activity of blood plasma in patients with peptic ulcer (PU) taking into account pathogenic Helicobacter pylori (Hp) strains. Materials and methods. 93 patients with PU were examined, of which 30 patients with PU and concomitant Hp cag cag A+/vac A+ (group I), 31 patients with PU and concomitant Hp cag A-/vac A- (group II), 32 patients with PU without concomitant HP infection (group III). The control group consisted of 30 healthy individuals. Fibrinolytic activity of blood plasma was investigated with the help of lysis of azofibrin (fibrin associated with the azo dye orange), which in the alkaline medium turns a bright red color. The level of total (ТFA), enzymatic (FFA) and non-enzymatic fibrinolytic activity (NFA) was evaluated. Proteolytic activity of blood plasma was determined by the lysis of azoalbumin, azocasein and azokol. Research results. The study of fibrinolytic activity of blood plasma showed that the total fibrinolytic activity of blood plasma (TFA) in all groups was significantly higher compared to the control indicators: in patients of group I by 61.5 %, in patients by 40.9 %, in patients of group III by 30.3 %, with a significant intergroup difference between the groups. The growth of TFA was mainly due to FFA. In patients of group I, FFA increased by 2.06 times (p < 0.05), and in patients of group II – by 1.79 times (p < 0.05), in patients of group IIІ – by 1.52 times (p < 0.05) compared with the control. In patients with group I, FFA increased by 12.5 % ​​(p < 0.05) compared with group II. In all patients examined, there was an increase in the proteolytic activity of blood plasma, in particular in group I, the lysis of azoalbumin, azocasein and azocolol increased significantly 2.94 times, 2.83 times and 1.90 times, respectively, and in the patients of group II the investigated indicators increased accordingly 1.87-fold (p < 0.05), 1.96-fold (p < 0.05) and 1.40-fold (p < 0.05), in patients of group III, respectively 1.55 times (p < 0.05), 1.59 times (p < 0.05) and 1.18 times, compared to these values ​​in almost healthy subjects. Significantly more significant changes in proteolysis were detected in the presence of pathogenic Hp strains. Conclusion. Increased proteolytic and fibrinolytic activity of blood plasma is observed in patients with PU. The presence of concomitant Hp in PU leads to more pronounced changes in proteolysis and fibrinolysis. Pathogenic strains of Hp cag cag A+/vac A+ cause significantly more abnormalities in hemostasis.

Recognition of Plasminogen Activator Inhibitor Type 1 as the Primary Regulator of Fibrinolysis.

The fibrinolytic system consists of a balance between rates of plasminogen activation and fibrin degradation, both of which are finely regulated by spatio-temporal mechanisms. Three distinct inhibitors of the fibrinolytic system that differently regulate these two steps are plasminogen activator inhibitor type-1 (PAI-1), α2-antiplasmin, and thrombin activatable fibrinolysis inhibitor (TAFI). In this review, we focus on the mechanisms by which PAI-1 governs total fibrinolytic activity to provide its essential role in many hemostatic disorders, including fibrinolytic shutdown after trauma. PAI-1 is a member of the serine protease inhibitor (SERPIN) superfamily and inhibits the protease activities of plasminogen activators (PAs) by forming complexes with PAs, thereby regulating fibrinolysis. The major PA in the vasculature is tissue-type PA (tPA) which is secreted from vascular endothelial cells (VECs) as an active enzyme and is retained on the surface of VECs. PAI-1, existing in molar excess to tPA in plasma, regulates the amount of free active tPA in plasma and on the surface of VECs by forming a tPA-PAI-1 complex. Thus, high plasma levels of PAI-1 are directly related to attenuated fibrinolysis and increased risk for thrombosis. Since plasma PAI-1 levels are highly elevated under a variety of pathological conditions, including infection and inflammation, the fibrinolytic potential in plasma and on VECs is readily suppressed to induce fibrinolytic shutdown. A congenital deficiency of PAI-1 in humans, in turn, leads to life-threatening bleeding. These considerations support the contention that PAI-1 is the primary regulator of the initial step of fibrinolysis and governs total fibrinolytic activity.

INFLUENCE OF HEPARINOID FROM PAEONIA LACTIFLORA ON HEMOSTATIC SYSTEM WITHIN CONDITIONS OF PRETHROMBOSIS

The search and development of direct and rapid anticoagulants used per os, is an urgent problem in physiological and medical science. A number of plants contain heparin-like components with a positive effect on the hemostatic system, both within normal and in some pathological conditions of the body.The aim of the work was to study the complex effect of fibrin, a heparin-like substance (heparinoid) from the roots of Paeonia lactiflora, on fibrinolytic, anticoagulant systems of the body and polymerization processes, when it is administered per os in animals within normal conditions and when modeling the state of prethrombosis.Materials and methods. To carry out the research, the roots of Paeonia lactiflora growing in the Botanical Garden of Moscow State University, and laboratory animals – male Wistar rats – were used. To study the antithrombotic effects of the extract from roots containing heparinoid, the state of prethrombosis was modeled in rats. The determined parameters of hemostasis were: anticoagulant activity according to the tests of activated partial thromboplastin time and thrombin time, fibrinolytic activity according to the test of total fibrinolytic activity, fibrin polymerization according to the test of fibrindepolymerization activity of blood plasma.Results. With repeated (every 24 hours within 3 days) oral administration of the extract containing heparinoid, in animals within normal conditions and with prethrombosis, the following anticoagulant effects were established in the blood: an increase in anticoagulant, fibrindepolymerization and fibrinolytic plasma activity. Possible mechanisms of the activating effect of heparinoid on fibrinolysis and anticoagulant properties of plasma due to the excretion of tissue plasminogen activator into the bloodstream from the endothelium, thrombin inhibition, and fibrin polymerization are described. Moreover, the anticoagulant effect of the use of the extract from the peony roots was equivalent to that of the reference drug of low molecular weight heparin from Celsus (USA). For the first time, it was revealed that when modeling experimental prethrombosis, the administration of heparinoid in rats at the dose of 37.5 IU/kg МЕ/кг body weight restored impaired hemostasis, which requires a further study.Conclusion. The ability of heparinoid from peony roots to normalize the functional state of the anticoagulant system during the development of prethrombosis in animals has been established. The restriction of fibrin polymerization during oral administration of heparinoid from peony in animals by increasing the enzymatic fibrinolytic and fibrindepolymerization activity of blood plasma was revealed. In the future, heparinoid can be used as an antithrombotic agent.

Breast tumour size as a predictor of hemostatic system status and endothelial function in dogs

The high level of morbidity of dogs from mammary gland tumours and deficiencies in the pathogenesis give relevance to study of the disorders of the hemostatic system for predicting the neoplasia process. Our research concerned the determination of markers of the hemostatic system and endothelial function at different sizes (≤ 2, 5–7 and >10 cm) of benign (n = 28) and malignant (n = 27) breast tumours in dogs, taking into account their histological structure. The progression of the tumour process was accompanied by a 1.24–1.81 times increase in the fibrinogen content in the benign forms and 1.39–2.38 times in malignant against the background of progressive excess accumulation of soluble fibrin. The results of coagulation tests indicate that the increase in blood coagulation, which correlates with the magnitude of tumours, occurred mainly externally. In malignant neoplasms reliable, compared to clinically healthy dogs, reduction of total fibrinolytic activity occurred due to deficient inhibition of tissue plasminogen activator by 1.62–2.03 times. Increase in the size of benign neoplasms was accompanied by increased activity of the α1-inhibitor of proteinases, and in malignant – only in tumours of the size of 5–7 cm, against the background of its decrease in dogs with small and giant neoplasia. The progression of the disease was characterized by an increase in the content of α2-macroglobulin in benign lesions from 1.19–1.24 times, malignant – from 1.25–2.03 times. At the same time, there was a deepening of oxidative stress, as evidenced by the excess accumulation in the blood of malondialdehyde by 1.29–1.70 times in benign tumours, and 1.58–2.73 times in malignant. Regardless of the pathomorphologic form, the magnitude of the neoplasia foci is directly correlated with excess accumulation of nitric oxide and the level of hypoproteinemia, and vice versa – the content of ceruloplasmin. Further research should be undertaken to study the hemostatic status in certain nosological forms of breast tumours in dogs, which will improve the diagnosis and development of effective treatment protocols.

ROLE OF FIBRINOLYTIC ACTIVITY OF BLOOD IN PATHOGENESIS OF NON-ALCOHOLIC FATTY LIVER DISEASE AND CHRONIC KIDNEY DISEASE (ORIGINAL RESEARCH)

The aim of the research − to find out of changes fibrinolytic activity of blood in patients with non-alcoholic fatty liver on the background of obesity, depending on the presence of comorbid chronic kidney disease. Material and methods of research: 444 patients were examined: 84 of them were with NAFLD and class I obesity (group 1), which contained 2 subgroups: 32 patients with non-alcoholic steatosis (NAS) and 52 patients with non-alcoholic steatohepatitis (NASH); 270 patients with NAFLD with comorbid class I obesity and CKD І–ІІІ stage (group 2), including 110 patients with NAS and 160 patients with NASH. The control group consisted of 90 patients with CKD of І–ІІІ stage with normal body weight (group 3). To determine the dependence of the NAFLD course on the form and stage of the CKD, the group of patients was randomized according to age, sex, degree of obesity, and activity of NASH. Research results. The study of fibrinolytic activity of blood showed that total fibrinolytic activity (TFA) of blood plasma in patients of all groups was significantly lower than the control indexes: in patients with NAS – by 7.1%, patients with NAS with CKD – by 14.9%, patients with NASH – by 17.2%, patients with NASH with CKD – by 18.9%, patients with CKD – by 10.6% (p <0.05) with the presence of a probable intergroup difference between groups with comorbidity and isolated course of CKD (p <0.05). The suppression of TFA occurred through the decrease of EF: in patients with NAS the index is significantly lower than that in the controls by 1.2 times, in patients with NAS with CKD – by 1.4 times, in patients with NASH – by 1.7 times, in the group of patients with NASH and CKD – by 1.9 times, while in the group of patients with CKD, the suppression of EF was registered – 1.3 times (p <0.05). At the same time, the NEF in patients of all groups increased in comparison with the AHP group: in patients with NAS – by 1.2 times, in patients with NAS with CKD – by 1.3 times, in patients with NASH – by 1.4 times, in the group of patients with NASH with CKD – 1.5 times, while in the group of patients with CKD the activation of NEF was registered 1.2 times (p <0.05), with the presence of a probable difference between the groups with comorbidity and isolated course of CKD (p <0.05). Conclusion. Analysis of hemostasis and fibrinolysis indices in examined patients with NASH, depending on the stage of CKD showed that with the growth of the CKD stage, the activity of the cohort increases, with the exception of the fibrinogen content (most likely due to coagulopathy consumption), the activity of the anti-coagulants decreases, the total and enzymatic activity of fibrinolysis is reduced, and non-enzymatic compensator increases. Thus, metabolic intoxication, oxidative stress, which accompany the flow of NAFLD with obesity and CKD, promote the activation of the kallikrein-kinin system, the formation of plasma and thrombin, with subsequent disturbance of equilibrium between them, the development of stasis, slag phenomenon, the formation of platelet and erythrocyte aggregates in blood circulation system. The consequence of significant activation of hemocoagulation against the suppression of total fibrinolytic activity (TFA) is the local clotting of blood in the arteries.

Лейцинсодержащие пептиды как перспективные антитромботические вещества

Here we study the gemostatic properties of leucine-containing regulatory oligopeptides Н-Pro-Leu-Pro-OH (PLP) and H-Pro-Leu-Pro-Ala-OH (PLPA) administered orally every 24 hours for 7 days to healthy animals (rats). Standard coagulologic techniques were applied. We demonstrated that 20 hours after the 7th administration of the peptides the anticoagulant activity in the blood plasma was increased by 10.3-30.6% according to the activated partial thromboplastin time data, all types of the fibrinolytic process — total plasma fibrinolytic activity by 16-24%, non-enzymatic fibrinolysis – by 9-11%, enzymatic fibrinolysis – by 28-49%, and the plasmin activity – by 29%. In contrast, the lysis time of the euglobulin fibrin clot (in absence of fibrinolysis inhibitors) was not affected. The activity of tissue plasminogen activator was either unchanged (PLP) or decreased under the effect of PLPA. The fibrinogen concentration either decreased by 18% (PLP), or remained at normal values (PLPA); the activity of factor XIIIa decreased by 17% (PLP) and 10% (PLPA). Both peptides suppressed the platelet aggregation by 31-33.5% (when used as an ADP inductor). The most prominent anticoagulant effect was observerd for the oligopeptide containing the additional alanine residue at the C-terminus. Since both peptided showed varying degrees of complex antiplatelet, anticoagulant, fibrindepolymerization, fibrinolytic activity, and moderate reduction of coagulation factor levels — fibrinogen and factor XIIIa — when administered orally to animals, they can be considered promising antithrombotic agents.

Pharmacological correction of the hemostasis system for the surgical treatment of bitches with tumours of the mammary gland

We carried out the clinical validation of pathogenetically substantiated protocols for the treatment of female dogs with tumour lesions of the mammary gland. The positive effect of corrective therapy was characterized by a decrease in the physiological level of fibrinogen: in benign tumours in 10 days, malignant – in 14 days against the background of its stable high level in control animals with malignant neoplasia. In the postoperative period in experimental animals, the functioning of the internal coagulation unit was restored, as evidenced by the normalization of the activated partial thromboplastin time for benign tumours by days 10–14, malignant – by 14 days. In control patients, these changes were detected only for benign neoplasms. Shifts of the external mechanism of blood coagulation in the experimental groups were eliminated in benign neoplasia cases in 10 days, malignant cases in 14 days against the background of severe disorders in this link in control patients throughout the observation period. The positive effect of complex treatment regimens is confirmed by the restoration of total fibrinolytic activity by days 10–14 due to the normalization in the same terms of the plasminogen activator and tissue plasminogen activator by days 3–7. In control animals, the total fibrinolytic activity was consistent with those of clinically healthy animals only for benign neoplasms. The balancing of coagulation and fibrinolytic mechanisms was accompanied by a decrease in the activity of proteolytic inhibitors: α2-macroglobulin in experimental groups with benign neoplasia by day 10, malignant – by days 10–14, and control – respectively, by days 3 and 14; α1-proteinase inhibitor in all groups by day 3. Concentration of nitric oxide after extirpation of benign tumors already at day 3 of the postoperative period in all groups corresponded to the indexes of clinically healthy dogs, with the removal of malignant tumours – in experimental animals at day 3, control – at day 14. The content of malondialdehyde and ceruloplasmin against the background of pharmacological correction was restored by days 10–14, whereas in control animals – only with benign neoplasms by day 14. Postoperative pharmacological correction of the hemostasis system can significantly improve the results of treatment in cancer patients.

Diagnosis and treatment of disseminated intravascular coagulation in burn patients

The aim — to study the violations of the hemostasis system in the acute period of burn disease in severely burned as a prophylaxis for thrombotic complications and provision of early surgical treatment.Materials and methods. The state of the blood clotting system was studied in 80 patients with deep dermal burns from 20 % to 60 % TBSA in the period from the first to the 14th day after the trauma. The age — 18 to 65 years. The following parameters of hemostasis were investigated: the number of platelets; serum fibrinogen, soluble fibrin, antithrombin III, protein C, partially activated thromboplastin time (APTT), prothrombin index, total fibrinolytic activity of blood plasma.Results and discussion. A moderately prolonged clotting time with the remaining level of platelets and fibrinogen were observed in all patient groups during the first day after the burn injury. The parameters of APTT were lowered on the 1st — 2nd day after the trauma in patients with IST for more than 90 units could have indicated the first phase of DIC syndrome; on the 3rd—5th day, an extension of the APTT was determined, which could have indicated a tendency for bleeding, especially during early operations. The level of antithrombin III was decreased in severely burned patients during the first 3 days after the burn injury, the plasma recalcification time decreased to 45 ± 5.4 s. Patients with extremely severe burns received 1.5 ± 0.3 L of fresh frozen plasma daily for 6.0 ± 1.5 days and also 0.6 ± 0.1 L of plasma for the following 10 days after the injury.Conclusions. Patients with deep common burns have are at high risk for thrombotic complications. Such include an increase in level of fibrinogen (> 5.5 g/L), soluble fibrin (> 4 mg/100 mL), prolongation of APTT (> 45 s), decrease in antithrombin III and protein C (< 75 %), fibrinolytic activity (< 65 %). The hemostasis system has the phase character of its functional state with widespread deep burns. Acceleration of blood coagulation was determined during the first two days after the trauma with the depression of physiological anticoagulants: antithrombin III and protein C. From day 3 — 4, intravascular coagulation corresponds to the hypocoagulation phase with the main inhibitors spending. Correction of the hemostasis system is achieved by fresh­frozen blood plasma prescription at a dose of 10 — 20 ml/kg and direct anticoagulants at a dose of heparin 200 — 400 un/kg until the PTT level reaches 40 — 60 s, the recalciation time is up to 2 minutes, the level of antithrombin III is above 60 — 70 %.

Особенности влияния гепариноида из пиона совместно с аргинином на растворение венозных тромбов у крыс

Цель исследования. Изучить влияние сочетанного применения гепариноида из пиона молочноцветкового, обладающего антикоагулянтным и фибринолитическим действием, и аргинина, улучшающего реологические свойства крови, на процессы фибрино- и тромбообразования, вызванные введением тромбопластина. Методика. Эксперименты выполнены на крысах-самцах Вистар (n = 36) массой 200-220 г. Процессы фибринообразования с последующим тромбообразованием вызывали внутривенным введением 0,4 мл 1% бычьего тромбопластина (модель тромбообразования - МТ). Для изучения эффективности сочетанного подкожного применения препаратов определяли антикоагулянтную и фибриндеполимеризационную активность каждого препарата в отдельности. За 30 мин до введения смеси гепариноида с аргинином (в соотношении 1:4 по весу) внутривенно вводили тромбопластин. Контролем служили животные, которым вводили физиологический раствор в том же объеме и в аналогичных условиях опыта. Результаты. Установлено в условиях in vitro, что препарат, включающий гепариноид из корней пиона молочноцветкового и аргинин при весовом соотношении 1:4, обладал антикоагулянтной и собственной фибриндеполимеризационной активностью. При подкожном введении этого препарата в минимальных дозах (0,05 мг на 200 г массы тела) в крови крыс повышались антикоагулянтная, суммарная, неферментативная и ферментативная фибринолитическая активность и снижалась до нормальных значений концентрация фибриногена. На модели крыс с тромбообразованием, вызываемым введением тканевого тромбопластина, показан антитромботический и тромболитический эффекты исследуемого препарата. Впервые показано, что применение этого препарата через 30 мин после начала тромбообразования у крыс препятствовало развитию процессов тромбообразования или даже способствовало растворению свежих образующихся фибриновых сгустков. Заключение. Сочетанное применение гепариноида из пиона с аргинином в минимально действующих дозах усиливает деполимеризацию фибрина и препятствует процессам тромбообразования. Исследование может служить экспериментальным обоснованием применения препаратов при тромботических осложнениях с целью благоприятного воздействия на растворение свежих только что образующихся тромбов.

Fibrinolytic activity of the blood plasma in gastroduodenopathies induced by non-steroi¬dal anti-inflammatory drugs in patients with osteoarthrosis, regarding pathogenic Helicobacter pylori strains

Objective. This research aims at estimating the changes of fibrinolytic activity of the blood plasma in gastroduodenopathies induced by non-steroidal anti-inflammatory drugs in patients with osteoarthrosis regarding the pathogenic Helicobacter pylori strains presence.Material and methods. The study involved 68 patients suffering from osteoarthrosis with concomitant Helicobacter pylori-associated gastro-duodenopathies induced by non-steroidal anti-inflammatory drugs. The patients were divided into groups depending on the presence of vacA and cagA Helicobacter pylori strains. Helicobacter pylori strains were detected in the stool, blood and biopsy samples using the technique of polymerase chain reaction. All the patients were investigated by means of general, clinical, laboratory and instrumental methods of examination. The computations were done using «Biostatistics» and «Statistica® 6.0» (StatSoftІnc., USA) programs. Results. The increase in the intensity of the fibrinolytic activity of the blood plasma was observed in all investigated patients. However, the patients with concomitant Helicobacter pylori pathogenic and ulcerogenic strain cagA+/vacA+ experienced an increase in total fibrinolytic activity by 13,14% and 36,4% respectively compared to the patients with cagA+/vacA- Helicobacter pylori strains and by 26,92% and 60,0% compared to the patients with cagA-/vacA+ Helicobacter pylori strains. Nonenzymatic fibrinolytic activity in patients with cagA+/vacA+ Helicobacter pylori strains tended to increase in comparison with cagA+/vacA- strains and cagA-/vacA+ Helicobacter pylori strains. Conclusion. A substantial increase in the fibrinolytic activity of the blood plasma in the examined patients with Helicobacter pylori vacA+/cagA+ compared to the patients with Helicobacter pylori vacA+/cagAand Helicobacter pylori vacA-/cagA+ has been reliably established.

Changes in blood parameters in the treatment of septic complications of diabetes with the use of ozone therapy

Objective. To study the influence of ozonotherapy on the course of purulent-inflammatory processes in diabetes mellitus in an experiment.Material and methods. Experimental studies were performed on 30 white old rats. The first group, which was administered subcutaneous fecal suspension (10 rats), the second group-fecal suspension and diabetes mellitus (10), the third group — simulated diabetes mellitus, fecal suspension and application of ozone (10). Diabetes mellitus was modeled by subcutaneous administration of alloxan (100 mg / kg body weight). Subcutaneously sterile 0.9% sodium chloride solution (1.0 ml per 100 g body weight) was injected to the control group (10). On the 14th day after administration of alloxan, animals were subcutaneously administered 10% fecal suspension (a fecal pool of 20 animals in 0.9% sodium chloride solution) at a dose of 0.5 ml per 100 g body weight. Rats of the first experimental group received subcutaneous administration of fecal suspension. Animals of the third group of the fecal mixture were subcutaneously injected and subsequently for three days, once a day, intraperitoneally administered 1.0 ml per 100 g of body weight of the animal once a day for 3 days, an ozonized 0.9% solution of sodium chloride was administered.Results. Application of ozonotherapy in the presence of soft tissue ulcers on the background of diabetes in the old rats was not accompanied by a significant improvement in the parameters of regulation of the aggregate state of blood. An increase in the manifestations of hypercoagulability was found: a decrease in the time of plasma recalcification, activated partial thromboplastin time, prothrombin time,thrombin time, Hageman dependent fibrinolysis, potential activity of plasminogen. The introduction of ozonized sodium chloride solution in purulent soft tissue processes against the background of diabetes in the old rats did not contribute to the tread properties, as indicated by the growth of lysis of azo-albumin, azocasein, total fibrinolytic activity, and Kunitz's protein activity in plasma. Ozonotherapy contributed to an increase in the degree of intoxication, in the presence of abscesses in diabetes in the old rats, as indicated by the increase in the concentration of medium molecules. Such changes in old rats can be considered as an aggravation of purulent-inflammatory process, an increase in intoxication, a decrease in tread properties, an increase in manifestations of hypercoagulation in the background of chronic CSD, caused by age-related changes. Conclusions. Application of ozonotherapy in conditions of purulent-inflammatory processes in diabetes mellitus in old rats does not show tread properties on hemostatic system and blood plasma proteolysis. Ozone therapy enhances manifestations of intoxication on the background of diabetes mellitus with suppurative and inflammatory proc treatmentesses in old rats, which causes the need for a restrained use of this method of treatment.

The role of factors of the hemostasis system and fibrinolysis in the development of gastroesophageal reflux in patients with chronic obstructive pulmonary disease

Gastroesophageal reflux disease (GERD) and chronic obstructive pulmonary disease (COPD) belong to common pathological conditions comorbid in 25–60 % of cases.Objective: to find probable mechanisms of GERD progressing against the ground of COPD by means of learning various links of the blood clotting system, anticoagulative blood activity and fibrinolysis for further improvement of the ways to correct the disorders found.Methods. 124 patients with COPD were examined including 90 with comorbid GERD. The control group included 24 practically healthy individuals of an appropriate age and sex. Clinical, biochemical, instrumental and statistical methods of examination were applied.Results. The patients were divided into the following groups: the 1st group – 34 patients with COPD, the 2nd group – 30 patients with COPD with endoscopically negative GERD, the 3rd group – 30 patients with COPD and endoscopically positive non-erosive GERD, the 4th group – 30 patients with COPD and endoscopically positive erosive GERD. The following results were found: while examining coagulation hemostasis – decrease of prothrombin time, fibrinogen content; while examining anticoagulation blood potential – decrease of thrombin time, activity of antithrombin III; while examining fibrinolytic blood activity – inhibition of the total fibrinolytic activity (at the expense of enzymatic activity decrease) and compensatory increase of non-enzymatic fibrinolytic activity, decrease of Hageman-dependent fibrinolysis activity.Conclusions. Disorders of coagulation hemostasis, anti-coagulation potential and fibrinolytic blood activity are intensified with the increase of severity of bronchial obstructive syndrome and promote the development of endoscopically positive erosive GERD

The neurokinin 1 receptor regulates peritoneal fibrinolytic activity and postoperative adhesion formation

BackgroundIntra-abdominal adhesions are a common source of postoperative morbidity. Previous studies in our laboratory have shown that a neurokinin 1 receptor antagonist (NK-1RA) reduces abdominal adhesion formation and increases peritoneal fibrinolytic activity. However, the cellular pathway by which the antagonist exerts its effects is unclear, as cultured peritoneal mesothelial cells exposed to the NK-1RA show increases in fibrinolytic activity despite having very low expression of neurokinin 1 receptor (NK-1R) messenger RNA and protein. Our aim was to determine whether the NK-1R plays an essential role in the adhesion-reducing effects of the NK-1RA, or if the NK-1RA is acting independently of the receptor. MethodsHomozygous NK-1R knockout mice and age matched wild-type mice underwent laparotomy with cecal cautery to induce adhesions. At the time of surgery, mice received a single intraperitoneal dose of either NK-1RA (25 mg/kg) or saline alone. Adhesion severity at the site of cecal cautery was assessed on postoperative day 7. In a separate experiment, peritoneal fluid was collected from wild type and NK-1R knockout mice 24 h after laparotomy with cecal cautery and administration of either NK-1RA or saline. Tissue plasminogen activator levels, representative of total fibrinolytic activity, were then measured in peritoneal fluid. ResultsIn wild-type mice, NK-1RA administration significantly decreased adhesion formation compared with saline controls. Among the NK-1R knockout mice, there was no significant reduction in adhesion formation by the NK-1RA. Fibrinolytic activity increased 244% in wild-type mice administered NK-1RA compared with saline controls; however, the NK-1RA did not raise fibrinolytic activity above saline controls in NK-1R knockout mice. ConclusionsThese data indicate that the NK-1R mediates the adhesion-reducing effects of the NK-1RA, in part, by the upregulation of peritoneal fibrinolysis, and suggest that the NK-1R is a promising therapeutic target for adhesion prevention.

UFEIII, a fibrinolytic protease from the marine invertebrate, Urechis unicinctus

A new serine protease with fibrinolytic activity from a marine invertebrate, Urechis unicinctus, was purified to electrophoretic homogeneity using column chromatography. SDS-PAGE of the purified enzyme showed a single polypeptide chain with MW ~20.8 kDa. Its N-terminal sequence was IIGGSQAAITSY. The purified enzyme, UFEIII, was stable at pH 6-10 below 60 °C with an optimum pH of 8.5 at approx. 55 °C. The enzyme activity was significantly inhibited by PMSF and SBTI suggesting that it was a serine protease. In fibrin plate assays, UFEIII was contained 1.46 × 10(3 )U (urokinase units) mg(-1) total fibrinolytic activity, which consisted of 692 U mg(-1) direct fibrinolytic activity and 769 U mg(-1) plasminogen-activator activity. Km and Vmax values for azocasein were 1 mg ml(-1) and 43 μg min(-1) ml(-1), respectively.