EVALUATION OF THE FUNCTIONAL STATE OF LIVER AND THE EFFICIENCY OF THERAPY FOR ENTEROPATHY OF CALVES.

Comprehensive study of hepatospecific biochemical blood markers and haemostatic system in calves which recovered from gastrointestinal pathology at age 2-7 days, was con- ducted. Implementation of a complex of tests for evaluation of the liver's functional state and the efficiency of therapeutic regimens shown that the experimental animals compared to control at the 30th day of life demonstrated significantly increased conjugated bilirubin concentration (1.6 times), aspartate- and alanin aminotransferase activity (1.2 times), gamma-glutamyl transpeptidase (1.5 times) and alkaline phosphatase (1.4 times), and content of soluble fibrin monomer complexes (4 times). Such deviation from the norm of the mentioned parameters of liver's functional state suggests the need to monitor the liver state even 3 weeks after their clinical health is confirmed. To prevent the development of secondary hepatopathology it is recommended to conduct further medical correction of the functional state of the liver. It is found that to stimulate recovery of liver function in case of toxic dyspepsia in newborn calves, it is advisable to implement a phospholipid containing supplement

[Evaluation of the blood coagulation system after surgeries on abdominal aortic aneurysms].

Basing on data of analysis of the hemostasis system state in the patients, suffering abdominal aorta aneurysm, a tendency for raising of postoperative soluble fibrin and D-dimer content in the blood plasm and reduction of these indices on the third day was noted. The abovementioned markers content depends on the aneurysm size, the fibrin deposits presence, the terms from clinical signs beginning to the certain therapy administration and anticoagulants application. Information about correlation between content of D-dimer and soluble fibrin in the treatment dynamics is important for determination of activation degree in the patients blood coagulation system and the thrombotic complications prognosis.

[Assessment of protein synthesizing function of the liver in experimental hepatitis].

Liver protein synthesis was estimated comparing the levels of prothrombin and its inactive form PIVKA-prothrombin. The latter indicates liver dysfunction. These diagnostic tests allow monitoring the effectiveness of the commonly applied preparation "Essentiale Forte" and that of the liposomal form of the biologically active additive (BAA) FLP-MD based on phospholipids of various origin.

[Effect of protein C activator on overall haemostasis potential in donor and hip arthroplasty patient plasma].

A method of overall haemostasis potential (OHP) determination for quantitative and rapid estimation of coagulation-fibrinolysis balance in plasma has been presented. The method is based on the analysis of the absorbance at 350 nm vs. time curve, which records the clot formation and dissolution in plasma in the presence of thromboplastin and t-PA. Three parameters of coagulation system--time, rate of formation and maximal turbidity of the clot, and three parameters of fibrinolytic system--half-, full-time and maximal rate of the clot dissolution, and main integral parameter--the area under the curve that characterizes the size and time of the clot existence and expresses OHP of plasma, can be determined by this method. It was shown that OHP value of patients plasma was 3.8 times more than that of the donor plasma. It is in concordance with elevated level of Fg (4.25 mg/ml), soluble fibrin (50 microg/ml), D dimer (630 ng/ml) and insufficient decrease of APC activity (93%) in patients. AcPC, added to donor and patient plasma, reduced OHP value 1.6 and 3.7 times, correspondingly. AcPC increased amidase activity of thrombin and APC in the donor plasma, and did not change that of plasmin. These data indicated that the effect of AcPC on OHP is mediated by formation ofAPC that helps to reduce the level of inhibitors in the investigated plasma.

[Influence of prothrombin cleavage products on platelet activation and aggregation].

Meizothrombin efficiently activates mechanically triggered platelets and enhance collagen-, ADP- and adrenalin-induced aggregation of platelet-rich blood plasma. Thus, in clotting system activation zone meizothrombin is able to enhance process of involving platelets in clotformation. Pre-thrombin 1 inhibits collagen-, ADP- and adrenalin-induced aggregation of platelet-rich blood plasma and so regulates not only plasm but platelet hemostasis by reverse negative relation.

[Process of complexation of prothrombine and fibrin E-fragment].

The study is devoted to research of the blood coagulation key proenzyme complexation process. It is prothrombin, and the E-fragment of fibrin can be a component in blood circulation. It is shown, that non-enzyme activation of prothrombin by the E-fragment proceeds as a result of formation of stable non-covalent prothrombin-E fragment complex. The cringle structures of prothrombin and the N-terminal site of beta-chain of E-fragment of fibrin are important for formation of the given complex. It has been defined, that other fragments of fibrin (D and DD) are not capable to induce amydolytic activity of prothrombin.

[Effect of soluble fibrin on the blood coagulation process and platelets aggregation].

The accumulation of soluble fibrin (SF) in the blood plasma causes acceleration of the final stage of blood coagulation. It increases functional activity of a hemostasis system platelet link, that is the precondition of thrombotic complication. Accumulation of SF in the blood plasma is accompanied by proportional reduction of coagulation time in ancistron and thrombin time tests, and also the intensification of platelets aggregation process. A conclusion was drawn that for early diagnostics of the DIC-syndrom it is expedient to carry out complex estimation of the hemostasis system with obligatory definition of the blood SF content, performance of ancistron and thrombin time tests, and also study of platelets aggregation.

[Characteristics of D-domain-containing specific inhibitors of polymerization on the fibrin self-assembly process].

The inhibitory effect of D- and DD-fragments and their mixture on plasma blood coagulation process under the action of thrombin has been studied. The significant increase of total efficiency of the inhibitory action has been shown. The inhibitors have influence on a lag-phase and lateral association of protofibrills of fibrin polymerization carried out by turbidimetry method in the model systems in vitro. The concentration of fragments which are close to parameters of fibrinogen/fibrin degradation products in pathological conditions was used. It is shown, that the covalent cross-link of D-dimer gamma-gamma-chains affects the inhibitory features and mechanism of interaction with fibrin monomer molecules. D-fragments inhibit the both phases of polymerization, whereas DD-fragments effectively inhibit the lateral association of protofibrill.

[Inhibition of the process of Glu-plasminogen activation by tissue activator with fibrin, DDE-complex, and D-dimer using alpha-2-antiplasmin].

The alpha-2-antiplasmin influence on the Glu-plasminogen activation by tissue activator both on fibrin and fibrin(ogen) fragments was investigated. The kinetics of activation was studied and velocity of this process in the absence and presence of the inhibitor was calculated. It was established that alpha-2-antiplasmin decreased the velocity of Glu-plasminogen activation on desAABBfibrin, DDE-complex and DD-dimer and did no influence upon proenzyme activation on fibrinogen fragment--Ho1-DSK. In the presence of fibrin plasminogen activation linear related to the amount added tissue activator in limit concentration from 5 before 50 units/ml. It was shown that alpha-2-antiplasmin reduced the activation velocity with used concentration of tissue activator. Fibrin hydrolysis by plasmin, forming on its surface during the plasminogen activation by tissue activator, was also inhibited with alpha-2-antiplasmin. The obtained results are explained by the influence of the inhibitor on formation of the triple complex between plasminogen, tissue activator and fibrin, and competition of the alpha-2-antiplasmin for lysine-binding sites of tissue activator kringle 2 or for binding sites of the activator on fibrin.

[The role of fibrin clot degradation products in the regulation of vital functions of PC-12 cells].

The products of the fibrin clot hydrolysis performed by PC-12 cells modulated dose-dependently the rate of cell proliferation and favored their survival when seeded in suboptimal density. Co-incubation of PC-12 cells with fibrin degradation products enhanced cell adhesion to tissue culture plastic, as well as the number of nicotinic acetylcholine receptor alpha3 and alpha5 subunits expressed. It was demonstrated that, in fact, such a heterogeneous and comprehensive influence was a sum of effects exerted by different fibrin fragments. Low molecular weight fraction (below 30 kDa), but not a purified alphaC-domain, stimulated PC-12 cell proliferation, diminished their adhesion to plastic, increased nicotinic receptor expression and caused processes outgrowth. On the contrary, high molecular weight products, in particular D, DD and E fragments, enhanced PC-12 adhesion to plastic and, as a result, slowed cell division. Both high and low molecular weight fragments favored the survival of PC-12 cells. These results showed that fibrin degradation products support the vital functions of neuron-like cells, favor their contacts with extracellular surrounding and act as neurotrophic factors.

[Effect of tissue-type plasminogen activator and its inhibitor in haemostasis system function in norm and pathology]. / Rol' tkanynnoho aktyvatora plazminohenu ta ioho inhibitora u funktsionuvanni systemy hemostazu za normy ta patolohiï.

The paper is dedicated to the study of structure and physiological functions of tissue type plasminogen activator and its inhibitor PAI-1, changes of these parametres in normal and pathology conditions. The interrelation of the coagulation and the fibrinolytic system during the range of pathologies was studied. It was demonstrated that simultaneous analysis of the coagulation and fibrinolytic systems analysis allow to diagnose the thrombotic complications.

[PC-12 cells hydrolyze the fibrin clot by producing both plasminogen and its tissue activator]. / Klityny PC-12 hidrolizuiut' fibrynovyi zhustok shliakhom produktsiï plazminohenu ta ioho tkanynnoho aktyvatora.

It has been shown that rat pheochromocytoma PC-12 cells degraded fibrin clots in vitro. SDS-PAGE performed in the gels of different density and Western blotting using monoclonal antibodies against DD- and D-fibrin fragments demonstrated that both high and low molecular weight degradation products similar to those of fibrin hydrolysis by plasmin had been formed. Enzyme electrophoresis, chromogenic assay and enzyme-linked immunosorbent assay using tPA-specific antibodies demonstrated that PC-12 cells constitutively secreted both plasminogen and tPA. The results obtained allow using PC-12 cells as a model to examine the interaction of nerve cells with the fibrin clot.

[Activation of key proenzymes of the blood coagulation system by the fibrin E fragment]. / Aktyvatsiia kliuchovykh profermentiv systemy zsidannia krovi frahmentom E fibrynu.

Plasminogen and prothrombin are key proenzymes of fibrinolytic and clotting system. It is known that they can be activated by indirect activators streptokinase and staphylocoagulase. In this paper it is shown that fibrin E fragment purified from DD-E complex induced catalytic activity in plasminogen and clotting activity in prothrombin. Streptokinase increased 2 times the rate of catalytic activity induction by E fragment in prothrombin. It's possibly that process is one of the factors providing for rethrombosis after thrombolytic therapy of myocardial infarction.

[Significance of the activity of indicators of the fibrinolytic system on assessment of the hemostatic state]. / Znachymist' deiakykh pokaznykiv fibrynolitychnoï systemy v otsintsi stanu hemostazu.

Complex analysis of pregnant women haemostasis system before and after caesarian section allowed find the coagulation system activation. It was shown the thrombotic markers accumulation, AT III and protein C levels decrease. Also change of ratio fibrinolytic system components was exposed. Definition of t-PA and PAI-1 activities can be used as prognostic markers of the fibrinolytic chain haemostasis system disfunction.

[Laboratory diagnosis of the status of the hemostasis system]. / Laboratorna diahnostyka stanu systemy hemostazu.

Activators of fibrinogen, prothrombin and protein C isolated from venoms of Agkistrodon halys halys and Echis multisquamatus may be used as a tool both for thrombosis investigations in the model systems and for diagnostic in the clinical practice. The complex of diagnostic tests developed on the base of these activators allows to characterize the haemostatic system at different pathologies and as well as to determine the unbalance between separate components of haemostasis. The tests are approved on plasma of patients with heart diseases, ulcers, nephrites, hestoses etc. The tests are sensitive, informative, easy to use and do not require an additional equipment. They have no analogues in Ukraine.

[A rapid turbidimetric micromethod in the determination of hemostatic and fibrinolytic disorders in arterial hypertension patients of different ages]. / Turbidymetrychnyi ekspres-mikrometod u vyznachennia porushen' hemostazu ta fibrynolizu u khorykh na Arterial'nu hipertenziiu riznoho viku.

Disorders of haemostasis and fibrinolysis were studied in 54 patients of various age with arterial hypertension by turbidimetric express micromethod. Activation of coagulative system and inhibition of fibrinolytic activity tending to get more pronounced with age were found in patients with hypertonic disease at II stage and systolic arterial hypertension. Turbidimetry has considerable advantages: it can be performed rapidly, requires as much as 0.1 ml of plasma for the assay and provides complete information about the state of haemostasis and fibrinolysis. The method may be important in detecting minor signs of disseminated intravascular coagulation in patients with hypertonic disease.

[Quantitative determination of protector activity of fibrinogen tryptic hydrolysates]. / Kil'kisne viznachennia protektornoi aktivnosti triptichnikh gidrolizativ fibrinogenu.

A method is developed for the quantitative determination of the protector activity. The method is based on registrating the coagulation time shortening for a mixture of fibrin monomer and fragment D. All determinations should be carried out at high concentrations of fragment D when its inhibitory effect on fibrin polymerization is very strong. It is necessary to control the exact conditions as to ionic strength and pH of the reaction mixture. Another approach to the protector activity estimation is demonstrated. It is based on determination of clot protein quantities after fibrin monomer clotting in mixtures with constant fragment D and different protector quantities.

[Biologically active low-molecular fragments of fibrinogen molecule]. / Biologichno aktivni niz'komolekuliarni fragmenti molekuli fibrinogenu.

A peptide fraction with a specific property of interaction with D fragment was found in the fibrinogen tryptic hydrolyzate. This fraction is called a protector. In the protector presence the time of the monomer fibrin polymerization prolonged with D fragment is greatly lowered. The protector was found in the first minutes of fibrinogen degradation. It was shown for low concentration of trypsin (enzyme-substrate ratio being 1:7500). The gel filtration of a tryptic fibrinogen hydrolyzate on Sephadexes G-10, G-15, G-25 showed that the protector has a low molecular weight. It was found that a preincubation of the protector with D fragment should take place before interaction of their mixture with a fibrin monomer. The interaction of the protector with D fragment is reversible.