[Effect of proteins from the Agkistrodon blomhoffii ussuriensis snake venom on platelets in the hemostasis system].

Influence of proteins from the Agkistrodon blomhoffii ussuriensis snake venom on platelet activation and aggregation was developed on different model systems in vitro. It was shown that novel disintegrin (Blomus-B) and phospholipase A2 (Blopholipase) from the venom, activated platelets and inhibited their aggregation. Fibrino(geno)lityc enzyme (Blomulyse) does not activate platelets and has no effect on their aggregation stimulated by collagen, but inhibit ADP and adrenalin-stimulated platelet aggregation. Thrombin-like enzyme (Ancistron-Bu) activates platelets but has no effect on their aggregation. Obtained proteins can be used under development of new antiplatelet agents and as instruments for detailed elaboration and deep investigation of processes which proceed with participation of platelets.

[Process of platelets activation by streptokinase].

This study concerns the influence of streptokinase and antistreptokinase antibodies on rabbits platelets in blood plasma depleted of plasminogen. The immune complex streptokinase-antibody causes platelets activation, whereas other investigated immune complexes didnot express such activity. Platelets aggregation wasnot detected in any case. It was determined that streptokinase induces platelets activation in the rabbit plasma with high titre of antistreptokinase antibodies in absence of plasminogen.

[Effect of streptokinase on the content of plasminogen activator inhibitor of type 1].

The influence of streptokinase on plasminogen activators inhibitor of type I (PAI-1) was investigated with the use of model systems in vitro and in vivo. It was defined, that intravenous streptokinase injection causes an increase in PAI-1 content in mammals' blood plasma. Experiments in vitro have shown that the increase in PAI-1 concentration takes place as a result of streptokinase action. It occurs due to platelets activation with subsequent PAI-1 secretion from their a-granules. It is established, that PAI-1 is secreted by platelets both in free and in complex forms. The data obtained in the work, allow to assume, that the simultaneous substantial increase in PAI-1 content with platelets activation, as a result of streptokinase influence, can lead to new thrombotic complications risk.

[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.

[Characterization of platelets aggregation inhibitor from Agkistrodon blomhoffii ussuriensis snake venom].

Platelet aggregation inhibitor--"Blomus-B" from Agkistrodon blomhoffii ussuriensis venom has been isolated by affinity and ion-exchange chromatography. The purified inhibitor is a novel non-enzymatic single-chain protein with molecular weigth of 13 kDa. "Blomus-B" causes a change of platelets shape and takes effect on ADP- and adrenalin-induced platelet aggregation.

[Plasminogen activator from Agkistrodon halys halys venom].

Plasminogen activator "Ahh-32" from Agkistrodon halys halys venom has been isolated and purified using affinity and ion-exchange chromatography. The purified enzyme consists of the single peptide-chain with molecular weigth of 32 kDa. It can convert free plasminogen into active form--plasmin. "Ahh-32" was inhibited by DFP and benzamidine. Besides, the enzyme influences significantly the activation of plasminogen by streptokinase without having effect on analogical process in case of usage of tissue tipe plasminogen activator. The obtained protein can be used as an instrument under investigation of protein-protein interactions in haemostasis system.

[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.

[Effect of streptokinase on the parameters of haemostasis in model systems in vivo]. / Vplyv streptokinasy na parametry systemy hemostazu v model'nykh systemakh in vivo.

The changes of the haemostasis system parameters were studied under treatment by streptokinase (Sk, thrombolytic agent of bacterial origin) with the use of model systems in vivo. We were especially interested in the study of the changing fibrinolytic system parameters such as tissue type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1), plasminogen, alpha2-antiplasmin activities. After treatment by Sk the activity and quantity of t-PA were significantly increased while the activity of PAI-1 was decreased. The parameters of haemostasis system evidenced for a disbalance which occurred under treatment by Sk: decrease of fibrinogen levels and appearance of soluble fibrin and accumulation of fibrin degradation products. Thus analysis of the data has displayed besides the well-known Sk function the influence of Sk on the change of fibrinolytic system potential probably due to its effect on the endothelial cells.

[Investigations of fibrin clots formation under the effect of thrombin and ancystron]. / Doslidzhennia formuvannia fibrynovykh zgustkiv za diï trombinu ta antsystronu.

It was shown that heterogeneous surfaces of two phases influence on fibrin films structures. The clot formed between two phases have the surface layer which is different from fibrin clot surface structure. Transmission electron microscopy revealed to determine the thickness of the surface layer for desAA- and desAABB-fibrin. The surface layer thickness is 25,8 and 29 nm for desAA- and desAABB-fibrin respectively.

[Binding of alpha-2-antiplasmin with fibrinogen/fibrin and their fragments independent of factor XIII]. / Zv'iazuvannnia alfa-2-antyplazminu z fibrynohenom/fibrynom ta ïkhnimy frahmentamy bez uchasti faktora XIII.

Possible interaction of alpha-2-antiplasmin with fibrinogen, fibrin and their fragments independent of factor XIII as well as the inhibitor effect on the Glu-plasminogen activation by tissue activator were studied. It was shown that alpha-2-antiplasmin is adsorbed on desAA- and desAABBfibrin films (Kd 69.0 +/- 1.0 nM 68.6 +/- 5.3 nM, respectively). Glu-Plasminogen has no effect on the inhibitor binding with desAABBfibrin. Alpha-2-antiplasmin shows strong affinity for fibrin D-dimer (Kd 65.0 +/- 4.0 nM) and D-fragment of fibrinogen (Kd 119.0 +/- 21.0 nM), but it does not interact with E-fragment. The inhibitor inside the fibrin clot decreases 10 times the activation rate of Glu-plasminogen by the tissue activator both is the presence and without factor XIII at physiological ratio of Glu-plasminogen, tissue activator, fibrin and alpha-2-antiplasmin. Thus we have shown that fibrinogen/fibrin binds alpha-2-antiplasmin independent of the factor XIII. Binding sites of the inhibitor are localized in D-fragment of fibrinogen and/or fibrin D-dimer. Alpha-2-antiplasmin inhibits the Glu-plasminogen activation by tissue activator on fibrin.

[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.

[Structure-functional analysis of peptide P3 identical to gamma370-383 binding sites with integrin alphaIIbbeta3 in gammaC-domain of fibrinogen]. / Strukturno-funltsional'nyi analiz peptydu P3, shcho ie identychnym do gamma370-383-dilianky zv'iazuvannia z intehrynom alphaIIbbeta3 u gammaC-domeni fibrynohenu.

The interactions between platelet integrin alpha IIb beta 3 and fibrinogen (Fg) mediate a range of adhesive reactions, which are necessary for platelet aggregation and fibrin clot retraction. The binding site for alpha IIb beta 3 resides in the gamma C domain of Fg. In our previous work we have identified a novel binding site in the gamma C domain, gamma 370-383 (P3), for integrin alpha IIb beta 3 and have demonstrated that the P3 sequence together with the C-terminal gamma C sequence 408AGDV411 accounts for the full binding of alpha IIb beta 3. In our present study, in order to define the amino acid residues in P3 involved in the interaction with alpha IIb beta 3, we have used SPOT-synthesis analyses. Libraries consisting of peptides covering P3 were created and probed with radiolabeled alpha IIb beta 3. Screening of the libraries showed that several positively charged residues may be critical for interaction of P3 with integrin alpha IIb beta 3.

[Specificity of endometrium phospholipids and their fatty acids changes in the development of hyperplasia]. / Spetsyfichnist' zmin vmistu fosfolipidiv i ïkh zhyrnykh kyslot v endometriï pry rozvytku hiperplaziï.

Quantitative changes in phospholipids (PL) and fatty acids content (FA) in patients with endometrium hyperplasia have been revealed. During observing endomentrium hyperplasia process we found out on the one hand reliable decrease in phosphatidylethanolamine (PEA), phosphatidylcholine (PC) content and on the other hand increase of its lysoforms. Changes in unsaturated fatty acids (PA n-6)--arachidonic (AA) content and docosahaenoic (DHA) acids content were also shown. The decrease of PC, PEA in patients with endometrium hyperplasia is interrelated with quantitative change of plasmalogenic phospholipids PL, which is too much sensitive to oxidative reactions and at the same time able to protect sells against oxidative stress. Considerable decrease in plasmenyl PEA content on the one side and plasmenyl PC content stability on the other was revealed in patients with endometrium hyperplasia. Above mentioned lipid metabolic derangements of the endometrium are specific with respect to different classes of phospholipids and their fatty acids composition.

[Effect of phospholipids containing omega-3 fatty acids on structural changes of microsomal lipids in cell membranes of functionally different cells]. / Vplyv fosfolipidiv, shcho mistiat' omeha-3 zhyrni kysloty, na strukturni zminy lipidiv mikrosom membran klityn, riznykh za funktsiieiu.

As a result of the experimental researches conducted it has been shown that administration of some normal animal marine phospholipids (PL) including in their structure omega-3 polyunsaturated fatty acids (PUFA) provides for quantitative changes of individual PL, fatty acids (FA) content and quantity in general and individual PL of liver, heart, brain and gonads microsomes. While estimating general microsomal PL fraction FA content under the action of PL omega-3 PUFA FA concentration change, unsaturation index (omega 6/omega 3) and relation of arachidonic acid to docosahexenic (AA/DHA) decrease have been identified. The decrease of AA/DHA relationship occurs due to AA and DHA quantitative changes. In the case of AA increase in some tissues there is observed the decrease of docosapentaenic acid and increase of DHA and eucosapentaenic (EPA) acidds. As a result of studying FA content in the individual PL composition it has been identified that certain PL classes characteristic for some tissues respond by changes of some certain FA. The relationship omega 6/omega 3 has been shown as decreasing in phosphatidilcholine (PC) all tissues microsomes (liver, gonads, heart, brain), in phosphatidilethanolamine (PEA) of liver and cardiac microsomes, in phosphatidilserine (PS) this relationship relationship decreases in the liver, brain and heart, for phosphatidilinositole (PI) the changes take place in liver, gonads, brain. Simultaneously, the decrease of AA/DHA relationship in the individual PL decrease of AA and increase of EPA and DHA depend on the tested tissues. The marine phospholipids might be supposed to render their effect on AA metabolism resulting in AA/DHA relationship in PEA and PS relationship displays itself as specific and depends on the tissues functions. The preference of PEA and PS use by certain tissues microsomes could be explained by their membrane protective capability.

[Pilot plant and experimental laboratory production. The role in biotechnology industry development]. / Pilotnyi zavod ta eksperymental'ne vyrobnytstvo. Rol' u rozvytku biotekhnologichnoï promyslovosti.

A stage-phase approach can contribute to unnecessarily long product development time. A simultaneous approach that integrates all development resources through an effectively managed pilot plant can significantly shorten the product development cycle. An intensive development of the domestic biotechnology manufacturing is impossible without creation of the real pilot plant market in Ukraine.

[Phospholipids of human spermatozoa and their role in ensuring fertility]. / Fosfolipidy spermatozoïdiv liudyny ta ïkh rol' u zabezpechenni fertyl'noï spromozhnosty.

The phospholipid composition of spermatozoa in healthy subjects (n = 13) and infertile men (n = 38) was studied. The level of inorganic phosphorus of total phospholipids decreased in spermatozoa of infertile men (relative and excretory infertility). It was found that the phosphatidyl ethanolamine amount in spermatozoa of infertile men fell. The level of phosphatidyl serine had certain tendency to decrease as well. Lyso-phosphatidyl serine was detected in some samples of infertile spermatozoa. Correlation between the amount of spermatozoa, its motility, vitality etc. and content of some phospholipids was found.

[Phospholipids of human seminal plasma and their role in ensuring fertility]. / Fosfolipidy semynal'noï plazmy liudyny ta ikh rol' zabezpechenni fertyl'noï spromozhnosty.

The phospholipid composition of seminal plasma from health (n = 13) and infertile men (n = 49) was studied. A significant decrease of lyso-phosphatidyl choline and lyso-phosphatidyl serine levels in seminal plasma of men with relative infertility was shown. Two-fold enhancement of phosphatidyl serine in seminal plasma of men with excretory infertility was found. A significant decrease of lyso-phosphatidyl choline in men with associated infertility was determined. An essential decrease of lyso-phosphatidyl serine and lyso-phosphatidyl choline in men with secretory infertility were shown. The level of phospholipids correlated with some morphological and functional characteristics of ejaculate.

[Lipid composition and fertility of human ejaculate]. / Lipidovyi sklad ta fertyl'na spromozhnist' liuds'koho eiakuliatu.

The morpho-functional indexes of ejaculates from 35 healthy and 145 infertile men were studied. The depression of functional sperm activity was found in the majority of infertile men. Phospholipid analysis of whole ejaculates from 12 healthy and 35 infertile subjects was performed. It was shown that inorganic phosphorus of total phospholipids decreased in ejaculates of men with secretory infertility. Lyso-phosphatidyl choline (LPC) was not detected in ejaculates of men with relative infertility. The amount of lyso-phosphatidyl ethanolamine (lyso-PE) and sphingomyelin in whole ejaculates of those patients decreased. The levels of LPC and lyso-PE decreased in ejaculates of persons with associated infertility. The level of phosphatidyl inositol, LPC and PE decreased in ejaculates of men with secretory infertility. The gas-lipid chromatography analysis of fatty acids composition of whole ejaculates of the infertile men showed significant changes in quantity of fatty acids. The quantity of docosahexaenoic acid in ejaculates of infertile men decreased and positively correlated with motility of spermatozoa.