Activity of coagulation and fibrinolytic system components in the vein thrombus.

PURPOSE: Behaviour of the vein thrombus is determined by the activity ratio of coagulation factors to factors of fibrinolytic system. The aim of the study is to evaluate activity of some coagulation and fibrinolytic factors in the vein thrombus. MATERIAL AND METHODS: The activity of platelets aggregating factors, tissue factor, thrombin, antithrombins, antiheparin factors, plasminogen activators, plasmin (plasminogen) and antiplasmins of the vein thrombus homogenate was determined using coagulative, fibrinolytic and caseinolytic tests. Retracted blood clot was a compared material. RESULTS: Tissue factor activity in the vein thrombus was above twofold higher and antiheparin activity was nearly twice higher in comparison to the blood clot. The vein thrombus contains also active thrombin. Plasminogen activators activity in the vein thrombus was twofold higher and activity of plasmin (plasminogen) was threefold higher than in the blood clot. High activity of the tissue factor, substances neutralizing heparin and presence of thrombin intensify the thrombus enlargement. However, the thrombotic tendency may be balanced by a high activity of plasminogen activators and high activity of plasmin (plasminogen). CONCLUSIONS: 1) Vein thrombus is characterized by high activity of tissue factor, presence of active thrombin and high antiheparin activity. 2) High coagulative potential of vein thrombus is balanced to a certain grade by high fibrinolytic potential: high activity of plasminogen activators and high activity of plasmin (plasminogen), as well as absence of antiplasmins activity.

Chemical compound content and enzyme activity in supernatant and sediment of liver homogenate.

Evaluation was performed of chemical compound contents and enzyme activities in the whole homogenate, its supernatant and sediment. Six rabbit livers were pulverized in liquid nitrogen and homogenized. After centrifugation, the contents of protein, haemoglobin, vitamin A, vitamin E, vitamin C, as well as the activities of cathepsin B, cathepsin D, superoxide dismutase, catalase, glutathione peroxidase and reductase were assessed in the whole homogenate, its supernatant and sediment. Protein, vitamin A, superoxide dismutase, catalase, cathepsin D, glutathione peroxidase and reductase reveal uniform localisation. Vitamin C and cathepsin B are localized in supernatant, whereas haemoglobin is localized mainly in sediment. Evaluation of chemical compounds and enzyme activities should be performed in the whole homogenate, supernatant and sediment to obtain a real interpretation of biochemical disturbances in the investigated material.

Some components of oxidative-antioxidative system in human blood plasma and serum.

Comparison of the concentrations and activities of components in the oxidative-antioxidative system between blood plasma and serum. Blood plasma and serum samples were obtained from 38 healthy adults to evaluate malondialdehyde concentration, the total antioxidative capacity, superoxide dismutase activity, protein and non-protein sulphydryl groups, ascorbate, haemoglobin, methaemoglobin and protein. Blood plasma shows higher activity of superoxide dismutase, as well as higher concentrations of low-molecular sulphydryl groups and ascorbate, when compared to those in blood serum. The total plasma antioxidative capacity is also higher than that assessed in blood serum. Processes of blood coagulation and blood clot retraction lead to antioxidant consumption. The evaluation of oxidative-antioxidative system for diagnostic purposes should be performed in blood plasma.

Haemoglobin of varicose vein, varicose vein with thrombophlebitis and in parietal thrombus of varicose vein.

Saphenous veins were taken for examination: unchanged, varicose with thrombophlebitis and varicose thrombus. The contents of haemoglobin and protein were determined in the homogenate of that material. Only small quantities of haemoglobin were found in walls of unchanged veins. Greater amounts of haemoglobin were observed in walls of varicose veins, especially in walls of varicose veins with thrombophlebitis. The varicose vein thrombus also contained marked quantities of haemoglobin.

Coagulative and fibrinolytic activity in parietal thrombus of aortic aneurysm.

Lumen of aortic aneurysm is usually filled with parietal thrombus. Behaviour of the parietal thrombus is determined by the ratio of coagulation factors to factors of fibrinolytic system. Activity of some factors of coagulation and fibrinolysis in the parietal thrombus of aortic aneurysm was determined using coagulative, fibrinolytic and caseinolytic tests. Retracted, blood clot was a comparative material. Tissue factor activity in the parietal thrombus of the aneurysm was above threefold higher and antiheparin activity was nearly twice higher in comparison to the blood clot. Activity of plasminogen activators in the parietal thrombus was higher than in the blood clot. The parietal thrombus contained fourfold more of the plasminogen. Antiplasmin activity in the thrombus was above twofold lower than in the blood clot. High activity of the tissue factor and substances neutralizing heparin may intensify the thrombus growth. Yet the thrombotic tendency may be balanced by a high activity of plasminogen activators and plasminogen.

Lysosomal cysteine proteinases and their significance in pathology.

The lysosomal cysteine proteinases are synthesized in a form of pre-proenzymes. They are submitted to posttranslational glycosylation and phosphorylation. These modifications make possible transport the modified proteins into Golgi apparatus and into lysosomes. Some disturbances of transport which occur mainly in tumour cells result in an increase of these enzymes activities in cytosol and in intercellular compartment. The activity of cysteine proteinases is regulated by specific inhibitors (cystatin, kininogen) which exist in some tissues and body fluids. The evaluation of activity and concentration of proteinases and inhibitors is important in clinical diagnostics.

Biological and diagnostic role of cathepsin D.

Biosynthesis, posttranslating modifications, intracellular transport and activation of procathepsin D are discussed. Active cathepsin D evokes degradation of cellular and extracellular proteins and it also activates proenzymes, prohormones and growth factors and inactivates their active forms. Impairment of lysosomes in hypoxia or necrosis evokes transition of cathepsin D to cytosol and body fluids. Increase of cathepsin D content in cytosol also evokes enhancement of the synthesis rate observed among others, in neoplastic tissues and regenerating organs. Increase of cathepsin D content and activity in cytosol and blood serum is of essential diagnostic and prognostic importance in some pathologic conditions.

Activity of enzymes of various subcellular localisation in cytosol obtained after cell organelle precipitation at pH 5.0.

Acidification to pH 5.0 of various organ homogenates prepared in 0.25 M sucrose causes aggregation of cell organelles. Aggregated organelles are removed through standard centrifugation. Cytosol obtained reveals only slight activity of membrane and lysosomal enzymes. The cytosol is useful for evaluation of changes in enzyme distribution in cell.

Disturbances of hemostasis during declamping shock.

The aorta, above or below renal arteries was clamped for 60 minutes, in a canine model. The blood was taken for testing from above the aorta bifurcation before clamping, after 30 minutes of its duration, directly after declamping and every 30 minute during next 4 hours. Irrespective of clamping level, the platelet count, clot retraction and prothrombin consumption do not undergo significant changes. However, the activity of platelet factor 4 is increased. Prothrombin time, recalcination time, kaolin-kephalin time and the activities of factors V, VII, XI and XII do not differ as well. Thrombin time is prolonged and antithrombin III activity is reduced. Euglobulin fibrinolysis time undergoes prolongation and antiplasmin content is increased. The observed changes show a variable tendency, regardless of clamping level and increase with the passage of experiment time. An increase in the coagulation activity and a decrease in the fibrinolytic activity of the blood plasma may be a resultant of the changes. Finally it may promote thrombus formation and indicates the preventive use of heparin.

Hemostatic activity of organs in declamping shock.

The aim of the study was to determine the haemostatic components activity of organs in declamping shock. The abdominal aorta was cross-clamped below or above renal arteries. No significant changes in tromboplastic and antithrombin activities were found in the kidney, liver, lung, heart and skeletal muscle. Renal cortex and medulla as well as the lungs show higher plasminogen activator activity and considerably higher antiplasmin activity. Diminished fibrinolysis in the kidney and the lung may promote thrombotic complications.

Activity of membrane, cytosol and lysosome enzymes in organs and blood serum during declamping shock.

The aim of the study was to evaluate the enzyme activity of cellular membranes (GGT), cytosol (ALT, AST) and lysosome (AP, AcP) in the cytosol, whole homogenate and blood serum during declamping shock, following release of abdominal aorta cross-clamping. The aorta was clamped for 60 minutes. An increase in GGT, AP and AcP activities in the cytosol and whole homogenate of the renal cortex, renal medulla, liver, lung, heart and the skeletal muscle occurs after declamping. Rise in the enzymatic activity, especially of acid phosphatase is higher when the aorta above renal arteries was clamped. However, its activity in the blood serum remains unchanged. Alterations in the distribution and the activity of the studied enzymes may indicate that aortic clamping damages the endoplasmic reticulum and lysosomal membranes. Yet, cellular membranes preserve their structural and functional integrity.

Cathepsin D activity and protein degradation products in organs and blood during experimental declamping shock.

Aortic cross-clamping in dogs for 60 min causes an increase in cathepsin D activity in the kidney, liver, lung, heart, skeletal muscle and the blood serum. It causes no changes in the content of protein and its degradation products of the examined organs, apart from the lungs, where the above parameters are higher. The intensity of the observed changes in the kidney and other organs does not depend on the level of aortic cross-clamping (above or below renal arteries).

[Effect of different vascular prosthesis materials on blood platelets, blood clotting and fibrinolysis]. / Wplyw tworzyw róznych typów protez naczyniowych na plytki krwi, uklad krzepniecia i fibrynolize.

Studies were performed on Dacron vascular prostheses (USA), Gore-Tex (Germany), polyester prostheses (Czechoslovakia), Lawsan (Russia), as well as on DALLON-standard and DALLON-double velour prostheses (Poland), non-impregnated or impregnated with collagen, with heparin and albumin, with antithrombotic and antibacterial preparation, and with antibacterial preparation. In vitro, all the materials of vascular prostheses cause an adhesion of platelets in different degree, increase the availability of platelet factor 3, release platelet factor 4, reduce the blood clot retraction, shorten clotting time of the whole blood as well as recalcination time of intact plasma and increase the prothrombin consumption. None of the materials induced platelet aggregation nor influenced the activity of fibrinolytic system. Prostheses impregnated with collagen or with heparin and albumin were the most thrombogenic. Those impregnated with antibacterial preparation or with antibacterial and antithrombotic preparations, and Gore-Tex prostheses showed the lowest thrombogenicity. Thrombogenicity of vascular prostheses not only depends on chemical structure of the material, but on the method of fibre manufacturing by individual producers as well.

[Changes in the activity of the clotting system and fibrinolysis components in the layers of double velour DALLON grafts at different periods after implantation into the aorta]. / Zmiany aktywnosci skladników ukladu krzepniecia i fibrynolizy w warstwach przeszczepów DALLON-dwustronny welur w róznym czasie po wszczepieniu do aorty.

During one year experiment, the thromboplastic, antiheparin, plasminogen activator, antithrombin and antiplasmin activities were evaluated in the homogenates of the intimal, medial and adventitial layers of polyester double velour DALLON prostheses implanted into the dog abdominal aorta. It was found, that 7 days after implantation the thromboplastic, antiheparin and plasminogen activator activities in DALLON graft neointima were high, whereas those of the antiplasmin and antithrombin were low. These changes facilitate the sealing of the prosthesis pores and at the same time prevents thrombosis. Four months after implanting, the activity of hemostatically active components in the various graft layers became similar to the activity of aorta components. Rapid decrease of the thrombogenic potential and increase of the fibrinolytic activity in layers of double velour DALLON grafts facilitate the maintenance of graft patency.