A new approach for creating double-stranded DNA biosensors.

The principle of 'sandwich'-type biosensors based on liquid-crystalline dispersions formed from [DNA-polycation] complexes is outlined. These biosensors will find application in the determination of a range of compounds and physical factors that affect the ability of a given polycationic molecule to maintain intermolecular crosslinks between neighbouring DNA molecules. In the case of liquid-crystalline dispersions formed from [DNA-protamine] complexes, the lowest concentration of hydrolytic enzyme (trypsin) detectable was approximately 10-(14) M.

Investigation of long-term hypocoagulability in rats after intratracheal administration of unfractionated heparin.

The comparative study of intratracheal and intravenous administration of heparin on blood coagulation and mast cell population was carried out in experiments in rats. Unlike the intravenous injection of heparin which induces fast short-time inactivation of all clotting factors, single intratracheal injection of heparin inactivated the intrinsic pathway of thrombin generation. Long-term hypocoagulability after intratracheal administration of heparin was related to the accumulation of heparin in mast cell.

Blood coagulation changes in pregnant women at risk of developing preeclampsia.

OBJECTIVES: A group of 50 pregnant patients were studied with regard to the time of onset of hypercoagulation, its character and mechanisms of initiation in the development of preeclampsia. METHODS: The blood coagulation characteristics, antithrombin (AT)-III and endogenous heparin levels were studied. All of the patients were designated at risk of developing preeclampsia at the time of registration at the antenatal clinic. RESULTS: Retrospective evaluation shows that endogenous heparin levels and AT-III activity decreased by nearly 50% within 15 days of the development of preeclampsia and were particularly low 1-7 days before the onset of clinical signs of preeclampsia. CONCLUSIONS: These findings indicate that abnormalities in AT activity are a marker of hypercoagulation and begin more than 2 weeks before the development of clinical signs of preeclampsia.

[Nucleic acid synthesis and spatial organization of DNA in the presence of heparin and the quaternary ammonium salt of the conidin oligomer-25]. / Sintez nukleinovykh kislot i prostranstvennaia organizatsiiaDNK v prisutstvii geparina i chetvertichnoi ammoniinoi soli oligomera-25 konidina.

A comparative study of polyconidin effects in the replicative and transcription activity of hepatocyte DNA in intact animals, and on peculiarities of spatial organization DNA in the DNA complex with oligomer-25 conidin was carried out. It was shown that polyconidin binding to DNA results in "cross-linking". This process is accompanied by formation of liquid-crystalline dispersion without abnormal optical activity. Liquid crystals possess high density packing of DNA molecules complexed with quaternary ammonium salt of oligomer-25 conidin. The addition of heparin to liquid-crystalline dispersions or phases, destroys the structure of the DNA complex with quaternary ammonium salt of oligomer-25 conidin. As a result "free" DNA molecules appear, they form the cholesteric liquid-crystalline phase. An increase in transcription activity and synchronization of DNA synthesis in hepatocyte was demonstrated. A correlation between chromatin modification and biological activity of chromatin after formation of the DNA--oligomer-25 conidin complex is proposed.

[The effect of synthetic polycation of the quarternary ammonium salts of polymetacryoil lupinin on the ionic permeability of erythrocyte membrane]. / Deistvie sinteticheskogo polikationa chetvertichnoi ammoniinoi soli polimetakriloil lupinina na ionnuiu pronitsaemost' membran éritrotsitov.

It is shown that synthetic polycation quarternary ammonium salt polymetacryloil lupinine (poly-MACL) effects on passive ion permeability of erythrocytes membranes. The amplitude of Cl-/OH- exchange decreased in the buffer with SDS-polycation complex and simultaneously with pH elevation K+ exflux increased. On the contrary, in presence of heparin co-effect of poly-MACL and SDS vanished. Thus, synthetic polycation in the complex with long-chain anion (SDS) injure the membranes and effect depends on lipophilic-hydrophilic balance of polycation-SDS complex.

[Morphometric analysis of a mast cell population]. / Morfometricheskii analiz populiatsii tuchnykh kletok.

A composite morphometric approach to the analysis of mast cell population is proposed including 4 criteria: the cytogram, the index of heparin saturation, degranulation index, a relative rate of degranulation form. The sensitivity and informative capacity of the composite morphometric analysis were tested in two experimental models: inoculation of heparin and inoculation of its antagonists. Subtle differences in the effect of mast cell degranulaton were found after inoculation of protamine sulphate and a synthetic polycation, 2.5-ionen. Important information on the consumption of exogenous heparin by mast cells was obtained. The simplicity of the analysis as well as the possibility of using its simplified version make the method available for both clinical and experimental pathology.

[Rheologic properties of the blood and the system of hemostasis in patients with circulatory encephalopathy treated by hemosorption]. / Reologicheskie svoistva krovi i sistema gemostaza u bo'lnykh dis- tsirkuliatornoi éntsefalopatiei pri lechenii gemosorbtsiei.

In patients with atherosclerotic encephalopathy (AE) the major indices of blood rheology, hemostasis and its cells' functional state were under investigation. Different patterns of blood rheology changes were detected with increased counts of pathological red blood cells and hypercoagulability. These disorders correlated with the rate of major neurological signs and syndromes, and with the disease stage as well. A multilevel positive effect of hemosorption was shown in different pathogenic links of AE related to disorders in lipid metabolism, blood rheology, shape and functional activity of erythrocytes, shifts in the homeostatic systems. This suggested the use of hemosorption as a tool in prevention of ischemic stroke.

[The blood coagulation system under overdoses of the copolymer of 1-vinyl-3-methylimidazole iodide with the diethyl ether of maleic acid demonstrating antiheparin activity]. / Issledovanie sistemy svertyvaniia krovi v usloviiakh peredozirovok sopolimera 1-vinil-3-metilimidazoliia iodida s diétilovym éfirom maleinovoi kisloty, proiavliaiushchego antigeparinovuiu aktivnost'.

The effects of the active heparin antagonist--1-vinyl-3-methylimidazolium iodide-maleic acid diethyl ether copolymer--on the blood coagulative system were investigated. The effect of polycation was shown to appear only when the therapeutical dose was significantly increased and to manifest itself in the development of hypocoagulation. Hypocoagulative changes occurred only by inhibiting the activity of clotting factors V and VIII. Also, there was a moderate reduction in platelet counts.

[The effect of the quaternary ammonium salt of conidine oligomer-25 (QAS CO-25) on the activity of prothrombin complex factors]. / Vliianie chetvertichnoi ammoniinoi soli oligomera-25 konidina (ChAS O-25K) na aktivnost' faktorov protrombinovogo kompleksa.

Quaternary ammonium salt of oligomer-25 conidine (QAS O-25C) was used for regulation of hypocoagulation effect of Syncumar--anticoagulant of indirect type of action. The experiments were carried out on male rats. Experimental methods included thromboelastography, determination of thromboplastin time (after Quick) and determination of activity of single coagulation factors. It was shown that injection of QAS O-25C reliably do not affect blood coagulation. The combined action of QAS O-25C and Syncumar resulted in a twofold increase in the activity of factors IX and X within 24 h after the injection and in complete recovery after 48 h. After Syncumar injection per os we observed deep-hypercoagulation which was maintained even within 48 after the injection. The effect of QAS O-25C modulating the Syncular anticoagulant activity is probably connected with intensification of protein synthesis in the liver and, specifically, with an increase in the yield of active precursors of the factors of prothrombin complex.

[The anticoagulant activity of fucoidans]. / Issledovanie antikoaguliantnoi aktivnosti fukoidanov.

The influence of the degree of fucoidan macromolecules sulfation on their ability to induce an anticoagulant effect was studied. In in vitro experiments with the basic stain and in in vivo experiments on rabbits with performance of Quick's test, APTT, and spectrophotometric determination of antithrombin III (AT-III) activity a direct dependence of the intensity and duration of the anticoagulant effect on the degree of fucoidan sulfation was shown. The degree of sulfation was increased by alkaline treatment and anion-exchange chromatography of the compound. The anticoagulant effect of the fucoidan fractions under study was realized practically without affecting AT-III activity. And this direct effect of inhibition of the coagulation factors could be immediately arrested by intravenous injection of a neutralizing dose of the heparin antagonist VIM-DEMA.

[Effect of the synthetic heparin antagonist, conidine oligomer 25, on the synthesis of nucleic acids in regenerating liver cells]. / Vliianie sinteticheskogo antagonista geparina oligomera-25 konidina na sintez nukleinovykh kislot v kletkakh regeneriruiushchei pecheni.

Localization of 14C-conidine oligomer (the synthetic antagonist of heparin) and of polyelectrolyte complex heparin-conidine oligomer 25 was studied in cells of regenerating liver tissue within 20 hrs after partial hepatectomy in rats. Effects of heparin, conidine oligomer 25 and of their polyelectrolyte complex on synthesis of nucleic acids were studied. Conidine oligomer 25 penetrated from circulation into hepatocytes and subcellular organelles of regenerating liver cells and was localized mainly in cytosol (75-88%). In nuclei and mitochondria of the hepatocytes 14C-conidine oligomer 25 was mostly found in the fraction of low density of the organelles. Polyanion heparin, polycation conidine oligomer 25 and their polymeric complex impaired replication and transcription processes.

[Changes in general hemostasis in patients with diabetic retinopathy]. / Izmeneniia obshchego gemostaza u bol'nykh diabeticheskoi retinopatiei.

The hemostasis system was analyzed in 106 patients with diabetes mellitus, including 86 patients with diabetic retinopathy. Parameters of coagulogram and thromboelastogram as well as platelets aggregation, stimulated by collagen, were examined in all patients. Besides, the fibrinolytic activity of blood plasma as well as the concentration of plasminogen, fibrinogen and of fibrin-degradation products were evaluated. Essential changes were detected, which correlated with an intensity of diabetic changes in the eye fundus, i.e. generalized hypercoagulation caused by a lower activity of the system antithrombin III-heparin, a higher activity of fibrinogen and activation of the fibrinolytic system. The obtained data denote an important role of impaired hemostasis in the pathogenesis of diabetic retinopathy.

[Heparin inhalations combined with intravenous laser exposure of blood in patients with diabetic retinopathy]. / Ispol'zovanie ingaliatsii geparina v sochetanii s vnutrivennym lazernym oblucheniiem krovi u bol'nykh s diabeticheskoi retinopatiei.

Heparin inhalations combined with intravenous laser exposure of the blood are effective in patients with nonproliferative, preproliferative, and proliferative diabetic retinopathy. Clinical effect consisted in decrease of edema in the macular area, partial resolution of hemorrhages, a tendency to decrease in the caliber of veins, improvement of visual acuity, and extension of visual field. Immunological studies revealed immunomodulating effect of heparin inhalations and intravenous laser exposure of the blood, manifesting by decreased levels of pathological circulating immune complexes and increased concentrations of immunoglobulins, mainly IgG and IgM. Diabetic hemophthalmia is to be treated by subtenon implantation of a collagen system with injection of a prourokinase thrombolytic in combination with preoperative preparation including heparin inhalations, intravenous laser exposure of the blood, and parabulbar administration of 0.5 ml 1% emoxipin for 5 days in order to normalize immune hemostatic and redox processes and create conditions for effective action of the thrombolytic.

[Heparin inhalations and laser exposure of blood in treatment of patients with open-angle glaucoma]. / Ispol'zovanie ingaliatsii geparina i lazernogo oblucheniia krovi pri lechenii bol'nykh s otkrytougol'noi glaukomoi.

Comparative assessment of methods aimed at amelioration of the immunohemostatic processes in patients with open-angle glaucoma suffering from chronic vascular diseases showed that the most remarkable improvement of the visual functions and decrease of the level of circulating immune complexes in the blood were attained in the patients treated by heparin inhalations combined with intravenous laser exposure of the blood as against patients treated by one of these methods alone or traditionally.