Method of controlling anticoagulant therapy
SUBSTANCE: invention relates to medical diagnostics and monitoring of medicinal agents and concerns method of measuring of combined activity of blood coagulation factors II and X for control of anticoagulant therapy and kit for implementation of this method. Method involves mixing of analyzed human plasma, undergoing analysis, with specially prepared plasma with deficiency in blood coagulation factors II and X at ratio of analyzed plasma sample and deficiency plasma from 1:1 to 1:20; addition of coagulation reagent and calcium to analyzed sample; determination of blood ability of coagulation. Kits of invention include coagulation reagent, calcium and specially prepared plasma with deficiency in factor II and factor FX. Kit components are accordingly lyophilized.
EFFECT: invention provides more accurate assessment of blood coagulation ability.
16 cl, 4 ex, 6 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: claimed invention relates to biotechnology, namely to obtaining infestin-4 mutants, and can be used for diagnostic purposes in the determination of characteristics of clotting of blood and its components. A polypeptide is characterised by a sequence of the infestin 4 mutant MutB SEQ ID NO: 1. The said sequence can have modifications outside inhibiting the loop part, which essentially preserve the activity of the said polypeptide. The polypeptide is used for the inhibition of contact activation in a tested sample of blood or its product in order to increase the time of the sample storage.
EFFECT: invention makes it possible to obtain the highly-selective inhibitor fXIIa, selectivity or activity of which is higher than in the native infestin-4 and Mut15.
25 cl, 7 dwg, 3 tbl, 4 ex
SUBSTANCE: invention refers to medicine, namely to haemocoagulogy, and can be used for detecting a group of individuals suffering a risk of haemocoagulation complications. Substance of the method: measuring initial blood coagulation trace amplitude, determining initial and final blood coagulation parameters and comparing them to corresponding normal blood coagulation parameters; if observing oppositely directed deviations, the disturbed functional state of the haemostasis system is diagnosed. A time constant is determined according to a calibration curve; the calibration procedure a priori covers two measured U1, U2 and known U01, U02 values of lower t1 and upper t2=kt1 limits of the adaptive range; the calibration curve U0i is a blood limit strength function compensating an uncertainty T* of the random time constant T0 and relating the reference Uri and measured Ui characteristics by normalising the measured values by the known ones the calibration curve U0i shows the actual values of the time constant T0 and blood limit strengthU0 which are used to draw the calibration curve of the blood limit strength, reference performance Uri
and to determine initial Ti and final Tf blood coagulation parameters wherein Ui, Uf are the normalised initial and final blood coagulation thresholds.
EFFECT: invention enables reducing a procedure error by tens orders, increasing a coagulation time accuracy by 4 orders, and reducing the efficiency three times as much that eventually provides higher metrological effectiveness of computer analysers applicable for the purpose of automating the process of detecting the individuals having a risk of haemocoagulation complications.
SUBSTANCE: invention relates to method of determining platelet resistance to acetylsalicylic acid (ASA) by impedance analysis of aggregation function of platelets in vivo, in which aggregation activity is analyses after incubation of biological material sample with ASA with application of aggregation inducer, and aggregation of platelets is induced by collagen in optimal concentration 2 mg/ml, and simultaneously with measurement of impedance determination of dynamics of platelet granules release is performed by luminescent method, where before carrying out aggregation samples are calibrated by means of adenosine triphosphate (ATP) standard, value of aggregation amplitude are determined in Ohms by obtained aggregatograms and obtained values are given points: values ≤6 correspond to 0 points, values 7-9 correspond to 1 point, values 10-12 correspond to 2 points, values >12 correspond to 3 points; after that, intensity of ATP release from platelet granules is determined in nmoles and obtained values are given points: values <0.5 correspond to 0 points, values 0.5-1.0 correspond to 1 point, values 1.0-1.5 correspond to 2 points, values 1.5 correspond to 3 points, and then resistance index (RI) is calculated by formula, value of IR parameter points to presence of aspirin-resistance of platelets.
EFFECT: carrying out fast complex diagnostics of platelet resistance to ASA with estimation of functional state of platelet granules before beginning treatment and possibility of preventing development of undesirable thrombotic or hemorrhagic complications.
2 dwg, 1 tbl, 2 ex
SUBSTANCE: invention represents a diagnostic technique for the disturbed thrombocyte aggregation accompanying mucoviscidosis in children involving a thrombocyte aggregation test using the Multiplate aggregometer inducers. Trays with a magnetic mixer and electrodes are added with NaCl 400 mcl at 37°C and immediately added with whole blood 400 mcl from a hirudin test tube, incubated in the chamber for two minutes; the tray is added with 30 mcl of an aggregation inducer specified in a group: soluble thrombin receptor - peptid-6, adenosine diphosphate, arachidonic acid. The thrombocyte aggregation rate is displayed on the screen in the form of a curve, and the sub-curve area U is automatically calculated; the sub-curve area U shows the thrombocyte aggregation state as compared to reference values in the group of healthy children; if the threshold area U has appeared to exceed the reference, the thrombocyte hyperaggregation, while the threshold area U being less than the reference, the thrombocyte hypoaggregation is stated.
EFFECT: invention provides the timely diagnosis of microcirculatory disorders accompanying mucoviscidosis.
2 ex, 1 dwg
SUBSTANCE: invention refers to medicine, particularly to clinical biochemistry, and aims at determining oxidative protein modification in a substance pool of an average molecular weight in a biological medium accompanying any pathological conditions by biochemical examination. The biological medium specified in the blood plasma, erythrocyte or urine is sampled; proteins are deposited by adding 10% trichloroacetic acid; if observing sedimentation, a centrifugation cycle follows at 1000 rpm for 15 minutes; thereafter, 2,4-dinitrophenylhydrazine 0.05 M in hydrochloric acid 2 M is added; the sample is centrifuged at 1000 rpm for 20 minutes; and if observing sedimentation, the sediment is washed twice in ethanol-ethylacetate (1:1), dried on a water bath for 10 minutes and dissolved in urea 8 M; the sample is kept in a boiling water bath for 10 minutes until dissolved completely; the prepared solution is analysed by spectrophotometry. The method is applicable both for single use, and for monitoring of the postoperative oxidation protein modification and average molecule levels.
EFFECT: method provides higher information value of biochemical tests, reducing consumption of the biological material.
9 tbl, 2 ex
SUBSTANCE: invention includes determination of content of soluble fibrin and D-dimers, formed in the process of fibrinolysis, activated in blood sample. In method, in accordance with the claimed invention, level of D-dimers, corresponding to destruction of soluble fibrin and level of D-dimers in sample with border values of the norm, are compared.
EFFECT: test in accordance with the claimed invention can be applied for determining whether resistance to blood coagulation in patient is sufficient.
4 tbl, 3 ex, 2 dwg
SUBSTANCE: method for thrombin activity test in an initially non-reacted mixture of thrombin and fibrinogen (versions) involving the stages: (a) reversible thrombin inhibition by adding an inhibitory solution having pH varying within the range of 8.5 to 11.5; (b) addition of the known amount of fibrinogen to the mixture (or the known amount of a chromogenic or fluorogenic thrombin substrate), (c) reversible thrombin activation by pH reduction to approximately 6.0 to less than 8.5, (d) enabling thrombin reacting with fibrinogen, (e) thrombin activity test initially found in the dry mixture. The method for fibrinogen functionality test in an initially non-reacted mixture of thrombin and fibrinogen (versions) involving the stages: (a) reversible thrombin inhibition by adding an inhibitory solution having pH varying within the range of 8.5 to 11.5; (b) addition of the known amount of thrombin to the mixture (or a thrombin-like enzyme), (c) reversible thrombin activation by pH reduction to approximately 6.0 to less than 8.5, (d) enabling thrombin reacting with fibrinogen, (e) fibrinogen functionality test initially found in the dry mixture.
EFFECT: group of inventions enables higher accuracy of thrombin and fibrinogen activity test.
32 cl, 1 dwg
SUBSTANCE: analyser has a revolving cuvette with a sample in which there is a control ferromagnetic ball, a magnet which can interact with said ball, a coagulation sensor which transmits signals from the ball and having a Hall sensor and a magnet, a signal processing device in form of a power supply unit, a Hall sensor, a microprocessor and a display device included in a common measuring circuit. The analyser is multi-channelled by fitting at least one additional revolving cuvette to form several channels. All cuvettes lie in a temperature-controlled unit included in the common measuring circuit. The longitudinal axis of each cuvette is inclined at an acute angle to the vertical. In the coagulation sensor, the magnet lies opposite the Hall sensor on the opposite side of the cuvette. The magnet is in form of a flat cylinder mounted with possibility of displacement along the cuvette. The analyser is also fitted with a unit for controlling rotation of the cuvettes and, included in the common measuring a circuit, a measurement parameter adjustment unit having rewritable read-only memory, and a timer configured to automatically switch on when a reactant is fed into the cuvette.
EFFECT: use of the invention increases reliability and broadens functional capabilities of the analyser owing to use of a multichannel measuring circuit, and simplifies the measuring procedure by automating the process.
4 cl, 2 dwg
SUBSTANCE: blood plasma is examined in 4 minutes after the beginning of spontaneous red blood cell aggregation for free red blood cell count and cell count in aggregates. A percentage of non-aggregated red blood cells (PNA RBC) by formula PNA RBC=FRBSC×100/(TRBCA+FRBSC) wherein FRBSC is the free red blood cell count, TRBCA are total red blood cells in aggregates. If the PNA RBC is 56 to 30%, I degree of severity is stated, 30% to 4% - II degree of severity, less than 4% - III degree of severity.
EFFECT: use of the invention enables objectifying and increasing precision of evaluation of red blood cell aggregation, evaluating an intensity of patient's microcirculation disorders in a relatively short time, and thereby ensuring well-timed adequate complex of therapeutic measures or corrected therapy.
SUBSTANCE: quantity of fibrin-monomers, dissolved in 0.5 N sodium hydroxide, is determined spectrophotometrically with application of ethanol test. Claimed method of quantitative determination of fibrin-monomers in blood makes it possible to reveal pathological process in organism with 95% reliability.
EFFECT: increase of determination accuracy.
2 tbl, 4 ex
FIELD: medicine, laboratory diagnostics.
SUBSTANCE: the suggested studying should be carried out on the glass simultaneously with several inductors by applying minimal inter-taking antilogarithms concentrations of aggregation inductors which correspond at double combination of inductors: ADP 5.0 x 10-8 M, adrenaline 3.0 x 10-9, collagen - dissolving the main suspension 1:8, thrombin 0.075 U/ml; at triple combination of inductors: ADP 10-9 M, adrenaline 10-9, collagen - dissolving the main suspension 1:9, thrombin 0.060 U/ml. The development of aggregation means thrombocytic activation in patients with arterial hypertension at metabolic syndrome. The method enables to evaluate the changes of thrombocytic functional state with combination of inductors more probably present in area of vascular lesion by applying minimal necessary concentrations that develops real conditions at hemostatic initiation in human vessels.
EFFECT: higher efficiency of studying.
3 dwg, 3 ex, 2 tbl
SUBSTANCE: method involves checking consciousness, blood coagulation state, peripheral blood leukocytes number, K+ ions, bilirubin, fibrinogen, hemolysis and hemoglobinuria availability, prothrombin index and exotoxic shock development. Each value is calculated in points as follows. Lucidity is evaluated as -2 points; depression - +3 points; coma - +6 points; lack of changes in blood coagulation system - -2 points; coagulation availability without clinical injuries - +2 points; coagulopathy with clinical manifestation signs - +19 points; K+ ions concentration being less than 3.0 mmole/l - +3 points, from 3.1 to 3.5 mmole/l - -5 points, from 3.6 to 5.0 mmole/l - 0 points, greater than 5.0 points - +7 points, failure in determining K+ ions concentration - 0 points; hemolysis availability - +6 points, its lack - -3 points; hemoglobinuria availability - +8 points, its lack - -1 points; leukocytes number being less than 12.0x109/l - -2 points, from 12,1 to 18.0x109/l - 0 points, higher than 18.0x109/l - +8 points; hourly urine output being less than 30 ml/h - +6 points, greater than 30 ml/h - -2 points; bilirubin content being less than 31 mcmole/l - -2 points, from 30.1 to 50.0 mcmole/l - 0 points, greater than 50.0 mcmole/l - +2 points, failure in determining bilirubin content due to hemolysis being available -+6 points; prothrombin index being equal to or less than 60% - +3 points, greater than 60% - 0 points, failure in determining prothrombin index due to hemolysis being available - +12 points; fibrinogen concentration in blood plasma being less than 2.1 g/l - +4 points, from 2.1 to 4.0 g/l - -1 point, from 4.1 to 6.0 g/l - +1 point, failure in determining fibrinogen concentration due to erythrocyte hemolysis being available - +13 points; exotoxic shock development - +9 points, its lack - -1 point. The points are summed up. The value being greater than +13, admission for treatment in resuscitation department is indicated. The value being less than -13, admission for treatment in therapeutics department is indicated. The value being from -13 to +13, resuscitation expert consultation is advised.
EFFECT: high evaluation accuracy.
FIELD: medicine, laboratory diagnostics.
SUBSTANCE: one should evaluate the time for clotting of plasma under testing in phospholipid-dependent test, moreover, one should apply high- and low-sensitive thromboplastin reagents to lupus anticoagulant to calculate the ratio of indices of prothrombin time prolongation and at its value being either equal to or above 1.1 one should diagnose APS.
EFFECT: shortened terms of research.
1 ex, 4 tbl
SUBSTANCE: method involves analyzing symptoms manifesting initial disseminated intravascular blood coagulation syndrome danger like burn area, availability of upper air passages burn, shock with its severity degree taken into consideration, sepsis development; clinical manifestations of disseminated intravascular blood coagulation syndrome like lung, kidney, liver function insufficiency, cerebral dysfunction, local and multiple hemorrhages, thrombosis, infarction; homeostasis system laboratory analysis data, hyper- and hypocoagulation based on chronometry test data, number of blood platelets, fibrin-monomer complexes, D-dimers, activity of antithrombin III, C and S proteins, XIIa-dependent fibrinolysis plasminogen content, availability of injured erythrocytes, combinations of laboratory tests for recognizing disseminated intravascular blood coagulation syndrome. Each sign under consideration receives a number of points corresponding to its diagnostic significance and integral value is calculated DIBCSIV=(X1+X2+…+Xn)/n, where n is the number of signs taken into consideration. DIBCSIV value equal to 1.0-1.5 units shows physiological norm. The value being between 1.6 and 2.5 units, light disseminated intravascular blood coagulation syndrome is diagnosed. The value being between 2.6 and 3.5 units, disseminated intravascular blood coagulation syndrome of medium severity is diagnosed; 3.6-4.5 points to one heavy severity degree; 4.6 and greater indicates highly severe case of disseminated intravascular blood coagulation syndrome.
EFFECT: high accuracy and objectiveness in differentiating syndrome severity degrees.
FIELD: medicine, diagnostics.
SUBSTANCE: one should study blood components to detect anticoagulant-fibrinolytic activity. Moreover, patient's blood should be sampled: in whole blood one should detect the presence of affected erythrocytes and evaluate the quantity of thrombocytes, in plasma it is necessary to study the activity of antithrombin III, XIIa-dependent fibrinolysis, the content of soluble fibrin-monomeric complexes, in blood serum of the sample taken one should detect the concentration of urea, creatinine, sodium, albumin, total cholesterol and the activity of aspartate aminotransferase, moreover, one should calculate integral value of renal-hepatic deficiency, to put corresponding point for the degree of parameters under testing, then one should calculate integral value of disseminated intravascular clotting (IVDIC) and at its value being 6.3 U and more DIC-syndrome should be diagnosed, moreover, at IVDIC value ranged 6.3-10.1 U it is possible to diagnose latent DIC-syndrome, at 10.2-14.6 - subacute DIC-syndrome and at 14.7 and higher - acute DIC-syndrome should be concluded.
EFFECT: higher accuracy and efficiency of diagnostics.
4 ex, 2 tbl
FIELD: medicine, obstetrics.
SUBSTANCE: the present innovation deals with predicting disadaptive processes in women in dynamics of menstrual cycle. During menstrual cycle beginning since the 1st d to the 21st d one should detect the dynamics for alteration in coefficient of activity of syntoxic adaptation programs (CASAP), calculated by the following formula:
where CST - concentration of blood serotonin, AAT-III - activity of antithrombin III, Aaoa - total antioxidizing activity of plasma, CCD8 + - concentration of T-suppressors, Cad - concentration of blood adrenalin, Cα2MG - concentration of α2-macroglobulin, CMDA - concentration of malonic dialdehyde, CCD4 + - concentration of T-helpers. Moreover, normally CASAP value alters two-fold against the first day of the cycle - since 0.70 up to 1.40 on the 21st d of the cycle, at no alterations in CASAP value one should diagnose female disadaptive alterations leading to failed pregnancy. The innovation enables to perform diagnostics of disadaptive processes in women in dynamics of menstrual cycle followed by prognostic conclusion upon future pregnancy.
EFFECT: higher accuracy of diagnostics.
SUBSTANCE: method involves determining spontaneous blood platelets aggregation and one induced by adrenalin and collagen, thrombocytospecific peptides activity of β-thromboglobulin and thrombocytic factor 4 in blood plasma.
EFFECT: high accuracy of diagnosis.
SUBSTANCE: method involves determining coagulating blood viscosity values like reaction period r, thrombin constant K, maximum amplitude MA, time T for forming fibrin-thrombocytic blood clot, spontaneous blood platelets aggregation intensity Ar, retraction and spontaneous clot lysis total FA. The r being within 5-7 min, Ar from -2 to -6 relative units, K being within 4-6 min, MA within 500-700 relative units, T within 40-60 min and FA equal to 10-20%, low inflammatory process activity is considered to be the case. The r being less than 5 min, Ar equal to -8 to -12 relative units, T less than 40 min and FA less than 10% with no changes in K and MA being observed, inflammatory process activity in chronic glomerulonephritis case is considered to be of high severity degree.
EFFECT: high accuracy of diagnosis; enhanced effectiveness of treatment method selection.
FIELD: medicine, clinical neurology, neurosurgery.
SUBSTANCE: one should study both activation and aggregation of thrombocytes in blood of carotid artery, at the quantity of thrombocytic active forms being above 70% and the number of aggregated thrombocytes being above 9.0% one should predict the development of cerebral ischemic lesion along with stable focal neurological symptomatology, and at the quantity of thrombocytic active forms being below 30% and the number of aggregated thrombocytes being below 8.0% it is possible to predict positive dynamics in the course of the disease mentioned without developing cerebral ischemic lesion.
EFFECT: higher accuracy of prediction.
FIELD: medicine, clinical neurology, neurosurgery.
SUBSTANCE: one should study the level of von Willebrand's factor in patient's carotid artery blood. At its content being below 105% one should predict the development of repeated AICH. The innovation improved information value of testing due to possibility to obtain reliable prediction in latent period, as well.
EFFECT: higher accuracy of prediction.
2 ex, 1 tbl