Method for inhibiting nuclear kappa b factor with using 2-ethyl-6-methyl-3-hydroxipiridinium l-2,6-diaminohexanoate in culture of cells
SUBSTANCE: invention relates to medicine, in particular to experimental pharmacology. Method is proposed for inhibiting the nuclear factor kappa B in a cell culture comprising adding a bacterial lipopolysaccharide at a concentration of 1 mcg/ml to a freshly isolated standard ficoll density gradient test for mononuclear blood cells of Wistar rats, characterized in that 2-ethyl-6-methyl-3-hydroxypyridinium L-2,6-diaminohexanoate is then added to this mixture at a final concentration of 35 mcg/ml.
EFFECT: technical result consists in the expressed inhibition of the nuclear factor kappa B in the cell culture and in the favorable safety profile.
1 cl, 1 tbl
SUBSTANCE: in a premature baby the concentration of neuron-specific enolase (NSE), concentration of a brain-derived neurotrophic factor (BDNF), concentration of a vascular-endothelial growth factor (VEGF) in umbilical blood and concentration of the vascular-endothelial growth factor (VEGF) in peripheral blood are determined on the basis of the enzyme immunoassay of umbilical and peripheral blood serum on the 7-th day of life, a prognostic index (PI) is calculated by formula: PI=-0.007×X1+0.006×X2-0.05×X3+0.0004×X4-3.9, where X1 is VEGF content in umbilical blood at birth (ng/ml); X2 is VEGF content in peripheral blood on the 7-th day of life (ng/ml); X3 is NSE content in umbilical blood (mcg/l); X4 is BDNF content in umbilical blood (ng/ml); Const=-3.9. When PI is higher than 0, a conclusion about the absence of risk of occlusive posthaemorrhagic hydrocephalus formation is made, and if PI is lower than 0, a high risk of the said pathology development is predicted.
EFFECT: invention makes it possible to increase the efficiency of prediction of occlusive posthaemorrhagic hydrocephalus formation in the premature children with an extremely low body weight at birth.
SUBSTANCE: fine-needle aspiration of nodular thyroid growths is controlled by ultrasonic examination. A puncture needle containing an aspirate is washed twice in normal saline 1 ml, centrifuged; a supernatant is selected, and thyroglobulin is measured by enzyme immunoassay. If the thyroglobulin content is less than 272.5 ng/ml, the absence of high differentiated cancer is stated; the value falling within the range of 272.5-355.5 ng/ml shows a risk of high differentiated thyroid cancer; if the thyroid value is more than 355.5 ng/ml, high differentiated thyroid cancer is suspected.
EFFECT: invention provides the pre-operative differential diagnostics of high differentiated cancer in the patients suffering nodular forms of thyroid diseases, and also enables the further selection of an adequate method of treating.
SUBSTANCE: invention refers to test systems for content determination for mycopolysaccharides and derivatives including sections similar to synthetic linear nona-β-(1→3)-D-glucoside in environment, in room dust as well, and can be applied for ambient object pyrogenicity assessment during ecological monitoring and residential and production facilities for determination of ambient pyrogenic dust load by inhibition immunoenzyme analysis in solid phase. Test system includes a tray with synthetic linear nona-β-(1→3)-D-glucoside conjugate with bull serum albumin, immobilized on the internal surface of craters, calibrators containing synthetic linear nona-β-(1→3)-D-glucoside solution; lyophilized specific polyclonal rabbit antiserum; anti-species antibody conjugate against IgG(H+L) of rabbit with horse radish peroxidase; 25-fold concentrate of 0.1M phosphate salt buffer solution with 0.05% Tween-20 (PSB-T) pH 7.2-7.4; solution for polyclonal antiserum dilution; solution for dilution of anti-species antibody conjugate against IgG(H+L) of rabbit with horse radish peroxidase; chromogene; stop reagent.
EFFECT: convenient and efficient system for ecological research.
2 tbl, 6 ex
SUBSTANCE: preoperative prediction of the complicated course of the early post-transplantation period is ensured by measuring the pre-transplantation blood plasma concentration of insulin-like growth factor-1 (IGF-1) and growth hormone (GH), ng/ml. That is followed by calculating a prediction index for the post-transplantation course (K) by formula: K=lg (IGF-1/GH). If K is less than minus 1.7, the complicated course of the early post-transplantation period is predicted.
EFFECT: using the method enables the objective prediction in the infants suffering congenital hepatobiliary diseases accompanying the early post-transplantation period manifested by transplant dysfunction.
FIELD: veterinary medicine.
SUBSTANCE: method of diagnostics of leptospirosis in farm animals, comprising determining the presence of antibodies to antigens of seven serogroups L.hebdomadis, L.pomona, L.tarassovi, L.grippotyphosa, L.canicola, L.sejroe and L.icterohaemorrhagiae in blood serum by the method of enzyme immunoassay, characterised in that the antigen is used as the antigenic proteins of Leptospira of three serogroups mixed in equal quantities - L.tarassovi, L.grippotyphosa and L.hebdomadis. The invention significantly simplifies and minimizes the diagnostics of leptospirosis, enables to implement the operational control of epizootic condition of the animals on leptospirosis.
EFFECT: method has a high specificity, sensitivity and commonality, provides creation of a safe labour conditions for the staff of diagnostic laboratories.
4 tbl, 4 ex
SUBSTANCE: technique according to the invention involves the two-stage immobilisation of S and R brucellosis antigens on a tray by a non-specifically adsorbed mouse's monoclonal antibodies 2H2 and 2H8 in wells of the immunological tray (the first stage), specific binding S and R brucellosis antigens (the second stage) thereto, introduction and incubation of the analysed material (blood serum or milk) combined with introduction of mixed mouse's monoclonal antibodies 2H2 and 2H8 marked by horseradish peroxidase. The brucellosis agent antibodies are detected by being competitive with the antigen in binding on the well surfaces of the tray between the antibodies of the analysed sampled and fragment-marked monoclonal antibodies. A decreased level of the enzymatic signal in the analysed sample as compared with the negative reference testifies to contamination of the animal. Introducing the analysed sample into the two wells of the tray with S and R brucellosis antigens enables differentiation thereof for antibody specificity to any given form of the disease agent.
EFFECT: method involving the enzyme immunoassay enables increasing the effectiveness of health-improving measures, reducing the length of health-improvement in livestock farms with the negative brucellosis situation, dropping the disease incidence.
3 tbl, 3 ex
SUBSTANCE: in order to predict severity of postoperative period course in patients with calculous cholecystitis level of hormone grelin is determined in patients' blood serum before operation. If value of grelin level is 0.8-1.5 ng/ml mild course of postoperative period is predicted, if value of grelin level is >1.5-10.0 ng/ml course of medium severity is predicted. And if grelin level in blood serum increases to >10.0 ng/ml, severe course of postoperative period is predicted.
EFFECT: method makes it possible to increase accuracy of prediction of postoperative period course severity in patients with calculous cholecyctitis in cholecystectomy in order to perform treatment correction.
FIELD: veterinary medicine.
SUBSTANCE: method includes the interaction of antigens with antibodies, with anti-species antibodies labeled with horseradish peroxidase, adding the substrate mixture and recording the results of the reaction on the colour intensity of the complex formed. In the reaction the plates are used with antigens of rotavirus, coronavirus, the virus of diarrhea preliminary sorbed on them, and after applying the test samples of serum 0.10-0.12 ml each. The plates were incubated for 1.5-2 h at 36-37°C and washed. Then total anti-species immunosorbent conjugate is applied of 0.10-0.12 ml consisting of enzyme-labeled antibodies against globulins of blood serum of cattle and the results of the reaction are recorded.
EFFECT: invention provides simultaneous diagnosis of rotavirus, coronavirus enteritis, viral diarrhea in cattle, sensitivity, specificity, and ease of the assay, and the ability to automate the process of research.
SUBSTANCE: for the purpose of prediction of developing atopic dermatitis in newborns, umbilical blood plasma is examined for a level of interleukin-18. If the related level is 34 pg/ml or lower, developing atopic dermatitis in the newborn is predicted.
EFFECT: use of the given method enables well-timed evaluation of a risk of developing atopic dermatitis in newborns and early preventive measures for prevention thereof.
SUBSTANCE: method under the invention provides that the complex immunoglobulin preparation containing a component of C3b complement is sorbed in microplate wells for the purpose of immunoassay. Then the microplate wells are added with a solution of an analysed sample containing human complement C1 inhibitor with the unknown activity. It is followed by incubation, drying and washing of the dish; the wells are added with a conjugate of enzyme with serine proteinase in the form of preparation of fibrinolysin and a substrate of this enzyme. The amount of the prepared enzymatic reaction product is used to derive the content of active C1 inhibitor. A kit for implementing the method comprises a flat-bottomed microplate with bound C3b, the conjugate of enzyme with serine proteinase, the substrate buffer of this enzyme and a reference for active C1 inhibitor.
EFFECT: use of the method under the invention enables determining activity of C1 inhibitor, simultaneously bound both with serine proteinase inhibition, and with binding inhibition of complement factor B and C3b complement component.
2 cl, 1 tbl, 1 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to chemical-pharmaceutical industry, and can be used in drug quality control centres and analytical laboratories when measuring anthocyanes in such crude drugs, as bilberry, black chokeberry, black currant, etc. The presented method for measuring anthocyanes involves recovering anthocyanes from crude drugs in alcohol water mixtures containing 0.5-1.0% muriatic acid with the ethanol concentration of less than 70% and measuring anthocyanes quantitatively by optical density in 95% ethanol containing 0.5% ammonium as compared to a reference solution at analytical wavelength of peak absorption within the range of 611-630 nm with using a reference solution of cyanidine-3-O-glucoside or with using its specific absorption in 95% ethanol containing 0.5% ammonium.
EFFECT: higher sensitivity and result reproducibility, as well as shortening the analysis procedure as compared to its prototype.
3 dwg, 7 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the field of biotechnology, namely, to obtaining inhibitors of adhesion and/or aggregation of platelets, and can be used in medicine. A polypeptide, used as a component of a pharmaceutical composition and in sets for screening of the inhibitors of platelet adhesion or aggregation, is obtained in a recombinant way with the application of a matrix of the salivary gland cDNA of Anopheles stephensi.
EFFECT: invention makes it possible to obtain the polypeptide, possessing inhibiting activity with respect to platelet aggregation and/or inhibiting activity with respect to platelet adhesion.
10 cl, 4 dwg, 5 ex
SUBSTANCE: method is based on measuring a flavonoid amount by differential spectral photometry, calculated as Cinaroside at wave length 400 nm, extracting in water and alcohol and using 70% ethanol as an extraction fluid; the total flavonoid amount calculated as Cinaroside and an absolutely dry raw material in percentage (X) is calculated by formula.
EFFECT: method enables measuring the total flavonoid amount as biologically active components having a basic therapeutic action.
3 ex, 14 dwg, 2 tbl
SUBSTANCE: method of Novocain extraction from water solutions includes the preparation of an aqueous salt solution of Novocain by its dissolution in a saturated solution of a salting-out agent, extraction and analysis of an equilibrium water phase, with the application as an extragent of a solvotrophic reagent solution in chloroform with the concentration of 10 wt %, for which purpose the aqueous salt solution of Novocain with pH 8.0±0.5 is prepared due to the application of a saturated ammonium sulphate solution as the salting-out agent and addition of an ammonium buffer solution, Novocain is extracted for 5-7 minutes with the solution of the solvotrophic reagent in chloroform with the ratio of volumes of the aqueous salt solution of Novocain and extragent of 5:1, then the aqueous saline phase is separated from the organic one and analysed by a method of UV-spectrophotometry at a wavelength of 291 nm, the concentration of Novocain in the water solution is found by means of a calibrating graph; the coefficient of distribution (D) and the degree of extraction (R, %) of Novocain is are calculated by formulae.
EFFECT: method for the extraction of Novocain from the water solution, which is characterised by expressiveness, makes it possible to realise the practically complete single extraction of Novocain from the aqueous salt solution and can be applied in the analysis of Novocain-containing water solutions.
SUBSTANCE: invention relates to analytical chemistry and can be used in a system for monitoring content of sodium thiosulphate in solutions. The method of determining sodium thiosulphate in solutions is characterised by adding an analysed sample into a reaction vessel containing a corresponding amount of photogenerated iodine, obtained by blowing with air for 1-2 minutes and irradiating the reaction mixture with a stabilised light source, the mixture consisting of 0.5 M potassium iodide solution, an acetate buffer solution with pH 5.6 and a sodium eosinate sensitising agent, by detecting change in current in a cell and upon achieving constant current, re-blowing the reaction mixture with air for 2-3 minutes and re-irradiating with the stabilised light source until achieving the initial amount of iodine in the vessel, measuring the iodine generation time spent on achieving the decrease thereof, determining the amount of sodium thiosulphate from a calibration curve from the change in current and generation time.
EFFECT: invention provides a simple method of determining sodium thiosulphate in solutions and avoids use of expensive equipment.
10 tbl, 5 dwg
SUBSTANCE: invention describes a method for lipoic acid measurement in biologically active additives by cathode voltammetry involving transferring a substance from a sample into a solution and taking voltammetric measurement; the cathode voltammetry is performed on a mercury-film electrode at potential -0.373 V of a relatively saturated silver-chloride electrode with borate buffer solution pH 9.18 at continuously current potential trace at 0.06 V/s with the determined lipoic acid content range from 4.5·106 to 1.1·10-3 mole/l.
EFFECT: improving sensitivity and expressivity of the method for measuring lipoic acid in tabletted BAAs by cathode voltammetry.
1 tbl, 1 ex, 3 dwg
SUBSTANCE: in agar plate which is in a Petri dish and inoculated with one of the fungal species of the genus Fusarium, circular holes are cut with the diameter of 8 mm, 0.05 ml of the working solution of the test preparation is applied into them, and placed in a thermostat at a temperature of 24.5-25.0°C, after 2-5 days depending on the type of fungus the diameter of the zone of growth inhibition of fungal mycelium is measured and the activity of the preparations is calculated according to the formula , and it is considered that when A=1 the fungicide is ineffective - the zone of growth inhibition is not formed (D=d), when A=2-3 the preparation activity is low, when A=4 it is average, and when A≥5 it is high.
EFFECT: invention provides accuracy of determining the activity of seed disinfectants and fungicides used in agricultural production.
SUBSTANCE: invention represents a method for preclinical study of cardiotropic antiarrhythmic drugs, involving determining the bioelectric parameters in isolated multicellular perfused preparations and measuring an action potential duration, differing by the fact that the isolated multicellular perfused preparations are presented by rat's pulmonary vein myocardium; the parameters are measured in three operation modes of the multicellular preparations; a resting potential is additionally measured; varying APD 90%, related APD 50%/APD 90%, a spontaneous shear velocity of the resting potential, the most positive membrane potential in the resting preparation, a spontaneous activity train repetition rate, spontaneous action potential train repetition and variability frequency, post-depolarisation number and intensity, as well as a shear membrane potential corresponding to the beginning of train activity are used to evaluate the signs of antiarrhythmic and arrhythmogenic action.
EFFECT: more reliable prediction of the antiarrhythmic action of the potential pharmacological agents and reduction of experimental phase time.
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: claimed is application of fat emulsion for parenteral feeding as solvent for compounds which are poorly soluble in water. Fat emulsion contains in 1 l of solution: 30 g of refined soybean oil, 30 g of triglycerides with the average chain length, 25 g of olive refined oil, 15 g of purified fish oil.
EFFECT: obtaining solvent for compounds, poorly soluble in water, which makes it possible to determine parameters and spectrum of biological activity of novel compounds of chemical nature at the stages of pre-clinical and clinical tests, which does not change basic biological constants and possesses biological inertness.
2 tbl, 2 ex
FIELD: veterinary medicine.
SUBSTANCE: method comprises selection of only living, mature females of Trichuris vulpis from colon, blind gut of wild and/or domestic carnivorous animals infected spontaneously with whipworms in the study with helminthological methods when autopsy, into separate tubes with officinal isotonic solution (0.9%) of sodium chloride (solutio Natrii chlorati isotonica) and the exposure of the tubes with the females of Trichuris vulpis at t = 37.5-39°C for 5 hours under conditions of a thermostat.
EFFECT: method enables to select a large amount of fertilised eggs of T vulpis, not contaminated with particles of undigested feed, the particles of destroyed tissue of genitals of female of T vulpis and secondary bacterial microflora.