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Method of growing colonies of microbial cells and device for its realisation Group of inventions relates to biotechnology. Claimed is a method of growing colonies of microbial cells on a surface of a porous plate. The method includes supply of a nutrient solution from bottom to top through the porous plate into zones of growth of colonies of the microbial cells on its upper surface, supply of a suspension of the microbial cells onto the upper surface of the porous plate, creation of controlled conditions for the colony growth, performing observation of the colony growth, separation of the grown colonies of the microbial cells from the zones of growth and their transfer into external means of identification. The nutrient solution is supplied into the zones of growth of the colonies of the microbial cells by creation of a pressure difference between the hole input and output. Holes are made in the plate from an anode aluminium oxide orthogonally to its large plane and are topologically coded. The said zones of growth are formed in them in the form of porous membranes. The porous membranes are located at the same level as the upper surface of the plate or with formation of a hollow and do not pass the microbial cells. After supply of the nutritional solution, the suspension of the microbial cells of a specified concentration is supplied onto the upper surface of the plate until their homogenous distribution is achieved. Between the zones of growth on the surface of the plate a film, preventing attachment of the microbial cells, is formed. Separation of the grown microcolonies from the zones of growth is performed by hydroblow. A hydroblow is directed from the side of the input of cylindrical holes of the plate and spreads along them and farther through the pores of the porous membranes with force, which does not destroy the microcolonies but is sufficient for their separation from the growth zones. Also claimed is a device for growing the colonies of the microbial cells by the claimed method. |
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Method of counting oil-oxidising bacteria in sea water Invention relates to microbiology and can be used in monitoring environmental-microbiological investigation of the quality of sea water to determine the amount of oil-oxidising microorganisms. The method involves preparing a mineral medium - bases containing NH4NO3, K2HPO4, KH2PO4, MgSO4, CaCl2, FeCl2, a concentrated solution, agar and distilled water in a given ratio, followed by addition of an oil product in a given amount, said product being bunker oil. Seeding sea water on the surface of the culture medium and incubating the seed for 3-4 hours enables to detect colonies of oil-oxidising bacteria. |
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Method of toxicity assessment of products from polymer and textile materials Method of toxicity assessment of products from polymer and textile materials is proposed. The method comprises the use of biosensor based on oxygen electrode, immobilisation of whole cells of bacteria E.coli K-12 on the surface of the oxygen electrode. The immobilisation is carried out using a semipermeable membrane. After immobilisation the respiratory activity of microorganisms is measured in the presence of the sample and standard samples of positive and negative control. Then the toxicity index is calculated and the sample toxicity is evaluated based on the value of the toxicity index. |
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Invention is a method of determining the nonspecific resistance of pathogenic microorganisms to antibiotics and the fact of the presence of bacterial biofilms on the basis of measurement of catalytic activity of phosphodiesterases cleaving the cyclic diguanosine monophosphate, with a threshold sensitivity of 50 pg/ml, comprising: 1) isolating the target-phosphodiesterase from lysed bacterial cells; 2) binding of phosphodiesterase with biotin-conjugated antibodies specific for non-catalytic domains of phosphodiesterase; 3) affinity purification of complexes formed by target-phosphodiesterase and biotin-conjugated antibody using paramagnetic particles containing neutravidin or its analogs that bind biotin; 4) interacting of the complexes of phosphodiesterase/biotin-conjugated antibody, immobilised on paramagnetic particles with complexes containing a-di-GMP in the form of G-quadruplex systems with intercalate dye, which is accompanied by decrease in the intensity while destruction of complexes of intercalate dye with c-di-GMP; 5) measurement of decrease of fluorescence upon hydrolysis with c-di-GMP and destruction of complex of c-di-GMP with intercalate dye, followed by quantitative estimation of the phosphodiesterase activity based on calibration curves made using known amounts of the recombinant enzyme of phosphodiesterase identical to the test target; 6) identification of increased level of phosphodiesterase activity detected by the test antibiotic-resistant bacterial strains capable of biofilm formation, as compared with the level of phosphodiesterase activity that can be detected for the control strains of bacteria of the same species not having the antibiotic resistance and the ability to form biofilms. |
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Method for in vitro determination of activation of plasminogen with bacteria Invention relates to medical microbiology and a method of determining activation of plasminogen with bacteria. The method involves adding protamine sulphate to a prepared supernatant fluid, incubating the obtained mixture, depositing cells by centrifuging, incubating the supernatant fluid with the protamine sulphate, depositing protein and detecting activation of plasminogen with bacteria from the amount of split arginine, content of which is determined by Sakaguchi method from the red colour of the sample. |
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Container for isolation and identification of microorganism Invention relates to biotechnology. Claimed is container for isolation and identification of microorganism. Container includes upper part, which has wide internal diameter, and lower part, which has capillary tube, middle conic part, connecting said upper and lower parts, and optic window on the bottom and/or on one or more than one wall of container. Optic window is less than 0.1 inch (2.54 mm) thick and is transparent for wavelength of near-infrared, visible and/or ultraviolet light spectrum. Window contains quartz, quartz glass, sapphire, acrylic resin, methacrylate, cyclic olefin copolymer, cycloolefin polymer or any their combination. Capillary tube has internal diameter from 0.01 inch (0.03 mm) to 0.04 inch (1.02 mm). |
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Method for quantitative assessment of bactericidal activity of disinfectants Invention refers to microbiology and biotechnology. Analysed bacterial strains are inoculated on a dense nutrient medium. Paper disks impregnated with a disinfectant are applied. They are incubated in a temperature chamber until the bacteria start growing. The bacterial growth inhibition areas are measured. A quantity of the grown colonies is counted to construct a dependence diagram of the bacterial growth inhibition area and the quantity of the grown colonies after the disinfectant reaction. The diagram and Shughart inspection sheet are used to assess the disinfectant activity on specific types of the microorganisms. The disinfectants with mean measurements of the bacterial growth inhibition area are above an upper control limit of the Shughart inspection sheet are considered to be high bactericidal activity agents. The disinfectants with mean measurements of the bacterial growth inhibition area are below a lower control limit of the Shughart inspection sheet are considered to be low bactericidal activity agents. The disinfectants with mean measurements of the bacterial growth inhibition area between the control limits of the Shughart inspection sheet are considered to be mean bactericidal activity agents in relation to all analysed agents. |
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Invention relates to biotechnology and can be used in biological treatment of waste water from electroplating plants from heavy metal salts. The method involves adding yeast biomass to waste water, said biomass being in form of brewery wastes containing a combination of yeasts of different strains of Saccharomyces cerevisiae with viability of 90-95% in a given amount. The biomass is mixed with the waste water to obtain a suspension. The obtained suspension is held for 8 hours at temperature of 10°C-29°C and solution pH of 5.5-8.0, followed by recycling spent yeast containing heavy metals by treating with lime Ca(OH)2, with the ratio of yeast biomass to lime of 1:5-8, to obtain a mixture. The obtained mixture is subjected to wet treatment at temperature of 90°C for 1 hour, followed by isolation of the obtained mixture, which contains heavy metals, in concrete paste. |
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Plasmid vector and method of detecting nonsense mutations and frameshift mutations in brca1 gene Present invention relates to molecular biology. Disclosed is a method of detecting frameshift and nonsense mutations in the BRCA1 gene, which involves construction of recombinant plasmids where the amplified gene fragment is located in a single translation frame with the alkaline phosphatase gene of E.coli (phoA). A plasmid vector pPhoA-frame, which contains a DNA sequence which encodes alkaline phosphatase of E.coli was constructed. A DNA fragment containing restriction endonuclease recognition sites BgIII, StuI, Apal, SacII and intended for cloning BRCA1 gene fragments (polylinker) was inserted into said DNA sequence. The amplified BRCA1 gene fragment is inserted into the plasmid vector pPhoA-frame in a single translation frame with phoA. The occurrence of mutations which violate reading frame integrity in the investigated gene fragment is evaluated visually from the absence of colour of E.coli. cell colonies transformed by the obtained recombinant plasmid on an indicator dish containing a substrate for alkaline phosphatase. The disclosed method enables to detect only mutations that are significant for development of pathology since it avoids detection of polymorphous versions which do not lead to stop codons and most cases have not significant effect on protein function. The method enables to detect any, including unknown, mutations which violate frame integrity. |
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Detection method of microfungi coccidioides posadasii 36 s and coccidioides immitis c-5 Detection method of microfungi Coccidioides posadasii 36 S and Coccidioides immitis C-5 in vitro involves pre-growth of culture in mycelial phase, preparation of a suspension corresponding to 5 units of activity of a standard opacity sample, possibility of spherules formation and detection of spherules filled with endospores. Culture in mycelial phase is grown during 3 days. Possibility of spherules formation is provided by infection of the one-day culture of cells of murine splenocytes, which is obtained on RPMI-1640, and further cultivation during 5 days at the temperature of 37°C, with content of CO2 in atmosphere of 5%. In order to detect spherules in the form of round double-outline formations filled with endospores, a sample is taken, deactivated with formalin and investigated by means of a light microscopy method. |
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Method involves preparation of bacterial suspension, separation of the obtained biomass of bacteria by centrifugation, dilution of the obtained biomass in physiological solution, preparation of monolayer of CaCo-2 cells, introduction of bacterial culture, cultivation of cells, flushing with physical solution, removal of monolayer with bacterial cells and counting of the amount of bacterial cells related to 1000 cells of CaCo-2; with that, bacteria refer to highly-adhesive ones if the number of bonded cells is 1010 to 3000, to average-adhesive ones of 210 to 1000, and to low-adhesive ones of 0 to 200. |
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Preparation for cleaning of soil from oil and oil products Preparation containing biodestructor of oil contamination represents centrate of cultural liquid of microbial mass of consortium of oil-oxidising microorganisms immobilised on peat carrier. Consortium composition includes the following: Rhodococcus eqvi SRI MCC B-1115 bacteria strain, Rhodotorula glutinis SRI MCC Y-1113 yeast and Rhodotorula glutinis ARSRI MCC Y-1114 yeast strain in the ratio of 1:1:1 respectively, which have been grown jointly. |
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Proposed chimeric protein with SEQ ID NO:02 is fluorescent biosensor, built on the basis of HyPer protein and mutant of PH-domain of Btk tyrosine kinase. |
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Method to detect number of microorganisms in air 1% sterile solution of glucose is prepared on the basis of a physiological solution, which is used as a nutrient medium. An absorber by Zaytsev is connected to an aspirator of "Briz-1" grade, into the flask of which 10 ml of the prepared 1% glucose solution is placed. The device is placed into the investigated room, and the aspirator is switched on for 15 min. Microorganisms that are in air go through the glucose solution and are retained in it. The solution is placed into a test tube and thermostatted at 37°C for 2 hours. Solution electroconductivity is measured with the help of a sensor KDS-1038. The number of microorganisms in air is determined according to the curve of empirical dependence of solution electroconductivity on the number of microbes, which is built in accordance with the produced values. |
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Method to protect yeast Saccharomyces cerevisiae against oxidising stress as a result of exposure to hydrogen peroxide includes growing of yeast culture under standard conditions till the end of the logarithmic or the start of the stationary phase of growth, incubation with a protective agent, exposure to hydrogen peroxide with subsequent determination of the number of survived cells. The protective agent is represented by soy inhibitors of proteases in concentration of 0.1-2.0 g/l. The time of incubation with the protective agent makes 5-6 hr, concentration of hydrogen peroxide is 700-750 mM. The invention provides for high extent of yeast cells production with no depressant action. |
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Composition for production of organosilicon sol-gel matrix for immobilisation of microorganisms in biosensor analysers is provided. The composition consists of 20% polyethyleneglycol solution in phosphate buffer solution, Tetraethoxysilane, and 0.2 mol/dm3 catalyst solution NaF, an additionally introduced hydrophobic additive - methyltriethoxysilane. The components are taken at a volume ratio of PEG:TES:MTES: NaF 4:(18-3.4) (2-16.6):1. |
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Method of detecting and counting viable legionella pneumophila microorganisms Present invention relates to microbiology and a method of detecting and counting viable Legionella pneumophila microorganisms in a sample. The described method involves: (1) contacting said microorganisms of said sample with at least one reducing compound which contains glutamate and pyruvate, and a culture medium which is a buffered charcoal yeast (BCYE) or GVPC agar culture medium, (2) incubating the product of step (1), and (3) detecting and determining the number of viable microorganisms. The reducing compound used directly or indirectly affects metabolism, reducing oxidative stress of the microorganism by reducing formation and/or breaking down reactive forms of oxygen. |
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Method of qualitative assessment of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys in operation of spacecrafts and the suspension of spore materials fungi of implementation of the said method is proposed. The test and control samples of aluminium and magnesium alloys are prepared. The prepared samples are dried, sterilised. Fungal cultures - strains of microorganisms Paecilomyces variotii Bainier of All-Russian collection of microorganisms F-4039D, Ulocladium botrytis Preuss of All-Russian collection of microorganisms F-4032D, Penicillium chrysogenum Thorn of All-Russian collection of microorganisms F-4034D, Aspergillus sydowii (Bainier et Sartory) Thorn et Church of All-Russian collection of microorganisms F-4037D, Cladosporium sphaerospermum Penz. of All-Russian collection of microorganisms F-4041D are inoculated for growing spores into tubes with a sloped agar medium of Czapek-Dox. The tubes are thermostated at a temperature of (29+2)°C for 14-28 days until appearance of mature spores. Spore materials suspensions of individual cultures of the said fungi are prepared, which are then mixed in equal proportions. And the concentration of spores of each fungi species in the suspension should be in the range of 1-2 mln/cm3. The resulting suspension is applied to the sterilised test samples. Then they are dried and placed as one sample in each Petri dish on the surface of agar medium of Czapek-Dox. The control samples not treated by the spore material are also placed on the surface of the agar medium of Czapek-Dox in the Petri dishes. The Petri dishes with the test and control samples are placed in different desiccators at the bottom of which water is poured to maintain the humidity more than 90%. The desiccators are closed and incubated in the thermostat at a temperature of (29±2)°C. The exposition of test and control samples is carried out for 40, 120 and 180 days. Then the samples are removed from the Petri dishes, and the corrosion products and mycelium are removed from the surface of the samples washing them in running water. The samples are soaked in 70% ethyl alcohol for 30 minutes, then they are washed with detergent and dried. Then, using a scanning electron microscope a qualitative assessment of biocorrosion damages is carried out. According to the change in the appearance of the surface of the samples the initial stages of biocorrosion and its type is evaluated, as well as the distribution of corrosion damage on the surface of the samples, and the dependence of the biocorrosion process on time is determined. |
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Apparatus for determining quality of organic and inorganic products Invention relates to means of controlling quality of organic and inorganic products, and can be used to evaluate safety of food and fodder products, natural water and waste water, ground, soil, determining the maximum allowable concentration of contaminants, as well as the impact of human activities on the environment, including oil extraction and refining products. The apparatus consists of a computer with a software system and a biodetector. The biodetector has a housing 1 inside of which there is a displacement controller for a board 2 with containers 3 for test objects, a light source - light-emitting diodes 4, an optical system with a television camera 5 and a lens 6 mounted on a support 7. The apparatus is provided with a light-proof casing 8 for closing the top of the board with containers for test objects, the inner surface of which is coated with white paint. The optical system with a television camera 5 and a lens 6 is mounted on the support 7 by a turning mechanism 9 in form of a ball head. Inside the housing 1 there is a stepper motor 10 for rotating the board 2 with containers 3. |
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Method to monitor microbiological activity in technological flows Device comprises a through cell equipped with holes, where at least one hole represents an inlet hole for intake of fluid medium from the specified technological flow, and at least one hole is an outlet hole for discharge of fluid medium from the specified through cell. To one of specified holes an RK probe is attached, possibly, an OVP probe, a cleaning accessory. The first pipeline is connected to the inlet hole. Possibly, the second pipeline is connected to the outlet hole. A valve is connected to the specified through cell. With the help of the specified devices and methods they measure volume microbiological activity and surface microbiological activity in a process flow of water by means of measurement of dissolved oxygen concentration. |
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Method includes the following stages: contact of a sample with a source of nutrition for cells, containing antioxidant, representing pyroracemic acid or its salt, and an inhibitor of cell proliferation, which is selected from ciprofloxacin and cefalexin; contact of the specified sample with fluorescent-marked oligonucleotide probes, capable of specific hybridisation at least with one section of ribosomal nucleic acids, which belong to microorganisms of Legionella pneumophila kind and type; and detection and quantitative determination of a fluorescent signal. |
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Agent for inhibiting cytokinin signal function Present invention relates to chemical industry and agriculture and is a plant growth stimulating agent. |
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Method for prediction of bacterial translocation into blood in generalised chronic periodontitis Method for prediction of bacterial translocation into blood in generalised periodontitis provides recovery of symbiont strains from gingival pocket biocenosis and comparison of their hemolytic activity (HA), antilysozyme activity (ALA) and growth. |
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Method for determining lysocyme activity in lachrymal fluid of infants Tear is taken by sterile disks made of a porous material to form a microbial lawn and to determine lysocyme activity by lysocyme diffusion from the disk into the microbial lawn. It is followed by incubation in a thermostat for at least 24 hours. A zone of microbial growth retardation is determined by measuring a diameter of the zone of microbial growth retardation. The diameter of the zone of microbial growth retardation in healthy infants is 28-30 mm, while the diameter of the zone of microbial growth retardation in infants suffering inflammatory eye diseases is 20-24 mm. |
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Method for evaluation of antioxidant activity of microorganisms Antioxidant activity of a broth culture of microorganisms or a cell suspension of microorganisms grown on a solid nutrient medium is evaluated. An oxidable medium is presented by a lecithin solution, while peroxide initiators are as follows: Staphylococcus aureus cells, ascorbic acid, ferric (II) ions (in the form of an aqueous solution of ferric sulphate). Antioxidant activity is evaluated by an ability to inhibit lipid peroxidation by adding concentrated orthophosphoric acid, and 1% 2-thiobarbituric acid in dimethyl sulphoxide or ethanol (1:1). The stained complex is extracted in n-butanol, and after a butanol phase separated, it is spectrophotometered with regard to transmission unit at 550 nm wherein a reference tray is a tray with n-butanol. It is combined with preparing two controls wherein test tubes containing an oxidation substrate - lecithin are added with known antioxidant activity, while in the other one initiated peroxidation is enabled with no antioxidant added. |
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Yersinia pseudotuberculosis 634 bacterial strain is recovered from a patient suffering pseudotuberculosis, deposited in the collection of the Russian Science and Research Antiplague Institute Microbe, No. KM 214 and is applicable as a test strain for differentiation of Yersinia pseudotuberculosis bacteria genetic group IA. The characteristic of the strain is the content of chromosomal gene of the Y.pseudotuberculosis YPMa/YPMc (urtA/C) superantigen and the YAPI (pilPQ) Yersinia adhesive pathogenicity island gene determined by PCR method. |
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Yersinia pseudotuberculosis 413 bacterial strain is recovered from a patient suffering pseudotuberculosis, deposited in the collection of the Russian Science and Research Antiplague Institute Microbe, No. KM 211 and is applicable as a test strain for differentiation of Yersinia pseudotuberculosis bacteria genetic group I. The characteristic of the strain is the content of chromosomal gene of the Y.pseudotuberculosis YPMa/YPMc (urtA/C) superantigen and the YAPI (pilPQ) Yersinia adhesive pathogenicity island gene, the pVM 82 plasmid of molecular weight 82 MDa determined by PCR method. |
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Yersinia pseudotuberculosis 563 bacterial strain is recovered from a patient suffering pseudotuberculosis, deposited in the collection of the Russian Science and Research Antiplague Institute Microbe, No. KM 212 and is applicable as a test strain for differentiation of Yersinia pseudotuberculosis bacteria genetic group IIA. The characteristic of the strain is the content of chromosomal gene of the Y. pseudotuberculosis YPMa/YPMc (urtA/C) superantigen determined by PCR method. |
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Yersinia pseudotuberculosis 1068 bacterial strain is recovered from a patient suffering pseudotuberculosis, deposited in the collection of the Russian Science and Research Antiplague Institute Microbe, No. KM 213 and is applicable as a test strain for differentiation of Yersinia pseudotuberculosis bacteria genetic group II. The characteristic of the strain is the content of chromosomal gene of the Y.pseudotuberculosis YPMa/YPMc (urtA/C) superantigen and the pVM 82 (dotO) plasmid of molecular weight 82MDa determined by PCR method. |
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Rapid assessment of upper airway state Sample from the upper airways contacts with a test strip. The test strip contains at least one wide-spectrum indicator which demonstrates a first spectral reaction in the presence of bacteria and a second spectral reaction in the presence of viruses. The wide-spectrum indicator is N-phenolate betaine. Besides, the test strip contains a mesh which comprises one differentiating indicator wherein the mesh demonstrates a third spectral reaction in the presence of one type of microorganisms and a fourth spectral reaction in the presence of the other type of microorganisms. |
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Invention concerns identifying toxigenic Vibrio cholerae O1 strains, determining their biovars and differentiating V.cholerae eltor biovars by typical isolates and changed versions. According to the invention, the multiplex polymerase chain reaction (PCR) is conducted in a single step in two reaction mixtures wherein each mixture contains a specially selected combination of primers: one - to the genes rfbO1, cas3 and ctxBclass another - to the genes rfbO1, rtxC and ctxBEltor. A test system for implementing the method comprises DNA recovery components, PCR components and result analysis components. The PCR components contain: the 10-fold buffer solution, pH 8.4, mineral oil, deionised sterile water, two positive controls, enzyme Taq-polymerase, mixed dNTP, mixed primers No.1 - rfbO1-F - rjbO1-R, cas3-F - cas3-R, ctxBclass-F- ctxBclass and mixed primers No.2 - rfbO1-F - rfbO1-R, rtxC-F - rtxC-R, ctxBEltor-F - ctxBEltor-R. |
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Method for simulating prevention of hospital strain formation Method involves the intramuscular introduction of the preparations Galavite 200 mcg/mouse and Lidocaine 8 mcg/kg of body weight once a day for three days running every 24 hours to experimental animals CBA line mice suffering a burning injury with underlying mycotic and bacterial infections accompanied by anti-infectious protection suppression. |
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Bacterial sensor for ph variation detection Invention may be used in medicine for early diagnosing of stomach cancer. A sensor element is a bacterial cell Helicobacter pylori containing plasmid pHP presented in dwg. 1 with the bacterial gene gfp under control of the inducible stress promotor PflaA. |
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Method of sampling lactobacillus bacteria to be included in formulations of probiotic preparations Method of sampling Lactobacillus bacteria to be included in formulations of probiotic preparations provides preparing isolates of said bacteria. The bacteria are sampled by activity of their enzymatic systems responsible for destruction of mycotoxins which are determined by an average geometry of activity of total dehydrogenases, peroxidases and cytochrome P-450-dependent oxidases. The bacteria having the average geometry of enzymatic activity 3.0 standard unit/1 g of nucleic acids and more are sampled. |
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Method for virus replication in avian embryo stem cells Method involves isolation, culture and replication of avian embryo stem cells in a complete culture medium containing all growth factors in the presence of an inactivated feeder layer, and added with animal serum; separation of the growth factors, serum and feeder layer; production of the non-adhesive embryo stem cells; proliferation of the produced cells in a suspension in the form of cell balls; infection of said cells with said virus; addition of the second serum-free medium to the cell culture; further culture of the infected cells for virus replication and collection of said virus. |
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For the purpose of prediction of developing bacterial complication of a staphylococcal aetiology following influenza, the pure culture S. aureus is recovered from nasopharyngeal mucosal microflora to determine hemolytic activity (HA), antilysozyme activity (ALA), then S.aureus is co-incubated in vitro with type B influenza; S.aureus is analysed again for hemolytic and antilysozyme activity, and increasing S.aureus HA by 1 mm and more and/or ALA by 1 mcg/ml and more after co-incubation, a developing bacterial complication is predicted. |
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Method of estimating degree of pseudomonas aeruginosa strain pathogenicity Estimating a degree of Pseudomonas aeruginosa strain pathogenicity is ensured by taking integral exometabolites of the night culture of Pseudomonas aeruginosa, freezing at minus 18°C for at least 1 hour, thawing and mixing with a rehydrated sensor that is a test culture of E coli lux+. The mixture is kept for 30 min at 20°C, and a degree of inhibition of test culture bioluminescence shows a potential pathogenicity index (PPI) calculated by formula PPI=(Ir-It)/lk·100% wherein Ik is the luminescence intensity of the reference sample, It is the bioluminescence intensity of the test sample. The value PPI≤30% shows a low degree of P. aeruginosa strain pathogenicity; the value PPI varying within 30% to 70% shows a moderate degree of pathogenicity, while the value PPI>70% presents a high degree of P. aeruginosa strain pathogenicity. |
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Method for estimating antibacterial activity of aerobic sporous microorganisms bacillus subtilis Antibacterial activity of aerobic sporous microorganisms Bacillus subtilis is estimated by test culture inoculation of the whole surface of meat infusion agar in Petri dishes. Zeolite granules of the Hongurinskoye deposit is prepared for 60 min at room temperature and processed with a suspension of bacterial antagonist strains Bacillus subtilis TNP-3-DEP and Bacillus subtilis TNP-5-DEP. They are placed on Petri dishes. The inoculation products are incubated and calculated by bacterial antagonist growth power after 72 h of incubation. |
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Method of evaluating ecological state of environment Method involves collecting samples of planktons inhabiting a water body, determining the contamination level through analysis thereof and evaluating analysis results, wherein the contamination level is determined by determining conservative and variable gene sequences of a plankton from the sample, followed by molecular physiological analysis to determine evolutional relationships of the analysed organism with other saprobionts. Analysis results are evaluated as follows: at high (more than or equal to 70%) value of bootstrap cluster support on the conservative and variable gene, including the analysed plankton and resistant saprobionts. The following conclusions are made. When the resistant indicator organisms of xeno- and/or xeno-oligosaprobic water bodies and the analysed plankton merge into a single cluster, the water body is in a safe ecological state and there is no threat of negative human impact. If the resistant indicator organisms of oligo- and/or oligo-mesosaprobic water bodies and the analysed plankton merge into a single cluster, the water body is in an unstable ecological state, is capable of self-restoration and does not need additional nature conservation activities. When the resistant indicator organisms of meso- and/or meso-polysaprobic water bodies and the analysed plankton merge into a single cluster, the water body is in a bad ecological state and needs nature conservation activities. When resistant indicator organisms of polysaprobic water bodies and the analysed plankton merge into a single cluster, there is a local ecological disaster and there is need for urgent remedial measures. |
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Method for screening of probiotic preparations for nocardioform actinomycetes Mice are infected by the oral introduction of a suspension 0.1 ml of a two-day swine Rhodococcus equi, pathogenic of culture containing 5×107 CFU in normal saline 1.0 ml. The infection is followed the oral five-fold introduction of a probiotic preparation 1-5×105-6 CFU in 1.0 ml every 24 hours; in 5 days after the infection, the mice are killed, and dense egg medium are inoculated of the internals suspensions. In 1-2 cultivation days at optimal temperature, but not later than in 5 days, the grown colonies are controlled by acid fast stain. Antagonist activity is evaluated by a growth block index that is a relation of Rhodococcus colony count grown at inoculation of the internals suspensions of the animals prescribed with no probiotic preparations to Rhodococcus colony count grown at inoculation of the internals suspensions of the animals prescribed with the probiotic preparations. The efficacy of the probiotic preparation is concluded if the value exceeds 3. |
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Method to determine toxicity of wastes and soils Method includes measurement of microorganisms test function level in presence and in absence of an analysed sample and calculation of toxicity based on the produced results. Dense samples are exposed to testing (wastes, soils). At the same time testing is carried out without preliminary procedure of producing an aqueous extract of the sample. A test item is a culture of a soil bacillus with dehydrogenase activity, and the test function is its dehydrogenase activity, which is identified using resazurin and recorded using a conventional measurement device for spectrophotometry. |
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Invention represents control strain of Neisseria gonorrhoeae, intended for intra-laboratory control over quality of analysis in carrying out laboratory diagnostics of gonococcal infection by culture method. Strain was isolated from patients with uncomplicated gonococcal infection before beginning their treatment with antimicrobial medications and is deposited in state collection of microorganisms of FSUE " Research institute of standardisation and control of biological preparations named after L.A. Tarasevich Rospotrebnadzor" under No 283. Obtained strain stably preserves its signs and properties and is the first our country reference-strain for intra-laboratory control over research quality in laboratory diagnostics of gonococcal infection. |
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Method of nanocarbon analysis for biotoxicity Sample is prepared: an additional weight of nanocarbon forms is dispersed in 1 ml of organic solvents with a degree of polarity smaller than that of water - dimethyl sulphoxide or ethanol. Then it is mixed and exposed to ultrasound for 30 minutes. The prepared nanocarbon suspension is transferred in an aqueous medium to the final concentration of the used solvent 2.5 %. The produced and control samples are added with a viable sensor recombinant luminescent Escherichia coli K12 strain with cloned luxCDABE genes of luminescent Photobacterium leiognathi system. It is followed by incubation for 60 - 180 minutes, measuring luminous intensity and evaluating optical properties of the analysed suspension simultaneously. A toxicity index (T) is calculated with evaluating an actual luminous intensity of the strain (Iact) in comparison with the control of the same concentration of the solvent, considering light absorbing properties of the analysed suspension (D) and an experimental luminescence level of the bacterial luminescent biosensor (Iexp). |
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Biosensor for detecting 2,4,6-trinitrotoluene Disclosed is use of a Yarrowia lipolytica VKPM Y-3492 yeast strain as a biosensor for detecting 2,4,6-trinitrotoluene in different media. Free or immobilised cells of Yarrowia lipolytica VKPM Y-3492 yeast are used to detect 2,4,6-trinitrotoluene in different media. Use of the Yarrowia lipolytica VKPM Y-3492 yeast strain enables to detect 2,4,6-trinitrotoluene in a wide range of concentration from 0.1 to 100 mg/l without using cultures, in a wide range of pH from 4.5 to 8.0 and temperature from +10°C to +33°C. |
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Method provides contact of said cell with a peptide nucleic acid (PNA) molecule and a photosensitising agent and cell exposure to light at wave length effective to activate the photosensitising agent where said PNA molecule is conjugated with positive peptide. Also, there are described compositions containing such conjugated PNA molecules, the cells produced with the use of the method, and application of the method. |
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Polypeptide, which is used in composition of pharmaceutical composition and in sets for screening of adhesion inhibitors of platelet adhesion or aggregation, is obtained in recombinant way applying matrix of cDNA of Anopheles stephensi salivary gland. |
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Method of phagocyte activation test Method provides phagocyte recovery from a cell mixture. The recovered cells are mixed with medium 199 in 18 bottles, and the cells are attached to glass at room temperature for 60 minutes. In 9 bottles, a culture medium containing medium 199, L-glutamine and mixed human serum heated for 30 min at 52°C is added to the cells in the preset amounts to produce cell samples. A culture medium of said composition containing 0.01% of zymosan particles is added to another 9 bottles with the cells to produce the cell samples. The prepared cell samples are divided into three portions. One one-third potion of zymosan and zymosan-free samples are frozen. The second portion of the zymosan and zymosan-free cell samples are cultivated at temperature 37°C for 4 to 6 h to be frozen, and the third portion of the zymosan and zymosan-free samples are cultivated at temperature 37°C for 18 to 24 h to be frozen respectively. All 18 bottles are exposed to simultaneous multiple freezing to -10°C and thawing at room temperature to complete recovery of intracellular lysozyme with determination of amount by a micromethod and calculation of synthesised lysozyme and a phagocyte activation value (PAV) by formula: PAV=Z Lsynth - Z-free Lsynth, mcg/ml, where Z Lsynth is a difference of lysozyme amount in the cultivated zymosan cells and lysozyme amount in the uncultivated zymosan cells, mcg/ml; Z-free Lsynth is a difference of lysozyme amount in the cultivated zymosan-free cells and lysozyme amount in the uncultivated zymosan-free cells, mcg/ml; the phagocyte activation value for long-term activation mechanism assessment is calculated by said formula by results of cell cultivation for 4 to 6 hours, and the phagocyte activation value for long-term activation mechanism assessment is calculated by said formula by results of cell cultivation for 18 to 24 hours. |
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Procedure for choice of strains of micro-organisms-destructors of oil and oil products Samples from oil contaminated surface are chosen for selection of strains of micro-organisms-destructors of oil and oil products. There are selected pure cultures of hydrocarbon containing bacteria and they are cultivated on dense growth medium. There is determined catalase activity of grown strains of micro-organisms. Further, they are used for preparation of one billion microbe suspension which is mixed with Raymond liquid medium at ratio 1:150. As a sole source of carbon there is added oil or oil products from a place of contamination at 1 cm3 per 1 dm3 of Raymond medium and there is carried out incubation during 12 days. Further, culture is sown on dense growth medium and cultivated. Upon completion of cultivation there is determined catalase activity of studied strains. At its decrease in comparison to a source at 30 % and more analysed strain of micro-organism is chosen as active destructor of oil and oil product. |
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Method for bioindication of water bodies Method for bioindication of water bodies involves collecting samples of planktons inhabiting in a water body, determining the contamination level by analysing said samples and assessing the analysis results. The contamination level is determined via phylogenetic analysis of ribosomal RNA genes (18S rRNA) of planktons in the sample. Phylogenetic trees built from the conservative 18S rRNA gene are determined and evolutionary relationships of the analysed object with other saprobionts are identified. Analysis results are assessed as follows: at high (over 85%) value of bootstrap support of clusters containing the analysed planktons and resistant saprobionts, the following conclusions are made: resistant indicator organisms xeno- or oligosaprobic (or exclusively xenosaprobic) of water bodies and the analysed plankton merge into one cluster, it is concluded that the water body is in a safe ecological state and there is no threat of negative anthropogenic action, if resistant indicator organisms oligo- and mesosaprobic (or exclusively oligosaprobic) of the water bodies and the analysed plankton merge into one cluster, it is concluded that the water body is in an unstable (transition from safe to unsafe state) ecological state, is under insignificant anthropologic load, is capable of self-recovery and does not need additional environmental protection measures, if resistant indicator organisms meso- and polysaprobic (or exclusively mesosaprobic) of water bodies and the analysed plankton merge into one cluster, it is concluded that the water body is in an unsafe state and is under considerable anthropologic load, natural capability of self-recovery is insufficient and the water body needs environmental protection measures, if resistant indicator organisms of polysaprobic water bodies and the analysed plankton merge into one cluster, it is concluded that there is a local ecological disaster and there is need for urgent recovery measures. |
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Immunoglobulin work solution migration time specific to each series and manufacturer is set; specific immunoglobulin is used as a reference mark (control). Further, an analysed sample suspected for the presence of sporous bacteria is prepared for an antigen-antibody reaction by adding the specific immunoglobulin work solution in a phosphate buffer to the sample. In reaction, a complex of bacterium in the sporous form and specific immunoglobulin is produced. It is followed with capillary electrophoresis, UV detection, complex migration time test in the capillary electrophoresis system to be compared with the migration time in the capillary electrophoresis system of the specific immunoglobulin (control). If the complex migration time exceeds the specific immunoglobulin migration time by 50 seconds, the presence of bacteria in the sporous form is detected in the sample. Also, the analysis conditions and parametres are described. |
Another patent 2513104.