Method for detecting glycogen in extract out of organs and tissues in bees |
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IPC classes for russian patent Method for detecting glycogen in extract out of organs and tissues in bees (RU 2256320):
      
 Method for detecting nerve fibers of adrenergic and cholinergic nature in tissue / 2256179
Method involves fixing material and setting histochemical reaction on detecting cholinergic nerve structures. The fixed material under study is incubated during 60-90 min in thermostat at 37°C in mixture containing acetylthiocholine iodide as substrate, washed, treated with alcohols in growing concentrations, cleared and enclosed into balsam. The material is fixed with 2% glyoxylic acid prepared on 0.1 M phosphate buffer solution having pH of 7,0. Histochemical reaction is set concurrently with material fixation for detecting adrenergic nerve structures containing neuromediators forming luminescent reaction products with glyoxylic acid, in which the material is incubated during 15-20 min at temperature of 20-25°C. Then it is dried and heated during 5-10 min in thermostat at 80°C, studied with luminescent microscope, subjected to morphometric study and photographed. Then, histochemical reaction is set on the same material for detecting cholinergic nerve structures.
Method for predicting clinical course of nonhealing peptic stomach and duodenal ulcer / 2255661
Method involves determining proportion of cyclic adenosine monophosphate to cyclic guanosine monophosphate, serotonin and histamine level and serotonin/histamine ratio coefficient, content of prostaglandin E2 and prostaglandin F2α and adenylatecyclase index. Cyclic adenosine monophosphate to cyclic guanosine monophosphate ratio being equal to 2.7-3.3, serotonin level of 4.5-6.6, content of prostaglandin E2 being equal to 670-1340 mg/g of protein and prostaglandin F2α to 320-960 mg/g of protein, label index being equal to 10.1-35,0% and adenylatecyclase index being equal to 19.2-30.0 rmole/g of protein/min, stomach and duodenal ulcer healing is predicted.
Method for differentiated prescription of hyperbaric oxygenation at pulmonary cancer / 2255338
Before carrying out hyperbaric oxygenation (HBO) one should detect the content of lipid peroxidation products in the condensate of expired air. If the content of TBC-active products is below 14 conventional units it is necessary to perform about 3-4 HBO seances at 1.3 absolute atmosphere for 40 min both in pre- and postoperative periods. If the content of TBC-active products varies 14-24 conventional units one should conduct about 3-4 HBO seances at 1.3 absolute atmosphere for 40 min in preoperational period only. If the content of TBC-active products is above 24 conventional units HBO séances should not be carried out . The method is considered to be noninvasive, of high information value and enables to increase efficiency to apply HBO in patients with pulmonary cancer planned for operative treatment.
Method for predicting encephalopathy in neonatals / 2254573
One should study placental tissue to prepare 10% homogenate and detect NADPH-oxidase activity according to reduction of 2.6-dichlorophenylindophenol at the presence of NADPH. Results of reaction should be specified spectrophotometrically at wave length being 600 nm. Method is atraumatic and enables to predict the development of encephalopathy in neonatals at high accuracy.
 Retentive chromatography method for separating analytes in sample / 2253116
Method involves producing substrate for detecting analytes in a sample by treating the sample at least under two selectivity conditions depending on adsorbent and eluate combinations. It enables one to retain analyte with the sorbent, to make identification by applying desorbing spectrometry. Method for setting diagnosis with the substrate is given.
Method for predicting peritoneal commissure disease in children / 2252711
Method involves carrying out blood serum examination. Infrared blood serum spectroscopy, infrared radiation transmission index and mean transmission coefficient value are calculated at the second day after surgical intervention. The mean transmission coefficient value being found less than 64.4 in 2120-1880 in 2120-1880 cm-1 bandwidth, peritoneal commissure disease is predicted to be the cases.
 Method for determining tissular plasminogen adjuvant / 2252421
Method involves incubating sample with plasminogen, plasminogen-to-plasmin transformation stimulator under the tissular plasminogen adjuvant action and chromogenic synthetic plasmin substrate and determining tissular plasminogen adjuvant activity from substrate hydrolysis product concentration colored with the plasmin. Lys-plasminogen pretreated with pancreatic trypsin inhibitor or Glu-plasminogen is used as the plasminogen. Specific HCO-Ala-Phe-Lys-p-ntroanilide is applied as the plasmin substrate. Soluble fibrin-monomer produced by means of thrombine-like enzyme ancistrone from snake poison of is applied as the plasminogen-to-plasmin transformation stimulator under tissular plasminogen adjuvant action. Incubation is carried out at pH=7.0-8.5. The most optimum concentration for Lys- or Glu-plasminogen is equal to 0.3 mcM, and it is equal to 0.6 mM for the specific HCO-Ala-Phe-Lys-p-ntroanilide substrate, and 0.025 mg/ml for the fibrin-monomer.
 Methods for detecting both components of binding pair / 2251111
Method involves making interaction of biological sample with solid phase reagent covered with catching antibodies showing affinity to component A binding to produce the first reacted particle. Making the particle to come in contact with labeled antibodies showing specific binding affinity with respect to A and B components results in the second reacted particle production. Labels are measured on the second reacted particle by applying flowing cystometry method.
Method for quantitative detection of mucin / 2250465
The quantity of salivary mucin should be detected due to spectrophotometric method by the difference of protein concentration in initial material and supernatant developed after acidic precipitation of mucin. For this purpose two experimental samples should be prepared: the first contains saliva and working reagent, the second - supernatant and working reagent; and standard sample containing aqueous albumin solution at 0.25 g/l concentration and working reagent; and a control sample containing distilled water and working reagent. The latter should be obtained due to mixing the solution of bromine phenol blue at concentration of 1.2 g/l and buffer solution (pH 3.0) containing 320 mM/l citric acid and 160 mM/l sodium phosphate at the ratio of 2:23. The content of each sample should be mixed and incubated for 10 min. One should detect optic density of experimental and standard samples against control one at wave length of 620 nm. Salivary protein concentration and that of supernatant should be calculated in g/l: C = (ODex : ODst)x 0.25, where ODex - optic density of experimental sample, ODst - optic density of standard sample. The method is very simple in its implementation, its conditions provide stability of stained product that enables to simultaneously investigate many samples.
Differential diagnosis method for detecting perinatal hypoxic nervous system injuries of newborns / 2249823
Method involves applying infrared spectroscopy approach for analyzing 0.1 ml of blood serum during 30 s in bandwidth of 3500-3200 cm-1 and 3500-3200 cm-1. Transmission coefficient is determined in each bandwidth interval and mean value is calculated. Mean value of transmission coefficient being from 2 to 4% in bandwidth of 3500-3200 cm-1, hypoxic nervous system injury is considered to be the case. The transmission coefficient being from 1 to 3% in bandwidth of 2120-1880 cm-1, traumatic injury is considered to be the case.
Method for predicting hereditary predisposition for the development of ovarian cysts in cows / 2255470
One should detect haptoglobin types in cow's blood serum. Animals should be tested at the age of 8-10 mo either before or after the first calving. Animals of Hp 2-2 type should be referred to those with hereditary resistance for ovarian cysts development, and animals of Hp 1-1 type and Hp 2-1 type should be considered to be hereditarily predisposed to pathology under investigation. Application of the present method enables to detect animals with hereditary predisposition to the development of ovarian cysts in earlier period.
Method for withdrawing lead out of poultry body / 2255469
One should daily introduce sodium selenite into chickens' diet at the quantity of 1.5 mg selenium/kg feedstuff. The method enables to decrease the content of lead in muscular tissue by 69.8-78.4%, in bones - by 74.4%.
 Method for preparing biohumus / 2255078
Method involves stacking organic substrate for layers formed by longitudinal distribution of fresh feeding consisting of non-pressed organic substrate followed by placing worm population to the substrate. After formation of substrate layers fresh feeding is applied on similar right or left lateral sides of all layers by all surface of each lateral side by a layer wherein compost worms migrate continuously and form active zone of vermicomposting and vermiculturing. Then after formation of biohumus the following feeding layer is applied on a layer wherein compost worms migrate continuously also and form the next active zone of vermicomposting and vermiculturing and a layer of ready biohumus is removed from the opposite left or right lateral side of each layer, respectively. Applying layers of feeding and gathering ready biohumus is carried out up to the complete using the parent distance between adjacent layers that provides the gradual transferring active zones of vermicomposting and vermiculturing and, therefore, all layers to a single site for the distance value between adjacent layers. Then feeding is applied on opposite initially taken lateral sides of all layers, and the process is repeated again providing the graduate transferring active zones of vermicomposting and vermiculturing and, therefore, all layers to another site and returning all layer to the initial point, and then all process is repeated. Method provides reducing cost in producing biohumus due to simplifying a method for its preparing and enhancing output. Invention can be used in utilization of waste in animal husbandry, residues of wastes and organic waste using rain worms and for preparing valuable organic fertilizer - biohumus.
 Method for preparing biohumus / 2255077
Method for preparing biohumus involves addition of manure worms to compost-containing substrate that is prepared by mixing beet pulp with lime in the ratio 10:1 by volume. Then the prepared mixture is mixed with garden earth in equal ratios by volume and a filling agent is added in the ratios 5-10% of volume of the latter prepared mixture. Cut straw is used as a filling agent. Invention provides enhancing the intensity process for processing waste and to reduce consumptions for preparing biohumus.
Method for production of acaricide and insecticide composition / 2254719
Suspension of entomopathogenic eelworms of genus Steinernema is mixed with antidesiccant. Said antidesiccant is obtained by subsequent extraction of biomass of Mortierella dichlotoma micromycete with non-polar extractant in above-critical state, water, alkali, water, acid, water, alkali, and water followed by conjugating of the first extract with the solid residue and is used in amount of 0.2-1 % based on composition mass.
Method for production of acaricide and insecticide composition / 2254718
Suspension of entomopathogenic eelworms of genus Steinernema is mixed with antidesiccant. Said antidesiccant is obtained by subsequent extraction of biomass of Pythium coloratum micromycete with non-polar extractant in above-critical state, water, alkali, water, acid, water, alkali, and water followed by conjugating of the first extract with solid residue and is used in amount of 0.2-1 % based on composition mass.
Method for production of acaricide and insecticide composition / 2254717
Suspension of entomopathogenic eelworms of genus Steinernema is mixed with antidesiccant. Said antidesiccant is obtained by subsequent extraction of biomass of Pythium insidiosum micromycete with non-polar extractant in above-critical state, water, alkali, water, acid, water, alkali, and water followed by conjugating of the first extract with solid residue and is used in amount of 0.2-1 % based on composition mass.
Method for manufacturing acaricide-insecticide composition / 2253990
The present innovation deals with manufacturing special preparations for plant protection against harmful insects and ticks. Nematodes' suspension of Steinernema sp. should be mixed with an antidesiccant obtained due to successive extracting Pythium catenulatum micromycete biomass with a non polar extracting agent in supercritical state with water, alkali, water, acid, water, alkali and water followed by combining the first extract with solid residue. Antidesiccant should be introduced into suspension of entomopathogenic nematodes at the quantity of 0.2-1% against composition weight. The method enables to shorten the expenditure of nematodes due to the increase of their survival rate in the composition suggested.
Method for manufacturing acaricide-insecticide composition / 2253989
The present innovation deals with manufacturing special preparations for plant protection against harmful insects and ticks. Nematodes' suspension of Steinernema sp. should be mixed with an antidesiccant obtained due to successive extracting Mortierella zychae micromycete biomass with a non polar extracting agent in supercritical state with water, alkali, water, acid, water, alkali and water followed by combining the first extract with solid residue. Antidesiccant should be introduced into suspension of entomopathogenic nematodes at the quantity of 0.2-1% against composition weight. The method enables to shorten the expenditure of nematodes due to the increase of their survival rate in the composition suggested.
Method for manufacturing acaricide-insecticide composition / 2253988
The present innovation deals with manufacturing special preparations for plant protection against harmful insects and ticks. Nematodes' suspension of Steinernema sp. should be mixed with an antidesiccant obtained due to successive extracting Mortierella marburgensis micromycete biomass with a non polar extracting agent in supercritical state with water, alkali, water, acid, water, alkali and water followed by combining the first extract with solid residue. Antidesiccant should be introduced into suspension of entomopathogenic nematodes at the quantity of 0.2-1% against composition weight. The method enables to shorten the expenditure of nematodes due to the increase of their survival rate in the composition suggested.
 Method for selecting first-calf heifers for milking herd / 2244419
The present innovation deals with dynamic loading onto cardio-vascular system in animals. Selection should be carried out by the following parameters:
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FIELD: veterinary medicine, biochemistry. SUBSTANCE: the present innovation deals with boiling an extract, cooling, centrifuging, dissolving a residue, cooling, centrifuging, dissolving a residue, adding sulfuric acid into a tube and 1%-condensate's solution followed by heating, cooling, photometry against the control at wave length being 315 nm, as a condensate one should apply resorcinol. EFFECT: higher accuracy and economy of detection. 2 ex, 1 tbl
The invention relates to the field of veterinary medicine, in particular biochemistry of animals. In veterinary medicine the determination of glycogen is one of the most important tests in identifying energy resource the body of bees. Glycogen is a polysaccharide, or animal starch, is synthesized by the body and deposited in all its organs and tissues. Glycogen is easily MobileTerminal backup form of glucose and is a branched polymer of glucose residues of blood. Know the definition of glycogen in the blood cytochemical CHIC - reaction method of Sabadash [see Clinical laboratory analyst Ed. Won. Volume 2. Private analytical technologies in the clinical laboratory. M - “Labelform” - RAMLA, 1999. S. 59-61], where they perform the preparation of blood smears, fixed in absolute alcohol and processing - incubation in a solution of Periodica and stained with Schiff's reagent, followed by staining with hematoxylin and drying, where glycogen is painted in cherry-purple color on a green background of the drug, and microscopy. Also known determination of glycogen in the blood [Horeysi J. Et al. Zaklady chemickeho vysetrovani v lekarstvi. Pppraha, 1957. Handbook of Biochemical research methods in the clinic. Edited by academician of Academy of medical Sciences Prof. Ash. Medicine. M: - 1969. The determination of glycogen in the blood kolori eticheskim method orcine (Horejsi), pages 230-231], where glycogen is precipitated by alcohol and hydrolyzing in an acidic medium to glucose and heated in sulfuric acid, which is converted to hydroxymethyl furfural and condenses orcine, forming a colored compound. All of the above was carried out by boiling the studied biological substrate in a concentrated sodium hydroxide solution in a centrifuge tube, followed by the addition of alcohol, cooling, centrifuging, the selection of the supernatant liquid, dissolving the precipitate in polysystem solution of sodium sulfate, and then further cooled by centrifugation and dissolved sediment, simultaneously adding to a centrifuge tube with a test biological substrate, a control test tube with water in a test tube with a standard glucose solution of 13 ml of sulfuric acid and 1 ml of 1% solution of orcine followed by heating, cooling, photomatrixovina against control and determination of glycogen by the formula
where X is the number of glycogen (mg%; Fabout- extenze experimental samples; Farticle- extenze standard solution, The disadvantages of the known methods is time-consuming lengthy process, is not sufficiently accurate determination of glycogen content in the blood, as well as the inability to determine the content of glycol is on in the tissues of the organs of bees. Technical solution to the problem is to increase the accuracy of diagnostics, reduce cost, time, material costs and use more of the available chemical reagents and the expansion of technological capabilities. This object is achieved in that in the method of determination of glycogen content in the extract from the organs and tissues of bees, including boiling the studied biological substrate in a concentrated solution of sodium hydroxide in centrifuge tube followed by the addition of alcohol, cooling, centrifuging, the selection of the supernatant liquid, dissolving the precipitate in polysystem solution of sodium sulfate, and then further cooled by centrifugation and dissolved sediment, simultaneously adding to a centrifuge tube with a test biological substrate, a control test tube with water in a test tube with a standard glucose solution of 13 ml of sulfuric acid and 1 ml of 1% solution of the condensation followed by heating, cooling photomatrixovina against control and determination of glycogen by the formula
where G is the number of glycogen (mg%; Fabout- extenze experimental samples; Farticle- extenze standard solution, as a biological substrate using extracts from organs and tissues of bees, is to boil - 30% sodium hydroxide solution, while cooling is carried out for 15 minutes, centrifuged at 3 thousand rpm for 15 minutes, add in a test tube 2 ml of 1% condensate, which is used as resorcin, and photometrate at a wavelength of λ=315 nm. The novelty of the proposed method due to the fact that is the most accessible and accurate, and allows the determination of glycogen in tissues of organs of bees in comparison with the known method of determining the glycogen content of the blood as a scientific and economic perspective. The proposed method allows to determine the content of glycogen in the extract of organs and tissues from bees. In addition, the inventive proposal is more economical, since it does not require the use of expensive chemical reagents. A specific example of the method of determination of glycogen content in the extract from the organs and tissues of bees Preparation of extract from organs and tissues of bees. For this, the bees removed the outer cover and intestine using tweezers and eye scissors. Tissues and organs obtained from ten bees were placed in a porcelain mortar and triturated with the pestle, then to the content added to the physiological solution at a ratio of 1:1, insisted during 1-1,5 hours in the refrigerator at a temperature of 5°and filtered through a paper filter. Preparation, the tion solutions For the preparation of 52% sulfuric acid by volume of 325 ml must take 225 ml of concentrated sulfuric acid and gently Prilepa, 100 ml of distilled water. To prepare 1% solution of resorcinol need to take 1 g of resorcinol and 99 ml of 52% sulfuric acid solution. To prepare a 30% solution of sodium hydroxide required to take 30 g of sodium hydroxide and 70 ml of distilled water. To prepare a standard solution of glucose take 10 mg of glucose and dissolved them in 300 ml of distilled water. Precipitation and hydrolysis of glycogen in the extract from the organs and tissues of bees The filtered extract from the organs and tissues of bees were placed in a centrifuge tube in the amount of 0.3 ml and there was made 0.2 ml of distilled water and 0.2 ml of 30% sodium hydroxide solution. Then the tube was placed in a water bath for 1.5-2 hours, after cooling the tube was introduced 1 ml of 96% vol. ethanol, suspended and cooled on ice for 15 minutes, then the solution was centrifuged at 3 thousand rpm for 15 minutes. The supernatant was carefully removed the Pasteur pipette, and the residue was added 1 ml of 96% vol. ethanol and cooled on ice for 15 minutes, then centrifuged at 3 thousand rpm for 15 minutes. After centrifugation the supernatant was carefully removed the Pasteur pipette, and the tank was mixed with 3 ml of distilled water, 13 ml of a 52% solution of sulfuric acid (mixed) and 2 ml of 1% resorcinol. In the control (pure tube) tube with standard glucose solution was poured in 3 ml of distilled water, 13 ml of a 52% solution of sulfuric acid (mixed), and 2 ml of 1% resorcinol. All test tubes were placed in a water bath at a temperature of 80°C for 20 minutes, then cooled on ice for 15 minutes. The tubes with the presence of glucose had a brown-yellow color. Photocolorimetry held against control KLF-3 in the selection of wavelength λ=315 nm. The calculation of the glycogen content in the extract from the muscles is conducted according to the formula
where G is the number of glycogen (mg%; Fabout- extenze experimental samples; Farticle- extenze standard solution. Example: the gray bees Caucasian breed Eabout- extenze experimental samples was 0,566 Earticle- extenze standard solution was 0,253 Thus, substituting in the formula, we get the following result:
On the basis of data given in the table, we can conclude that the breed bees Prioksky has the largest energy resource in comparison with other breeds of bees. This breed bees resistant to adverse weather conditions and has the ability to reproduce offspring with increased immunity. Method for determination of glycogen content in the extract from the organs and tissues of bees, including boiling test substrate in a concentrated solution of sodium hydroxide, cooling, centrifugation, dissolving the precipitate in a solution of sodium sulfate, cooling, centrifugation, dissolution of the precipitate, the simultaneous addition of the test tube with the test biological substrate, a control test tube with water in a test tube with a standard glucose solution of 13 ml of sulfuric acid and 1% solution of the condensation followed by heating, cooling, photometry against control and determination of glycogen by the formula: G=Eabout·100/Earticlemg%, where Y is the quantity of glycogen in mg%; Fabout- extenze experimental samples; Farticle- extenze standard process is and, for boiling the substrate using 30%sodium hydroxide, cooled to 15 min, centrifuged at 3 thousand rpm./min for 15 min, add in a test tube 2 ml of 1% solution of the condensate, which is used as resorcinol, and photometrate at a wavelength of 315 nm.
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