Method for in vitro recovery of experimental tuberculous granuloma in cultures
SUBSTANCE: method for in vitro recovery of experimental tuberculous granulomas in culture involves induction of granulomatous inflammation by infecting experimental animals with BCG mycobacteria, recovery and mechanical disintegration of spleen tissues containing granulomas over a period adequate to form them in an animal's body, homogenisation of spleen tissue in a solution by agitation, purification of the homogenate from coarse tissue fragments by spontaneous deposition in a culture medium, removal of the deposits, recovery of the granulomas from a supernatant, washing in a new portion of the culture medium by centrifugation at acceleration 15-28 g at least three times and transfer of the deposited granuloma in the culture medium.
EFFECT: effective granuloma integrity and higher purity of the recovered granulomas.
4 cl, 3 ex
The invention relates to experimental biology and medicine, and in particular to methods of explantation and cultivation of granulomas in vitro, and can be used when developing and evaluating the effectiveness of the treatment of tuberculosis complicated with tuberculous granulomatous process (necrosis, fibrosis), as well as in the development of new immunotropic drugs that regulate the functional activity of cells of granulomas and enhance their anti-TB activity.
There is a method of allocating chistosortnykh granulomas, including the induction of granulomatous inflammation by intraperitoneal infection of experimental animals (mice CBA/J) cercariae schistosomes (Schistosoma mansoni), the selection of the liver over a period of time sufficient for the formation of granulomas (8 weeks), preparation of homogenate of liver tissue by mechanical disruption using a Waring blender at low speed, the laundering of the homogenate several times in buffer solution RPMI 1640 by spontaneous deposition, the allocation of granulomas from the homogenate by centrifugation cooled to +4°C. the homogenate at 350 g for 5 min, the location of sediment containing granulomas in a nutrient medium for cultivation of granulomas (1). The disadvantage of this method is its inapplicability to highlight tuberculous granulomas. This is due t the m unlike chistosortnykh granulomas, the cells are firmly linked together by collagen fibers, tuberculosis granulomas do not possess such strength and are destroyed when chopping liver tissue or captured together with areas of tissue with which they are closely linked.
There is a method of separating and explantation chistosortnykh granulomas in culture in vitro, including the induction of granulomatous inflammation by intraperitoneal infection of experimental animals (mice C57BL/6) cercariae schistosomes (Schistosoma japonicum), the selection of the liver over a period of time sufficient for the formation of granulomas (6 weeks), preparation of homogenate of liver tissue by mechanical disintegration within 15 seconds using a blender in 10 volumes of buffer Hanks solution, the laundering of granulomas in cold buffer Hanks solution, the selection of granulomas from the homogenate by centrifugation cooled homogenate at 200 g up until the supernatant becomes transparent, the location of granulomas in the nutrient medium for cultivation (2). The known method have the same shortcomings.
Closest to the claimed method is the selection of tuberculous granulomas, including the induction of granulomatous inflammation by vnutribruchinnogo infection of experimental animals (mice) the Mycobacterium is M. avium, removing the liver through a period of time sufficient for the formation of granulomas, mechanical disintegration of the liver tissue by gently rubbing it between two subject chilled glasses (between their polished surfaces), preparation of homogenate of liver tissue in buffered Hanks solution, cleaning homogenate from large pieces of fabric by spontaneous deposition, removal of sediment, the allocation of granulomas from the supernatant of the suspension by subsequent triple centrifugation at 150 g with sequential removal of the supernatant and dissolve the precipitate with granulomas in the new portion of the buffer Hanks solution; the location of the selected granulomas in the medium for cultivation (3). The disadvantage of this method is that using the liver as a source of granulomas does not allow to distinguish "pure granulomas without impurities. This is due to the significant integration of granulomas in the liver tissue, the high density of the liver that prevents the release of even the most durable and compact vibrazioni tubercular granuloma intact, because the process of disintegration must be very hard. For this reason, in the known method using glass slides with polished surface as a means of mechanical disintegration of tuberculous granulomas that PR is leads to significant structural damage granulomas, in terms of the violation of their integrity. In addition, due to the high degree of connection of hepatocytes of the liver among granulomas in their mechanical selection there is a large number of fragments of liver tissue, comparable in size with granulomas, which makes the process of fractionation of pure suspension of granulomas very problematic. Another disadvantage of this method is the presence of a dedicated granulomas impurities in the form of a large number of erythrocytes, lymphocytes and macrophages. This is because the centrifugation of the homogenate (after deposition of a large piece of fabric) at 150 g leads to the deposition of these cells together with granulomas, which makes to get a clean granuloma
The problem to which the invention is directed to a more efficient preservation of the integrity of granulomas, increasing the purity of their selection.
The solution of this problem is achieved by the fact that the selection of granulomas use tissue of the spleen, the supernatant suspension containing granulomas, centrifuged in a gradient accelerations 15-28 g; deposition of large fragments of material from the homogenate and breeding besieged granulomas after each centrifugation is carried out in a culture medium; a mechanical disintegration of the tissue of the spleen produced by grinding into fragments of size 0.5-1 mm; it is each centrifugation the supernatant suspension is carried out for 2-5 min; the selection of the spleen are carried out through 30-90 days after infection;
Description of the invention
The method of obtaining experimental tuberculous granulomas in vitro as follows.
Experimental animals such as mice, intravenous Mycobacterium BCG vaccine for the induction of systemic tuberculosis inflammation. After the formation of experimental tuberculous granulomas (after 30-90 days after infection) are slaughtered in compliance with the rules of work with experimental animals with the use of anesthesia. Remove the spleen observing aseptic conditions and verify a sample of the spleen in the presence of induced granulomas, for example, by fluorescent or histochemical analysis. This is followed by mechanical disintegration of the tissue of the spleen by grinding into fragments of size 0.5-1 mm using a cutting tool such as a scalpel. Transfer minced tissue of the spleen in the medium for cultivation and homogenized by shaking, for example, on a shaker, within 1-3 minutes Spend cleaning homogenate from large fragments of tissue spleen by spontaneous sedimentation within 1-2 min in culture medium. Remove sediment. To highlight granulomas obtained supernatant, the suspension is centrifuged at an acceleration in the drop 15-28 g for 2-5 minutes The precipitate, containing granulomas, resuspension in a new piece of culture medium, repeat the procedure with the specified mode centrifugation at least two times. This allows you to clear granulomas from impurities of different cell types (erythrocytes, lymphocytes, excess macrophages). The precipitate, containing granulomas, diluted in medium for cultivation, cultivated under standard conditions at 37°C in atmosphere containing 5% CO2.
The advantage of using the spleen as an organ for excretion of granulomas is that granulomas are less integrated in the tissue of the spleen, which gives the opportunity to avoid their destruction at the stage of extraction and mechanical disintegration of the tissue of the spleen. In addition, the use of spleen allows you to select BCG-induced granuloma at all stages of their development, including prior to the beginning of fibrosis.
The selection of granulomas appropriate conduct through 30-90 days after infection. Early (30 day) in the spleen fully formed granulomas, in this period of adhesive contacts between cells granuloma sufficient to preserve the integrity of granulomas when you select them (they are not "crumble"). The increase in the period of formation of granulomas can be carried out to study the morphogenesis and development of granulomas in the later stages of vplo the ü to the stage of fibrosis.
Replacement homogenization by grinding between the glass (see the description of the prototype) on the mechanical grinding into fragments of size 0.5-1 mm contributes to the preservation of granulomas and increased purity of their allocation, because, on the one hand, their sizes from 60 to 125 μm, significantly smaller than these pieces of cloth; on the other hand, chopping into chunks of a specified size allows the release of granulomas from large pieces of tissue of the spleen.
The choice of gradient acceleration 15-28 g and duration of centrifugation experimentally chosen so as to precipitate granuloma of the homogenate earlier than in the sediment will fall in the homogenate of erythrocytes, lymphocytes and macrophages, the latter can be removed with the supernatant liquid, and does not precipitate with granulomas, as in the method prototype, which significantly increases the purity of the selection of granulomas. The speed of rotation of the centrifuge rotor when the same acceleration depends on the model of a centrifuge, or rather the radius of the rotor, and is determined by the well-known formula G=(2·π·f)2·R, where R is the radius of rotation of the rotor or the orbit of the body), f is the rotation speed in revolutions per second. For example, when using a centrifuge rotation speed corresponding to the acceleration 15 g, 400 rpm.
Method for assessing the viability of the cells based on p is jiznennoi colouring Trifanova blue it is shown that 99.5% of the cells in the granulomas remain viable. According to a computer morphometry digital photos granulomas, explanted into culture, it is shown that the culture of granulomas does not contain red blood cells and lymphocytes, and the number of macrophages is very small.
The number of free macrophages in suspension granulomas when released by the claimed method varies from 2-3 to 10-15 per selected granuloma or from 20 to 150 macrophages in 1 ml of the suspension depends on the number of ottavani granulomas by centrifugation in a solution of culture medium and the individual characteristics of the animal. Increasing the number of these procedures it is possible to achieve almost complete absence of macrophages in the composition of the granulomas. However, the presence of a small number of macrophages contributes to the improvement of culture conditions granulomas and increases the likelihood of their acceptance in the culture.
Using the inventive method culture medium (instead of buffer solution Hanks in the prototype) at the stage of spontaneous deposition of large fragments of material from the homogenate and on stage centrifugation promotes the viability of granulomas.
Another advantage of the claimed method is the possibility of its implementation without the use of expensive reagents, which makes extensive use of the ü it in practice research immunopathological mechanisms granulomatosa in the development of tuberculosis, the study of the immunological mechanisms that regulate the formation and evolution of granulomas in infectious granulomatous inflammatory processes, search, development and study of new anti-TB drugs.
Examples of specific performance
The mice BALB/c mice were injected intravenously Mycobacterium BCG vaccine (0.5 mg in 0.25 ml saline). 30 days after injection in mice was induced by chronic systemic inflammation, simulating generalized tuberculous inflammation. The mouse was scored after light ether anesthesia method of cervical dislocation. The spleen was removed, observing aseptic conditions for obtaining biological tissue samples for cultivation (in a laminar box "Clinic-572"), and were placed in a sterile plastic Petri dish (diameter 40 mm), which was pre-made and 1 ml of culture medium, namely the medium of alpha-MEM containing 5% serum of cow embryos.
Small fragments of spleen (1-2 mm) were cut with a scalpel for subsequent morphological analysis. Typical BCG-induced granulomas in the spleen was verified by fluorescent analysis (on the microscope AxioI-mager Z1, Zeiss) fragment spleen stained with acridine orange, as well as in the subsequent analysis of the sample villages is the eye the standard method of histological analysis (according to the traditional scheme, including formalin fixation, obtaining paraffin sections, dewaxing, painting with hematoxylin and conclusion in balm).
The contents of the spleen were released from the capsules and conducted preliminary mechanical homogenization of the splenic tissue directly in the Petri dish in 1 ml of the specified environment for cultivation by grinding with a scalpel into fragments of a size of 0.5-1.0 mm Further pre gomogenizirovannogo tissue of the spleen was placed into the flask with a working volume of 20 ml, was made in 9 ml of the specified environment for the cultivation and were shaken on a shaker at a frequency of 20 Hz for 1 min. Then the homogenate in the form of a suspension containing red blood cells, splenocytes, granulomas and larger fragments negozinegoziodisnay splenic tissue was transferred into a centrifuge tube 10 ml, left 1 min for deposition of large fragments negozinegoziodisnay splenic tissue. The supernatant suspension containing red blood cells, splenocytes, granulomas in the volume of 10 ml was centrifuged at acceleration 15 g for 5 minutes the Supernatant was decanted, the precipitate was reasponsible in a new portion of the specified culture medium. The procedure was repeated twice, and the duration of the second centrifugation was 5 min, the third - 3 minutes the precipitate containing granulomas, bred in an environment e is I cultivation and made in plastic tablets for cultivation, in which pre-placed glass cover. The cultivation was carried out on the basis of standard procedures: at a temperature of 37°C in atmosphere containing 5% CO2.
Dimensions isolated from spleen granulomas, explanted in culture, varied from 50 to 70 microns in diameter, reflecting the various stages of the granulomatous process.
The number of granulomas in the suspension amounted to a value of 19 granulomas/ml without morphological signs of violation of their integrity (integrity - 100%), while erythrocytes and lymphocytes were absent, and the content of macrophages was small (85 cells per 1 ml of suspension).
The claimed method was carried out analogously to the description in example 1 with the difference that the spleen was isolated after 90 days after infection. Each centrifugation was performed three times when starting acceleration 28 g for 2 minutes
Method for assessing cell viability, based on the in vivo colouring Trifanova blue, shows that 99.5% of the cells in the granulomas remain viable. According to a computer morphometry digital photos granulomas, explanted into culture, sizes isolated from spleen granulomas, explanted in culture, varied from 60 to 125 microns in diameter, reflecting the various stages of the granulomatous process.
The number of granulomas in suspense and amounted to the value of 21 granulomas/ml, the share of granulomas without morphological signs of integrity - 86%, the content of macrophages - moderate (175/ml suspension).
The claimed method was carried out analogously to the description in example 1 with the difference that the spleen was isolated in 60 days after infection. Each centrifugation was performed 4 times during acceleration 15 g for 5 minutes It was possible to isolate a pure granuloma, almost not containing macrophages, invalid splenocytes and other related cells, which may be necessary for future research in the study of factors that produced the actual granulomas.
The number of granulomas in the suspension amounted to a value of 20 granulomas/ml suspension, the share of granulomas without morphological signs of integrity violation - 90%, the content of macrophages is very low (15/ml suspension).
Sources of information
1. The method of obtaining experimental tuberculous granulomas in vitro culture, including the induction of granulomatous inflammation by infection of experimental animals with Mycobacterium BCG, selection and mechanical disintegration of the tissues of the body, containing granulomas, through a period sufficient for their formation in the animal body, homogenization of tissue in the solution by shaking, cleaning homogenate from the available fragments of tissue spontaneous deposition in a physiologically acceptable solution, removal of sediment, the allocation of granulomas from the supernatant suspension and laundering in a physiologically acceptable solution by centrifugation at least three times; the transfer besieged granulomas in the environment for cultivation, characterized in that the selection of granulomas use tissue of the spleen, the supernatant suspension containing granulomas, centrifuged at acceleration 15-28 g; deposition of large fragments of material from the homogenate and breeding besieged granulomas after each centrifugation is carried out in a culture medium.
2. The method according to claim 1, characterized in that the mechanical disintegration of the tissue of the spleen is produced by grinding the fragments of 0.5-1 mm.
3. The method according to claim 1, characterized in that each centrifugation the supernatant suspension is carried out for 2-5 minutes
4. The method according to claim 1, characterized in that the selection of the spleen carried out in 30 to 90 days after infection.
SUBSTANCE: lachrymal fluid is examined for extracellular peroxidase activity (EPA) both pre-therapy, and on the 7th therapeutic day. The EPA variation index is calculated as the relation of the EPA value on the 7th therapeutic day to the pre-therapy EPA value. If the EPA variation index exceeds 0.87, an unfavourable clinical course is predicted.
EFFECT: use of the method enables early prediction of the clinical course of bacterial conjunctivitis.
1 tbl, 5 ex
SUBSTANCE: prepared biological material is fixed in 10% neutral formalin, imbedded in paraffin; sections are made in a microtome, thereafter placed on a slide surface and chemically purified from paraffin with using xylene with collagen fibre maturity being evaluated by two independent criteria: a degree of allowed cross striation (D-periodicity) and a collagen fibre hardness wherein the degree of allowed cross striation is evaluated as a pitch height of a slot and an overlap of separate microfibrilles by Fourier analysis (Image Analysis 3, NT-MDT) on the basis of axially transverse and longitudinal profile analysis on topographic images pre-processed by a non-linear high-frequency filter with the use of Image Analysis 3 program (NT-MDT), while the collagen fibre hardness is evaluated as a failed wave phase of a cantilever of a scanning probe microscope. The sample is tested in the air medium.
EFFECT: along with simplification of the method, higher result reliability is ensured.
SUBSTANCE: reagent for thrombocyte measurement containing Nile blue hydrosulphate as a reagent for thrombocyte staining. A reagents kit for thrombocyte measurement containing Nile blue hydrosulphate as a first reagent, and a buffer solution as a second reagent. A method for thrombocyte measurement wherein Nile blue hydrosulphate is used as the first reagent, and the buffer solution - as the second reagent. A reagent for thrombocyte measurement containing Nile blue and an acid as a staining agent for thrombocyte staining. The reagents kit for thrombocyte measurement containing Nile blue as the first reagent, and a buffer substance as the second reagent. The method for thrombocyte measurement wherein Nile blue and the acid are used as the first reagent, and the buffer substance - as the second reagent.
EFFECT: reagents described above provides higher accuracy of thrombocyte evaluation.
13 cl, 15 dwg, 10 tbl, 48 ex
SUBSTANCE: chemiluminescence analysis is used to assess functional activity of white blood cells. Additionally, the parameters of chemiluminescence kinetics induced by a suspension of a clinical strain of methicillin-resistant Staphylococcus aureus, and if the curve peaking time values ranging within 2287 to 3520 seconds, the maximum signal intensity ranging within 4257 to 9516 standard units, while the light sum value is within 169000 to 362000 standard units, a normal functional ability of phagocytes is stated, while the different values enables recording a disturbed functional ability of phagocytes.
EFFECT: use of the declared method enables higher sensitivity for assessing the functional ability of phagocytes and higher specificity for detecting phagocyte dysfunction.
1 ex, 4 tbl
SUBSTANCE: method involves DNA recovery from peripheral venous blood, analysis of lymphotoxin α gene polymorphism, and a homozygous genotype shown by high-producing allele (+250GG) Ltα enables to predict a risk of multinodular lesions accompanying uterine leiomyoma.
EFFECT: higher prediction accuracy.
SUBSTANCE: method for evaluation of cephalosporin antibiotics both in oral fluid and whole blood that involves sampling of oral fluid and whole blood containing an antibiotic that is followed by sample preparation to remove solid elements, and to settle and remove proteins, measurement of the intrinsic absorption UV-spectra of the antibiotic in the specific range as shown by a calibration curve constructed by a reference medium with the calibration curve constructed to comply with the measured optical densities from the antibiotic concentration in a reference solution; using whole blood as a biological medium requires blood corpuscles to be removed for the purpose of sample preparation.
EFFECT: method provides cutting time for evaluation of cephalosporin antibiotics with a reduced detection level, higher detection accuracy, lower uncertainty of the result.
2 cl, 6 dwg, 1 tbl, 4 ex
SUBSTANCE: diagnostic technique for latent prostatitis consists in the fact that laser provocation by low-intensity laser is used at wave length 0.82 mcm and power 10 mWt to cover a perineum in a projection of prostate for 10 minutes daily within a course of 5 procedures with a prostate secretion used thereafter. If the prostate secretion analysis shows leukocytes > 20 in a visual field and/or a pathogenic flora growth in titre 1*104 CFU/ml, latent prostatitits is diagnosed.
EFFECT: use of the declared technique enables higher effectiveness of diagnosing latent prostatitis.
1 ex, 1 tbl, 1 dwg
SUBSTANCE: method for nominal evaluation of ethanolemia by the ethanol concentration in corpse's muscular tissue involves measuring the ethanol concentration in corpse's muscular tissue with regard to the prescription of death, death type and a period of storage of sampled muscular tissue, and the ethanolemia is calculated by formula: where Cnominal is the nominal ethanolemia, ‰; DNS is the prescription of death, days; Dx is the period of sample storage, weeks; Cmuscle is the ethanol concentration in muscle, ‰; K is a conversion coefficient selected on the strength of the death type: 0.665 - strangulated asphyxia; 0.675 - death caused large blood loss; 0.692 - death caused by hypothermia; 0.721 - pulmonary death type; 0.645 - other death types in view of the borders limiting the target ethanolemia by formula: 0.953xCnominal -0.307≤Cx≤1.047xCnominal +0.307, where Cnominal is the nominal ethanolemia, ‰; Cx is the actual ethanolemia, ‰.
EFFECT: method enables evaluation of the ethanolemia by the ethanol concentration in corpse's muscular tissue in case of incomplete putrid biotransformation, and impossible use of the instrumental examination.
SUBSTANCE: method for evaluation of ethanolemia by the ethanol concentration in corpse's muscular tissue involves measuring the ethanol concentration in corpse's muscular tissue, and the minimum potential ethanolemia is calculated by formula: cmin = 1.388 × cmusk - 0.181 where Cmin is the minimum potential ethanolemia, ‰; Cmusk is the ethanol concentration in corpse's muscule, ‰, while the maximum potential ethanolemia is calculated by formula: cmax = 1.583 × cmusk + 0.181 where Cmax is the maximum potential ethanolemia, ‰; Cmusk is the ethanol concentration in corpse's muscule, ‰.
EFFECT: invention provides more precise evaluation of the minimum and maximum ethanolemia.
SUBSTANCE: invention relates to field of molecular biology, clinical biochemistry, medicine, veterinary, pharmacology, endocrinology and oncology. To diagnose prostate cancer, patient's blood is sampled. Sample preparation of blood is performed aimed at its deproteinisation. Sedimentation of proteins from analysed fluid is carried out with methanol, after which direct mass-spectrometric analysis of fractions of analysed substances - markers - is carried out by quadrupole-time-of-flight mass-spectrometry.
EFFECT: method makes it possible to diagnose prostate cancer.
1 dwg, 3 ex, 1 tbl
SUBSTANCE: method involves insulating a capillary section where the detection window is located with radical polymerisation in the quartz capillary. Radical polymerisation of the reaction mixture in the capillary is initiated via electron-beam processing by exposing the reaction mixture to accelerated electrons at absorbed dose values ranging from 25 to 75 kGy. The outer polyimide layer is removed with drops of hot sulphuric acid while simultaneously washing the column with a cooling liquid or a liquid-gas mixture.
EFFECT: high reliability and a simple method of making a detection window.
2 cl, 2 ex
FIELD: machine building.
SUBSTANCE: proposed system comprises first assembly to direct gas sample from sampling site to gas characteristics measurement site. First assembly contains components to get in contact with gas sample in operation and gas sampling tube. System includes sample analyser area to measure characteristics of, at least, one component in gas sample, and second assembly outside of the first assembly during all time of operation so that all components of second assembly do not get in contact with gas sample. Note here that second assembly comprises pump connected with said tube to feed gas there through into analyser area without contact with gas sample. Second assembly comprises flow measuring device to get data on gas flow sample, gas pressure gage, or whatever combination of pump, flow measuring device or gas pressure gage.
EFFECT: preventing contamination of reusable components, lower production costs, ease of maintenance.
33 cl, 9 dwg
SUBSTANCE: probe is a sealed container which is connected by a conducting rope to a control unit and has a system of containers connected in parallel by hoses for gathering water samples and a sampling tube, one end of which is connected through an electromagnetic valve to a system of containers, and the second end of which has holes for inlet of void water and is fitted with a filter. The void water container is in form of syringes whose piston rods are locked by spring-loaded locks so as allow their unlocking and filling the syringes with water depending on the immersion horizon of the sampling tube in the sediments. The probe is fitted with a mechanism for immersion of the sampling tube and a system for monitoring its immersion depth in the sediments.
EFFECT: design of the sampling tube enables to increase the depth of water bodies from which samples can be collected in situ, increases efficiency and quality of the collected samples and cuts the time spent on collecting samples.
7 cl, 1 dwg
SUBSTANCE: invention refers to medicine, particularly to dentistry. Diagnosis of generalised periodontitis is enabled by examining crevicular fluids by V-dehydration technique. The presence of three exactly localised facies regions makes it possible to state normal condition of periodontium tissues. The presence of arched cracks in an edge region provides diagnosing slight generalised periodontitis. The presence of arched cracks in the edge region and a band of the lower wave edge region, the absence of an intermediate region and a typical crystalline structure in the central region provides diagnosing a moderate degree. If observing a tendency to fusion of all facies regions and the presence of beam-like cracks in the edge region enables diagnosing severe generalised periodontitis.
EFFECT: technique enables diagnosing generalised periodontitis and determining its severity.
SUBSTANCE: invention relates to investigation of protective properties of packets of filter materials for skincare agents based on activated carbon-containing sorbents in dynamic conditions. The method is realised by using methyl salicylate which mimics the penetrating power of ypertite in an air stream at temperature 26±1°C, concentration of methyl salicylate vapour in air stream of 0.05±0.01 mg/l, relative humidity of 65±5% and pressure gradient on the thickness of the packet with surface area 22 cm2 equal to 49 Pa, with subsequent analytical determination of the minimum amount of methyl salicylate penetrating the packet and the minimum time for protective action, calculation of the external effective dose of DDS, which the packet of the filter protective material protects from based on the given relationship.
EFFECT: high safety, reliability and information content of the evaluation process.
2 cl, 1 tbl, 2 dwg
SUBSTANCE: method for accurate determination of the displacement factor and relative permeability involves conducting field pressure transient tests and taking geophysical measurements. An injection well from which drilling has been stopped or was previously used for oil extraction, having a vertical or deviated hole which penetrates the formation from top to bottom is used for investigation. Before injection, geophysical investigations are conducted in the well in order to determine the initial distribution profile of the oil saturation factor. The working fluid is then injected into the well. Bottom-hole pressure, the injectivity profile of the working fluid through section of the formation, as well as the distribution profile of the water saturation factor are measured at different time instances. Measurements are then stopped at the n-th phase when the water saturation factor profile is the same as that measured at the (n-1)th phase. Results of said monitoring are then used to determine the oil displacement factor with the working fluid, as well as the relative permeability function for each characteristic interval of the section of the formation.
EFFECT: high accuracy of determining the displacement factor and relative permeability not at one point but on the entire section of a formation.
FIELD: machine building.
SUBSTANCE: proposed device comprises core holder and gage intake container arranged there below, high-permeability mixer arranged atop tested specimen and coupled with feed puncheon wherein two chambers are made to contain two diverse fluids. Note here said feed puncheon is made up of two tight chambers with preset fluid flow rate fitted one into another concentric about device axis. Note also that inner chamber space is provided with bottom shutoff valve to form circular clearance around its working lateral surface. Inner chamber bottom is provided with, at least, four radial channels with their outlets communicated with said circular clearance while their inlets are communicated with inner space of another chamber and furnished with needle restrictors threaded surface of which are coupled with appropriate surfaces of circular skirt made on outer cylindrical surface of inner chamber bottom part. Note here that high-permeability mixer represents small-height perforated sleeve. Two sector thrusts for mounting feed puncheon are arranged top said sleeve. Concentric about device axis. Besides, sleeve is fitted in case lateral surface to seal lateral surface of tested specimen. Note that vertical and radial intercommunicated channels are made in feed puncheon lateral wall and bottom to communicate spaces above upper boundaries of diverse fluids in puncheon and mixer.
EFFECT: higher homogeneity of filtered flow, simplified design, easier replacements.
SUBSTANCE: method of determining number of ferromagnetic particles in a liquid or air stream, involving installation of a detachable element with a magnetic system perpendicular to a pipe, through which passes a portion of the stream moving through the pipe, characterised by that the liquid or gas passes through the detachable element, the liquid undergoes controlled collection with ferromagnetic particles for subsequent weighing, wherein the detachable element consists of a cylindrical housing with flanges for inlet and outlet of liquid or gas, and the inlet flange is hermetically connected to the collection pipe lying inside the pipe and moving in its cross-section from the horizontal axis to the inner surface and for synchronous joint movement with a permanent magnet which is joined through a telescopic rod. The magnetic system also consists of a permanent magnet which moves vertically on the outer surface of the cylindrical housing of the detachable element and ferromagnetic liquid which forms a film upon contact with the permanent magnet, which, in form of a piston, divides the cylindrical housing into a lower chamber for collection and a top chamber for subsequent weighing of the liquid or glass with ferromagnetic particles.
EFFECT: low power consumption and high accuracy of determining the number of ferromagnetic particles in a liquid or gas stream moving in a pipe.
2 cl, 1 dwg
SUBSTANCE: invention relates to analytical methods of measuring impurities in a gas, which is based on converting impurity molecules into aerosol particles and subsequent detection of the aerosol, and can be used in high-sensitivity gas analysers used to solve environmental problems, as well as in diagnosis of diseases by analysing exhaled air. The method of measuring trace elements in an air stream involves converting impurity molecules into a condensation nucleus. The nuclei undergo two-step enlargement to size of aerosol particles in supersaturated vapour of the detecting substances in thermodiffusion-type condensation devices and light-scattering of the obtained aerosol is measured. To perform conversion, the stream is heated to 200-600°C, and the detecting substance used at the first enlargement step is semi-volatile organic acids with ionisation constant in water of 4·10-2-2·10-1.
EFFECT: high sensitivity and selectivity of detecting ammonia and organic amines in atmospheric air.
1 tbl, 6 dwg
SUBSTANCE: dynamic coefficient of external friction between two samples lying on each other and moving relative each other is determined, where the flat working surface of the bottom sample has a fixed angle of inclination with respect to the horizontal. The top sample is suspended using a hinge joint. Relative movement of the samples takes place on the horizontal until obtaining a steady-state turning angle of the hinge joint relative the direction of movement. The dynamic coefficient of external friction is determined using a formula.
EFFECT: possibility of high-accuracy determination of dynamic coefficient of external friction with measurement of only geometrical parameters of the system without measurement of frictional force.
2 cl, 2 dwg
SUBSTANCE: disclosed is a method of cultivating Bacillus brevis strain 101 for producing gramicidin S. Submerged cultivation of a culture is carried out on a synthetic culture medium. The medium contains yeast autolysate and casein hydrolysate in concentration of 0.1 g/l and 0.2 g/l on amine nitrogen, glycerine in concentration of 20 ml/l, edible 40% lactic acid 2.0-4.0 ml/l, ammonium phosphate-chloride 0-3.4 g/l, di-substituted ammonium phosphate 0.85-4.5 g/l, mono-substituted potassium phosphate 0-1.0 g/l, magnesium sulphate heptahydrate 0.9 g/l, sodium citrate 1.0 g/l. Content of amine nitrogen in the initial medium is equal to 1.3-1.6 g/l. When concentration of amine nitrogen falls to less than 1.4 g/l, a concentrated culture solution is added to the medium until achieving concentration of 1.75 g/l. The concentrated culture solution contains glycerine, edible 40% lactic acid, di-substituted ammonium phosphate and chloride and magnesium sulphate with ratio of concentration of glycerin, lactic acid, nitrogen, phosphorus and magnesium equal to 1:(0.008-0.032):(0.027-0.036):(0-0.008):(0.002-0.008). During growth, the rate of stirring is increased from 200 to 500 rpm. pH is kept at 6.5-6.8 by adding potassium and sodium hydroxide. The process is stopped 2-6 after the onset of a stationary phase.
EFFECT: method enables reproducible production of a large amount of gramicidin S.
10 tbl, 5 ex