Method of determining fractions of modified lipoproteins in blood
SUBSTANCE: patient's blood serum is treated with 7% solution of polyethylene glycol-6000, incubated and with dye Sudan B at 40°C for 1 hour, separated electrophoretically in agarose gel. After that, additionally, before treatment of blood serum with 7% PEG-6000 to 0.6 ml of sample 0.2 ml of 0.1% tritone X-100 solution is added, incubated for 15 minutes at 20°C, after which mixture is mixed by shaking 120 times per 1 minute. Application of method makes it possible to detect additional intensive minor fraction of modified LP(a).
EFFECT: increase of diagnostics accuracy.
3 dwg, 1 tbl
The invention relates to medicine and can be used in cardiology or therapy.
Known methods of fractionation of lipoproteins of blood by the method of analytical ultracentrifugation (Angelov, Ngiculela. Lipids, lipoproteins, and atherosclerosis. - Peter, the press, p.98-102).
The known method of fractionation PL polyacrylamide gel (ACA. Biochemical studies in the assessment of the cardiovascular system. In kN. "Research methods in pathology", M, 1988, p.95-97).
There are also known methods of fractionation PL by electrophoresis in agarose gel (lab. Research methods / edited by Riv. M.: Medicine, 1987, s-249), SU 1720015 And 15.03.1992. EN 2063040 C1, 27.06.1996. EN 2115121 C1, 10.07.1998. EN 2097038 C1, 27.11.1997. EN 2060034 C1, 20.05.1996. EP 0074610 A, 23.03.1983.
The disadvantage of these methods is that they do not allow to identify all the PL fraction of the blood, including chylomicrons (HM), high density lipoprotein (HDL), low-density lipoprotein (LDL), very low density lipoproteins (VLDL) and LP(a), and serve to divide into fractions HMM, HDL, LDL, VLDL from the complex of albumin with neeterificirovannah fatty acids, and also fail to detect minor fraction of modified lipoproteins in the blood, namely the modified abnormal Lipoprotein (Lp(a) or LP(a)).
There is a method of determining the fractions l is poproteins the blood of American authors, NV, Fedorova N.A., Perova NV, Hargreaves EXAMPLE, Lanovoy A.A., science, A.N., American A.V., Pokraev E.N. EN 2210079 C2 G01N 33/92, 10.08.2003 bull. No. 22.
This method is closest to the proposed to the technical essence and the achieved result and selected as a prototype.
The disadvantage of this method is that it is not possible to identify a minor fraction of the modified LP(a). This is due to the fact that a minor fraction of LP(a) is the most atherogenic. Therefore, early diagnosis is especially important to identify minor fraction of LP(a) along with the most complete definition of the other factions PL of blood.
The aim of the invention is to improve the accuracy of the method.
This goal is achieved by the fact that the serum is treated with a 7% solution of PEG 6000, and then incubated with PEG-6000 - precipitate serum with dye Sudan B at 40°C in the dark thermostat for 1 h and contribute to electrophoretic studies in magnified 2 times by volume of the well of the agarose gel. Before treatment serum 7% PEG 6000 to 0.6 ml of sample add 0.2 ml of 0.1% solution of Triton X-100 and incubated at 20°C for 15 min, stirring the mixture by the shaking method 120 times in 1 min and reveal minor fraction of modified lipoproteins in the blood.
New in this method is that the test Siva otci the patient's blood is treated with the detergent Triton X-100, incubate 15 min at 20°C, stirred the mixture with the shaking method 120 times in 1 minute, which allows to additionally reveal intense minor fraction of modified lipoproteins in the blood.
Therefore, only a comprehensive modernization of the prototype method allows to get the desired result.
To obtain PEG-6000 precipitate serum was used 7%solution of PEG-6000. It is known that the 7%PEG-6000 is the ultimate concentration to obtain precipitates serum containing immune complexes and contains no coarse serum proteins. The use of a 6%aqueous solution of PEG-6000 does not completely precipitate the immune complexes in the blood serum. Therefore, to improve the accuracy of the experimental method was chosen by 7%concentration of PEG-6000, which allows, on the one hand, completely precipitate the immune complexes from serum to guarantee the absence coarse (high molecular weight) serum proteins, and on the other hand, to fully precipitate circulating immune complexes (CIC)present in the serum. Thus, if the concentration of the CEC in the serum, equal to 2.5 g/l, this concentration of the CEC were detected in 7%of cases the precipitates serum (+3%), which is associated with an increase in the number of manipulation rather than change koncentracijski in the precipitates serum.
Incubation of 7% PEG-6000 precipitate analyzed serum with dye Sudan B increases the affinity of the dye to the PL of precipitates of blood and a large amount of taken for studies of the blood sample allows to detect modified LP(a)contained in the blood of patients with coronary heart disease (CHD) in very low concentrations. Additional treatment serum Triton X-100 with stirring of the mixture by the shaking method 120 times in 1 min improves the accuracy of the method and to obtain the desired result. The proposed method allows to identify the following minor fraction of the modified LP: HDL, LDL, VLDL and LP(a).
Each newly introduced in the claims the sign performs the function of increasing the accuracy and efficiency of the method: processing of 0.6 ml samples of blood serum of a patient with 0.2 ml of 0.1% solution of Triton X-100, incubation of the samples at 20°C for 15 min, stirring the mixture by the shaking method 120 times in 1 min and an additional detection of minor fractions of modified lipoproteins.
Research different PL classes in the diagnosis of coronary heart disease (CHD) is recommended in all-Russian scientific society of cardiologists according to the position of the recommendations of the European society for the study of atherosclerosis- "Diagnosis and correction of lipid metabolism disorders prevention and treatment is therosclerosis" (Moscow, 2005; Clinical laboratory diagnostics, No. 10, 2008., p.21-32).
At present promising research methods lipids with a detergent /Triton X-100 and others/. In laboratory practice increasingly used direct or homogeneous methods for the determination of lipoprotein (LP) and lipids. Such methods are based on the use of different detergents that can block or solubilisate classes of pharmaceuticals for specific allocation of HDL, LDL and other factions PL.
When using such methods isolation of other classes of PL does not require additional operations and the concentration of cholesterol (LDL) in PL classes can be defined directly in the serum of conventional enzymatic methods in the same cell ("Clinical laboratory diagnostics, No. 10, 2008., p.21-32).
Increasing the concentration of PL or abnormal LP (a) in blood is considered an independent risk factor for atherosclerosis. When blood levels of LP(a) more than 300 mcg/ml at norm 0-300 µg/ml the risk of coronary atherosclerosis increases twice, and while increasing the level of LP(a) cholesterol and LDL - 5 times (J.A. M.A. - 2001. - Vol.285, - p.2486-2497, Eur. Heart. J. - 2003. - Vol.24, - p.1601-1610).
Currently, special attention is paid to the study of minor fractions of the modified LP(a), and therefore developed methods of laboratory diagnostics, allowing to investigate this is t lipoprotein blood. He refers to APO-B-containing lipoproteins, rich in cholesterol (LDL). LP(a) is identical to the "sinking" of pre-β-PL (sinking pre-β-Lp)with electrophoresis mobility pre-β-PL. LP(a) contains 27% protein, 8% carbohydrates and 65% of lipids, of which AHS make up 59%, NAHS 14%, PL 14%.
Protein component of LP(a) is vysokopotentsirovannye polypeptide-APO(a)with close structural similarity to plasminogen - one of the factors of the coagulation system - protivosvertawate blood. With increasing concentration of LP(a)and its modified forms in the blood impairs processes of microcirculation in blood arteries with the possible formation of microthrombi.
Due to the presence in the structure of APO(a) sialic acids LP(a) is more negatively charged compared to the β-PL in an electric field, it is better soluble in water, can interact with metal ions (calcium). This lipoprotein and its modified forms heterogeneous. All this testifies to the special role of LP(a) and modified LP(a) in atherogenesis.
LP(a) can interact with LDL receptors, exerting little influence on the activity of MMC-COA reductase, a cholesterol esterification. The half-life of LP(a) is longer than that of LDL and is 3.3 days. The content of LP(a) blood in norm does not exceed 30 mg/l At high concentration in blood LP(a) is detected in the defeat of cosudow area concentrations of fibrinogen. Higher concentration of LP(a) is often combined with IIaIIbtypes of hyperlipoproteinemia which often identifies modified PL. Therefore, in clinical practice it is extremely important determination of LP(a) simultaneously with the determination of proteins of the acute phase of inflammation. It was found that most of the lipid-lowering drugs do not affect elevated levels of LP(a).
Fraction of LP(a) heterogeneous. It is established that during electrophoresis LP(a) are in the field of β-globulins, but up to 5% of LP(a) can be detected in α-globulin. The reason for this pronounced heterogeneity is difficult to assess during electrophoresis entire faction LP(a), and especially its minor faction that may remain on the start line, if the size of the particles is large enough. Therefore, the use of common research in the last fifty years of the detergent Triton X-100 for processing serum, namely LDL, blood, leading to partial delipidization PL and increase their mobility during electrophoresis allows to detect minor fraction of LP(a). In the chemical industry uses a variety of detergents (izgotovlenie washing powder, detergents etc), but when working with biological material is used mainly Tritonx-100. Processing mode samples the Triton X-100 were selected on the again of experimental studies empirically. This was done with a different dilution of Triton X-100, different temperatures and different exposure time in minutes. When using different concentrations of a solution of Triton X-100, low concentrations (below 0.1%) did not increase the allocation of minor factions LP, including LP (a)and high concentrations (greater than 0.1 per cent) led to a complete delipidization and fracture patterns PL. The results of the study are presented in table 1.
Processing 0.3 ml serum for the study of lipoproteins of different concentrations of a solution of Triton X-100 during a different time of incubation of the sample at different temperatures.
Therefore, the optimal conditions for treatment of serum with a solution of Triton X-100 concentration of 0.1% in volume of 0.1 ml at a temperature of 20 degrees Celsius for 15 minutes.
The improvement of the method relates to mechanical mixing of the sample with a frequency of 120 times in 1 min on a laboratory shaker for preparation of reagents. More frequent or prolonged shaking leads to the increase of the temperature of the sample and the emergence of remantic forms of lipoproteins in the blood, which prevents further research and distorts the electrophoretic analysis of the sample. Less than a short period of shaking, equal to 30 seconds, does not improve the results.
Because LP(a) Nai is more heterogenen, it is very important in the early stages of the disease to identify the maximum content of LP(a) in the blood of each patient. This allows us to diagnose coronary heart disease before the stage of significant changes in other clinical and laboratory parameters increases the accuracy of diagnosis of the disease. In turn, these patients early is assigned pathogenetically substantiated therapy. No less important is the identification of minor factions LP(a) to assess the effectiveness of therapy of disease and predicting the course of coronary heart disease.
All of the above indicates the extreme importance of developing methods of laboratory diagnostics, allowing you to identify LP(a), modified forms and their minor faction.
Currently in widespread clinical laboratory practice does not use enough of the ways of determining LP(a). At the same time, the popularity of the method of electrophoretic separation of fraction PL of blood in the agarose gel due to its high sensitivity, simplicity of implementation and adequate adequacy of the obtained results (lab. case, 1980, No. 5, page 287-290, Clinical laboratory diagnostics, No. 10, 2008, p.21-32).
The essential features characterizing the invention, shown in the inventive combination of new properties that are not explicitly derived from the prior art in this region the STI and are not obvious to the expert.
Identical set of features not found in the study of patent and scientific literature.
This invention can be used in medical practice to improve the accuracy of diagnosing the degree of atherosclerosis in ischemic heart disease.
Thus, it should be considered the present invention with the relevant conditions of patentability: novelty", "inventive step", "industrial applicability".
The method is based on the electrophoretic mobility of modified lipoproteins in the blood.
The invention will be clear from the following description of the attached drawings.
Figure 1 presents the results of electrophoretic separation in the PL fraction of the blood serum and in the control group, the method of the prototype; b - in the control group proposed method;
In figure 2 a, b, C presents the results of electrophoretic separation of fraction PL of serum in the group of patients with coronary artery disease by the method of the prototype;
Figure 3 presents the results of electrophoretic separation in the PL fraction of the blood serum of the patient B: a - method-prototype; b - the proposed method.
The method is carried out in stages as follows:
1) preparation of a solution of Sudan B: 400 mg of Sudan B was dissolved in 20 ml of ethylene glycol in a boiling water bath for 50 minutes, filter the comfort, store in a glass container;
2) preparation of agarose gel: 320 mg of agarose And "Sigma" are dissolved in 20 ml of water by boiling, and then placed in a thermostat at 55°C; add 20 ml albumin (1 g of albumin in 200 ml veronal-medialog buffer, pH to 8.6). 3 ml of agarose gel is applied on the skim horizontally installed the scanner glass, place a metal rod steel-bar 20 mm×4 mm (height 10 mm), which after solidification of the gel is removed by a magnet;
3) to 0.6 ml samples of blood serum of the patient add 0.2 ml of 0.1% solution of Triton X-100, incubated for 15 min at 20°C, stirred the mixture with the shaking method 120 times in 1 min, then add 40 ml of 7% of polyethylene glycol (PEG)-6000 and incubated for 1 h at 20°C, centrifuged for 40 min at 25000 g. The precipitate washed twice.
4) 0.25 ml of precipitate used for electrophoretic studies. To 0.25 ml of the precipitate serum add 0.15 ml of a solution of Sudan B, placed for 1 h in the dark in a thermostat at 40°C, then add 0.2 ml of a hot solution of agarose gel, mixed, heated at 55°C and heated contribute in the groove of the agarose gel;
5) glass slide is placed in a chamber for electrophoresis layer of agarose down, electrophoresis is carried out in the course of an hour in the refrigerator at 4°C at a voltage of 100 V and current 40-45 mA;
6) electrophoregram fixed in 5% is aStore acetic acid for one hour, then dried between sheets of filter paper, continuously wetting 96% ethanol;
7) densitometry performed on microphotometer MT-4.
To confirm the performance of the proposed method and obtain a technical result were examined in the control group (n=12) and group of patients with CHD (n=24). Both groups were examined using the proposed method and the prototype method.
In the control group, 7% PEG-6000 precipitates serum in the case of using the prototype method and the proposed method, the PL is not revealed (figa, b).
In the group of patients with coronary heart disease in 7% PEG-6000 precipitates serum, using the prototype method identified LDL and VLDL and traces of LP(a) (figa, b, C).
In the group of patients with coronary heart disease in 7% PEG-6000 precipitates serum proposed as an invention than fractions of LDL, VLDL in the area between LDL and VLDL, revealed a minor fraction of the modified LP(a) in 18 patients, and in 6 patients in this group faction LP(a) were detected in low concentrations.
Patient, 55 years. Case history No. 194. Was admitted to the Department of CHD and atherosclerosis research Institute of cardiology, Tomsk scientific with complaints of pain in the sternum and the heart that occurs during physical activity; pain stymied by nitroglycerol. The patient is concerned about shortness of breath during physical activity. HELL 160/90 mm Hg, heart rate is acoe 66 beats per minute.
Diagnosis: ischemic heart disease, angina FC II.
Laboratory results: total cholesterol of 8.8 mmol/l; triacylglycerol 1.6 mmol/l; HDL cholesterol 0.9 mmol/L.
The results of electrophoretic studies PL blood of the patient by the method prototype presented in (figa), and the proposed method (figb). The identified modified LDL, HDL, VLDL, LP(a) in low concentration.
The proposed method allows to reveal in CHD patients with severe hypercholesterolemia presence of minor fractions of the modified LP(a) (m PL(a)) by the method of electrophoresis of pharmaceuticals in the agarose gel.
The application of the proposed method of fractionation PL blood unlike the prototype method allowed us to identify additional intensive minor fraction of the modified LP(a), which increases the accuracy of the method.
Thus the proposed method is easy to use and interpretation of the results.
Table 1. Investigation of the influence of the detergent Triton X-100 to identify lipoprotein fractions at different time of incubation and different temperatures, i.e., the determination of optimal conditions;
Figure 1. Presents the results of electrophoretic separation in the PL fraction of the blood serum and in the control group, the method of the prototype; b - in the control group proposed method;<> Figure 2 (a, b, C). Presents the results of electrophoretic separation of fraction PL of serum in the group of patients with coronary artery disease by the method of the prototype;
Figure 3. Presents the results of electrophoretic separation in the PL fraction of the blood serum of the patient B: a - method-prototype;
b - the proposed method.
|Detergent||Concentration||The incubation period||Temperature, degrees Celsius||Identifying fractions LP(a)|
|Triton X-100||0.1 ml of 0.01%||10 minutes||15||-|
|Triton X-100||0.1 ml of 0.01%||15 minutes||20||-|
|Triton X-100||0.1 ml of 0.01%||20 minutes||25||-|
|Triton X-100||0.1 ml of 0.1%||10 minutes||15||traces|
|Triton X-100||0.1 ml of 0.1%||15 minutes||20||+|
|Triton X-100||0.1 ml of 0.1%||20 minutes||25||traces|
|Triton X-100||0.1 ml of 0.15%||10 minutes||15||-|
|Triton X-100||0.1 ml of 0.15%||15 minutes||20||-|
|Triton X-100||0.1 ml of 0.15%||20 minutes||25||-|
The method for determining the fractions of modified lipoproteins of blood by treating serum 7%solution of polyethylene glycol-6000, incubation with the dye Sudan B at 40°C for 1 h followed by electrophoretic separation in agarose gel, characterized in that it further before treatment serum 7% PEG-6000 to 0.6 ml of sample add 0.2 ml of 0.1%solution of Triton X-100, incubated for 15 min at 20°C and then stirred the mixture with the shaking method 120 times in 1 min
SUBSTANCE: invention relates to medicine, in particular to hepatology, and is intended for prediction of probability of chronic hepatitis C recurrence in patients after three months since beginning of antiviral therapy. For this purpose after three months since beginning of antiviral therapy in patient determined are biochemical parametres of blood: albumin in %; beta-lipoproteids in g/l, by which by means of certain nomogram probability of disease recurrence is determined.
EFFECT: method ensures increased accuracy of chronic hepatitis C outcome prediction.
2 ex, 1 dwg
SUBSTANCE: invention relates to the field of medicine, in particular to ophthalmology, and may be intended for treatment of hemodynamics abnormalities in visual nerve vessels in case of their atherosclerotic lesion. Medical therapy is administered with established treatment regimen. At the same time lipid spectrum of blood plasma is investigated, and if level of total cholesterol is more than 5.1 mmole/l, treatment with pentoxifylline in tablet form, 0.2 g 3 times a day after food intake, piracetam in pills, 40 mg 3 times a day before food intake, simvastatin, 10 mg per day, in tablet form, once, in the evening time, treatment is administered for 6 months. Afterwards hemodynamic indices are investigated by means of ultrasound Doppler sonography.
EFFECT: improved blood circulation in vessels of carotid system and regional vessels of visual nerve.
3 tbl, 1 dwg, 1 ex
SUBSTANCE: invention refers to medicine, namely to gastroenterology and cardiology, can be used in medical institutions for determination of severity level of metabolic disorders and risk of metabolic syndrome in the patients suffering chronic cholecystitis. For risk evaluation of metabolic syndrome in the patients with chronic cholecystitis by the severity of immune-metabolic disorders, the following significant diagnostic characters are used: fasting glucose, apoprotein atherogenicity coefficient (apoB/anoAl), high density lipoprotein cholesterol, T-helpers and T-suppressors ratio, phagocytic index. The derived values are entered in an arithmetic formula to evaluate the risk of metabolic syndrome by the severity level of immune-metabolic disorders by total value expressed in standard units.
EFFECT: method facilitates work of a doctor, allows to evaluate the patient's status in a relatively short time, to predict the risk of metabolic syndrome and to prescribe the well-timed therapy to prevent the progression of metabolic syndrome and cardiovascular complications in the patients with chronic cholecystitis.
1 tbl, 3 ex
SUBSTANCE: invention relates to determining cholesterol in remnant-like particles without separation of analysed sample components.
EFFECT: obtaining higher reactivity to RLP-cholesterol in samples of type III hyperlipidemia and accurate determination of RLP-cholesterol in samples of type III hyperlipidemia.
12 cl, 14 ex, 8 dwg, 13 tbl
SUBSTANCE: level of LP(a) is determined by additional processing of 0.3 ml of blood serum with 0.1% solution of Triton X-100 with 15 min incubation at 20°C. After that blood serum is incubated with solution of Sudan B for 1 hour in dark thermostat at 40°C and sample is introduced into hole in agarose gel with area 4×20 mm for LP electrophoresis with further fixation of electroforegrams, their drying, densitometry. In case of 40% or higher reduction of LP(a) level in comparison with initial level treatment of coronary heart disease is estimated as efficient.
EFFECT: increased accuracy of predicting coronary heart disease.
2 ex, 6 dwg
SUBSTANCE: 0.6 ml of blood serum are treated with 0.2 ml of 0.1% Triton X-100 solution and incubated for 15 min at 20°C with further addition of 7% solution of polyethylene glycol 6000 with further electrophoretic separation. If LP(a) level reduces on 40% and more, and total cholesterol level reduces on 20% and more in comparison with the initial level, prediction of disease course is considered to be favourable, contributing to transition of exertional angina FC III-IV into FC I-II. If level of modified LP(a) reduces less than on 40%, total cholesterol on less than 20% in comparison with the initial level prediction of disease course is considered to be unfavourable.
EFFECT: increased accuracy of CHD course prediction.
2 ex, 1 tbl, 6 dwg
SUBSTANCE: invention relates to medicine, namely to traumatology and orthopedics and can be used for early diagnostics of endoprosthesis instability development after total endoprosthetics. Examination of primary, secondary and final isopropanol-soluble products of lipid peroxidation and heptane-soluble products of lipid peroxidation in terms 3 and more months after the operation is carried out. Endoprosthesis stability or instability after total endoprosthetics of hip joint caused by osteoarthrosis is diagnosed by increase of the level of said indices with respect to the norm.
EFFECT: method is simple, available and efficient.
SUBSTANCE: invention concerns medicine, namely to paediatric gastroenterology. To diagnose the primary form of hypercholesterolemia in infants and preschoolers with gastroenterological diseases, total cholesterol level and homocysteine level are determined by the biochemical blood analysis. If total cholesterol exceeds 5.2 mm/l, while homocysteine is more than 8.5 mcM/l, primary hypercholesterolemia is diagnosed.
EFFECT: higher diagnostic accuracy of primary hypercholesterolemia in infants and preschoolers.
FIELD: chemistry, biochemistry.
SUBSTANCE: invention refers to medicine, namely to biochemistry, can be used for assessment of state of erythrocytes during medical treatment of patient with somatic diseases. Erythrocyte resistance assessment method involves blood sampling, extraction of erythrocytes and determination of their spectral characteristics; at that, extracted erythrocytes are washed in sodium-potassium buffer. After that, 31R-NMR spectra of erythrocyte suspension are taken; there determined is content of 2,3-diphosphoglycerate (2,3-DFG) and inorganic phosphate (IP) in terms of percentage; on the basis of obtained values, there determined by formula Per=2,3-DFG/IP is energy erythrocyte resistance parametre (Per), and when this value is less than 4.0, energy erythrocyte resistance is considered to be altered.
EFFECT: invention allows using Per parametre both for assessment of medical treatment effectiveness and for dynamic control of patient's condition after he/she has been discharged from the hospital.
5 ex, 1 tbl
SUBSTANCE: α-spectrin, β-spectrin and 4.1 band protein are defined by disc electrophoresis method in stepped gradual 10% polyacrylamide gel with sodium dodecylsulphate in erythrocyte membranes of healthy pregnant and pregnant with 1:6400 and 1:12800 antibody titre to herpesvirus, and if α-sprectrin percentage falls from 11% to 15.5%, β-spectrine percentage falls from 10.7% to 14.6%, 4.1 band protein percentage falls from 32.5% to 33.2%, then erythrocyte membrane instability is diagnosed.
EFFECT: assessment of erythrocyte membrane cytoskeleton alteration in the period of herpesvirus infection episode of pregnant.
SUBSTANCE: method of determination of triterpene saponins in vegetable raw material and medications includes dissolution of saponin-containing fraction in mixture water-ammonium buffer, determination of its optic density and calculation of saponin content in terms of oleanolic acid, under specified conditions.
EFFECT: claimed method represents express method, facilitates analysis and increases degree of reliability of obtained results.
SUBSTANCE: in order to estimate efficiency of treating ischemic nephropathy in newborn babies in early neonatal period activity of gamma-glutamyltransferase and cholinesterase in child's urine is determined in dynamics of treatment. If activity of said enzymes decreases with respect to initial level, treatment is estimated as efficient, if activity increases or does not change - as inefficient.
EFFECT: application of method makes it possible to increase accuracy of estimation of ischemic nephropathy treatment in newborns, carry out correction of therapeutic measures in due time and improve disease outcome.
1 tbl, 3 ex
SUBSTANCE: in a first trimester of pregnancy, the microalbuminuria level is determined. If the value is 45 mg\l and more, placental insufficiency is predicted.
EFFECT: method enables early prediction of placental insufficiency by a simple quantitative estimation and thereby ensures early adequate preventive treatment.
SUBSTANCE: detecting expression of GalNac-T14 molecules enables to predict sensitivity or indicates that a tissue or cell sample is sensitive to apoptosis inducing agents, such as DR4 or DR5 agonist antibodies. The information obtained by the analysis aimed at detecting GalNac-T14 expression in the mammal's tissue or cell sample can provide a hospital doctor with data which can be used for prescribing an optimal treatment schedule for patients suffering such diseases as pancreas cancer, lymphoma, non-small cell carcinoma of lung, colon cancer, rectal cancer, melanoma or chondrosarcoma.
EFFECT: expanded scope of the compounds.
16 cl, 53 dwg, 14 tbl
SUBSTANCE: invention relates to field of medicine, in particular toxicology and resuscitation science and can be used for early prognosis of course of acute poisoning with psychotropic drugs. At the time of patient's admission to hospital albumin fraction of blood serum is isolated. After that, general level of reduced thiols is determined, and if its value is lower than 220 mcmol/l development of negative disease dynamics is predicted.
EFFECT: method allows to increase efficiency of performed treatment in said category of patients.
1 tbl, 8 ex
SUBSTANCE: invention refers to chlitin derivative photoproteins and to application thereof both as intracellular calcium indicators, and in cellular studies. Said proteins are produced by mutagenesis of a coding sequence of chlitin. Also, there are offered nucleic acids coding said protein, a vector containing said nucleic acids and a host cell carrying the vector. They can find application in genetic communication technologies for monitoring the cellular events associated with signal transmission and gene expression. Besides, photoproteins of the present invention can be used as intracellular calcium indicators in diagnostic techniques based on calcium concentration measurement in response to the various effects.
EFFECT: produced proteins exhibit enhanced bioluminescence, high affinity to calcium and prolonged light emission.
19 cl, 16 dwg, 5 ex
SUBSTANCE: in order to realise the method level of ferritin in blood serum and bile bladder tissue, rang point is defined, and ferritin values in serum from 0 to 10 ng/ml, and bile bladder from 0 to 0.25 mg/l - are taken for 1 point, respectively, level of ferritin in serum 70 ng/ml is taken as 7 points, and in bile bladder tissue 0.75 mg/l is taken as 3 point, and if sum is 10 points, conclusion is made about non-destructive cholecystitis, and if sum is from 10 points conclusion about destructive cholecystitis is made.
EFFECT: application of invention allows to increase accuracy of diagnostics of bile bladder tissue destruction in acute cholecystitis.
SUBSTANCE: invention relates to pharmacology. Medication for brain cell protection contains dominant protein extract from placenta tissue, obtained by chromatographic separation of protein fraction of 8-10 components with total molecular weight from 15 to 200 kDa, 70-80% of fraction consisting of protein with molecular weight about 70 kDa. In order to obtain said medication powdered placenta is extracted by means of alkalescent buffer in presence of inhibitors of proteolytic protein cleavage and centrifuged. Centrifugate is passed through chromatographic column with anion-exchange carriers balanced by means of the same buffer, column is washed from unbound material, proteins are eluted with neutral salt solution. Collected material is diluted with water bringing pH to subacid value and diluted substrate is successively passed through column with anion-exchange carrier and column with cation-exchange carrier, balanced with buffer solutions to pH about 6.0. Final product is obtained after sterilisation.
EFFECT: method application allows to obtain efficient medication of high purity, normalising nervous system functions.
3 cl, 1 tbl, 4 ex
SUBSTANCE: invention relates to field of medicine, namely to surgery. Blood is sampled from cubital vein in patients with acute pancreatitis on 1-3 and 7-10 day from disease beginning. Level of blood serum myoglobin is determined by method of performing reaction of passive hemagglutination. If level of blood serum myoglobin on 1-3 day increase from 95 to 128 ng/ml, acute fatty pancreatic necrosis is diagnosed, if myoglobin level in blood serum is higher than 128 ng/ml, hemorrhagic pancreatic necrosis is diagnosed, and if level of myoglobin in blood serum on 7-10 day increases higher than 256 ng/ml, hemorrhagic pancreatic necrosis in stage of infection of pancreatic necrosis nidus is diagnosed.
EFFECT: method allows to correct drug therapy and individually ground tactics of treating patients with acute pancreatitis depending on severity of disease form and presence of infection.
1 tbl, 3 ex
SUBSTANCE: there is offered an arteriosclerosis diagnostic technique based on the detection of various combinations of specific polypeptide markers. To detect the presence or absence of polypeptide marker, capillary electrophoresis, gaseous-phase ion spectrometry and/or mass spectrometry are used.
EFFECT: method allows early high-probability minimally invasive arteriosclerosis detection.
13 cl, 1 ex, 5 tbl
FIELD: medicine, ophthalmology.
SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.
EFFECT: higher accuracy of prediction.