Method of predicting development of critical stenosis of coronary arteries in patients with ischemic heart disease
SUBSTANCE: in the patient's blood serum the concentration of matrix metalloproteinase - 9 (MMP-9) and/or procollagen 1 C-terminal propeptide (PICP) is determined. If the value MMP-9 is larger than 101.8 ng/ml and/or PICP 195.6 ng/ml the unfavourable course of IHD is predicted, if lower than these values - the favourable course without the formation of critical stenoses in the coronary arteries.
EFFECT: increased accuracy and specificity of the method.
The invention relates to medicine, in particular to therapy, cardiology, namely to the early diagnosis of development of critical stenosis in the coronary arteries of patients with coronary heart disease (CHD).
Today, CHD is the leading cause of mortality and disability worldwide, and its prevalence is expected to increase in the coming years. The results of multicenter studies led to a better understanding of the pathogenesis of coronary artery disease. If traditional risk factor assessment was focused on parameters such as age, gender, high blood pressure, cholesterol, heredity, Smoking, in the last decade, there are new risk factors - biological and genetic markers, which are detected in the serum and tissues. They change the approach to risk stratification, their definition becomes available, as an important tool in the assessment and treatment of CHD.
A known method for predicting risk of ischemic heart disease: a method of calculating the probability of occurrence of coronary heart disease through the study of lipids of blood serum, characterized in that it further determine before and after treatment modified LP(A) (Cannes N. In., Fedorova N. And., 2008). This method has a high degree of sensitivity and specificity.
The technical result of the image�etenia is to increase the sensitivity and specificity of the method of predicting risk of development of critical stenosis in patients with coronary artery disease.
The problem is solved in that in a patient serum determine the level of markers of myocardial fibrosis - matrix metalloproteinase-9 (MMP-9) and/or C-terminal propeptide of procollagen type 1 (PICP), a value of MMP-9 more than 101; 8 ng/ml and/or PICP more 195,6 ng/ml determine an increased risk of developing coronary heart disease.
The method is carried out as follows. Blood sampling in a volume of 5 ml is carried out in sterile conditions according to standard procedures. Blood samples are immediately centrifuged, plasma was frozen at -20°C. Storage of plasma at temperature-20-70°C not continue for more than 6 months. For analysis do not use hemolized or lipemic samples. The levels of MMP-9, PICP in the blood is measured using an enzyme immunological test system.
To assess the level of MMP-9, use a set of reagents "Human MMP-9 ELISA" (Bender MedSystems, Austria). The analysis is performed in several steps. Initially, human MMP-9 present in the test samples of serum, associated with monoclonal antibody to human MMP-9, adsorbed in the cells of the microplate. "Detection" polyclonal antibodies to MMP-9 binding molecules of the human MMP-9 captured by the first antibody. After incubation by washing of the cells are removed, not second bound anti-MMP-9 antibodies. Later in the cell add the conjugate monocle�exponentially mouse intential and after incubation and washing, remove not bound enzyme conjugate. The next phase in the cell add the substrate solution, giving the interaction with the enzyme complex staining. The reaction is stopped by adding acid and the intensity of the color measured concentration of human MMP-9. The limit of sensitivity of this set is 0.8 ng/ml In serum samples of healthy people, chosen at random, the levels of MMP-9 are in the range of 20.3-77,2 ng/ml, the average level of 43 ng/ml.
The definition of PICP is carried out using a diagnostic kit "Metra CIPC EIA Kit (Quidel Corporation, USA) by ELISA. The method is based on the "sandwich" immunoassay analysis in a microplate format, using monoclonal anti-PICP antibody immobilized in the wells of the microplate, rabbit anti-RSR antiserum, conjugate goat antigalactic antibodies with alkaline phosphatase and pNPP substrate for the quantitative determination of PICP in human serum. Antibodies to PICP have 100% cross-reactivity with serum PICP human blood. The minimum detectable using the set level is 0.2 ng/ml. In the analysis of serum samples of adults older than 25 years, with the help of this set, the values obtained are in the range 69-163 ng/ml.
When carrying out statistical processing of the material were found statistically significant differences between record�mi fibrosis depending on the presence of coronary artery disease and severity of coronary artery lesion.
Building a model of the development of IHD performed on the entire array of patients involved in the study. CHD was diagnosed in 108 patients from 216. The analysis of multiple regression shows that valuable predictors to predict the development of CHD are MMP-9, PICP. The risk of developing CHD increases with increase in the concentration of MMP-9 and/or PICP. For the practical use of these indicators as predictors of CHD development was evaluated their diagnostic efficiency and identifies specific thresholds. To this end performed the ROC analysis and the graphs of ROC curves.
The study of these parameters was performed in relation to their significance in terms of the development of significant stenosis (>75%) in patients with coronary artery disease. The area of the ROC curve for MMP-9 amounted to 0.662 (p=0,0024), indicating a good model.
Index Adena amounted 0,306, and associated criterion more at 101.8 ng/ml.
The area of the ROC curve for PICP amounted 0,680 (p=0.0007). Index Udena was 0.345 and an associated criterion more 195,6 ng/ml.
The odds ratio for MMP-9 more at 101.8 ng/ml, provided that the value of PICP remains constant is 3,41 (confidence interval 1,38-of 8.47; p=0.007). The odds ratio for PICP more 195,6 ng/ml, provided that the value of MMP-9 remains constant, is 4,66 (confide�individual interval 1,68-12,9; p=0.003).
Using the proposed method improves the accuracy of early diagnosis of critical coronary stenosis in patients with coronary artery disease.
Sensitivity 98%, specificity of 93%.
EXAMPLES of SPECIFIC APPLICATIONS
Example No. 1. Patient A., age 69, teacher. History of angina of II FC, hypertension, atrial fibrillation paroxysmal form. Complaints of episodes of increased blood pressure up to 160/100 mm Hg.CT., accompanied by headache, weakness, dizziness, nausea. Pain behind the breastbone notes during BP and during exercise (distance 300 m). Taking Enap 10 mg 2 times a day, amlodipine 10 mg 1 tab. for the night. An objective examination of the condition is satisfactory, the lungs breath held, heart sounds are clear, correct rhythm, no noise. The abdomen is soft, painless, the liver is on the edge of the costal arch. HELL 150/90 mm Hg.PT. Pulse 68 / min, regular rhythm. In General, the analysis of blood from 18.06.2013: erythrocytes of 4.0×1012/l, hemoglobin 142 g/l, platelets 322×109/l, leukocyte count of 4.1×109/l, uh - 1% p - 2%, and 53%, l - 37%, m - 6%, ESR 2 mm/h. Total blood count, lipid profile and biochemical analysis of blood without deviation from the norm. When conducting Holter ECG - single SVPBS, VES. When conducting VEM - increase in blood pressure to 160/90 mm Hg.CT., ischemic changes were revealed. The level of MMP-9 122,58 ng/ml. the Diagnosis of coronary artery disease. Angina of II �K. Hypertension stage III, 2 degrees AD risk 4. Complications: Paroxysmal form of atrial fibrillation. During the coronary angiography stenosis was found, PMA 80%) and ACP 75%.
Conclusion. Determination of MMP-9 has a high probability to speak before the coronary angiography that the patient has ischemic heart disease, functional class III, there is a critical stenosis.
Example No. 2.
Patient P., 56 years old, the chef, was admitted with complaints of chest pain and oppressive aching, faults in work of heart and increase in blood pressure to 150/90 mm Hg.PT.
History of arterial hypertension, angina of II FC. Complaints of episodes of increased blood pressure to 170/105 mm Hg.CT., accompanied by headache. Pain behind the breastbone notes when walking 200-300 meters, stopped after 10 minutes when you stop. Taking ACE inhibitors (co-Renitec 20/12,5 mg 1 tab. in the morning, Concor 5 mg 1 tab. in the morning). An objective examination of the condition is satisfactory, the lungs breath held, heart sounds are clear, correct rhythm, no noise. The abdomen is soft, painless, the liver is on the edge of the costal arch. HELL 130/90 mm Hg.PT. The pulse is 62 / min, regular rhythm. In General, the analysis of blood from 18.06.2013: erythrocytes of 3.8×1012/l, hemoglobin 124 g/l, platelets 210×109/l, leukocyte count of 5.6×109/l, e - 3% p - 1%. with - 55% l - 35%, m - 7%, ESR 12 mm/h. Total blood count and biochemical analysis of blood without deviation �t standards. In lipid spectrum pays attention to the increase of total cholesterol to 6.3 mmol/l, LDL 3.5 mmol/l When performing ECG Holter monitoring - frequent SVPBS (up to 70 in h), by GEN. When conducting VEM - increased blood pressure up to 170/90 mm Hg.CT., ischemic changes were not revealed. The level of PICP - 218 ng/ml. the Diagnosis of coronary artery disease. Angina of II FC. Hypertension stage III, 2 degrees AD risk 4.
Subsequent coronary angiography revealed stenosis of the RCA to 85% and PMA 90%.
Conclusion. The patient identified coronary heart disease, which was confirmed by stress test and subsequent determination of the PICP. Later this was confirmed by the data of KBR. The definition of PICP has a high probability to say that the patient has a critical stenosis of the coronary arteries with that, when you receive the diagnosis of angina II FC only.
Example No. 3. Patient P., 44, Builder. History of arterial hypertension, angina of II FC. Complaints of episodes of increased blood pressure to 180/95 mm Hg.PT. Pain behind the breastbone notes under load (distance 200 m). Taking Concor 10 mg 1 time per day, amlodipine 10 mg 1 tab. for the night. An objective examination of the condition is satisfactory, the lungs breath held, heart sounds are clear, correct rhythm, no noise. The abdomen is soft, painless, the liver is on the edge of the costal arch. HELL 140/90 mm RT.PT. The heart rate of 55 per minute, regular rhythm. General �analysis of blood, lipid profile and biochemical analysis of blood without deviation from the norm. When conducting ECG Holter monitoring - frequent SVPBS, VES, episode ST depression of 1 mm to 5 minutes. When conducting VEM - increase in blood pressure to 180/90 mm Hg.CT., ischemic changes were not revealed. The level of MMP-9 to 133.5 ng/ml, the level of PICP 235,2 ng/ml. the Diagnosis of coronary artery disease. Angina of III FC. Hypertension stage III, 2 degrees AD risk 4. During the coronary angiography stenosis was found, PMA 90%, VTK-1 75%, FE 58%.
Conclusion. Determination of MMP-9 and PICP has a high probability to speak before the coronary angiography that the patient has coronary artery disease with critical stenosis.
Example No. 4. Patient A., 49, housewife. History of angina of FC I-II arterial hypertension. Complaints of episodes of increased blood pressure to 150/100 mm Hg.CT., accompanied by headache, weakness. Pain behind the breastbone notes during BP and during exercise (walking 500 m). Takes Renitec 10 mg 2 times a day, amlodipine 10 mg 1 tab. for the night. An objective examination of the condition is satisfactory, the lungs breath held, heart sounds are clear, correct rhythm, no noise. The abdomen is soft, painless, the liver is on the edge of the costal arch. HELL 140/90 mm RT.PT. Pulse 88 / minute, regular rhythm. Total blood count, lipid profile and biochemical analysis of blood without deviation from the norm. When conducting Holter ECG - rare VAW�With, VES. When conducting VEM - increase in blood pressure to 160/90 mm Hg.CT., ischemic changes were not revealed. The level of MMP-9 is 12.58 ng/ml, PICP to 44.8 ng/ml. the Diagnosis of coronary artery disease. Angina of FC I-II. Hypertension stage II, 1 degree AD risk 3. During the coronary angiography stenosis was found, PMA 20% and ECT-1 is 25%.
Conclusion. Determination of MMP-9 and PICP has a high probability to speak before the coronary angiography that the patient has ischemic heart disease, functional class I-II, no critical stenosis.
Thus, our studies reveal the criteria on the basis of which it is possible to predict the clinical course of CHD, in particular the development of critical coronary stenosis. The obtained data allow cardiologists in the early stages of diagnosing coronary artery disease. A significant advantage of this method is the small amount of blood required for testing, high diagnostic accuracy and specificity. Furthermore, the method is not traumatic and has no contraindications. The method was tested on clinical material and find application in the practice of cardiologists, physicians.
A method of predicting risk of development of critical coronary stenosis in patients with coronary artery disease with blood, characterized in that in the serum of the patient determine concentrationtime metalloproteinases - 9 (MMP-9) and/or C-terminal propeptide of procollagen type 1 (PICP) and MMP-9>a 101.8 ng/ml and/or RSR>195,6 ng/ml predict the development of critical coronary stenosis in patients with coronary artery disease.
SUBSTANCE: invention relates to medicine, namely to immunology, and describes a method of determining the functional activity of the human complement factor B by its influence on target cells in a calcium-free medium, which contains magnesium ions, in the presence of the reagent RB, representing human blood serum, selectively deprived of the factor B activity, as the target cells selected are infusoria Tetrahymena pyriformis, the suspension of which together with the tested sample, which contains the determined factor B, and the reagent RB is introduced in measuring cells of the device for the automated calculation of the number of alive infusoria with the following determination of the number of alive cells in each minute, and the concurrence of dynamics of the change in the number of alive cells in time for the tested sample and control one, which represents a pool of 10 serums of healthy donors, supposes the equality of activities of the complement factor B in the said samples.
EFFECT: invention ensures the more accessible target for the action of the complement which infusoria Tetrahymena pyriformis represent.
2 cl, 1 dwg, 1 ex
SUBSTANCE: children are divided into 4 groups; the first group includes the children having the following symptoms: short period of breastfeeding in the medical history, frequently ill family members, no complications of respiratory infections, IgA from 0.7 to 1.1 mg/ml, IgG from 10.6 to 16.2 mg/ml; the second group: mother's gestosis during the pregnancy, threatening miscarriage in the medical history, frequent complications of respiratory diseases - sinusitis, otitis; alpha2-globulin from 9.4 to 11.8 g/l, gamma-globulin from 17.8 to 22.6 g/l, TNF-α from 8.7 to 10.1 pg/ml, IgG from 16.7 to 23.7 mg/ml, CD3+ from 51 to 56.2%, CD4+ from 27.2 to 29.4%, CD16+CD56+ from 6.5 to 8.5%, nitro blue tetrazolium reduction test from 25 to 32.8%; the third group: mother's anaemia during the pregnancy, child's positive endocrine heredity, respiratory diseases frequently complicated with bronchitis, Candida seeding in throat swabs, gamma globulins from 12.1 to 14.9 g/l, IgG from 10.9 to 15.5 mg/ml, CD19+ from 7.6 to 9.6%, nitro blue tetrazolium reduction test from 25.5 to 31.7%; the fourth group: allergic reactions in the child's medical history, positive allergic heredity, respiratory diseases frequently complicated with obstructive bronchitis, gamma globulins from 18.1 to 23.1 g/l, IgE from 67 to 261 IU/ml. Acute respiratory diseases are prevented by Multitabs, a vitamin mineral complex, and one of the immunocorrective preparations: in the first group with IRS-19, a preparation of bacterial lysates; in the second group with a preparation of glucosaminyl muramyl dipeptide (Licopid); in the third group with a preparation of azoximer bromide (polyoxidonium); in the fourth group the immunocorrection is not recommended.
EFFECT: using the given method enables selecting a variant of immunocorrection depending on the presence or absence of diagnostic signs.
2 ex, 1 tbl
SUBSTANCE: invention refers to medicine, namely to paediatrics, and can be used to predict hospital-acquired intestinal infection in an infant. That is ensured by quantitative measurement of the sIgA content in coprofiltrate at admission to hospital. If the sIgA level is less than 23.2-63.5 mg/l, a secondary hospital-acquired intestinal infection is predicted.
EFFECT: using the given method enables detecting the infants suffering changes in local intestinal immunity at admission to hospital and conducting an adequate therapy added with immunocorrective agents for the purpose of preventing the hospital-acquired intestinal infection.
SUBSTANCE: invention consists in providing a method of preserving an immunoperoxidase conjugate using protein stabiliser solution based on an aqueous emulsion of hen egg white as a stabilising medium of drying while lyophilisation and after dissolving the product with preserving the stability of physical and immunochemical indicators.
EFFECT: invention enables to maintain the stability of physical and immunochemical indicators of the immunoperoxidase conjugates during the long period.
1 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to immunology. What is presented is a completely human monoclonal antibody, which binds insulin-like growth factor-II (IGF-II) and has a cross responsiveness to IGF-I, as well as its antigen-binding fragment. There are disclosed a nucleic acid molecule coding an antibody according to the invention, a vector and a host cell for the expression of the antibody according the invention. There are described a pharmaceutical composition, as well as conjugates for treating and diagnosing malignant tumour, using the antibody according to the invention in preparing the therapeutic agent and a method for determining IGF-II and IGF-I levels in a patient's sample.
EFFECT: present invention can find further application in cancer therapy.
16 cl, 27 ex, 18 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: presented group of inventions concerns fused proteins, nucleic acids coding these proteins, an expressing cartridge providing nucleic acid expression, a vector comprising this cartridge, a diagnostic technique for in vitro borreliosis, a kit for this diagnostic technique, which use these proteins, as well as a vaccine composition for preventing borreliosis containing these proteins. The characterised fused proteins contain (i) at least one sequence of DbpA protein of the species Borrelia specified in B. afzelii, B. burgdorferi sensu stricto and B. garinii, and (ii) least one sequence of OspC protein of the species Borrelia specified in B. afzelii, B. burgdorferi sensu stricto and B. garinii.
EFFECT: presented group of inventions enables performing more sensitive and specific analyses related to the presence of certain pathogenic species Borrelia.
11 cl, 8 tbl, 7 ex
SUBSTANCE: group of inventions relates to the field of biochemistry. Claimed is a device for nucleic acid sampling, application of the device of nucleic acid sampling, a set for nucleic acid amplification, as well as a method of nucleic acid amplification. The device includes a probe for holding the nucleic acid sample and a manipulator, connected to the probe with an account of a possibility of the probe manoeuvring. The probe includes a multitude of probe elements, which are capable of travelling with respect to each other between approximated and mutually remote configurations. Each probe element has a nozzle. When the probe elements move into an approximated configuration, each nozzle is joined with other nozzles to form a composite nozzle. The composite nozzle is capable of contacting with nucleic acid in order to deliver its sample. The method of amplification of nucleic acid from higher eukaryotes includes the contact of the sampling device with a source of nucleic acid from higher eukaryotes, introduction of the sampling device or its part into a reaction vessel to carry out the reaction of nucleic acid amplification without preliminary material processing and realisation of the reaction of nucleic acid amplification.
EFFECT: inventions provide portability, easiness of the application of device for nucleic acid sampling, as well as carrying out the analysis in the short period of time.
43 cl, 27 dwg, 2 tbl, 12 ex
SUBSTANCE: present invention refers to immunology. Presented is an antibody able to bind to an amplified epidermal growth factor receptor (EGFR) and to de2-7 EGFR, a truncated version of EGFR, and characterised by sequences of variable domains. There are also disclosed a kit for diagnosing a tumour, an immunoconjugate, pharmaceutical compositions and methods of treating a malignant tumour based on using the antibody according to the invention, as well as a single-cell host to form the antibody according to the present invention.
EFFECT: invention can find further application in diagnosing and treating cancer.
43 cl, 98 dwg, 20 tbl, 26 ex
SUBSTANCE: group of inventions refers to medicine. Presented are diagnostic technique for chronic obstructive pulmonary disease (COPD) and differentiating stages I/II and III/IV COPD in a human involving measuring heat-shock protein 27 (HSP27), 70 (HSP70) and 90 alpha (HSP90 alpha) and measuring interleukin-1 (ST2) receptor 4 or histon-related DNA fragments in a sample. The COBD is diagnosed, when the measured amount of HSP27, HSP70, HSP90 alpha, ST2 and histon-related DNA fragments appears to increase as compared to that in the healthy people. The stage I/II COBD is diagnosed, when the measured amount of HSP27 in the sample of the above individual falls within 2,600 and 3,300 pg/ml, the amount of ST2 is more than 160 pg/ml, while the amount of HSP70 and HSP90 alpha is at least 40% increased as compared to that in the healthy people. The stage III/IV COBD is diagnosed, when the amount of HSP27 in the sample falls within 3,400 and 5,500 pg/ml, while the amount of HSP70, HSP90 alpha and histon-related DNA fragments is at least 40% increased as compared to that in the healthy people.
EFFECT: group of inventions provides the effective diagnostic techniques for the COPD and differentiating between the stages I/II and III/IV COPD in the individual.
11 cl, 20 dwg, 2 tbl, 2 ex
SUBSTANCE: invention relates to laboratory diagnostics, namely to clinical immunology. Method of determining C3-convertase stabilisation of classical way of human complement activation is realised by carrying out reaction of lysis of ram erythrocytes with 0.8% of human blood serum for 10 min. Reaction is stopped by addition of buffer, which contains 10 mM ethylendiamintetraacetic acid, degree of lysis of erythrocytes in control sample is determined. Experimental sample is additionally incubated for 30 min at 37°C, degree of lysis is determined, activity of C3-convertase is calculated as difference between experimental and control samples, with difference higher than 10% it is evaluated as autoimmune pathological condition.
EFFECT: application of simple and fast in realisation method makes it possible to detect preclinical condition of immunodeficiency, can serve as specific marker for immunomodulating therapy, carry out profound research of pathogenesis of autoimmune diseases.
3 tbl, 3 ex
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.
FIELD: medicine, medicinal microbiology.
SUBSTANCE: method involves growing microorganism culture to be studied in solid nutrient medium followed by preparing microbial suspension and its incubation in the presence of lactoferrin. Control sample is prepared in parallel series. Control and experimental samples are incubated, supernatant is removed from bacterial cells and lactoferrin concentration is determined in supernatant of experimental and control sample by immunoenzyme analysis. Then anti-lactoferrin activity is calculated by difference of concentrations of residual lactoferrin in experimental and control samples. This method provides enhancing the sensitivity and precision in carrying out the quantitative evaluation of anti-lactoferrin activity in broad spectrum of microorganisms that is urgent in diagnosis and prognosis of diseases with bacterial etiology. Invention can be used in determination of persistent indices of microorganisms for assay of their etiological significance in pathological processes.
EFFECT: improved assay method.
3 tbl, 3 ex
FIELD: medicine, biology.
SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.
EFFECT: improved an valuable properties of nutrient medium.
FIELD: medicine, cardiology.
SUBSTANCE: in peripheral blood one should detect the level of CD95(+) and CD16(+) neutrophilic granulocytes and at combination of increased level of CD95(+) neutrophilic granulocytes by 4 times and more and CD16(+) neutrophilic granulocytes by 0.6 times against the norm with ECG signs of myocardial infarction one should predict lethal result of large-focal myocardial infarction.
EFFECT: higher accuracy of prediction.
FIELD: medicine, parasitology.
SUBSTANCE: one should carry out immunoenzymatic assay to detect diagnostic optic density and that of labeled immune complex in a plot's hole with tested serum measured in conventional units at wave length being 492 nm. One should calculate coefficient of antibodies concentration measured in conventional units by the following formula: CAC = (Odtsh - Odd) x 100, where CAC - coefficient of antibodies concentration, Odtsh - optic density of the hole with tested serum, Odd - diagnostic value of optic density, 100 - coefficient of serumal dilution. By CAC value one should detect the titer of antibodies to Lamblia intestinalis antigens to interpret results of the trial. The method enables to study the dynamics of disease flow.
EFFECT: higher efficiency and accuracy of diagnostics.
1 ex, 1 tbl
SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.
EFFECT: higher accuracy of detection.
FIELD: medicine, immunology.
SUBSTANCE: one should carry out reaction of blast-transformation, detect proliferation of T-lymphocytes activated with antibodies to CD3 in the presence of interleukin-7 (ACT IL-7) and in the presence of interleukin-7 and dexametazone (ACT IL-7 D), calculate the index for dexametazone action as the ratio of ACT IL-7 to ACT IL-7 D, moreover, the value of dexametazone action index being above 1.2 indicates increased production of cytokins that suppress T-lymphocytes in neonatals. The method enables to detect functional defect of immune system that characterizes neonatal period.
EFFECT: higher efficiency of detection.
SUBSTANCE: method involves measuring forced exhalation volume per 1 s (FEV1) in l, full right ventricle evacuation time (RVE) in ms and angiotensin II value (AII) in ng/l. Discriminant relationship is built as D=0.504·RVE+3.038·FEV1 - 2.0·AII. D being less than 83.88, pulmonary hypertension occurrence is predicted within 1 year. D being equal to or greater than 83.88, no pulmonary hypertension is predicted to occur.
EFFECT: enhanced accuracy of prediction.
FIELD: medicine, medicinal immunology.
SUBSTANCE: method involves determination of heterophilic antibodies in human serum blood by the Paul-Bunnel's method relatively the level of circulating immune complexes, complement-activating properties of heterophilic antibodies by incubation of standardized ram erythrocytes with 0.8% serum for 30 ± 5 min and the following measurement of the erythrocytes lysis degree. The measurement of the effector function coefficient of heterophilic antibodies is carried out by the complement system Keff.f.h.a.-c.s. by the formula: Keff.f.h.a.-c.s. = Y/Tg.a. wherein Y means a lysis degree, %; Tg.a. means a reverse titer of heterophilic antibodies to ram erythrocytes. The damage assay is carried out by comparison of the immune status with the relative level of circulating immune complexes in serum. Method provides detection of preclinic from of immunodeficiency and autoimmune diseases that opens the possibility for their prophylaxis at most early stages of development. Invention can be used for assay of damage in the immune status in human serum blood.
EFFECT: improved method for assay.
5 tbl, 1 ex
SUBSTANCE: method involves concurrently examining anti-inflammatory IL-4 level in blood serum and lacrimal fluid. The value being within the limits of 60-70 pg/l in blood serum and 5-15 pg/l in lacrimal fluid, disease prognosis is considered to be unfavorable. The IL-4 concentration being within the limits of 90-100 pg/l in blood serum and 20-30 pg/l in lacrimal fluid, disease prognosis is considered to be favorable.
EFFECT: high accuracy of diagnosis.