Method for determination of preparation influence on interaction of complement with antigen-antibody complex

FIELD: chemistry.

SUBSTANCE: invention relates to medicine, namely biological chemistry, and can be used to determine the effect of drugs on the classical pathway of complement activation by studying the hemolytic activity of complement. To do this, 50 mcl of blood serum from ½ to 1/128 of blood serum are added to 50 mcl each before dilution in a saline solution to a concentration of 10-2 M, preliminary incubation is carried out at a temperature of 4–6 °C for 30 minutes, then add 50 mcl of sensitized erythrocytes of ram and veronal buffer. Then the cells are re-incubated, the supernatant is taken out and its optical density is determined at 450 nm, and complement fixation (C%) with sensitized erythrocytes in the presence of the studied preparations is calculated by the formula: C%=(Eo-Ec)/Eo-100, where: Eo – optical density in the presence of the preparation, Ec – the optical density of the control, which is used as a veronal buffer. Value of C is judged on the effect of the drug on the C1q subcomponent of the first component of the classical complement pathway in terms of the degree of hemolysis of the sensitized erythrocytes of the sheep.

EFFECT: use of this method makes it possible to make a comparative quantitative assessment of the degree of influence of the tested drugs on the classical pathway of complement activation.

1 cl, 2 dwg, 2 ex

 



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to transfusion medicine, and is applicable in selecting blood plasma donors. That is ensured by sampling blood plasma to be analysed for antimicrobial peptides defensins. If the male defensin titre is from 180 to 545 ng/ml, and the female one is from 128 to 386 ng/ml, a donor is considered to be suitable for blood plasma drawing.

EFFECT: invention enables using the donor plasma in treating the patients suffering from infectious diseases and postoperative, injury and burn complications.

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to pediatrics, and can be applied for predicting reduction of child's health level at the age of 12-16. For this purpose an hour after meal and 15 minutes before mouth fluid sampling oral cavity is washed with 0.9% sodium chloride solution. Analysis of child's mouth fluid with determination of ratio of immunoglobulin A concentration to immunoglobulin G concentration is performed. Analysis is carried out twice with difference in 14 days. In case of 1.4 fold and higher reduction of immunoglobulin A concentration to immunoglobulin G concentration ratio during second analysis child is allocated to group of risk of health level reduction.

EFFECT: application of claimed method makes it possible to carry out screening in the process of carrying out prophylactic medical examination to plan health-improving procedures in groups of dispensary observation.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to gynaecology, and concerns a method for the prediction of endometrial hyperplasia in the patients with hysteromyoma following uterine artery embolisation (UAE). Substance of the method: all the patients undergo a pre-UEA diagnostic and therapeutic uterine scrapping; 6 months after the UAE, an endometrial pipelle biopsy followed by a pathomorphological study of the biopsy material is conducted. If the study shows no pathology, the biopsy materials are analysed by means of an immune histochemical study using anti-Ki-67 monoclononal antibodies; a percentage of Ki-67 proliferation marker in the uterine gland epithelium (coefficient A) and in the endometrial stroma (coefficient B) is determined; a prognostic index D1 is calculated in initial(pre-UEA) non-atypical hyperplasia, and a prognostic index D2 - in initial (pre-UEA) endometrium in the late proliferation phase: D1=A*0.05-0.74 and D2=B*0.02-0.3. If D1 is 0 or more, recurrent endometrial hyperplasia is predicted, whereas D1 less than 0 enables stating a low risk of recurrent endometrial hyperplasia. If D2 is more than 0, a low risk of developing endometrial hyperplasia is stated, whereas D2 of 0 or less shows developing endometrial hyperplasia.

EFFECT: presented method enables the high-accuracy prediction of the post-UEA endometrial hyperplasia that makes it possible to prescribe the preventive hormone therapy in due time.

4 ex

FIELD: medicine.

SUBSTANCE: claimed invention relates to medicine, namely to immunology, and can be applied for detection of contaminants in glucose polymers. For this purpose analysis of inflammatory response is carried out in vitro with application of cell line, which represents cell line of either macrophages or differentiable in macrophages, or cell, expressing one or several toll-like receptors (TLR) or NOD-like receptors, selected from TLR2, TLR4 and NOD2. Analysis contains the following stages: (a) placing macrophages in presence of preparation of glucose polymers, which can contain anti-inflammatory contaminants, and measuring cytokine RANTES production, with production of RANTES cytokine indicating to the fact that preparation contains contaminants capable of initiating inflammatory reaction, and (b) placing cell line, which makes it possible to detect activity of innate immunity receptor or several receptors of innate immunity, selected from TLR2 and NOD2, in presence of preparation and detection of signal of reporter gene, bound to said receptor, with detection of said activity or said signal indicates presence in preparation of contaminant, which represents receptor agonist.

EFFECT: application of claimed method makes it possible to detect contaminants of glucose polymers, and addition of components, such as MDP or LPS, in tested sample makes it possible to act synergistically with contaminants, which increases sensitivity and reduces threshold of detection, with synergetic response being registered for RANTES.

28 cl, 5 ex, 23 dwg, 5 tbl

FIELD: medicine.

SUBSTANCE: peripheral leukocyte blood values before and after a loading test are measured with the use of the gradual submaximal exercise. The method involves the differentiated recovery of different types of leukocytes from the blood to produce a preparation; a total T-cell (Tt) and active T-cell (Ta) count is determined; a Tt/Ta ratio is derived; mononuclear cells are incubated with granulocytes; a granulocyte-binding lymphocyte index (GLI) is determined in accordance with a granulocyte rosette formation (GRL - contact bound to three granulocytes) to granulocyte contact lymphocytes (GCL - contact bound to one granulocyte) ratio in the preparation; leukocyte indices: lymphocyte index (LI), immune reactivity index (IRI), adaptation index, ("CПHP") are determined; immune functional state adaptation coefficients (K) are derived for each value. Total immune functional body state TIFBS is calculated by formula TIFBS = K"спнр"+Kli+Kiri+Kgli+K"итл" before the exercise - TIFBS1 and after the exercise - TIFBS2. A specific immune functional state coefficient SIFSC is calculated by formula SIFSC = (TIFBS1+TIFBS2)/5, and a level of the immune functional SIFSC reserve is determined. If the SIFSC values are above +1.0, the level of the immune functional reserve is considered to be optimal; the SIFSC values falling within the range of 0 to 1.0 show the satisfactory reserve, whereas the SIFSC values below 0 shows the unsatisfactory reserve.

EFFECT: method enables assessing the functional body reserves with the use of combined characteristics including the adaptation body potential, immune reactivity and immune cell interaction.

1 ex

FIELD: medicine.

SUBSTANCE: DNA is recovered from peripheral venous blood. Genetic polymorphisms of tumour necrosis factor α (-308 G/A TNFα), tumour necrosis factor 1 receptor (+36 A/G TNFR1), interferon-inducible T-cell chemoattractant (A/G I-TAC), interleukin 1A (-889 C/T IL-1A), lymphotoxin α (+250 A/G Ltα) are typed by polymerase chain reaction. A high risk of developing hyperplastic processes of endometrium is predicted if detecting a combination of alleles -308 G TNFα, +36 A TNFR1, A I-TAC, -889 T IL-1A and/or a combination of alleles +36 A TNFR1, A I-TAC, -889 T IL-1A and/or a combination of alleles -308 G TNFα,+250 G Ltα, -889 T IL-1A.

EFFECT: higher accuracy.

2 dwg, 1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: chronic infectious-inflammatory diseases (CIIDs) are diagnosed. Clinical blood analysis and bacteriological tests are conducted. A sensibilisation index (SI) and an immune responsiveness index (IRI) are calculated; total microbial count per 1 m3 of the working space air is measured, and the total microbial number (TMN) is derived. If the TMN is less than 500 CFU/m3 with no CIIDs diagnosed accompanied by the SI of less than 1.08 standard units and the IRI of less than 13 standard units, the immunoassay is considered to be inadvisable. If the TMN falls within the range of 500-2,500 CFU/m3 with one CIID diagnosed accompanied by the SI from 1.08 to 1.3 standard units and the IRI from 13.1 to 15.7 standard units, the immunoassay with the first-level tests seems advisable. Whereas the TMN exceeding 2,500 CFU/m3 with at least two CIIDs accompanied by the SI of 1.4-1.5 standard units and the IRI of 15.8-18.3 standard units, the immunoassay with the second-level tests is thought expedient.

EFFECT: invention enables detecting the workers in need of further examination for the purpose of timely immune correction in the setting of mass routine examinations.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: DNA from peripheral venous blood is extracted. An analysis of a combination of genetic versions of polymorphous markers of genes of cytokines of the gene regulator of the activity of normal expression and secretion of T-cells (-403 G/A RANTES), macrophage protein -1β (+1931 A/T MIP 1β), factor of stromal cells (-801 G/A SDF1), interleukin -1 (-511 C/T IL-1B), monocyte chemoattractant protein -1 (C/G MCP-1), interleukin -4 (-590 C/T IL-4) is performed. An increased risk of development of a combination of uterine myoma with endometriosis and hyperplastic processes of the endometrium is predicted if the combination of alleles 403 A RANTES, G MCP-1,+1931 A MIP 1β, -590 C IL-4 or the combination of alleles -403 A RANTES,+1931 A MIP 1β, -801 G SDF1, -511 C IL-1B is identified.

EFFECT: application of the claimed method makes it possible to detect a group of patients with a risk of developing a combination of proliferative reproductive system diseases, which makes it possible to prescribe an adequate therapy to prevent further progressing of the diseases.

3 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: invention deals with method of predicting level of arterial pressure in women of Russian nationality, born in Central Black Earth region of Russia. Method includes separation of DNA from lymphocytes of peripheral venous blood and analysis of genetic polymorphisms. +46G/A ADRB2 and 4a/4b eNOS by method of polymerase chain reaction Level of systolic arterial pressure in women in late pregnancy is predicted by results of multiple regression equation of the following type: Y1=15,455+2,544x1+9,946x2+0,736x3+4,716x4+0,185x5, where x1 is genetic variant in locus - 4a/4b eNOS, namely 4b4b=1; 4a4b=2; 4a4a=3; x2 is presence of preeclampsia in relatives: yes=0, no=1; x3 is level of systolic arterial pressure before pregnancy, mm Hg; x4 is presence of cardiovascular system pathology: yes=0, no=1; x5 is woman's weight before pregnancy, kg Level of diastolic arterial pressure in women in late pregnancy is predicted, for which purpose multiple regression equation of the following type is used: Y2=14,200+7,768x1-2,877x2+7,500x3+0,414x4+3,668x5, where x1 is genetic variant in locus - 4a4b eNOS, namely 4b4b+4a4b=1, 4a4a=0; x2 is genetic variant in locus +46G/A ADRB2, namely GG+GA=1, AA=0; x3 is presence of preeclapsia in relatives:yes=0, no=1; x4 is systolic arterial pressure before pregnancy, mm Hg; x5 is presence of cardiovascular system pathology: yes=0, no=1.

EFFECT: invention makes it possible to realise early prediction of increase of arterial pressure level in women in late pregnancy, will make it possible to form of women at the stage of pregravidal preparation and at early terms of pregnancy groups of high risk of developing hypertension in late pregnancy, as well as realise required therapeutic-preventive measures aimed at prevention of development of said pregnancy complication in due time.

2 dwg, 2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method is based on contacting a membrane test strip with an analysed fluid sample and initiating thereby a motion along the test strip membranes of reagents being parts of the sample or coating the membrane, and forming the immune complexes to be detected in the course of reactions in the membrane pores or on the surface thereof. A distinguishing feature of the presented method for antigen detection is that the test strip is coated additionally within the test sample contact area with a certain amount of specific antibodies, which react to the detected antigen expected to be found in the sample, when a fluid front moves and block a certain number of binding sites. The number of the coating free antibodies is specified so that the low content thereof in the analysed sample being of no diagnostic importance ensures blocking the binding sites completely that prevents the antigen from binding in the analysed area of the test strip and from developing a destructive staining in the analysed area.

EFFECT: presented approach enables reliable diagnosing based on the detection results of the antigens of gastrointestinal disorders, avoiding the achievement of positive test results for the low-antigen samples, which testifies to no development of the disease in an individual.

1 tbl, 2 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.

2 ex

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.

3 ex

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

FIELD: medicine.

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.

2 ex

FIELD: medicine.

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

FIELD: medicine.

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.

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