Diagnosis of stress cardiomyopathy
FIELD: medicine, diagnosis, in particular earlier diagnosis of heart disorders in sportsmen.
SUBSTANCE: claimed method includes detecting of cardial antigen presence in blood serum using reaction of passive hemagglutination inhibition, and when titer of antimiocardial test-serum in presence of serum of tested subject by to ore more times stress cardiomyopathy is diagnosed.
EFFECT: diagnosis of stress cardiomyopathy in earlier steps.
The proposed method relates to medicine, particularly cardiology, and can be used for early (pre-clinical) diagnosis of heart disease in athletes.
The most frequent disease of the heart in athletes is degeneration of the myocardium due to physical stress (1, 2). In the international classification of diseases (before revision) this disease is called stress cardiomyopathy, therefore, in recent years in our country instead of the term "degeneration of the myocardium due to physical stress" began to use the term "stress cardiomyopathy" (3, 4). The use of this term is justified also by the fact that patients is not only a physical and nervous stress.
The primary method of detection of stress cardiomyopathy is an ECG study. About the development of degenerative process in the myocardium show changes of repolarization and the emergence of rhythm and conduction. These changes are usually accompanied by a drop in performance. However, degeneration of the myocardium can be both diffuse and focal in nature, so in the latter case, changes on the ECG may be missing.
Therefore, the main disadvantage of ECG diagnosis is that it is not possible to identify early stages of development of stress CT is homeopatie before any changes the recorded ECG focal changes.
In connection with the foregoing, is warranted search different information criteria, indicating the development of myocardial degeneration, even when there are no ECG changes and does not suffer the performance, i.e. the pre-clinical stage of disease. Such early diagnosis of myocardial degeneration will allow to carry out treatment and preventive measures. This, in turn, gives the opportunity to maintain health, high performance and to prevent the development of disease.
The closest technical solution to the claimed method is a method described in the work Gavrilova E. "Stress cardiomyopathy in athletes: (dystrophy of the myocardium of physical stress)". Abstract. Prof. Diss., SPb., 2001.
According to this method, one of the criteria for the diagnosis of stress cardiomyopathy is the detection in serum of antibodies to the antigen of the myocardium in the reaction (TPHA) passive haemagglutination.
The objective of the invention is to increase the effectiveness of the treatment of stress cardiomyopathy by identifying it on the preclinical stages of development.
However, this method confirms the presence of myocardial degeneration by clinical stage, but you can't identify it at the preclinical stage, when netizenry on ECG, that's when you need to prevent that did not develop clinical stage: to reduce the volume and intesively training loads and to prescribe the drugs.
This object is achieved due to the fact that in the known method for the diagnosis of stress cardiomyopathy by serological analysis of blood serum, blood serum to determine the presence of cardiac antigen with response inhibition passive haemagglutination and at lower titers of therapy for anti-myocardial test serum in the presence of serum of the studied persons in two or more times diagnosed with stress cardiomyopathy.
The proposed method for the diagnosis of stress cardiomyopathy is based on serological analysis of blood, which detect the presence of myocardial antigen by the statement of response inhibition passive haemagglutination (RTPA).
When it runs in the blood serum of the subject (in the case of destructive processes in the myocardium) serologic method to detect the presence of antigen, the presence and the level of which indicates the degree of myocardial damage.
The method is based on the fact that the antigen of the myocardium, which may be present in the blood during stress cardiomyopathy, inhibits the interaction of the test antigen with antibodies in response inhibition passive haemagglutination. When it is determined the lower titers of diagnostic antimonial serum after incubation with the test serum. In these cases, the reduction of the titer of the antisera to one degree or more (for example, from 1:160 to 1:80 and over) mean detection in blood antigen and is a sign of destruction of the myocardium. The lower titers of test antisera on 2 stages and more allows you to assess the level of detectable antigen and, therefore, the degree of destruction of the myocardium.
The presence in the serum antigens myocardium indicates the presence of stress cardiomyopathy, even in cases when there are no clinical manifestations of the disease, or they are very poor, and when there are no ECG changes.
The proposed method preclinical diagnosis of stress cardiomyopathy is as follows.
1. Cooking myocardial antigen
The tissue of the myocardium from a person with 0 (I) blood group who died from accidental causes, used fresh or after storage in the frozen state.
A portion of the fabric is shredded with scissors, placed in a gauze bag, washed for one hour running water. The washed fabric is thoroughly crushed in a porcelain mortar with quartz sand, add saline, passed through one layer of cheesecloth to remove large particles. The resulting material is washed 5 times with saline by centrifugation at 18-20 thousand rpm for 1 minute. The precipitate is mixed with phosphate buffer solution (pH of 7.2) and incubated at 4°With during the day. After that, the liquid is centrifuged for 20 minutes at 3000 rpm./min, then adosados centrifuged at 18 tycobrahe 30 minutes at 4°C.
The supernatant was adjusted to a concentration of 2 mg protein in 1 ml and stored in the refrigerator with the addition of merthiolate in a dilution of 1:10000.
2. Getting therapy for anti-myocardial test serum with antibodies to myocardial antigen
Rabbits weighing not less than 2.5 kg are subjected to immunization with antigen from the myocardium according to the following scheme. In the pads of the hind legs of a rabbit injected with a mixture of antigen with complete adjuvant's adjuvant in a volume of 0.4 ml: 0.2 ml antigen containing 2 mg protein in 1 ml + 0.2 ml adjuvant. After 3 weeks antigen in a total dose of 10 mg injected directly into the enlarged lymph nodes. 10 days after the last immunization take the blood sample, determine serum antibody titer. When the titer of 1:320 and above blood samples were taken, prepare serum, which is heated at 37°C, poured into sterile vials and stored at 4°s to use.
3. Preparation of sera examined persons
The blood serum of patients with inactivating 30 minutes at 56°and drain the washed erythrocytes of sheep: 0.5 ml serum, 0.5 to sediment red blood cells, washed with physiological saline containing 1% krolic is her serum. Incubation for 2 hours at 37°and 18-20 hours at 4°C. the Supernatant is used for the given reaction.
4. Treponema pallidum haemagglutination assay. Preparation of red blood cells
Fresh, washed 3 times with saline, the sheep erythrocytes at a concentration of 50% is mixed with an equal volume of chromium chloride (3,8×10 M, 1 mg/ml), prepared ex tempore, with the addition of 100 μg/ml of protein.
After 5-minute incubation at room temperature the interaction is stopped by adding 20-50 volumes sabotinova saline solution and washed by centrifugation 4 times. After that erythrocytes resuspended in physiological solution containing 1% normal rabbit serum, receiving a 0.5% suspension of erythrocytes.
5. The setting reaction of the passive haemagglutination (TPHA)
Reaction set in tablets. Subjects serum of patients diluted by a factor of 2 in the wells containing 0.05 ml of 1% rabbit serum. Later in the wells add 0.025 ml of the suspension loaded with the antigen of red blood cells, incubated at 4°18-20 hours. Consider the reaction of agglutination of red blood cells in the negative control: 1% rabbit serum or 1% rabbit serum + normal human serum.
6. Formulation of response inhibition passive haemagglutination (RTPA)
0.05 ml of whole test blood serum of people who add in the hole, containing 0.05 ml of therapy for anti-myocardial test serum diluted 1:10. The mixture was incubated for 2 hours at a temperature of 37°and then 18-20 hours at 4°C. After that, the serum is diluted by a factor of 2, add to each well loaded with antigen, the red blood cells, determine the final dilution at which agglutination occurred, in comparison with dilutions therapy for anti-myocardial test serum serum, where the test material was added. The lower titers of therapy for anti-myocardial test serum by one step (for example, from 1:160 to 1:80) estimate as a likely indicator of the presence of the antigen of the damaged myocardium, decreased by 2-4 degrees as indicators of high antigen content of the myocardium, which indicates the presence of stress cardiomyopathy.
In control studies, the reduction of the titer of agglutination at 1 stage ensured the detection of 3 µg protein antigen, 2 stage - 6 µg, 4-speed - 12 µg protein antigen.
Patient K. History no._____.
Professional athlete, master of sports of international class 18. Asked the sports medicine center complaining of reduced tolerance to physical activity, decreased athletic performance, no other complaints.
According to clinical examination and objective signs of disease were absent. Heart t is on rhythmic, correct, no noise, rhythm heart rate = 62 beats./min, BP = 120/70. In the lungs vesicular breathing, wheezing no. Peripheral edema no. ECG is normal. Pathological changes. Biochemical and clinical analysis of blood without pathological changes. An ultrasound of the heart pathology was not found.
The presence of cardiac antigen carried out by the statement of response inhibition passive haemagglutination. Results - lower titers of therapy for anti-myocardial test serum at stage 1 (from 1:160 to 1:80), which corresponds to the presence in the blood antigen damaged myocardium.
Diagnosed with stress cardiomyopathy
The patient is assigned therapy: Neoton 4 grams per day, Mildronate 10% - 10 ml/day, papansin 10 ml/day. therapy was carried out for seven days.
After the end of treatment the patient began intensive training. Tolerance to physical load fully recovered. Significantly increased athletic performance.
The example says that the patient had stress cardiomyopathy, pre-clinical stage. In this period did not reveal clinical manifestations and diagnosis made only by means of serological blood tests.
Studies suggest that the proposed method has) the public importance and can be used for the diagnosis of preclinical stage of stress cardiomyopathy. The proposed method can be used in medical practice for the diagnosis of stress cardiomyopathy its preclinical stage.
Thus, the proposed method allows to diagnose stress cardiomyopathy and to take urgent measures for future medical treatment that will prevent the development of disease and to maintain health and high performance. The advantage of this method is its sensitivity, high information content and specificity.
Sources of information
1. Butchenko L.A., Kushakovskij MS Dystrophy of the myocardium in athletes. - M.: Medicine. - 1980.
2. Dembo A.G. Medical control in sport. - M.: Medicine. - 1988.
3. Zemtsovsky EV Dystrophy of the myocardium in athletes // Cardiology. - 1994, No. 9. - P.65-74.
4. Zemtsovsky EV, Bondarev S.A., M. Lobanov Connective tissue dysplasia, tolerance to physical activity and degeneration of the myocardium physical overexertion // journal of sports medicine Russia. - 1999. - V.24, n 3. - P.27.
5. Gavrilova E.A. Stress cardiomyopathy in athletes: (dystrophy of the myocardium of overtraining), abstract. Prof. Diss., St. Petersburg,. 2001.
A method for the diagnosis of stress cardiomyopathy by serological analysis of blood serum, characterized in that the serum to determine the presence of cardiac antigen with response inhibition passive haemagglutination and at lower titers of therapy for anti-myocardial test serum in the presence of serum of the studied persons in two or more times diagnosed stress cardiomyopathy.
SUBSTANCE: method involves determining triiodothyronine content in peripheral blood serum of children of the second half-year of life having perinatal central nervous system injury manifestations and no signs of hypothyroidism. The value being equal to 0.69±1.34 ng/ml, the children are predicted to have small height to the third year of life.
EFFECT: high accuracy and specificity of the method; enhanced effectiveness in selecting appropriate prophylaxis measures.
SUBSTANCE: method involves determining interleukine 1α content in peripheral blood by applying immune enzyme assay method. Interleukine 1α concentration being equal to 21.8±0.21 pg/ml, the child is judged to belong to group threatened by hypoxic ischemic encephalopathy development.
EFFECT: high accuracy of the method; enhanced effectiveness in selecting appropriate therapy at early stage.
SUBSTANCE: method involves calculating survival period duration in months from a formula being linear combination of several variables like morphological tumor parameters, that is, tumor cell area, mean ploidity of tumor cell nuclei, p53 expression level and special area of spontaneous necroses.
EFFECT: high accuracy of prognosis.
SUBSTANCE: method involves determining anti-inflammatory cytokines IL-1β and TNF-α and anti-inflammatory IL-4 content. K coefficient is calculated from formula K=(ln IL-1β + ln TNF-α)/ ln IL-4. K value being found greater than 2, recurrent clinical course of diffuse toxic goiter is to be predicted.
EFFECT: high prognosis accuracy.
SUBSTANCE: method involves carrying out ophthalmic examination. Posteroanterior axis is determined by applying ultrasonic echobiometry method. Availability and spread of peripheral chorioretinal dystrophy is ophthalmologically estimated in grades with Goldman lens as total over the whole eyeball perimeter. Immunoregulation index ratio (IRI=CD4 count/CD8 count), circulating immune complex content, and immunoglobulin IgA content are determined in blood. Posteroanterior axis being equal to or greater than 26.0 mm, peripheral chorioretinal dystrophy spread being greater than 180° and greater than eye ball perimeter, IRI being reduced 1.3 times as small and more, circulating immune complex content being increased by 60% and more and IgA being 1.5 times as high and more, complications development risk is considered to be high after Lasik operation.
EFFECT: high accuracy of prognosis.
SUBSTANCE: the present innovation deals with laboratory methods for studying skin diseases by detecting the content of inflammatory process markers in blood serum at the moment of disease exacerbation and in the end of therapy. As markers of inflammatory process one should apply alpha-1 acid glycoprotein and nitrogen oxide. At decreased content of acid glycoprotein in blood serum ranged 1.19-1.21 g/l up to 0.78-0.82, and as for nitrogen oxide ranged 44.5-54.4 mcM/l up to 24-29.2 mcM/l one should evaluate psoriasis activity to be high. At decreased content of alpha-1 acid glycoprotein ranged 1-1.04 g/l - up to 0.76-0.8 g/l, and nitrogen oxide ranged 37.9-43.5 mcM/l up to 20.6-27.4 mcM/l the psoriasis activity should be evaluated as low. The innovation provides higher significance of diagnostics, decreases labor intensity and period for carrying out the experiments mentioned that enables to develop a number of individual curative procedures for every patient.
EFFECT: higher accuracy of evaluation.
FIELD: medicine, perinatology.
SUBSTANCE: one should sample the waters during the period of delivery in pregnant women from the risk group at verified urogenital infection and/or the presence of fetoplacental insufficiency to detect the value of TNF-α and neopterin. At TNF-α being above 88.1±7.9 pg/ml and neopterin being above 17.3±1.6 ng/ml one should predict hypoxic-ischemic lesion of neonatals' CNS. The innovation enables to indicate earlier curative procedures.
EFFECT: higher accuracy of prediction.
3 ex, 1 tbl
SUBSTANCE: method involves calculating life duration in months from a formula being linear combination of several variables defined as morphological tumor parameters like carcinoma differentiation degree, tumor cell area, mean tumor cell nuclei ploidity level, p53 expression level, PCNA expression level and progesterone receptors expression level.
EFFECT: high accuracy of survival period prognosis.
FIELD: medicine, oncology.
SUBSTANCE: the present innovation deals with predicting the relapse of serous ovarian cancer after radical therapy before its clinical manifestation. The suggested innovation could be applied in daily practice of pathologoanatomic departments of hospitals, oncological dispensaries, specialized centers and research Institutes. The risk for the development of relapse is represented by a dichotomous variable calculated by the formula determined by logistic regression that includes the combination of several variables being morphological parameters of tumor, that is the area of tumor cell, average ploidy of tumor cells' nuclei, the level of p53 expression the level of expression of estrogen receptors, the level of PCNA expression. The innovation enables to predict repeated cancer growth according to morphological characteristics of primary tumor till clinical manifestation of the relapse.
EFFECT: higher accuracy and efficiency of prediction.
FIELD: medicine, vaccines.
SUBSTANCE: disclosed is method for identification, isolation and production of antigens interacting with hyperimmune serum from specific pathogen useful in vaccines for human or animals. Claimed method includes providing of antibody preparation from serum pool of certain animals or human or individual serums with antibodies against certain pathogen; production of at least one expression library of certain pathogen; data screening of said at least one expression library using abovementioned antibody preparation; identification of antibodies binding to antibodies in antibody preparation during screening; screening of identified antigens using individual antibody preparations from individual serums obtained from subjects having antibodies against said certain pathogen; identification of reacting with hyperimmune serum antigen part of identified antigens than bind to relevant part of individual antibody preparations from individual serums optionally isolation of reacting with hyperimmune serum antigens and production thereof by chemical or recombinant methods.
EFFECT: improved method for identification of effective antigens for certain pathogen useful as preferable candidates of antigen vaccines.
20 cl, 11 dwg, 10 tbl, 7 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.