Instant diagnostic techqniue for mixed herpes viral and bacterial infections in children

FIELD: medicine.

SUBSTANCE: sequential staining by Romanowsky-Giemsa and conducting an immunocytochemistry reaction are used to analyse destructive processes in epitheliocytes nuclei, specifically changed lymphoid and epithelial cells, high colonisation activity of opportunistic and pathogenic microorganisms, and phagocytic activity. If observing brown-black inclusions in cells under optical microscopy ensured by specific staining of the viral antigens in the cell structures, mono- or mixed herpes viral infection Herpex simplex 1, 2, EBV, CMV are diagnosed.

EFFECT: using the technique enables higher accuracy of etiological diagnosis, expressivity and reduced injures of material sampling for research.

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The invention relates to a new medical biotechnology and can be used in clinical practice, as a method of early diagnosis of mixed herpes and bacterial infections in children.

Viral disease of mixed etiology is one of the urgent problems of modern clinical medicine. A special place among them is herpes virus infection associated with respiratory viruses groups of pathogenic and conditionally pathogenic bacteria, fungi and protozoa. The presence of mixed-herpes virus infection contributes to long-term treatment of respiratory diseases, chronic pulmonary and ENT pathology, as well as the involvement of other organs and systems. Over the last 10-15 years has increased the number of patients with recurrent herpes infections. Among the examined preschool children attending organised groups, the majority (80-90%) are infected with any of the herpes viruses. Representatives of the family Herpesviridae have the ability for life to persist in the body of an infected person. Along with adverse exogenous factors in the persistence and reproduction of herpes in immunocompetent cells determine the formation of secondary immune deficiency and contribute to the development of chronic relapsing course of infectious disease is Oia, as well as high frequency layers of concomitant viral and bacterial infections. This leads to the formation of chronic pathology of the respiratory, cardiovascular and other systems, disrupting the harmonious development of the child.

Currently, there are 8 clinically significant types of viruses of the family Herpesviridae: herpes simplex viruses type 1 and 2 (HSV 1, 2), varicella-zoster virus (herpes zoster), Epstein-Barr (EBV), cytomegalovirus (CMV), herpes viruses types 6, 7 and 8.

The current level of laboratory diagnostics, allowing to identify a large number of pathogens or specific antibodies of different classes, marking their presence is not always possible to characterize the etiological significance of identified pathogens, which requires a detailed study of the microbial landscape of the upper respiratory tract pathogens and colonization properties of microorganisms. Differential diagnosis of latent and chronic herpes virus infections in patients carriers of pathogenic and conditionally-pathogenic bacterial flora difficult. This is due to the low immunogenity of herpes viruses, which are causative agents of opportunistic infections and reactivated under the influence of many factors in the presence of defects in the links of antivirus protection.

Known for the manual detection of herpes viruses in tissue samples and biological fluids (urine, saliva) - cytological method.

The method is based on the detected under a light microscope with immersion in smears taken from the affected areas of the mucous membrane and stained by Romanovsky-Giemsa, multinuclear giant cells and intranuclear inclusions (for suspected HSV infection), atypical mononuclear cells (suspected EBV infection) and cells in the form of "owl eyes" (for suspected CMV infection). Cytological method is simple technique of obtaining a large number of informative material. However, although cytological method is technically available and different speed setting, it has a low specificity and it is not possible to diagnose a mixed bacterial-herpesvirus infection (Microbiological aspects of pertussis infection in children. Dissertation of candidate of medical Sciences Oselinsky. S.-Petersburg, NIDI, 2006).

Also known a method for estimating the resistance of the mucous membranes, based on the definition of key indicators of anti-infective drug resistance in bacterial infections in children (Skripchenko, NV, Talikova E.V., Zhelezova LI, Martynova I.A., Kalinogorskaya O.S algorithm for estimating the resistance of the mucous membranes in predicting outcomes of infectious diseases in children: a training manual, St. Petersburg, 2002. - 14 C.). The presence of smears taken sporogenic areas of the mucous membrane, signs of inflammation: polymorphonuclear leukocytes (PMNL), mucus strands of fibrin, as well as micro-organisms, morphologically similar to the etiological significance (pneumococci - lanceolate, diplococci with the capsule, staphylococci, fungi etc), confirms the role of the bacterial component in the development of a mixed infection. To determine the prognosis of bacterial infection in addition use the method of its estimation based on integrated analysis of indicators of colonization activity of the pathogen and nonspecific resistance of the organism, the index of adhesion (IA) - the average number of microbial cells on one involved in the adhesive process epitheliocyte, the index of infection (AI) - the percentage of epithelial cells with adhered cells of the microorganism, phagocytic activity (FA) - the percentage of active phagocytes of the total number of poly - and mononuclear (50 or 100), phagocytic index (PI) is the average number of microbial cells, absorbed one active phagocyto. However, this method provides a screening diagnosis of bacterial infection (mono - and mixed), but does not allow you to diagnose herpes virus infection, including (CMV, EBV etc)that ultimately does not provide a proper diagnosis.

Known molecular genetic method of diagnosis of herpes infection, based on m is agarrate election copying (amplification) of a particular stretch of DNA by enzymes in vitro (in vitro) - polymerase chain reaction (PCR). The method allows on the basis of the detection of the amplified DNA fragments of herpes viruses in the blood is an indication of the activity of the infection. The frequency of detection of DNA viruses in the blood during acute herpes infections ranges from 11% to 62% depending on the severity of the disease. However, with its chronic course, when the pathogen is mainly intracellular, PCR is not efficient enough. In addition, PCR reveals not only the DNA can replicate actively virus, but the virus residing in the phase of latency that is not possible to determine the involvement of viruses in the development of the inflammatory process and does not guarantee the accuracy of diagnosis.

Closest to the proposed method of rapid diagnosis of herpes infection is immunocytochemical method (Everymove, Wavin, Evenascence, Rahasyam, IVA, Asimina. "Clinical and etiological features of infectious mononucleosis in children of early age, Children's infections, 2009, No. 1), allowing to determine the antigens of herpes in blood lymphocytes using monoclonal antibodies to latent membrane protein EBV-LPM-1, structural viral protein late stage replication of CMV PP 65 and polyclonal antibodies to early antigens of HSV type 1 and 2. The results of the method which allows to determine semiquantitative expression of antigens in peripheral blood lymphocytes ("+" - slabopolozhitelnym, "++" positive "+++" - strongly positive). However, the method provides diagnostics only herpes virus infection, and not combined viral-bacterial infection, which reduces the accuracy of diagnosis. The disadvantage of the method is also traumatic way of obtaining material for research (intravenous blood sampling).

To eliminate these drawbacks will help proposed by the authors of the method of rapid diagnosis of mixed herpes and bacterial infections in children by sequential staining Romanovsky-Giemsa and setting reaction of immunocytochemistry. The authors first propose a method of simultaneous detection of significant etiological microorganisms and herpes in the brush-biopsy of the mucosa of the oropharynx sick children with staining of the drug Romanovsky-Giemsa, followed by the staging of the reaction of immunocytochemistry. The presence in a destructive processes in the nuclei of epithelial cells, specifically modified lymphoid and epithelial cells (atypical mononuclear cells, multinuclear giant cells and cells in the form of "owl eyes"), high colonization activity conditionally pathogenic and pathogenic microorganisms and phagocytic activity in relation to them establish a mixed viral-bacterial infection. Then the drug Obasi is more, discolor, applied to selected areas of the slide-specific monoclonal serum to antigens of EBV and CMV and polyclonal HSV 1, 2. When the cells under light microscopy inclusions brown-black color due to specific staining of virus antigens in the cell structures diagnose mono - or mixed-herpesvirus infection of HSV 1, 2, EBV, CMV.

The technical result of the present invention is to improve the accuracy of the etiological diagnosis, expressiveness and in reducing the morbidity of biopsy specimens for research.

The authors first proposed effective rapid laboratory method for the diagnosis of mixed bacterial and herpes virus infection in children. The method has several differences and advantages. Use brush-biopsies from the mucous membrane of the oropharynx eliminates the need for invasive method of selection of material, as the sample is collected by the brush of the brush. Conducting research using citrobactereae and setting reaction of immunocytochemistry performed in one sample on the same slide. The method ensures the integrity of the cell material, which is concentrated in one place, which greatly increases the efficiency of the presented method and saves expensive serum and time, and also extends titolo the AI, already in the early stages of infection allowing for the etiological diagnosis.

The authors solved this problem by claiming a method of detecting bacterial flora and antigens of herpes in the brush-biopsy of the mucosa of the oropharynx sick children using color smear Romanovsky-Giemsa and the subsequent raising of the reaction of immunocytochemistry.

Advantage research of smears is non-invasive nature of the sampling for the study, which is particularly important in pediatric practice. The simplicity of the fence material allows for the observation period to monitor the presence of antigens of herpes viruses.

Immunocytochemical method of diagnosis of herpesvirus infections, which uses monoclonal antibodies to EBV, CMV and polyclonal to HSV-1, type 2, has a high specificity, the ability to visualize the antigenic complex of viruses into target cells of patients with any form of herpes infection.

The proposed method is as follows. The patient produce the fence brush-biopsies of the mucosa of the oropharynx with the brush from the endoscope mounted on the holder, put on a glass slide, dried in air, fixed in acetone (t +4°C) (20 min), pour a solution of paint Romanovsky-Gimza in the amount of 5-8 drops (30 min), add to the paint smears, Feb the number of drops of neutral distilled water, heated to 50°-60°C (3-5 min), wash off the paint with a jet of neutral distilled water, dried with filter paper. On the dried smear assess the anti-infective resistance of the mucosa of the oropharynx, calculating the index of adhesion (IA) - the average number of microbial cells on one involved in the adhesive process epitheliocyte, the index of infection (AI) - the percentage of epithelial cells with adhered cells of the microorganism, phagocytic activity (FA) - the percentage of active phagocytes of the total number of poly - and mononuclear (50 or 100), phagocytic index (PI) is the average number of microbial cells, absorbed one active phagocyto. Then discolor the product: immersion oil on the slide dissolved in 96% ethanol, immersed in distilled water, treated for 5-7 min with 2% hydrochloric acid, washed with 3 portions of distilled water. Then, given the tropism of herpes viruses to columnar epithelial cells, spend immunocytochemical research on antigens of herpes in obespechennim smear. Smears were fixed in acetone for 3-5 minutes, then washed in phosphate-buffered saline (pH 7.2-7.4) for 3-4 minutes, and placed in 3% hydrogen peroxide solution for 5 minutes to remove endogenous peroxidase. Then smears were washed in two portions phosphate is olevaga buffer for 5 minutes each, inflicted on them specific antibodies with a ready breeding production company BioGenex (USA) VEB (clone 1108-1), CMV (clone VM), and herpes simplex I, II types (polyclonal serum), and incubated at t 37°C for 30 minutes. After incubation were washed in two portions fosfato-salt buffer for 5 minutes each, and subsequently used the test system QD420-YIK company BioGenex (USA) Super sensitive Polymer-HRP Detection System". Initially, the strokes inflicted reagent Super Enhancer and incubated at room temperature for 20 minutes, washed in two portions of phosphate-saline buffer for 5 minutes each, then sprayed reagent "Poly-HRP Reagent", incubated at room temperature for 30 minutes, washed in two portions of phosphate-saline buffer for 5 minutes each, were treated in an aqueous solution containing 0.05% 3,3-diaminobenzidine tetrachloride (DAB) and 0.025% solution of hydrogen peroxide for 3 minutes, and finally stained with hematoxylin. To obtain reliable results and to exclude possible non-specific reactions were performed quality control of reagents and specificity of the antisera. To do this, instead of specific antibodies inflicted non-immune serum or buffer, while receiving negative results. The nonspecific binding of serum was eliminated by the maximum dilution of highly concentrated antibodies and shortening the incubation time. Thus, the formulation of the immunocytochemical reaction took about 3-4 hours.

Assessment criterion ICH reaction serve as the intensity of the color (light, dark brown staining until black), its localization (cytoplasm, nucleus) and the degree distribution on the cell. In each smear was evaluated at least 100 cells in one field of view, and examined not less than 3 zones of the glass. In the presence of from 5% to 30% of lymphocytes containing the antigen, the reaction is slabopolojitelen (+). In the presence of lymphocytes with positive reaction in an amount of from 30 to 70%, the reaction is considered positive or moderately positive (++), and staining of 70% or more of the lymphocytes is strongly positive (+++). The results ICH studies allow semiquantitative estimate the level of expression of antigens in peripheral blood lymphocytes. It is held at a known scale McCarthy, which implies taking into account the intensity of staining cells in points (1, weak staining, 2 - moderate staining, 3 - severe 4 - very strong staining).

Example No. 1.

Last name, first name Daniel A.

Paul M

Date of birth / age 17.12.2008, 1 year 1 month

no case history No. 15628

Diagnosis directions: sickly child, examination

Antibodies to EBV, CMV: IgM (VCA) with the WEB will believe., IgG (EA) WEB put., IgM CMV - neg., IgG CMV - neg.

PCR liquid is x: saliva: WEB I will., CMV - neg.; blood: EBV - neg., CMV - neg.; urine: EBV - neg., CMV - neg.

ICH smear from fauces: moderately positive reaction on the WEB

Seeding flora - S.pneum. 105

Morpho-functional study of the mucosa of the oropharynx:

Compensated form of mucosal dysbiosis of the II degree.

The microstructure of the mucous neurotology and laryngo-tracheal secretion:

Resident microflora presents saprophyte neisseriae and viridans streptococci (Streptococcus viridians) - 103

Morphofunctional state of the mucous naso-oropharynx:

Epithelial cells: with the destruction of the core is 20%, the threads of fibrin 2+.

AI index infection, % - percentage of epithelial cells colonized significant etiological microorganisms is highly virulent strain of pyogenic Streptococcus - less than 20%/

IA (index of adhesion, the average number of cells pyogenic Streptococcus adhered on the 1st epitheliocyte) - 10-20 microbial cells.

Cellular elements of blood seen on the mucous neurotology (average number of cells per field):

- polimorfismo-nuclear leukocytes (PMNL) - 20 cells in field of view

lymphocytes 3-6 in the field of view

- eosinophils - 2-3 in the field of view.

- others do not.

The state of phagocytosis: FA - phagocytic activity (number of blood cells in % with phagocytic the activity relative to cells pyogenic Streptococcus - 40%

PHI - phagocytic index (the average number of absorbed cells of the microorganism per 1 fagozyt) - 3-5 ml/fagozyt.

The level of local nonspecific immunity:

SIgA - level General non-specific secretory immunoglobulin a in the reaction of indirect immunofluorescence (lg) - 4 lg.

Thus, using the proposed method is a non-invasive method revealed a mixed viral-bacterial infection (EBV + CMV + pneumococcal) on the background of a dysbacteriosis of the mucous membrane of the II degree and reduce anti-infective resistance of mucous. The etiology of herpes infection is confirmed by serological method (IgM VCA) with the WEB will believe.) and molecular biology (PCR saliva on the WEB put.).

Example No. 2.

Last name, first name Daniel P.

Paul M

Date of birth / age 30.11.2008, 1 year 1 month

no case history No. 15589

Diagnosis directions: sickly child, examination

Antibodies to EBV, CMV: IgM (VCA) with the WEB will believe., IgG (EA) WEB put. IgM CMV - put., IgG CMV - neg.

PCR fluids: saliva: EBV - neg., CMV - neg; blood: EBV - neg., CMV - neg; urine: EBV - neg., CMV - neg.

ICH smear from fauces: moderately positive response to EBV and CMV.

Seeding flora: neg.

Morpho-functional study of the mucous neurotology:

Decompensated form of cisbio is and mucous III degree.

The resident microflora is virtually absent: register a single cell non-pathogenic Neisseria and "viridans" streptococci (Streptococcus viridas) - 101.

Pathogenic flora: Streptococcus pneumoniae 104the biofilm!

Epithelial cells: degradation of cores - 15%, strands of fibrin 1+

AI index infection, % - percentage of epithelial cells colonized significant etiological microorganisms is highly virulent strain of pneumococcus - 20-30%

IA (index of adhesion, the average number of pneumococcus cells adhered on one epitheliocyte) - an average of about 20 cells pneumococcus (on the surface of epithelial cells in the biofilm!)

Cellular elements of blood seen on the mucous neurotology (average number of cells per field):

- polymorphonuclear leukocytes (PMNL) - 2-5 cells in field of view

lymphocytes 2-5 in the field of view

- eosinophils - none

- other - atypical mononuclear cells, 2-3 in sight!

The state of phagocytosis: FA - phagocytic activity (number of blood cells in % with phagocytic activity against pneumococci - 10%

PHI - phagocytic index (the average number of absorbed cells of the microorganism per 1 fagozyt) - 3-5 ml/fagozyt. Incomplete phagocytosis.

The level of local nonspecific immunity:

SIgA - level General non-specific secretory to immunoglobu the ina As in the reaction of indirect immunofluorescence (lg) - 2 lg

Thus, using the proposed method is a non-invasive method revealed a mixed viral-bacterial infection (EBV + pneumococcal) on the background of a dysbacteriosis of the mucous membrane of III degree and reduce anti-infective resistance of mucous. The etiology of herpes infection is confirmed by serological method (IgM (VCA) with the WEB will believe., IgG (EA) WEB put. IgM CMV - put.). Molecular biological analysis of blood, saliva and urine were uninformative (PCR blood, saliva and urine DNA EBV and CMV neg).

Example No. 3.

Last name, first name Vladimir D.

Paul M

Date of birth / age 17.07.2007, 2 g 6 months

no case history No. 15374

Diagnosis directions: sickly child, examination

Antibodies to EBV, CMV: IgM (VCA) with the WEB will believe., IgG (EA) EBV - neg. IgM CMV - neg., IgG CMV - believe.

PCR fluids: saliva: EBV - neg., CMV will believe; blood: EBV - neg., CMV - neg; urine: EBV - neg., CMV - neg.

ICH smear from fauces: expressed positive reaction on the WEB and moderately positive for CMV.

Seeding flora: St.aureus 103

Morpho-functional study of the mucosa of the oropharynx:

Decompensated form of dysbiosis of the mucosa of the oropharynx III degree. Resident microflora presents saprophyte neisseriae and viridans streptococci (Streptococcus viridians) - 10 3conditionally-pathogenic (S.aureus) - 105individual in the field of view of the threads of actinomycetes.

Epithelial cells: with the destruction of the cytoplasm and nucleus of 5-10%, the threads of fibrin 1+.

AI index infection, % - percentage of epithelial cells colonized significant etiological microorganism - S.aureus) - 40%.

IA (index of adhesion, the average number of cells of Staphylococcus aureus, adhesionand on one epitheliocyte) - 30-40 ml on the surface 1 of epithelial cells.

Cellular elements of blood seen on the mucous neurotology (average number of cells per field):

- polymorphonuclear leukocytes (PMNL) - 1-2 cells in field of view

- lymphocytes - 3-6 in the field of view

- others do not.

The state of phagocytosis: FA - phagocytic activity (number of blood cells in % with phagocytic activity compared to cells important etiological microorganisms is not registered.

PHI - phagocytic index (the average number of absorbed cells of the microorganism per 1 fagozyt) - not registered

The level of local nonspecific immunity:

SIgA - level General non-specific secretory immunoglobulin a in the reaction of indirect immunofluorescence (lg) - 4 lg.

Thus, using the proposed method is a non-invasive method revealed a mixed viral-bacterial infection (EBV + CMV + staphylococcal) in the region of dysbiosis of the mucous membrane of III degree and pronounced reduction of anti-infective resistance of mucous. The etiology of herpes infection is confirmed by serological method (IgM (VCA) with the WEB will believe., IgG CMV - put.). Using PCR detected only CMV DNA in the saliva.

Clinical observations suggest that the suspected disease herpes virus causes a typical and particularly atypical forms of infectious mononucleosis both acute and chronic) justified advanced diagnostic examination of a patient, aimed at the detection of viruses of the family Herpesviridae, and bacterial agents.

The method is distinguished by the simplicity and accessibility, can be used in the practice of any bacteriological laboratory, does not require expensive reagents and equipment.

The method of rapid diagnosis of mixed herpes and bacterial infections in children through painting Romanovsky-Giemsa and setting reaction of immunocytochemistry, characterized in that the charge brush-sampling of the mucosa of the oropharynx, put on a glass slide, fixed in acetone and in the presence of destructive processes in the nuclei of epithelial cells, atypical mononuclear cells, high colonization activity conditionally pathogenic and pathogenic microorganisms and phagocytic activity in relation to them establish a mixed viral-bacterial infection, then the drug is degreased, discolor, put on vydeleny the e zone slides specific serum to antigens of HSV-1, 2, EBV, CMV, and upon detection of the drug in light microscopy specific brown-black staining of virus antigens in the cell structures diagnose mono - or mixed bacterial-herpes virus infection (EBV, CMV, HSV 1, 2,).



 

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FIELD: medicine.

SUBSTANCE: recipient of a cadaver kidney graft is examined to compare recipient's and donor's HLA alloepitopes. The alloepitopes found in the donor, but not found in the recipient are recovered. It is followed by comparing the recipient's HLA-DR antigens and the recovered alloepitopes, the derived combinations are compared with the following combinations: A9C-DR7; A10C- DR3; A10C-DR6; A28C-DR1; Bw4-DR6; B8C-DR3; B12C-DR2; B27C-DR5. Detecting said combinations enables stating an adequacy of the preventive application of monoclonal antibodies blocking an alpha-chain of an interleukin-2 receptor molecule. The preventive application of the monoclonal antibodies blocking an alpha-chain of the interleukin-2 receptor molecule shall be indicated to the patients with said combinations.

EFFECT: improved outcome of the cadaver kidney grafting that is combined with reducing the price of the method.

1 tbl, 2 ex

FIELD: medicine.

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1 ex

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SUBSTANCE: invention describes a method for prediction of reduced transport function of band-3 protein of red-cell membranes of newborns delivered by mothers suffered aggravated herpes viral infection in the third trimester of pregnancy wherein newborn's peripheral blood is examined for the TNFa content; band-3 protein is detected in the newborn's red-cell membranes by disk electrophoresis in polyacryl amide gel (10%) with sodium dodecyl sulphate; a discriminant function is calculated by formula Df=(5.167×band-3)+(2.196×TNFα), wherein Df is the discriminant function, band-3 is red-cell membrane stromal protein, TNFα is newborn's blood interleukine, and if Df is more than 218.67, reduced transport anion function by protein band-3 through newborn's red-cell membranes is predicted.

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1 ex

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SUBSTANCE: peripheral venous blood of the patients with autoimmune thyroiditis is examined for the absolute transferring receptor (CD71+) T-lymphocyte count and the interleukin-2 (IL-2) concentration. If the CD71+ T-lymphocyte count exceeds 0.3×109 cell/l, and the IL-2 concentration is less than 25 pg/ml, a favourable clinical course of the disease is predicted, while the CD71+ T-lymphocyte count less than 0.3×109 cell/l and the IL-2 concentration exceeding 25 pg/ml shows an unfavourable clinical course of autoimmune thyroiditis.

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3 ex

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2 ex

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1 tbl, 2 ex

FIELD: medicine.

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1 tbl, 1 ex

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1 ex

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1 ex

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1 ex, 4 tbl

FIELD: medicine.

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1 tbl, 2 ex

FIELD: medicine.

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2 ex, 2 tbl

FIELD: chemistry.

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EFFECT: higher sensifility of the analysis.

2 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: histological sections of internal organs are examined under a microscope; the histological examination is preceded by preparing a solution of diazotised aniline colour for staining the histological sections to detect indole, phenol, skatole; the histological section is stained and microphotographed; digital microphotos of the histological sections are processed on a computer. An average degree of staining with the diazotised aniline colour is estimated by the RGB colour grade system, and if observing the degree of staining within 250 to 100 RGB colour grade units, a mild severity of endogenous intoxication is diagnosed; the degree of staining varying from 100 to 50 RGB colour grade units shows a moderate severity of endogenous intoxication, while severe endogenous intoxication is indicated by the degree of staining being 50 to 0 RGB colour grade units.

EFFECT: using the technique enables higher toxicity and specificity of post-mortem diagnosis of endogenous intoxication based on the immediate detection of toxic metabolites collected in the internal organs.

1 tbl

FIELD: medicine.

SUBSTANCE: method for keeping viable blood cells for purposes of chemoluminescence analysis involves blood sampling and preserving with the blood sampled in disposable plastic test tubes and Faglucide® used as a haemopreserving agent in the ratio 1:6; the prepared blood is kept at temperature 2 to 6°C.

EFFECT: prolonged period of keeping viable leukocytes for purposes of chemoluminescence analysis, maintained the morphofunctional integrity and the rheological blood properties in cattle for 168 hours and the pH value at a physiological level that enables blood transportation for diagnostic researches from distant regions to laboratories.

2 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely cardiology, and concerns prediction of the developing life-threatening ventricular arrhythmias in the patients with no structural cardiac changes. That is ensured by determining a blood serum interleukin 6 level by a quantitative method, and the value > 14.9 pg/ml enables predicting the developing life-threatening ventricular arrhythmias in the given category of patients.

EFFECT: method provides specifying the indications to well-timed antiarrhythmic therapy.

3 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to pediatrics, and can be used for selection of drug therapy of atherogenic dyslipidemia in children and teenagers with abdominal-visceral obesity. For this purpose index of body weight (IBW) is determined. By results of blood serum lipidogram calculation of atherogenity index (AI) is performed. If IBW is higher than 90-th percentile set for said age group, and AI level is from 2 to 4 including anthropometric parameter - sagittal diameter of abdomen (SDA) is additionally measured. If SDA index equals or is higher than 24.0 cm, state is estimated as requiring drug therapy with chophytol.

EFFECT: method ensures increase of drug therapy efficiency and reduction of risk of early development of atherosclerosis in said group of patients due to possibility of fast and objective determination of degree of expression of risk of early atherosclerosis development, individualisation of indications for drug therapy and its early administration.

1 tbl, 2 ex

FIELD: measurement equipment.

SUBSTANCE: invention refers to measurement equipment and can be used in medicine for various diagnostic purposes. System of biosensors determining the concentration of investigated substance as per output signal generated with oxidation-reduction reaction of the investigated substance is proposed. System of biosensors introduces the correction to relationship in order to determine concentrations of the investigated substance as per output signals at one and the same temperature for determination of concentrations of the investigated substance as per output signals at the other temperature. Relationship with correction to temperature between concentrations of the investigated substance and output signals at reference temperature can be used for determination of concentrations of the investigated substance as per output signals at the specimen temperature.

EFFECT: improving the accuracy of determination of the investigated substance concentration.

29 cl, 12 dwg

FIELD: medicine.

SUBSTANCE: invention relates to oncology, immunology, laboratory diagnostics and is intended for prediction of dissemination of tumour process in patients with stomach cancer. Absolute content of monocytes and concentration of tumour necrosis factor alpha (TNF-α) are determined in peripheral venous blood of examined people. If content of monocytes is lower than 0.35×109 cell/l and TNF-α is higher than 55 pg/ml, high probability of dissemination is predicted.

EFFECT: method makes it possible to reduce time and simplify methods of prediction, as it makes it possible to use materials, reagents and methods of analysis, widely used in standard clinical-laboratory practice.

3 ex

FIELD: medicine.

SUBSTANCE: endothelial dysfunction is diagnosed if observing the twofold increase of a rat's blood glucose level. The technique involves determining antioxidant system values, a concentration of total NO metabolites, eNOS activity, a micro- and macrohaemodynamic status, a MDA concentration and Na+ K+ ATP-ase activity in myocardial and hepatic cell membranes. The values variations enables stating the presence of endothelial dysfunction in alloxan diabetes. Such dysfunction is corrected by the subcutaneous introduction of the preparation Ubiquinone (Coenzyme Q10) 0.11 mcl/100 g of animal's weight once a day for 30 days.

EFFECT: more accurate and reliable diagnosis of the vascular disorders accompanying said type of diabetes, and effective correction of such disorders also provides reproducibility, ease, availability, safety and low cost of performing the experiment.

2 cl, 2 tbl, 6 dwg, 1 ex

FIELD: medicine, psychiatry.

SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.

EFFECT: more objective prediction of disease development.

3 ex

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