[Creation of registers as one of the mechanisms for improving medical care for patients with chronic heart failure. Problem state].

The annual mortality of patients with clinically pronounced symptoms of chronic heart failure in the Russian Federation reaches 26-29%, i.e., from 880 to 986 thousand patients with heart failure die in the country in one year, which is comparable to the population of a large city. Providing care for patients with heart failure places a heavy burden on the country's health care system, making a significant contribution to mortality rates, hospitalization rates, including readmissions, which in turn requires considerable costs. The article presents an overview of registry studies that are devoted to assessing the effectiveness of diagnostics, the completeness of examinations, as well as the adequacy of ongoing drug treatment.

[Monoclonal antibodies to hemagglutinin of influenza A/H7N3 virus (Orthomyxoviridae: Alphainfluenzavirus: Influenza A virus)].

INTRODUCTION: Variants of influenza virus A/H7 have the same high pandemic potential as A/H5. However, the information about the antigenic structure of H7 hemagglutinin (НА) is considerably inferior in quantitative terms to similar data for H5 НА.The aims of the study were development and characterization of the monoclonal antibodies (MAbs) panel for HA subtype H7 of the influenza A virus. MATERIAL AND METHODS: Viruses were accumulated in 10-day-old chicken embryos. Purification and concentration of the virus, determination of protein concentration, preparation of MAbs and ascitic fluids, hemagglutination and hemagglutination inhibition (HI) tests, assessment of antibodies' activity in indirect enzyme-linked immunosorbent assay (ELISA), as well as determination of MAbs isotypes and neutralization reaction (NR) were carried out by standard methods. RESULTS: The obtained MAbs to А/mallard/Netherlands/12/2000 (H7N3) strain were studied in HI test with a set of strains of different years of isolation belonging to different evolutionary groups. MAbs had a reduced reactivity compared to the immunogen-virus for all the studied strains. Cross-interaction of MAbs 9E11 and 9G12 in HI test with influenza A/H15 virus has been observed. DISCUSSION: Influenza A agent with H7 HA variant could serve as a potential cause of a future pandemic. Development of the MAbs panel for subtype H7 HA is an urgent task for both veterinary medicine and public health. CONCLUSION: The obtained MAbs can be used not only for epitope mapping of the H7 HA molecule (currently insufficiently studied) and as reagents for diagnostic assays, but also for determining common («universal¼) epitopes in HA of different strains of this subtype.

[Modeling influenza virus infection in mature Wistar rats].

It has now been established that blood vessels are target for influenza, but the mechanism by which the influenza virus affects the cardiovascular system is unknown. The aim - adaptation of influenza virus A/St. Petersburg/48/16 H1N1(pdm09) to mature Wistar rats, as these animals are the main experimental model for studying the pathology of the cardiovascular system. MATERIAL AND METHODS: Passage of influenza A virus (IAV) in embryonated chicken eggs, intranasal inoculation of rats with virus-containing material s, production of pulmonary homogenate, determination of IAV titer in embryonated chicken eggs, detection of histological changes in lung and pulmonary vessels. RESULTS: The article presents the results of the adaptation of influenza virus A/St. Petersburg/48/16 H1N1(pdm09) to mature Wistar rats. The infectious titer of the virus in the homogenates of infected rats lungs at the last stage of adaptation was 7.0 lg EID50/ml. Histological studies revealed pronounced changes in the respiratory tract (spasm of bronchioles, submucosal edema, desquamation of ciliated epithelium of bronchioles) and pulmonary vessels (spasm, desquamation and swelling of endotheliocytes, dissociation and swelling of the elastic membrane and media). In order to identify IAV in blood vessels and lung tissues, an immunohistochemical study was performed using monoclonal antibodies to NP antigen of IAV. CONCLUSION: The data obtained allow us to conclude that the strain of influenza virus A/St. Petersburg/48/16 H1N1(pdm09) was adapted to mature Wistar rats maintaining virulent properties. The infectious titer of the virus at the last stage of adaptation was 7.0 lg EID50/ml. IAV identification is confirmed by immunohistochemical examination.

[Monoclonal antibodies to hemagglutinin of influenza b viruses victoria evolutionary lineage.]

Co-circulation of two evolutionary distinct lineages of influenza virus in one epidemic season has led to development specifc reagents for rapid identifcation and typing of new isolates. Panel of MAbs to hemagglutinin of influenza virus B/Brisbane/46/15 belonging to Victoria evolutionary lineage was developed. All MAbs reacted in ELISA with B/Victoria-like strains only. There were no interactions with heterologous influenza viruses of B/Yamagata lineage, seasonal and potentially pandemic influenza A viruses. All MAbs reacted in hemagglutination inhibition and virus neutralization. MAbs interacted in hemagglutination inhibition only with B/Victoria-like viruses, but did not interacted B/Yamagata-like strains. Neutralization and hemagglutination inhibition studies of viruses isolated before 1983 with MAbs revealed that MAbs 6E11, 9G5, 9B5 and 6A4 had the ability to interact with the virus B/ Russia/69 which may evidence that B strains of early isolation period (before lineage separation) have common epitope with recent Victoria lineage viruses. MAbs 7C8, 7G9, 7H8 and 8D11 were directed to a conserved epitope (or epitopes) specifc for influenza hemagglutinin viruses of B/Victoria group. The presence of differences in the effectiveness of the interaction of MAbs 6A9, 7G9 and 8A8 in hemagglutination inhibition test allows the identifcation and differentiation of strains isolated in chicken embryos and MDCK cell culture. Thus, the developed MAbs can be successfully used for identifcation and antigenic analysis of B/Victoria-like strains.

GENETIC AND ANTIGENIC CHARACTERISTICS OF RESPIRATORY SYNCYTIAL VIRUS STRAINS ISOLATED IN ST. PETERSBURG IN 2013-2016.

Antigenic and genetic characteristics of Russian RSV isolates are presented for the first time. Of the 69 strains isolated in St. Petersburg, 93% belonged to the RSV-A antigenic group. The antigenic variations in the F-protein RSV were analyzed using a panel from 6 monoclonal antibodies by the method of micro-cultural ELISA. Depending on the decrease in the effectiveness of interaction with monoclonal antibodies (relative to the reference strain Long), RSV-A isolates were divided into 4 antigenic subgroups. The results of 24 isolates sequencing showed that more than 60% of them had substitutions in significant F-protein sites compared to the ON67-1210A reference strain of the current RSV genotype ON1/GA2. The most variable were the signal peptide and antigenic site II. When comparing the results of ELISA and sequencing, it was not possible to identify any specific key substitutions in the amino acid sequence of the F-protein that affect the interaction of the virus with antibodies. The nucleotide sequence of the F-gene from 19 of the 24 characterized isolates was close to that of ON67-1210A reference virus and was significantly different from RSV-A Long and A2 viruses. A separate group consisted of 5 strains, in which the F-protein structure was approximated to RSV Long.

[THE APPLICATION OF DOT-TECHNIQUE FOR DETECTING ANTIGENS OF ADENOVIRUS IN CLINICAL SAMPLES].

The article substantiates possibility of application of point enzyme-linked immunosorbent assay (dot-technique) for detecting viral antigens in samples from patients. To diagnose adenovirus infection conjugate of virus-specific monoclonal antibodies and peroxidase of horse-radish were used The chromatographic rectification of conjugate from free peroxidase permits diminishing background coloring of nitrocellulose membrane and therefore to increase sensitivity. The application of direct conjugates on the basis of virus-specific monoclonal antibodies increases specifcity of dot-technique and significantly shortens time period of analysis. As in case of application of direct conjugates on the basis of polyclonal serum, samples from patients require preliminary processing with detergent for preventing non-specific reactions. The dot-technique demonstrates good coincidence with data of polymerase chain reaction and after clinical trials it can be used in diagnostic of human viral infections.

[The development of immune enzyme test-systems for detecting potentially pandemic influenza viruses subtype A(H2), A(H5), A(H7) and A(H9).]

PURPOSE OF STUDY: To develop diagnostic test-systems constructed according principle of "sandwich" enzyme-linked immunosorbent assay to detect antigens of influenza viruses with pandemic potential: A(H2), A(H5), A(H7), A(H9).The panels of subtype-specific monoclonal antibodies interacting with molecule of hemagglutinin of potentially pandemic viruses of influenza are obtained and characterized. The viruses of every sub-type were supplied with selected pairs monoclonal antibodies/monoclonal antibodies' conjugate with peroxidase of horse-radish, interacting most effectively with virus-immunogen and lacking non-specific activity related to influenza viruses of geterologic types. Considering established threshold values, the sensitivity of test-systems varied depending on sub-type and strain of viruses within limits of 4 - 30 ng/ml at evaluation of viral purified concentrates and 0.3 - 8 hemagglutinin units at analysis of virus-containing allantoic fluid. The specificity of immune-enzyme test-system manifested in conditions of absence of non-specific interactions with both seasonal and potentially pandemic geterologic viruses. The technique can be applied for fast identification of viruses of influenza non-typeable in conditions of routine functioning of practical laboratories.

[Epitope analysis of the hemagglutinin molecule of the Victoria lineage influenza B viruses].

A panel of five monoclonal antibodies (MAbs) to the HA1 molecule of the influenza B virus of the Victorian lineage with high virus-neutralizing activity was developed. For identification of the virus neutralizing epitopes in HA1 escape mutants (EM) of the influenza BIShandong/07/97 and B/Malaysia/2506/04 virus were selected using virus- neutralizing antibodies (MAbs). Three EMs had single, two--double and one--triple amino acid substitutions (AAS) in HA1 (H122N, A202E, K203T, K2031, K203N or A317V). In addition, AAS N197S was detected in three EMs. A correlation of AAS identified with peculiarities of interaction of EMs with Mabs was discussed.

[Influenza virus reproduction in the endothelial cells of human blood vessels].

The current epidemic strains of the influenza subtypes Y5N1, H3N2, and H1N1 can be reproduced in the cultured human endothelial EAhy926 cells with an infective activity of 3.0-4.5 Ig TCD50. These findings were confirmed by the analysis of the autopsy material from patients who had died from influenza during the 2009-2010 epidemic, which showed influenza virus HA and NP proteins in the pulmonary blood vascular endothelium. The results obtained reflect a new aspect of the pathogenesis of influenza, which is important for the design of antiviral agents and for the development of combination therapy.

[Development of competitive enzyme immunoassay for the detection of influenza A(H5) virus antibodies].

An indirect enzyme immunoassay (EIA) using the purified fraction of surface viral glycoproteins (GP) as an antigen for solid phase sensitization was not shown to be a specific method for the differential detection of influenza A(HS) (HS-Ab) virus antibodies (Abs) due to total conservative epitopes in the structure of GPs of influenza A(H5) and A(H1NI) viruses. The cross activity of some monoclonal Abs (MAbs) to influenza A(H5) and A(HIN1) viruses, which had been obtained at the Research Institute of Influenza, was proof of the presence of total immunodominant determinants in the structure of influenza H1 and H5 virus hemagglutinin (HA). In this connection, an EIA, which was based on the competition of influenza A(H5) H5-Ab virus HA-specific MAbs in the test sera for an association with influenza A(H5) virus, was proposed for the subtype-specific detection of H5 Ab. Comparison of the results of competitive EIA (cEIA), microneutralization (MN) test and HA inhibition test (HAIT) (using equine red blood cells) in the examination of sera obtained from 44 volunteers immunized with inactivated vaccine containing influenza A/Indonesia/5/2005 (H5N1) virus showed the high sensitivity and specificity of cEIA in detecting H5-specific Abs. The effectiveness of cEIA for the sera strictly positive for the content of H5 Abs was close to that of MN test and was 9-34% higher than HAIT (depending on those used in the analysis of H5 virus antigens). cEIA may be proposed to assess new influenza vaccines as an additional laboratory test. Since the infectious virus is not used during cEIA, it may be recommended for the serodiagnosis of influenza A(H5) at practical virological laboratories.

Gasless transanal endoscopic surgery for rectal adenomas and early carcinomas.

Transansal endoscopic microsurgery (TEM) allows the excision of rectal tumors but is difficult and complex. A new TEM modification simplifying the procedure and reducing its cost is required. Between 1999 and 2004, 128 patients (78 women and 50 men) underwent gasless transanal endoscopic surgery (GTES) without use of a completely closed system by means of laparoscopic equipment. Histological examination revealed 112 adenomas, including 15 with neoplastic changes (6 cases of Tis and 9 cases of T1G1G2), 12 adenocarcinomas (1 case of Tis, 8 cases of T1G1G2, and 1 case of T2G1G2) and 3 carcinoid tumors. There was no operative mortality. One patient had postoperative bleeding. Adenomas recurred in 9 patients (8.3%). The adenocarcinomas demonstrated neither local recurrences nor distal spread. GTES may be considered a safe and effective minimally invasive treatment for patients with large adenomas and early carcinomas of the rectum.

[Contemporary approaches to the treatment of arterial hypertension and ischemic heart disease].

Prognostic advantage of a combination of calcium antagonist and angiotensin converting inhibitor (ACEI) over combination of beta-adrenoblocker with thiazide diuretic was proved in a large randomized study of long term treatment of arterial hypertension. In another randomized study administration of ACEI for 12 months caused substantial retardation of cardiac remodeling in elderly patients with normal left ventricular function in postinfarction period. These results are important for a practical physicians as they concern treatment of wide circle of in- out-patients.

[Transanal endoscopic surgical method of removing rectal tumors]. / Transanal'nyi éndokhirurgicheskii metod udaleniia opukholei priamoi kishki.

Experience in endosurgical removal of rectal tumors in 96 patients is presented. Performed from 1999, open non-gas method and closed method of transanal endomicrosurgery (TEM) have reduced rate of adenoma recurrences to 8.8%. Development of the open method increased it accessibility for hospitals using endoscopic techniques.

[Development of immunoenzyme assay for subtype-specific detection of antibodies to influenza viruses A (H1N1) and A (H3N2)]. / Razrabotka immunofermentnykh test-sistem dlia podtipospetsificheskoi detektsii antitel k virusam grippa A (H1N1) i A (H3N2).

Conditions were developed for obtaining surface viral glycoprotein (GP) fraction intended for solid phase sensitization with the aim of constructing enzyme immunoassay test systems (EIATS) for detection of subtypical IgG and IgG to influenza A (H1N1) and A (H3N2) viruses. New variants of test systems were compared with the traditional methods for serological diagnosis of influenza. GP-based EIATS more often diagnosed influenza than EIATS based on purified whole-virion (WV) suspensions, hemagglutination inhibition and neutralization tests. Evaluation of conversions of subtypical IgG showed that the results coincided with the findings of neutralization and hemagglutination inhibition tests in 83-90% (EIATS-GP) and 50% (EIATS-WV). Cross-detection of antibodies to both virus subtypes in EIATS-GP and EIATS-WV was observed in 4 and 31% cases, respectively.

[The indices of the fibrinolysis, blood coagulation and lipid transport systems in hypertension patients]. / Nekotorye pokazateli sistem fibrinoliza, svertyvaniia krovi i lipidnogo transporta u bol'nykh gipertonicheskoi bolezn'iu.

Lipid spectrum, basal insulin concentration, the activity of coagulation factor VII and inhibitor of plasminogen tissue activator, content of plasminogen tissue activator antigen were assessed in 54 hypertensive patients with left ventricular hypertrophy (mean age 48.9 +/- 0.9 years, mean duration of the disease 15.7 +/- 1.1 years) after discontinuation of hypotensive therapy. A correlation was found between a rise in thrombogenic, atherogenic and a fall in fibrinolytic potential of blood. These findings suggest an essential role of insulin in regulation of thrombogenesis and fibrinolysis in hypertensive patients.

[Do glycoproteins IIb/IIIa participate in activation of human platelet s by low density lipoproteins?]. / Uchastvuiut li glikoproteidy IIb/IIIa v aktivatsii trombotsitov cheloveka lipoproteidami nizkoi plotnosti?

Low density lipoproteins (LDL) enhance the entry of Ca2+ and other bivalent cations via receptor-operated channels. The use of a synthetic peptide and antibodies blocking the glycoproteins IIb and IIIa interaction with ligands revealed that fibrinogen receptor antagonists prevent platelet aggregation without any effect on the ability of LDL to increase the cytoplasmic Ca2+ level. These data indicate that glycoproteins IIb-IIIa are not involved in the activation of the second messenger system by LDL but play a role in the proaggregant effect of LDL by providing cell-to-cell interactions.

Efficacy and tolerability of cilazapril in patients with essential hypertension.

This study investigated the effect of the angiotensin-converting enzyme (ACE) inhibitor cilazapril on blood pressure in patients with essential hypertension [defined as a sitting diastolic blood pressure (DBP) between 95-120 mm Hg]. The study group comprised 43 patients (24 men and 19 women; mean age, 51 +/- 10 years) with uncomplicated essential hypertension. In an open single-center design, all subjects were treated with a once-daily dose of cilazapril for 6 weeks, following a 1-week washout period. The initial dose was 2.5 mg/day. In nonresponders, this dose was doubled after 3 weeks of treatment. Thirty-eight patients completed the study. One was withdrawn due to symptomatic hypotension, and four others due to noncompliance. Treatment produced a useful and significant reduction in blood pressure (systolic from 155 +/- 3 to 138 +/- 3 mm Hg, p < 0.01; diastolic from 103 +/- 1 to 90 +/- 2 mm Hg, p < 0.001). No significant variations in mean heart rate were recorded. Normalization of DBP, or its reduction by > 10 mm Hg, was found in 28 patients (efficacy in 74% of cases). Cough was reported by one patient. There was no significant variation in laboratory profiles during the trial. Cilazapril proved to be a well-tolerated antihypertensive drug effective on a once-daily regimen in a high percentage of patients with essential hypertension.

[Evaluation of the efficacy and tolerance of the new delayed-action angiotensin-converting enzyme inhibitor cilazapril in patients with hypertension (preliminary report)]. / Otsenka éffektivnosti i perenosimosti novogo ingibitora angiotenzinprevrashchaiushchego fermenta prolongirovannogo deistviia silazaprila u bol'nykh gipertonicheskoi bolezn'iu (predvaritel'noe soobshchenie).

Cilazapril [correction of Silazapril], a new inhibitor of long-acting angiotensin-converting enzyme, was given to 39 hypertensive subjects aged 24-68 with initial diastolic pressure 95-120 mm Hg. Of them 16 were females and 23 were males. The course of the treatment lasted 8 weeks, the drug being taken daily in a single dose 2.5 mg in the morning. Checkups, measurements of arterial pressure (AP), blood counts, urinalyses were made before the treatment, on the treatment week 4 and 8. In one patient the drug was discontinued 4 weeks after the treatment start because the AP fall to 110/63, in 23 patients the dose was raised to 5 mg, the rest continued on 2.5 mg/day. The treatment ended in the systolic pressure decrease by 14.9% (from 161.0 +/- 2.9 to 137.8 +/- 2.8 mm Hg, p < 0.0001) and diastolic pressure fall by 13.5% (from 104.5 +/- 1.4 to 90.1 +/- 1.7 mm Hg, p < 0.005). No side effects causing the drug withdrawal were registered. It is concluded that the drug is effective in essential hypertension stage IIA-IIB as monotherapy.