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The article presents data for creating technological line for the cultivation of modified lines of mammalian cells expressing recombinant proteins. This line is composed of three sections: for the creation of the modified lines of mammalian cells using modern molecular biological tools; for the implementation of scalable cultivation of genetically modified mammalian cells lines; for separation and purification of recombinant polypeptides using validated methods. Within the framework of the experimental study, the cells of СНО-S line, producing recombinant protein GP of the Ebola virus, have been grown up in the mixer reactor and in the wave-type fermenter with the use of suspension technology. It is established, that the wave-type fermenters have the advantage in the accumulation of cells and necessary protein. The recombinant polypeptide GP with purity exceeding 95 % has been isolated and purified from the cultural liquid. During the study of recombinant protein GP by ELISA and Western blot methods, it was detected that it's structure coincides with that of the native glycoprotein of the Ebola virus causative agent.

Ebola outbreak in eastern parts of the Democratic Republic of the Congo in 2018–2020 proved that the virus remains highly hazardous for humans, and the outbreak in West Africa in 2014–2016, which was the largest Ebola outbreak in history, showed that it could be imported to other continents, including Russia. In 1993 the Federal State Budgetary Institution “48th Central Scientific Research Institute” of the Russian Ministry of Defence developed a specific equine immunoglobulin for emergency prophylaxis of Ebola in risk groups. The evaluation and improvement of the product’s properties is an important area in the development of biological defence technologies.The aim of the study was to examine the properties of the equine anti-Ebola immunoglobulin which had been stored for a long time at 2–8 °C.Materials and methods: the authors studied batches of heterologous anti-Ebola immunoglobulin that had been stored for 17–22 years. The properties of the product were evaluated according to the requirements of the State Pharmacopoeia of the Russian Federation, 14th ed. (Ph. Rus. 14 ed.). The specific activity of the product was determined in a plaque reduction neutralisation test using Ebola virus and African green monkey kidney cells (GMK-AH-1(D)). Immunoglobulin molecular parameters were determined by size-exclusion high-performance liquid chromatography using the test methods described in the European Pharmacopoeia 9.6 and Ph. Rus. 14 ed.Results: the storage of anti-Ebola immunoglobulin for 17–22 years at 2–8 °C resulted in a four-fold reduction of the level of virus-neutralising antibodies against Ebola, decrease in the proportion of monomers from 98 to 74–90%, increase in the proportion of dimers and polymers, and formation of immunoglobulin molecules’ fragments. Signs of toxicity for mice were observed in one of the three product batches. Conclusions: the obtained results suggest the need to perform more studies to test the quality of antiEbola immunoglobulin batches that were stored for shorter periods of time in order to assess the stability of their initial characteristics.

Haemorrhagic fever caused by the Ebola virus is a highly hazardous infectious disease with a mortality rate of 50– 90 %. Heterologous immunoglobulins with a high virus-neutralizing titer are an important element of the WHO-endorsed set of measures for emergency prevention and treatment of the disease. Specific activity of these products is largely determined by their fractional composition, and, in particular, by molecular mass distribution (MMD). The size-exclusion-high-performance liquid chromatography (SEC-HPLC) has traditionally been used for determination of the MMD of the target protein in human immunoglobulin-based products. The use of this method for evaluation of molecular parameters of heterologous immunoglobulin requires confirmation of its specificity, accuracy and precision, and establishment of the chromatographic system suitability criteria in the context of a new test object. The aim of the study was to test the applicability of the SEC-HPLC method to the assessment of molecular parameters of anti-Ebola immunoglobulin derived from horse serum. Materials and methods: three batches of purified equine anti-Ebola immunoglobulin were used in the study. Normal equine and human immunoglobulins of the IgG isotype were used as reference standards. The HPLC test procedures described in the European Pharmacopoeia 9.6 and State Pharmacopoeia of the Russian Federation, 14th ed., were used for determination of monomers and other immunoglobulin fractions. An Agilent 1260 Infinity (Agilent, USA) HPLC system with a diode array detector and an Agilent Bio SEC-3 HPLC column were used for quality evaluation of the tested products. Results: the resolution factor between IgG monomer and dimer peaks (1.69 and 2.10), and the chromatographic column efficiency (>2000) make it possible to use the SEC-HPLC system for evaluation of molecular parameters of heterologous immunoglobulin. The study demonstrated reproducibility of the test procedure. Conclusions: the study confirmed the applicability of the SEC-HPLC procedure for evaluation of molecular parameters of anti-Ebola immunoglobulin derived from horse serum. It demonstrated the compliance of the purified immunoglobulin to the national and international quality requirements in terms of «Molecular parameters».

Analysis of methodological approaches to preclinical evaluation of sensitizing activity of horse serum products shows that the anaphylactogenicity scale recommended in national regulatory documents is imperfect as it is not capable of fully predicting the safety of the clinical use of products. The article demonstrates feasibility of another methodological approach to preclinical evaluation of sensitizing activity of heterologous serum products which is based on calculation of a sensitizing dose inducing anaphylactic reactions in 50 % of guinea pigs. This approach will make it possible to more effectively predict side effects of already authorized heterologous immunoglobulins and drugs currently under development, and to select a clinical use strategy on a case-by-case basis.

The aim of the study: to examine the indicators of relapse-free survival and evaluate prognostic factors that had the greatest impact on relapse- free survival in patients with rectal cancer complicated by obstructive obstruction and undergoing emergency surgery.Materials and methods. The analysis of the immediate and long-term results of treatment – relapse-free survival of patients undergoing emergency surgery for rectal cancer complicated by obstructive obstruction in general surgical and specialized hospitals in Smolensk from 2001 to 2017 is presented. Relapse-free survival was analyzed by the Kaplan–Meier method. To determine the influence of potential risk factors on the rate of occurrence of the studied event, Cox regression was used.Results. Postoperative complications of the III–IV degree according to Clavien–Dindo were more common in patients undergoing emergency surgery for complicated rectal cancer in general surgical hospitals (p = 0.0056). In specialized hospitals, anastomosis leakage in 5.3 % (1 of 19 cases), in general surgical hospitals – 16.3 % (9 of 55 cases). Five-year relapse-free survival in general surgical hospitals at the IIB stage was 18.3 %, median survival – 32 months; in specialized hospitals at stage IIB 5‑year relapse-free was 45.8 %, median survival – 52 months (p = 0.028 and 0.011, respectively). A multivariate analysis confirmed the influence of the following factors on the performance of a 5‑year relapse-free survival: hospital specialization (risk ratio (RR) 1.35; 95 % confidence interval (CI) 1.18–1.55; p

The article presents the 2-year results of prospective study, which included 67 heart transplantations for end-stage heart failure, performed in Krasnodar, in Clinical regional hospital No 1. Survival rate was 89,1% (59 patients). Causes of death were septic complications, thromboembolia of the pulmonary artery, severe pancreatonecrosis, and transplant rejection due to non-compliance with immunosuppression therapy, heart rhythm disturbances. The most common complications were pneumonia (19%), heart graft rejection (10%), steroid-induced diabe- tes (10%). In a year after heart transplantation chronic heart failure, II stage, was diagnosed in 29 patients (43%), I stage – in 30 (44%) patients. Monitoring and therapy are carried out by cardiologists domiciliary and by spe- cialists of polyclinic of Krasnodar clinical hospital. Orthotopic heart transplantation is an effective treatment for end – stage heart failure. It is possible to provide surgical treatment and further management of patients in multisectoral regional medical hospital.