Aim. To evaluate dynamic changes in the lungs, hemostasis system, immune system in different terms after coronavirus pneumonia. Materials and methods. Ventilation-perfusion single-photon emission computed tomography/computed tomography (CT), functional methods of lung investigation, evaluation of hemostasis system, immune status and specific humoral immune response were performed and evaluated in different terms after coronavirus pneumonia. A total of 71 patients were examined according to this protocol. We examined patients with the lesion volume not less than 50% according to chest CT. All patients were divided into 2 groups depending on the distance from the acute stage of coronavirus pneumonia. Group 1 included patients who were examined early (3060 days after hospital discharge), group 2 included patients who were examined later (61180 days after hospital discharge). Results. We obtained gradual regression of pathologically-modified tissue from 67.3% during the inpatient phase to 30.9% during the early period and to 19.7% during the late period of examination, according to CT scan of the chest organs. The same tendency was demonstrated by diffusion capacity of the lungs. Perfusion scintigraphy data showed a decrease in perfusion deficit from 26.012.8% during the early period of examination to 19.46.2% during the late period of examination. On the contrary, ventilatory scintigraphy demonstrates the increase of isotope passage time through the alveolar-capillary membrane over time (from 48.231.3 minutes in the early period to 83.637.2 minutes in the late period). An increase in D-dimer was detected in 24% of patients in the early group. The levels of inflammatory markers, indices of immune status, and specific humoral immune response did not differ in the two described groups. Conclusion. The results demonstrate gradual regression of pathological changes caused by coronavirus infection.

Using an electrochemical method based on the use of an optically transparent working electrode, it was shown that, under conditions of hypothermic (4°C) storage of erythrocytes, the degree of reversibility of changes in their shape decreases when exposed to direct electric current. With cathodic polarization of an optically transparent working electrode (–0.5 V), the proportion of erythrocytes converted to spheroechinocytes decreased from 99.6 ± 0.4 (day 1) to 42.8 ± 14.5% (day 42) (p < 0.05), and with anodic polarization (1.2 V), the proportion of erythrocytes converted to stomatocytes decreased from 80.6 ± 6.7 (day 1) to 35.4 ± 5.6% (day 42) (p < 0.05). At a later storage period, an increase in the number of cells that underwent destruction under the influence of direct electric current was observed in the samples, the presence of which in the transfused erythrocyte suspension can lead to an increase in free hemoglobin in the recipient’s blood after transfusion. The results obtained can form the basis for the development of new methods for assessing the quality of blood components.

АКТУАЛЬНОСТЬ. Темпы развития пандемии новой коронавирусной инфекции COronaVIrus Disease 2019 (COVID-19) и высокая летальность обусловили необходимость поиска всех возможных методов лечения. Реконвалесцентная плазма (РП) стала ценным эмпирическим ресурсом для поддержки здравоохранения, особенно на начальном этапе пандемии COVID-19. Она по-прежнему остается объектом внимания многих ученых, в то время как данные относительно эффективности РП являются достаточно противоречивыми. ЦЕЛЬ ИССЛЕДОВАНИЯ. Целью настоящего обзора является анализ результатов современных исследований по эффективности и безопасности клинического применения РП. МАТЕРИАЛЫ И МЕТОДЫ. Обзор литературы был проведен в электронных базах данных PubMed, MedRxiv, Cochrane Library, Cochrane COVID-19 study registry с 7 июня 2021 г. по 20 декабря 2021 г. РЕЗУЛЬТАТЫ. В данной работе выполнен обзор основных сравнительных ретроспективных или проспективных исследований по применению РП. Применение РП известно с 1880-х гг. при лечении дифтерии, испанского гриппа, кори, полиомиелита, а в XXI в. РП использовали при эпидемии лихорадки Эбола, вируса гриппа H1N1, других коронавирусах. Терапия РП основана на концепции пассивной иммунизации и включает в себя введение антител от доноров-реконвалесцентов, обладающих вирус-нейтрализующей активностью (ВНА) к данному вирусу. В целях минимизации рисков гемотрансмиссивных инфекций используются технологии редукции патогенов. Исторические и текущие данные об использовании РП подтверждают безопасность ее использования. Были рассмотрены критерии эффективности применения и сроки назначения РП. Представлены клинические данные, подтверждающие эффективность РП у определенных групп пациентов. ВЫВОДЫ. Применение РП является безопасным и целесообразно у серонегативных больных с COVID-19 на ранних сроках заболевания или при наличии иммунодефицитного состояния. Наибольшей эффективностью обладает РП с высокими титрами ВНА. У тяжелых больных на поздних сроках болезни терапия РП не приводит к регрессии заболевания и увеличению выживаемости.

INTRODUCTION. High mortality during the pandemic of COVID-19 conducted the research for all available treatments. Convalescent plasma (CP) has become valuable empirical resource for health support, especially during the initial phase of COVID-19 pandemic. It still draws attention of many scientists, while data on the effectiveness of CP is rather controversial. OBJECTIVES. The purpose of this review is to analyze the results of modern studies on the efficacy and safety of the clinical use of CP. MATERIALS AND METHODS. The search for publications was carried out in electronic databases PubMed, MedRxiv, Cochrane Library, Cochrane COVID-19 study registry from June 7, 2021 to December 20, 2021. RESULTS. The article analyzes data from recent retrospective and prospective studies related to CP therapy. The use of CP is known since the 1880s in the treatment of diphtheria, Spanish flu, measles, polio. In the 21st century CP has been used in the epidemic of Ebola, H1N1pdm09, other SARS1 and MERS coronaviruses. CP therapy is based on the concept of passive immunization and includes the infusion of antibodies from convalescent donors with virus-neutralizing activity (VNA). Pathogen reduction technologies are used to minimize the risks of transfusion-transmitted infections. Historical and current data confirm the safety of CP use. The criteria for effectiveness and timing for transfusion of the CP were considered. Clinical data is presented confirming the effectiveness of CP in certain groups of patients. CONCLUSIONS. The use of CP is safe and reasonable in seronegative patients with COVID-19 on the early stages of the disease or in the presence of an immunodeficiency. CP with high VNA titers has the highest efficiency. CP therapy in severe patients on the late stages of the disease does not provide disease regression and increased survival.

Introduction. Among the most common congenital coagulopathies are haemophilia and Von Willebrand disease. These illnesses are often mimicked by orphan hereditary coagulopathies, including combined coagulation factor V and VIII deficiency.Aim — description of a clinical presentation, hampered diagnosis and choice of haemostatic therapy in a surgical patient with combined blood coagulation factor V and VIII deficiency.Main findings. We describe a clinical case of congenital combined factor V and VIII deficiency and detail the aetiology, frequency, localisation and intensity of haemorrhages. Comorbidity and surgical indications are demonstrated to require an inter-specialty medical involvement.

Profound immunological dysfunction is the key factor determining the development of infectious complications in chronic lymphocytic leukemia (CLL). The aim of this work is to assess the features of the subpopulation composition of T-lymphocytes (T-helpers (Th), cytotoxic T-lymphocytes (Tcyt), T regulatory cells (Treg), T-NK cells, naive Th, Th-memory, activated T-lymphocytes, TCRγδ cells) and NK cells in peripheral blood of patients with newly diagnosed chronic lymphocytic leukemia (CLL) and receiving ibrutinib therapy. Hematological and immunophenotypic studies have been performed in 30 patients with previously untreated CLL, 122 patients on ibrutinib therapy and 20 healthy donors. The subpopulation composition of T-lymphocytes (Th, Tcyt, Treg, T-NK, naive T-helpers, memory T-helpers, TCRγδ cells, activated T-lymphocytes) and NK cells has been assessed on flow cytometer (FACSCanto II (BD)) using the following panel of monoclonal antibodies: CD45, CD19, CD3, CD4, CD5, CD8, TCRγδ, CD127, CD16, CD56, CD57 CD45RA, CD45R0, HLA-DR, CD25. Compared to controls all CLL samples were found to have higher the absolute number of T-lymphocytes, NK cells and their subpopulations, T-helpers (especially of memory T-cells), cytotoxic T-cells, regulatory T-cells, TCRγδ T-cells, activated T-lymphocytes, increased cytotoxic potential of NK cells in previously untreated CLL patients. Patients who received ibrutinib therapy have registered a positive trend towards recovery of the subpopulation composition of T-lymphocytes and NK-cells. CLL patients have been found to have quantitative and functional changes in the subpopulations of T-lymphocytes and NK cells, indicating dysregulation of the immune response, and a high risk of developing infections. Monitoring of immunological parameters for ibrutinib therapy make possible to estimate impact of ibrutinib on the adaptive anti-CLL immune response.

BACKGROUND AND OBJECTIVES:COVID-19 convalescent plasma is an experimental treatment against SARS-CoV-2. The aim of this study is to assess the impact of different pathogen reduction methods on the levels and virus neutralizing activity of the specific antibodies against SARS-CoV2 in convalescent plasma. MATERIALS AND METHODS:A total of 140 plasma doses collected by plasmapheresis from COVID-19 convalescent donors were subjected to pathogen reduction by three methods: methylene blue (M)/visible light, riboflavin (R)/UVB and amotosalen (A)/UVA. To conduct a paired comparison, individual plasma doses were divided into 2 samples that were subjected to one of these methods. The titres of SARS-CoV2 neutralizing antibodies (NtAbs) and levels of specific immunoglobulins to RBD, S- and N-proteins of SARS-CoV-2 were measured before and after pathogen reduction. RESULTS:The methods reduced NtAbs titres differently: among units with the initial titre 80 or above, 81% of units remained unchanged and 19% decreased by one step after methylene blue; 60% were unchanged and 40% decreased by one step after amotosalen; after riboflavin 43% were unchanged and 50% (7%, respectively) had a one-step (two-step, respectively) decrease. Paired two-sample comparisons (M vs. A, M vs. R and A vs. R) revealed that the largest statistically significant decrease in quantity and activity of the specific antibodies resulted from the riboflavin treatment. CONCLUSION:Pathogen reduction with methylene blue or with amotosalen provides the greater likelihood of preserving the immunological properties of the COVID-19 convalescent plasma compared to riboflavin.

Актуальная тема сегодняшнего дня — терапия новой коронавирусной инфекции. Важной особенностью пандемии COVID-19 является отсутствие стандартной терапии, применение всех имеющихся средств носит экспериментальный характер. Опыт предшествующих эпидемий вирусных заболеваний дает основания предположить, что использование плазмы выздоровевших будет эффективным при COVID-19. В данной работе представлены результаты анализа первого опыты клинического применения плазмы выздоровевших для лечения COVID-19 в стационарах Департамента здравоохранения г. Москвы (ДЗМ). ЦЕЛЬ ИССЛЕДОВАНИЯ Провести анализ первого опыта клинического применения плазмы реконвалесцентов при COVID-19 в стационарах ДЗМ. МАТЕРИАЛ И МЕТОДЫ Свежезамороженную плазму реконвалесцентов, полученную методом афереза и подвергнутую патогенредукции, вводили 226 пациентам со среднетяжелым и тяжелым течением COVID-19. Для сравнения ретроспективно сформирована контрольная группа — 226 пациентов со сходными характеристиками. РЕЗУЛЬТАТЫ Выявлено статистически значимое снижение смертности на 10-е (5,3 по сравнению с 17,2%) и 28-е (14,2 по сравнению с 22%) сутки в основной группе, более выраженное у тяжелых пациентов, не достигших по степени дыхательной недостаточности перевода на искусственную вентиляцию легких (ИВЛ) — относительный риск смерти 0,369 (95% ДИ 0,159—0,859). ВЫВОДЫ Подтверждена эффективность применения плазмы реконвалесцентов в комплексе терапии COVID-19 в целом, а также показана оптимальность ее использования у пациентов с клинически значимой дыхательной недостаточностью, но не достигших перевода на ИВЛ.

Background. The pandemic of the new coronavirus infection has challenged the medical community for quickly finding and implementing effective methods of treatment. In the absence of a vaccine or specific therapy with proven effectiveness, the usage of convalescent plasma can be the one of perspective methods. An important aspect of this technology is the efficient and safe preparation of convalescent plasma. To date, in the world literature there are practically no publications about donor recruitment and the specifics of the preparation of convalescent plasma. Purpose of the research. Presentation of the experience of organizing a workflow for recruiting donors and stockpiling of convalescent plasma with a high titer of virus-neutralizing antibodies to SARS-CoV-2. Methods. The analysis of the work of the Blood Service of the Moscow Department of Health for stockpiling of COVID-19 convalescent plasma has been executed. In total it has been stockpiled 1240 doses. The normative documentation has been developed by a working group on the basis of the current federal legislation of Russian federation and been approved by the Moscow Department of Health. The titer of neutralizing antibodies (VNA) has been determined as the basic method for assessing the immunological viability of convalescent plasma. The main characteristics of donors, the characteristics of the disease course, the results of preliminary testing for the presence of specific antibodies by ELISA and CLIA methods has been compared with VNA titers in the stockpiled convalescent plasma. Results. Due to a Moscow Health Departments order No. 325 dated 01.04.2020 (a basic local regulatory document) it has been developed a regulation for the stockpiling, examination, storage, safety and transfering of fresh frozen pathogen-reduced plasma of COVID-19 convalescent donors to medical organizations of the Moscow Health Department. For arranging an interaction with donors it has been created a call-center. For effective preliminary selection, it has been formed a donor characteristics list, which has been combined with screening of specific antibodies by ELISA and CLIA methods. Conclusions. Developed a system of recruiting donors and procurement process of convalescent plasma for treatment СOVID-19, which includes the necessary regulations, algorithms for the selection and recruitment of donors, the registry of donors and recipients, algorithms, efficiency and safety of convalescent plasma.

Background. The application of convalescent plasma (CP) is currently seen as a feasible therapeutic approach in the treatment of COVID-19.Aim. To analyze the experience of recruiting a donor cohort from COVID-19 convalescents for banking of CP as part of a pilot project at the Moscow Healthcare Department.Materials and methods. A retrospective research included 493 COVID-19 convalescents as potential CP donors, all examined at the Sklifosovsky Research Institute for Emergency Medicine. CP was banked using the plasmapheresis method. Only those donors with a documented medical history of COVID-19, which was confi rmed by polymerase chain reaction of SARS-CoV-2 RNA in pharyngeal swabs, and no sooner than 14 days after complete recovery were eligible for donation. Viral neutralizing activity (VNA) was chosen as the key characteristic of the immunological viability of CP. All the donors having VNA titers were characterized in terms of gender, age, time interval since the disease onset, regression of clinical symptoms and clinical features of the COVID 19 course.Results. Effective (1:160 or more) and acceptable (1:80) VNA titers were found in 21.1 % and 24.75 % of donors, respectively. Signifi cant predictors for a donor having a high VNA titer included: male sex, age over 36 years and verifi ed viral pneumonia. The absence of a signifi cant body temperature response (38.5 °С) can be considered as a negative marker of a potential donor.

Background. The application of convalescent plasma (CP) is currently seen as a feasible therapeutic approach in the treatment of COVID-19. Aim. To analyze the experience of recruiting a donor cohort from COVID-19 convalescents for banking of CP as part of a pilot project at the Moscow Healthcare Department. Materials and methods. A retrospective research included 493 COVID-19 convalescents as potential CP donors, all examined at the Sklifosovsky Research Institute for Emergency Medicine. CP was banked using the plasmapheresis method. Only those donors with a documented medical history of COVID-19, which was confi rmed by polymerase chain reaction of SARS-CoV-2 RNA in pharyngeal swabs, and no sooner than 14 days after complete recovery were eligible for donation. Viral neutralizing activity (VNA) was chosen as the key characteristic of the immunological viability of CP. All the donors having VNA titers were characterized in terms of gender, age, time interval since the disease onset, regression of clinical symptoms and clinical features of the COVID 19 course. Results. Effective (1:160 or more) and acceptable (1:80) VNA titers were found in 21.1 % and 24.75 % of donors, respectively. Signifi cant predictors for a donor having a high VNA titer included: male sex, age over 36 years and verifi ed viral pneumonia. The absence of a signifi cant body temperature response (38.5 °С) can be considered as a negative marker of a potential donor.

Disorders in the erythrocyte morphology and structure of their membranes during long-term storage of erythrocyte suspension (30 days at 4°C) were studied by atomic force microscopy. The morphology and nanostructure of erythrocyte membranes, biochemical parameters, ion exchange parameters, and hemoglobin spectra were recorded. The transformation of erythrocyte morphology and destruction of their membranes were observed throughout the storage period. Irreversible forms of spheroechinocytes and their fragments formed by the end of storage. The concentrations of potassium ions and lactate in solution of the blood preservatiive increased, while pH value decreased. Hemolysis detected by the erythrocyte “leakage” effect was observed starting from days 16–23 of storage.

Objective: to reveal the patterns of blood biochemical changes, a trend in poikylocytosis, and impairment in the ultra-structure of red blood cells of the whole blood stored in citrate-phosphate-dextrose-adenine-1 during long-term storage. Material and methods. The whole blood that had been obtained from 5 donors aged 25±3 years of different sexes and stored at +2 to +4°С was examined. On storage days 1, 7, 14, 21, and 30, the morphological composition of a red blood cell population was assessed in monolayer blood smears (1000 cells per smear), by using an Olympus BX-500 microscope, computer, and ImageScopeM program. The surface of the red blood cell membranes was analyzed using an IntegraPrima NT-MDT atomic force microscope (AFM). Blood biochemical composition was examined using an i-stat 1® analyzer M: 300. Statistical processing was made with a STATISTICA 7 program. Results. A change in the biochemical composition of the whole blood was revealed within just the first 24 hours of storage. Morphological changes were observed in the red blood cells 3 days later. At 21 days, discocytes were 31.3±3.27% as compared to the control (in the first 24 hours) (p<0.0001). After 30 days of storage, altered red blood cells amounted to more than 80%, among them echinocytes prevailed as compared to the control (79.6±2.74%; p<0.0001). The examinations using AFM illustrated that the nanostructure of the red blood cell membranes tangibly changed with storage, they showed local lesions (excrescences of different sizes) growing into large spicules. Conclusion. The performed studies of the biochemical parameters of the blood, the morphological characteristics of the red blood cells and their membranes suggest that their changes evolved in just the first 24 hours of blood storage. The findings are essential for the development of new methods for storing blood and its components.