Genetically modified animals for use in bio‑pharmacology: from research to production

In this review, the analysis of technologies for obtaining biologically active proteins from various sources is carried out, and the comparative analysis of technologies for creating producers of biologically active proteins is presented. Special attention is paid to genetically modified animals as bioreactors for the pharmaceutical industry of a new type. The necessity of improving the technology of development transgenic rabbit producers and creating a platform solution for the production of biological products is substantiate

Effect of the secretome of multipotent mesenchymal stromal cells induced by dexamethasone in vitro on the expression of phospho-NF-κB p65 and Ki-67 in mononuclear cells

To investigate the influence of secretomes from native and dexamethasone-treated adipose-derived multipotent mesenchymal stromal cells (MMSC) on the proliferation of mononuclear cells (MNCs) and on their expression of phospho-NF-κB p65 in vitr

Immunohistochemical evaluation of surfactant-associated protein in fibrosis-cavernous pulmonary tuberculosis

Background. Despite the progress and development of scientific directions in various fields of medicine, the problem of tuberculosis and its morphological manifestations remains relevant and is not fully disclosed due to complex pathogenesis, the presence of various clinical forms, therapeutic pathomorphosis, torpid to therapy, the presence of relapses. It is known that a surfactant system occupies a special place in the system of local lung protection.Aim. To study the condition of surfactant-assotiated protein A in the foci of specific destruction and in the surrounding intact lung tissue to assess its functional status, degree of respiratory failure and possible dissemination of tuberculous inflammation.Material and Metods. An analysis of 163 lung fragments of the dead or operated on for cavernous pulmonary tuberculosis with active bacterial excretion of 89 fragments and with clinical abacilation – 74 was carried out. Results. A morphological study revealed stereotypical dynamic depression of surfactant-associated protein A in all the samples studied, both in the areas of cavernous destruction and pericavernouse zone, and in intact lung tissue. The maximum intensivity of the immunohistochemical expression of this surfactant protein was recorded in the alveolar macrophages, which indicated intensive recycling and utilization of the components of the surfactant.Conclusion. Minimizing the production of surfactant components and its active utilization in intact lung tissue leads to a collapse of the alveoli with subsequent progression of respiratory failure

Молекулярно‑генетическая характеристика колоректального рака

Colorectal cancer ranks third among all cancers. More than 1 million cases are registered annually in the world, half of which are fatal. The investigation of the molecular genetic mechanisms of tumors’ development is relevant, which is an important contribution to the prospect of determining the prognosis and treatment tactics. This review presents the current classification of the mechanisms of tumor progression in colorectal cancer.Колоректальный рак занимает третье место по заболеваемости среди всех онкологических заболеваний. Ежегодно в мире регистрируется свыше 1 млн случаев, половина из которых приводит к летальному исходу. На сегодняшний день актуально изучение молекулярно-генетических механизмов возникновения опухолей, что является важным вкладом в перспективу определения прогноза и тактики лечения. Представленная обзорная статья раскрывает современную классификацию механизмов опухолевой прогрессии при колоректальном раке

Изучение противовоспалительной и иммунотропной активности секретома мультипотентных мезенхимальных стромальных клеток, индуцированных эритропоэтином, вальпроевой кислотой или дексаметазоном in vitro

The aim of this study was to evaluate the effect of treatment with valproic acid, erythropoietin, and dexamethasone on the anti-inflammatory and immunosuppressive activity of the secretome of adipose-derived multipotent mesenchymal stromal cells (MMSCs) in an in vitro experiment.Materials and methods. MMSCs were isolated from the fat of 6 healthy donors. The cells were grown in the culture up to passage 4. Then they were treated with valproic acid, erythropoietin or dexamethasone for 3 hours, washed from preparations, and incubated in a serum-free medium for 48 hours. Some of the cells were not treated with preparations. Supernatants from the cell cultures were concentrated by ultrafiltration, and protein standardization was performed using a nanophotometer. Then the supernatants were sterilized and added to mononuclear cells from peripheral blood of 8 healthy donors. The mononuclear cells were isolated by Ficoll density gradient centrifugation according to the standard protocol. Concentrations of TNFα, IL-2, IL-4, IL-6, IL-10, and IFNγ cytokines in 24-hour cultures and IL-9, IL-10, IL-17A, and IL-21 cytokines in 48-hour cultures were determined using multiplex analysis.Results. The production of IL-2, IL-6, TNFα, and IL-10 was reduced by the secretome of MMSCs treated with valproic acid. The production of IL-2, IL-6, and TNFα decreased during incubation of the mononuclear cells with the secretome of MMSCs treated with erythropoietin. The secretome of dexamethasone-treated MMSCs suppressed the production of IFNγ, IL-1β, IL-1ra, IL-2, IL-6, IL-9, IL-10, and IL-17A. No statistically significant differences were revealed in the production of IL-4, IL-5, IL-9, and IL-21.Conclusion. Among the studied inducers, dexamethasone enhanced the anti-inflammatory and immunosuppressive activity of MMSCs the most, which was manifested through the effect of their supernatants on peripheral blood mononuclear cells.Цель. Исследовать влияние обработки вальпроевой кислотой, эритропоэтином и дексаметазоном на противовоспалительную и иммуносупрессивную активность секретома мультипотентных мезенхимальных стромальных клеток (ММСК) жировой ткани в эксперименте in vitro.Материалы и методы. ММСК выделяли из жира шести здоровых доноров. Клетки растили в культуре до четвертого пассажа, затем обрабатывали вальпроевой кислотой, эритропоэтином или дексаметазоном в течение 3 ч, отмывали от препаратов и инкубировали в бессывороточной среде в течение 48 ч. Часть клеток не обрабатывали препаратами. Супернатанты от культур клеток сконцентрировали ультрафильтрацией, стандартизировали по содержанию белка с помощью нанофотометра, стерилизовали и добавляли к мононуклеарам из периферической крови восьми здоровых доноров. Мононуклеары выделяли в градиенте плотности фиколла по стандартному протоколу. Концентрации цитокинов фактора некроза опухоли альфа (TNFα), интерлейкина (IL) 2, IL-4, IL-6, IL-10, интерферона гамма (IFNγ) в суточных культурах и IL-9, IL-10, IL-17A, IL-21 в 48-часовых культурах определяли с помощью мультиплексного анализа.Результаты. Продукция IL-2, IL-6, TNFα, IL-10 снижается под действием секретома от обработанных вальпроевой кислотой ММСК. Продукция IL-2, IL-6, TNFα уменьшается при инкубации мононуклеаров с секретомом клеток, обработанных эритпропоэтином. Секретом обработанных дексаметазоном ММСК подавляет продукцию IFNγ, IL-1β, IL-1ra, IL-2, IL-6, IL-9, IL-10, IL-17A. Статистически значимых различий по изменению продукции IL-4, IL-5, IL-9, IL-21 не выявлено.Заключение. Среди изученных индукторов дексаметазон показал себя более активным в усилении противовоспалительной и иммуносупрессивной активности ММСК, выраженной через влияние их супернатантов на мононуклеары периферической крови

Иммунофенотип макрофагальной популяции при фиброзно-кавернозном туберкулезе легких

Objective: to study the immunophenotype of the macrophage population and the mechanisms of their vectorial redistribution in fibrous cavernous pulmonary tuberculosis.Materials and methods. The material for the study was fragments of the fibrous cavern wall and pericavernous lung tissue of the dead or surgical patients diagnosed with fibrous cavernous tuberculosis (n = 163). All patients were divided into 2 main groups: patients with active bacteria excretion (MTB+, n = 84) and patients with clinical abacillation (MTB–, n = 79) for immunohistochemistry with a panel of markers for: macrophages and histiocytes – CD68; vascular growth factor A – VEGF-A; T-helpers – CD4, and T-cytotoxic lymphocytes – CD8.Results. Following the analysis of CD68 expression, the population heterogeneity of macrophages was revealed depending on the intensity of the cytoplasmic reaction, functional activity, localization and quantitative characteristics. Three groups were identified: highly active, moderately active and weakly active. Based on the reaction with vascular growth factor A, it was determined that VEGF+ cells correspond to weakly active CD68+ macrophages and are located on the border between the specific granulation tissue and fibrous layer as well as in the pericavernous zone and intact lung tissue with a statistically significant predominance in patients with MTB– (p < 0.05). Regardless of the scope of bacterial secretion, the number of VEGF+ cells in the lymphoid follicle zone directly correlates with that of CD68+ macrophages in the pericavernous zone (R = 0.68) and indirectly correlates with the number of diffusely scattered VEGF+ cells in the fibrous capsule (R = –0.75). In the meantime, CD68+/VEGF+ are visualized in the zone of CD8+ T-lymphocytes, and CD68+/VEGF- – in the zone of CD4+ cell clusters. Such correlation indicates the redistribution of macrophages into type 2, which has a remodeling effect on the surrounding tissues with the potentiating participation of lymphoid cells.Цель исследования: изучение иммунофенотипа макрофагальной популяции и механизмов их векторного перераспределения при фиброзно-кавернозном туберкулезе легких.Материалы и методы. Материалом для исследования явились фрагменты стенки каверны и перикавернозной легочной ткани прооперированных по поводу фиброзно-кавернозного туберкулеза (ФКТ) легких (n = 163). Все пациенты были разделены на две основные группы (с активным бактериовыделением (МБТ+, n = 84) и клиническим абациллированием (МБТ–, n = 79)) для проведения иммуногистохимического исследования с панелью маркеров: макрофагов и гистиоцитов – CD68; сосудистого фактора роста А – VEGF-A; Т-хелперов CD4, цитотоксических Т-лимфоцитов CD8.Результаты. При анализе экспрессии маркера CD68 установлена популяционная неоднородность макрофагов в зависимости от интенсивности цитоплазматической реакции функциональной активности с различными локализационными и количественными характеристиками: высокоактивные (тип 1), умеренно активные (тип 2) и слабоактивные (тип 3). На основании реакции с сосудистым фактором ростом A определено, что клетки VEGF+ соответствуют слабоактивным макрофагам CD68+ и локализуются на границе перехода специфической грануляционной ткани в фиброзный слой, перикавернозной зоне и интактной легочной ткани со статистически значимым преобладанием у пациентов с МБТ– (р < 0,05). Независимо от активности бактериовыделения число клеток VEGF+ в зоне лимфоидных фолликулов прямо коррелирует с количеством макрофагов CD68+ в перикавернозной зоне (R = 0,68) и обратно – с числом VEGF+ диффузно рассеянных клеток в фиброзной капсуле (R = макрофагов 0,75). При этом CD68+/VEGF+ визуализируются в зоне CD8+ Т-лимфоцитов, а CD68+/VEGF- – в зоне скоплений клеток CD4+. Такой характер корреляционной взаимосвязи свидетельствует о перераспределении макрофагов во второй тип, обладающих ремоделирующим действием на окружающие ткани при потенцирующем участии клеток лимфоидной популяции

Effect of the COVID-19 pandemic on surgery for indeterminate thyroid nodules (THYCOVID): a retrospective, international, multicentre, cross-sectional study

Background Since its outbreak in early 2020, the COVID-19 pandemic has diverted resources from non-urgent and elective procedures, leading to diagnosis and treatment delays, with an increased number of neoplasms at advanced stages worldwide. The aims of this study were to quantify the reduction in surgical activity for indeterminate thyroid nodules during the COVID-19 pandemic; and to evaluate whether delays in surgery led to an increased occurrence of aggressive tumours.Methods In this retrospective, international, cross-sectional study, centres were invited to participate in June 22, 2022; each centre joining the study was asked to provide data from medical records on all surgical thyroidectomies consecutively performed from Jan 1, 2019, to Dec 31, 2021. Patients with indeterminate thyroid nodules were divided into three groups according to when they underwent surgery: from Jan 1, 2019, to Feb 29, 2020 (global prepandemic phase), from March 1, 2020, to May 31, 2021 (pandemic escalation phase), and from June 1 to Dec 31, 2021 (pandemic decrease phase). The main outcomes were, for each phase, the number of surgeries for indeterminate thyroid nodules, and in patients with a postoperative diagnosis of thyroid cancers, the occurrence of tumours larger than 10 mm, extrathyroidal extension, lymph node metastases, vascular invasion, distant metastases, and tumours at high risk of structural disease recurrence. Univariate analysis was used to compare the probability of aggressive thyroid features between the first and third study phases. The study was registered on ClinicalTrials.gov, NCT05178186.Findings Data from 157 centres (n=49 countries) on 87 467 patients who underwent surgery for benign and malignant thyroid disease were collected, of whom 22 974 patients (18 052 [78 center dot 6%] female patients and 4922 [21 center dot 4%] male patients) received surgery for indeterminate thyroid nodules. We observed a significant reduction in surgery for indeterminate thyroid nodules during the pandemic escalation phase (median monthly surgeries per centre, 1 center dot 4 [IQR 0 center dot 6-3 center dot 4]) compared with the prepandemic phase (2 center dot 0 [0 center dot 9-3 center dot 7]; p<0 center dot 0001) and pandemic decrease phase (2 center dot 3 [1 center dot 0-5 center dot 0]; p<0 center dot 0001). Compared with the prepandemic phase, in the pandemic decrease phase we observed an increased occurrence of thyroid tumours larger than 10 mm (2554 [69 center dot 0%] of 3704 vs 1515 [71 center dot 5%] of 2119; OR 1 center dot 1 [95% CI 1 center dot 0-1 center dot 3]; p=0 center dot 042), lymph node metastases (343 [9 center dot 3%] vs 264 [12 center dot 5%]; OR 1 center dot 4 [1 center dot 2-1 center dot 7]; p=0 center dot 0001), and tumours at high risk of structural disease recurrence (203 [5 center dot 7%] of 3584 vs 155 [7 center dot 7%] of 2006; OR 1 center dot 4 [1 center dot 1-1 center dot 7]; p=0 center dot 0039).Interpretation Our study suggests that the reduction in surgical activity for indeterminate thyroid nodules during the COVID-19 pandemic period could have led to an increased occurrence of aggressive thyroid tumours. However, other compelling hypotheses, including increased selection of patients with aggressive malignancies during this period, should be considered. We suggest that surgery for indeterminate thyroid nodules should no longer be postponed even in future instances of pandemic escalation.Funding None.Copyright (c) 2023 Published by Elsevier Ltd. All rights reserved

Гістологічна характеристика зіскрібків порожнини матки у жінок з гіперпролактинемією різного генезу, ускладненою перерваною вагітністю

The main morphological feature in spontaneous abortions in hyperprolactinemia of various origin is reduced activity of invasive cytotrophoblast. At the same time a decrease in utero-placental barrier and increased hypoxia, resulting in dystrophic and necrotic processes in decidual tissue and chorionic villi. Delay differentiation of chorionic villi in the form of primary cystic edematous and secondary avascular villi, characteristic mainly for early pregnancy suggests genetically deterministic pathogenesis of hyperprolactinemia. The distinctive morphological feature of scrapings of the uterus in women with missed abortion of miscarriage, is the presence of severe inflammatory reaction and fibrinoid necrosis.Основным морфологическим признаком при самопроизвольном выкидыше при гиперпролактинемиях различного генеза является снижение уровня инвазивной активности цитотрофобласта. При этом наблюдается уменьшение маточно-плацентарного барьера и усиление гипоксии, что приводит к дистрофическим и некротическим процессам в децидуальной ткани и ворсинах хориона. Задержка дифференцировки хориальных ворсин в виде первичных кистозно отечных и вторичных бессосудистых ворсин, характерных, в основном, для ранних сроков беременности, свидетельствует о генетически детерминированном генезе гиперпролактинемии. Отличительной морфологической чертой соскобов полости матки у женщин с несостоявшимся выкидышем от самопроизвольного выкидыша является наличие выраженной воспалительной реакции и фибриноидных некрозов.Основною морфологічною ознакою при мимовільному викидні при гіперпролактинемії різного ґенезу є зниження рівня інвазивної активності цитотрофобласта. При цьому визначено зменшення матково-плацентарного бар’єра і посилення гіпоксії, що призводить до дистрофічних і некротичних процесів у децидуальної тканині і ворсинах хоріона. Затримка диференціювання хоріальних ворсин у вигляді первинних кістозно набряклих і вторинних безсудинних ворсин, характерних, в основному, для ранніх строків вагітності, свідчить про генетично детермінований ґенез гіперпролактинемії. Відмінною морфологічною рисою зіскрібків порожнини матки у жінок з викиднем, що не відбувся, від мимовільного викидня, є наявність вираженої запальної реакції і фібриноїдних некрозів

Immunophenotype of the macrophage population in fibrous, cavernous pulmonary tuberculosis

Objective: to study the immunophenotype of the macrophage population and the mechanisms of their vectorial redistribution in fibrous cavernous pulmonary tuberculosis.Materials and methods. The material for the study was fragments of the fibrous cavern wall and pericavernous lung tissue of the dead or surgical patients diagnosed with fibrous cavernous tuberculosis (n = 163). All patients were divided into 2 main groups: patients with active bacteria excretion (MTB+, n = 84) and patients with clinical abacillation (MTB–, n = 79) for immunohistochemistry with a panel of markers for: macrophages and histiocytes – CD68; vascular growth factor A – VEGF-A; T-helpers – CD4, and T-cytotoxic lymphocytes – CD8.Results. Following the analysis of CD68 expression, the population heterogeneity of macrophages was revealed depending on the intensity of the cytoplasmic reaction, functional activity, localization and quantitative characteristics. Three groups were identified: highly active, moderately active and weakly active. Based on the reaction with vascular growth factor A, it was determined that VEGF+ cells correspond to weakly active CD68+ macrophages and are located on the border between the specific granulation tissue and fibrous layer as well as in the pericavernous zone and intact lung tissue with a statistically significant predominance in patients with MTB– (p < 0.05). Regardless of the scope of bacterial secretion, the number of VEGF+ cells in the lymphoid follicle zone directly correlates with that of CD68+ macrophages in the pericavernous zone (R = 0.68) and indirectly correlates with the number of diffusely scattered VEGF+ cells in the fibrous capsule (R = –0.75). In the meantime, CD68+/VEGF+ are visualized in the zone of CD8+ T-lymphocytes, and CD68+/VEGF- – in the zone of CD4+ cell clusters. Such correlation indicates the redistribution of macrophages into type 2, which has a remodeling effect on the surrounding tissues with the potentiating participation of lymphoid cells

Renal cell carcinoma drug and cell therapy: today and tomorrow

Today, considerable progress in the renal cell carcinoma (RCC) treatment has been made due to development of targeted and immunotherapeutic approaches to the RCC treatment, especially in metastasising carcinoma. In the early stages of RCC, it is possible to use partial or total surgical nephrectomy, but in metastases development, the range of efficient treatment methods is dramatically limited. Appearance of targeted drugs like PD-1 and CTLA-4 receptors and their ligands' inhibitors in clinical practice has significantly increased the total survival rate of patients with renal cell carcinoma. Emergence of adoptive cell therapy has opened new possibilities and prospects in RCC treatment. Previously activated in vitro cells are used there, which provides antineoplastic activity. For example, it could be antigen-specific cytotoxic T-lymphocytes (CTL), lymphokine-activated natural killers (LAK-NK-cells) and tumour-infiltrating lymphocytes (TILs). In this review, the authors specified the main molecular markers, associated with RCC; and signalling pathways (VEGFR- and EGFR-signalling pathway), which directly take part in carcinogenesis. The paper also looks at clinically applicable targeted immune drugs and the principle of their effect on tumorous cells. Besides, modern clinical studies of cell drugs have been considered. At the moment, there are a number of variants of targeted and immune drugs for the metastatic RCC treatment. Patients have no opportunity to use all the available agents because of their cost and toxicity level. For the most efficient treatment of patients with diagnosed metastatic RCC, it is necessarily to carry out risk stratification and prognostic factors for the response to treatment