Антивозрастной белок Клото как новый потенциальный супрессор опухолевого роста

-Isomer of Klotho Protein has been actively studied over the past 20 years. It has been found that -Klotho is closely associated with aging and has anti-aging properties. Besides, it is involved in development of malignant tumor diseases. Its universal influence on many biological mechanisms leading to hyperplastic changes in cells and tissues has been proven experimentally. Recent studies demonstrated inhibitory effect of -Klotho on the classical carcinogenic pathways such as insulin-like growth factor 1 (IGF1), fibroblast growth factor (FGFs), transforming growth factor 1 (TGFb1), Wnt, P53/p21, oxidative stress response pathways. Impeding effect of -Klotho protein on the development of such types of malignant tumors as breast, pancreatic, lung, stomach, hepatocarcinoma, and others is proven in a number of cases. This review describes the role of -Klotho in oncogenesis, its involvement and influence on the main signaling pathways known for their role in the development of malignant tumors, and the possibility of regulating its expression.С момента своего открытия 20 лет назад белок Клото активно изучается во всем мире. Выявлено, что его изомер -Клото тесно связан с процессами старения и обладает антивозрастными свойствами. В экспериментальных работах было показано, что -Клото универсально воздействует на многие биологические механизмы, ведущие к гиперпластическим изменениям клеток и тканей, ингибирует известные канцерогенные пути с вовлечением инсулиноподобного фактора роста 1 (IGF1), фактора роста фибробластов (FGFs), трансформирующего фактора роста 1 (TGFb1), Wnt, P53/p21, а также воздействует на путь окислительного стресса (oxidative stress response). Все больше научных данных указывает на то, что белок Клото влияет на развитие таких видов злокачественных опухолей, как рак молочной и поджелудочной желез, легких, желудка, гепатокарциномы и др. В данном обзоре рассматривается роль -Клото в онкогенезе, его влияние на основные сигнальные пути развития злокачественных опухолей и возможности регуляции его экспрессии

Analysis of status of preterm infants with bronchopulmonary dysplasia

Objective: the combination frequency of BPD in premature infants, assessment of the degree of functional lesion of the respiratory tract and the efficiency of respiratory support. Material and Methods. The survey included 36 women and 38 preterm infants (two twins). Results. In 99% of the surveyed women pregnancy was complicated by chronic intrauterine fetal hypoxia, 2.1% — RH-conflict. At 43.2% of cases acute respiratory viral infection during pregnancy was revealed, 15.3% of bad obstetric history, 5% of women who had bad habits (smoking, alcohol intake). In 2015 there were 5 deaths (13.2%). 14 children (36.8%) were transferred to other hospitals for further treatment, the remaining 19 (50%) were discharged from the hospital. One child — full-term, the other— premature. At 87.7% of children with BPD, the diagnosis was RDS, in 4.2% — aspiration of meconium, 8.1% — congenital pneumonia. 15 children (30.5%) received therapy with surfactant once, including 9 newborns (23.7%) received medication twice. All newborns were carried out intensive care, including respiratory support. Mechanical ventilation in 14 children had been conducting for 8.4±2.3 day, in 13 cases for 17.3±3.8 day, in 11 cases for 23.4±4.1 day. Nasal CPAP was conducted in 23 newborns: from 4 to 6.8±1.9 day, 13 — less of 16.8±2.9 per day, 6 children have had more than 23.2±3.8 day. Conclusion. There is necessity for prevention during pregnancy with glucocorticoids, the prolongation of pregnancy; preterm neonates — surfactant therapy; adequate provision of resuscitation care in the delivery room and respiratory support. The reduction of time of mechanical ventilation and the expansion of the indications for non-invasive methods of respiratory therapy reduced the incidence of BPD, the severity of the disease and improve the prognosis.</p

Climate variations and the state of zooplankton in the Barents Sea

We present data on zooplankton biomass distribution in August-September 2005-2006 obtained in the integrated ecosystem system survey for the Barents Sea performed by Russian and Norwegian vessels. In order to establish the general regularities of plankton biomass, data collected in the central latitudinal zone of the Barents Sea in individual years (cold 1987, moderate warm 1989, anomalous warm 2002, 2004, 2005) were analysed. Considerable annual differences in the species and age structure of plankton community in water masses of different origin depending on dynamics of the ice cover in that part of the sea were found. In 2002, owing to the northerly position of the ice edge in summer, the plankton community was characterized by its mixed composition (Atlantic and Arctic species) and high abundance. In 2005 when the advective processes were weak and the eastern areas became free from ice later, a considerable reduction in the abundance of warm-water species was observed. When the position of the ice edge was most southerly in 2004, the bulk of the plankton community was made up of Arctic species and a lack of warm-water species was recorded. In 1987 and 1989, high horizontal gradients of water temperature in the areas of Atlantic and interactions of Arctic water masses were, together with anomalous ice distribution, the main factors influencing the state of zooplankton

Invitroexperimental rewiring of 4 neutrophilic granulocyte subsets from the pro-inflammatory to the anti-inflammatory phenotype in children with surgical purulent infection of soft tissue [ЭКСПЕРИМЕНТАЛЬНАЯ ПЕРЕОРИЕНТАЦИЯ INVITRO ФЕНОТИПА 4 СУБПОПУЛЯЦИЙ НЕЙТРОФИЛЬНЫХ ГРАНУЛОЦИТОВ ИЗ ПРОВОСПАЛИТЕЛЬНОГО К ПРОТИВОСПАЛИТЕЛЬНОМУ У ДЕТЕЙ С ХИРУРГИЧЕСКОЙ ГНОЙНОЙ ИНФЕКЦИЕЙ МЯГКИХ ТКАНЕЙ]

Treatment of young children with atypical or recurrent purulent soft tissue infections (PSTD) that do not respond well to surgery and antibiotics is most challenging. PSTD occurs against the background of impaired functioning of the immune system and, first of all, the system of neutrophilic granulocytes (NG). The vector effect of immunotropic therapy on a specific NG subsets may allow the correction of NG dysfunctions without compromising host protection, including strategies to enhance, inhibit or restore their functions. The aim of study: to evaluate in vitro the modulating effects of arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine (HP) on the transformed phenotype of 4 NG subsets, as well as on the functional activity of NG in children with purulent-inflammatory soft tissue diseases. We studied samples of peripheral blood (PB) from young children 2-4 years old: 17 children with atypical acute PSTD and 10 apparently healthy children. At stage I, a comparative assessment of the content and phenotype of 4 NG subsets CD16+CD62L+СD63-, CD16+CD62L+СD63+, СD64-CD16+CD32+CD11b+, СD64+CD16+CD32+CD11b+, phagocytic and microbicidal functions of NG was carried out. At stage II, the in vitro system determined the effects of HP on NG in children with PSTD according to the studied parameters. By the method of flow cytometry (FC500 “Beckman Coulter” (USA), conjugates of MkAT “Beckman Coulter International S.A.” (France)), the relative number of NGs of the studied subsets and the density of receptor expression (MFI) were determined. To assess the phagocytic function of NG a microbiological method was used to assess the completeness of phagocytosis with S. aureus (strain 209). The activity of NG NADPH oxidase was investigated in the NBT-spontaneous test (NBTsp.) and in the in vitro NBT-induced test (NBTind.). A comparative study of PB samples from conventionally healthy children and children with PSTD made it possible to identify various variants of transformation of the phenotype of the studied NG subsets, associated with defects in their functional activity. In the in vitro system the effects of HP were demonstrated, manifested by a decrease in the amount of CD16+CD62L+CD63+NG and an increase in CD16+CD62L+CD63-NG, modulation of the negatively altered phenotype of subsets CD64-CD32+CD16+CD11b+NG and CD64+CD32+CD16+CD11b+NG, aimed at restoring phagocytic function and maintaining the tension of NADPH oxidases. As a result of the study it was found the immunomodulatory effects of HP, which is manifested in the reorientation of NG from the pro-inflammatory phenotype to the anti-inflammatory one, which can be used in the future when creating personalized targeted immunotherapy aimed at correcting defective functioning NG in early children, suffering from PSTD. © 2021 Russian Association of Allergologists and Clinical Immunologists, St. Petersburg Regional Branch (SPb RAACI). All rights reserved

IN VITRO EXPERIMENTAL REWIRING OF 4 NEUTROPHILIC GRANULOCYTE SUBSETS FROM THE PRO-INFLAMMATORY TO THE ANTI-INFLAMMATORY PHENOTYPE IN CHILDREN WITH SURGICAL PURULENT INFECTION OF SOFT TISSUE

Treatment of young children with atypical or recurrent purulent soft tissue infections (PSTD) that do not respond well to surgery and antibiotics is most challenging. PSTD occurs against the background of impaired functioning of the immune system and, first of all, the system of neutrophilic granulocytes (NG). The vector effect of immunotropic therapy on a specific NG subsets may allow the correction of NG dysfunctions without compromising host protection, including strategies to enhance, inhibit or restore their functions.The aim of study: to evaluate in vitro the modulating effects of arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine (HP) on the transformed phenotype of 4 NG subsets, as well as on the functional activity of NG in children with purulent-inflammatory soft tissue diseases.We studied samples of peripheral blood (PB) from young children 2-4 years old: 17 children with atypical acute PSTD and 10 apparently healthy children. At stage I, a comparative assessment of the content and phenotype of 4 NG subsets CD16+CD62L+СD63- , CD16+CD62L+СD63+, СD64- CD16+CD32+CD11b+, СD64+CD16+CD32+CD11b+, phagocytic and microbicidal functions of NG was carried out. At stage II, the in vitro system determined the effects of HP on NG in children with PSTD according to the studied parameters. By the method of flow cytometry (FC500 “Beckman Coulter” (USA), conjugates of MkAT “Beckman Coulter International S.A.” (France)), the relative number of NGs of the studied subsets and the density of receptor expression (MFI) were determined. To assess the phagocytic function of NG a microbiological method was used to assess the completeness of phagocytosis with S. aureus (strain 209). The activity of NG NADPH oxidase was investigated in the NBT-spontaneous test (NBTsp.) and in the in vitro NBT-induced test (NBTind.). A comparative study of PB samples from conventionally healthy children and children with PSTD made it possible to identify various variants of transformation of the phenotype of the studied NG subsets, associated with defects in their functional activity. In the in vitro system the effects of HP were demonstrated, manifested by a decrease in the amount of CD16+CD62L+CD63+NG and an increase in CD16+CD62L+CD63- NG, modulation of the negatively altered phenotype of subsets CD64- CD32+CD16+CD11b+NG and CD64+CD32+CD16+CD11b+NG, aimed at restoring phagocytic function and maintaining the tension of NADPH oxidases.As a result of the study it was found the immunomodulatory effects of HP, which is manifested in the reorientation of NG from the pro-inflammatory phenotype to the anti-inflammatory one, which can be used in the future when creating personalized targeted immunotherapy aimed at correcting defective functioning NG in early children, suffering from PSTD.Лечение детей раннего возраста с атипичными или рецидивирующими гнойными инфекциями мягких тканей (ГЗМТ), которые не демонстрируют хорошего ответа на хирургическое лечение и антибактериальные препараты, является наиболее сложным. ГЗМТ возникают на фоне нарушений функционирования иммунной системы и в первую очередь системы нейтрофильных гранулоцитов (НГ). Векторный эффект иммунотропной терапии на конкретную субпопуляцию NG может позволить корректировать дисфункции NG без ущерба для защиты хозяина, включая стратегии усиления, подавления или восстановления их функций.Цель исследования: оценить в системе in vitro модулирующие эффекты влияния аргинил-альфа-аспартил-лизил-валил-тирозил-аргинин (ГП) на трансформированный фенотип 4 субпопуляций НГ, а также на функциональную активность НГ детей с гнойно-воспалительными заболеваниями мягких тканей. Исследованы образцы периферической крови (ПК) детей раннего возраста 2-4 лет: 17 детей с нетипично протекающими острыми ГЗМТ и 10 условно здоровых детей. На I этапе проведена сравнительная оценка содержания и фенотипа 4 субпопуляций НГ CD16+CD62L+СD63- , CD16+CD62L+СD63+, СD64- CD16+CD32+CD11b+, СD64+CD16+CD32+CD11b+, фагоцитарной и микробицидной функции НГ. На II этапе в системе in vitro определены эффекты ГП на НГ детей с ГЗМТ по изучаемым параметрам. Методом проточной цитометрии (FC500, Beckman Coulter, (США), конъюгаты МкАТ Beckman Coulter International S.A. (Франция)), определялось относительное количество НГ исследуемых субпопуляций и плотность экспрессии рецепторов (MFI). Для оценки фагоцитарной функции НГ использован микробиологический метод с оценкой завершенности фагоцитоза со S. аureus (штамм 209). Активность NADРН-оксидазы НГ исследовали в НСТ спонтанном тесте (НСТсп.) и в нагрузочном в системе in vitro (НСТст.). Сравнительное изучение образцов ПК условно здоровых детей и детей с ГЗМТ позволило выявить различные варианты трансформации фенотипа изучаемых субпопуляциях НГ, сопряженных с дефектами их функциональной активности. В системе in vitro были продемонстрированы эффекты ГП, проявляющиеся снижением количества CD16+CD62L+CD63+НГ и повышением CD16+CD62L+CD63- НГ, модуляцией негативно измененного фенотипа субпопуляций CD64- CD32+CD16+CD11b+НГ и CD64+CD32+CD16+CD11b+НГ, направленные на восстановление фагоцитарной функции и поддержания напряженности NADPH-оксидаз. В результате исследования установлено иммуномодулирующее действие ГП, которое проявляется в переориентации НГ с провоспалительного фенотипа на противовоспалительный, что может быть использовано в будущем при создании персонализированной таргетной иммунотерапии. Направлена на коррекцию неполноценного функционирования НГ у детей раннего возраста, страдающих ГЗМТ

Effects of recombinant IFN-α2b on the phenotype of neutrophil granulocyte subpopulations in patients with COVID-19

Investigation of molecular mechanisms associated with interferon (IFN) production and receptor function of neutrophil granulocytes (NGs) in COVID-19 is highly relevant because it can be promising in the search for new therapeutic strategies targeting NGs and their reactivity to restore and strengthen the innate immune response against SARS-CoV-2. Objective. To assess the effects of recombinant IFN-α2b on the phenotype of CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119–, and CD16+IFNα/βR1+CD119+ NGs from peripheral blood of patients with COVID-19 in an in vitro experiment. Patients and methods. We analyzed blood samples from 31 patients with a mean age of 61 years (range: 57;71 years) with moderate COVID-19. We assessed the number of CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119–, and CD16+IFNα/βR1+CD119+ NGs, receptor density (FC 500, ‘Beckman Coulter,’ USA), phagocytic activity of NGs before and after incubation with recombinant IFN-α2b. We also measured serum levels of several cytokines, including IFNα, IFNγ, IL-6, IL-8 (ELISA, ‘Vektor-Best’ LLC). The control group comprised 22 adult healthy individuals with a mean age of 58 years (range: 57; 70 years). Results. Patients with moderate COVID-19 demonstrated low serum levels of IFNα and IFNγ along with elevated levels of IL-6 and IL-8. We observed transformation of 3 phenotypes among NG subpopulations: CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119-, and CD16+IFNα/βR1+CD119+. We observed positive remodulating effects of recombinant IFN-α2b on the number and phenotype of NG subpopulations and their phagocytic activity in our in vitro experiment. Conclusion. Recombinant IFN-α2b demonstrated positive effects in in vitro experiments; therefore, it can be considered in the future as a potential therapeutic tool for moderate COVID-19. Restoration of type I IFN might be an effective treatment option for COVID-19, because it can promote faster virus elimination, restore normal functioning of the IFN system, and have positive regulatory effects on the phenotype of NG subpopulations. © 2022, Dynasty Publishing House. All rights reserved

PHENOTYPE REMODELING IN NEUTROPHILIC GRANULOCYTE SUBSETS CD64(-)CD32(+)CD16(+)CD11B(+)NG, CD64(+)CD32(+)CD16(+)CD11B(+)NG IN DE NOVO EXPERIMENTAL MODEL OF VIRAL-BACTERIAL INFECTION IN VITRO

A search for new targeted therapeutic strategies based on examining immunopathogenetic mechanisms for emerging co-infections is relevant and may further contribute not only to optimizing choice of immunotropic drugs, but also to achieving positive clinical and immunological remission for abnormal infectious processes. Previously, our studies found that recurrent viral-bacterial respiratory infections are associated with dysfunction of neutrophilic granulocytes (NG) with varying degree of intensity in altered effector properties. NG dysfunctions are often associated with diverse phenotypic profiles characterized by varying density for expression level of functionally significant trigger receptors. The aim of the study was to pinpoint phenotype transformation in CD64(-)CD32(+)CD16(+)CD11b(+), CD64(+)CD32(+)CD16(+)CD11b(+) neutrophilic granulocytes in experimental model of viral-bacterial infection in vitro. We examined 52 peripheral blood samples collected from 13 healthy adult volunteers. Viral, bacterial and virus-bacterial infection was modelled in vitro by incubating blood-derived cell samples with formyl-methionyl-leucyl-phenylalanine (fMLP), double-stranded RNA (dsRNA) or in combination followed by assessing changes in immunophenotyping of CD64(-) CD32(+)CD16(+)CD11b+NG C.D64(+)CD32(+)CD16(+)CD11b(+)NG by using using MAbs CD16-ECD, CD64-FITC, CD32-PE, CD11b-PC5 conjugates (Beckman Coulter International SA, France). It was demonstrated that NGs from healthy adult volunteers were dominated by CD64-CD32(+)CD16(+)CD11b(+)NG as well as minor subset.D64+CD32+CD16+CD11b(+) NG varying in expression density of membrane molecules. Percentage of the minor subset.D64+CD16+CD32+CD11b(+) NG was significantly increased after exposure with dsRNA, fMLP and dsRNA(+)fMLP compared to untreated samples. Comparative analysis re vealed that various immunotropic agents differed in affecting expression of surface receptor molecules CD16, CD32 and unidirectional effects, but of varying magnitude altering CD11b marker both in major and minor subsets. Preincubation with dsRNA followed by adding fMLP allowed to find that they co-stimulated expression of surface receptors in both NG subsets. We generated an experimental model of viral-bacterial co-infection in vitro by using fMLP and dsRNA and observed types of phenotype transformation in CD64(-) CD32(+)CD16(+)CD11b(+) NG and.D64+CD32+CD16+CD11b+ NG subsets. This model can be used to evaluate transformation of other NG subset phenotypes, NG functional activity, features of NET formation as well as impact of various immunotropic agents on NG

Phenotype remodeling in neutrophilic granulocyte subsets CD64-CD32+CD16+CD11B+NG, CD64+CD32+CD16+CD11B+NG in de novo experimental model of viral-bacterial infection in vitro

A search for new targeted therapeutic strategies based on examining immunopathogenetic mechanisms for emerging co-infections is relevant and may further contribute not only to optimizing choice of immunotropic drugs, but also to achieving positive clinical and immunological remission for abnormal infectious processes. Previously, our studies found that recurrent viral-bacterial respiratory infections are associated with dysfunction of neutrophilic granulocytes (NG) with varying degree of intensity in altered effector properties. NG dysfunctions are often associated with diverse phenotypic profiles characterized by varying density for expression level of functionally significant trigger receptors. The aim of the study was to pinpoint phenotype transformation in CD64-CD32+CD16+CD11b+, CD64+CD32+CD16+CD11b+ neutrophilic granulocytes in experimental model of viral-bacterial infection in vitro. We examined 52 peripheral blood samples collected from 13 healthy adult volunteers. Viral, bacterial and virus-bacterial infection was modelled in vitro by incubating blood-derived cell samples with formyl-methionyl-leucyl-phenylalanine (fMLP), double-stranded RNA (dsRNA) or in combination followed by assessing changes in immunophenotyping of CD64-CD32+CD16+CD11b+NG, CD64+CD32+CD16+CD11b+NG by using using MAbs CD16-ECD, CD64-FITC, CD32-PE, CD11b-PC5 conjugates (Beckman Coulter International SA, France). It was demonstrated that NGs from healthy adult volunteers were dominated by CD64-CD32+CD16+CD11b+NG as well as minor subset CD64+CD32+CD16+CD11b+ NG varying in expression density of membrane molecules. Percentage of the minor subset CD64+CD16+CD32+CD11b+ NG was significantly increased after exposure with dsRNA, fMLP and dsRNA+fMLP compared to untreated samples. Comparative analysis revealed that various immunotropic agents differed in affecting expression of surface receptor molecules CD16, CD32 and unidirectional effects, but of varying magnitude altering CD11b marker both in major and minor subsets. Preincubation with dsRNA followed by adding fMLP allowed to find that they co-stimulated expression of surface receptors in both NG subsets. We generated an experimental model of viral-bacterial co-infection in vitro by using fMLP and dsRNA and observed types of phenotype transformation in CD64-CD32+CD16+CD11b+ NG and CD64+CD32+CD16+CD11b+ NG subsets. This model can be used to evaluate transformation of other NG subset phenotypes, NG functional activity, features of NET formation as well as impact of various immunotropic agents on NG.Поиск новых таргетных терапевтических стратегий, базирующихся на изучении иммунопатогенетических механизмов возникновения коинфекций, является актуальным и может в дальнейшем способствовать не только оптимизации выбора иммунотропных лекарственных средств, но и достижению позитивной клинико-иммунологической ремиссии нетипично протекающих инфекционных процессов. Ранее нашими исследованиями было установлено, что возвратные вирусно-бактериальные респираторные инфекции ассоциированы дисфункциями нейтрофильных гранулоцитов (НГ) с разной степенью выраженности нарушений их эффекторных свойств. Зачастую дисфункции НГ сопряжены с различными фенотипическими профилями, характеризующимися разными уровнями и плотностью функционально значимых триггерных рецепторов. Целью исследования было уточнение вариантов трансформации фенотипа субпопуляций CD64-CD32+CD16+CD11b+Hr, CD64+CD32+CD16+CD11b+Hr в созданной экспериментальной модели вирусно-бактериальной коинфекции in vitro. Исследовано 52 образца периферической крови 13 здоровых взрослых добровольцев в возрасте от 21 до 32 лет. Для воспроизведения условий вирусной, бактериальной и вирусно-бактериальной инфекции образцы инкубировали с формил-метионил-лейцил-фенилаланин (fMLP), двухцепочечной РНК (дцРНК) и совместно, затем определяли фенотипические характеристики субпопуляций CD64-CD32+CD16+CD11b+Hr, CD64+CD32+CD16+CD11b+Hr с использованием конъюгатов МКАТ CD16-ECD, CD64-FITC, CD32-PE, CD11b-PC5 (Beckman Coulter International S.A., Франция). Анализ полученных данных продемонстрировал, что НГ здоровых взрослых лиц представлены мажорной субпопуляцией CD64-CD16+CD32+CD11b+ НГ и минорной субпопуляцией CD64+CD16+CD32+CD11b+ НГ с разной плотностью мембранных молекул. Минорная субпопуляция CD64+CD16+CD32+CD11b+ НГ значительно увеличилась под влиянием дцРНК, fMLP и дцРНК + fMLP по сравнению с интактными образцами. Сравнительный анализ моновлияния иммунотропных субстанций позволил выявить их разные эффекты в отношении действия на поверхностные рецепторные молекулы CD16, CD32 и однонаправленные, но разной интенсивности на CD11b, как в мажорной, так и в минорной субпопуляциях. Преинкубация с дцРНК с последующим добавлением fMLP в группе исследования позволила выявить эффекты совместного стимулирующего влияния субстанций на уровни поверхностных рецепторов обеих субпопуляции НГ. Нами была создана экспериментальная модель вирусно-бактериальной коинфекции в системе in vitro с использованием fMLP и дцРНК и установлены варианты трансформации фенотипа субпопуляций CD64-CD32+CD16+CD11b+Hr и CD64+CD32+CD16+CD11b+Hr Данная модель может быть использована для оценки вариантов трансформации фенотипа других субпопуляций НГ, изучения функциональной активности НГ, особенностей формирования NET, влияния на НГ различных иммунотропных субстанций