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The aim of the study was to examine specifics of changes in the level of stomach- and pancreas-released blood hydrolases in chronic viral hepatitis B and analyze the mechanisms underlying such changes. We assessed serum markers of HBV infection, liver enzymes tests as well as gastric and pancreatic hydrolase level. The patients examined were divided into three groups: control (healthy) and two study groups — HBV post-infection and chronic HBV infection. Patients with HBV post-infection had no significant deviations from normal range for blood level of gastric and pancreatic hydrolases. Patients with chronic HBV infection were found to contain increased blood level of amylase and lipase, which may evidence about increasing pancreatic functional activity and development of covert pancreatitis. At the same time, decline in the concentration of serum pepsinogen-1 below 40 μg/l could indicate about prominently decreased secretion of hydrochloric acid and development of atrophic gastritis, and it was found that the major factor contributing to development of such disorders was the short-chain peptide CCK-8, which utilization declines in patients with chronic HBV infection. CCK-8 can play a pivotal role in inhibiting stimulation of gastric acid secretion and controls gastric acid, plasma gastrin and somatostatin secretion. Cholecystokinin has been found to inhibit acid secretion by activating CCK type A receptors as well as via somatostatin-involving mechanism. The secretion of gastric somatostatin-14 increased by fivefold due to CCK-8 alone, but was blocked by the CCK-A receptor blocker loxiglumide. These data show that CCK-8 directly inhibits acid reactions by stimulating the release of gastric somatostatin indirectly through the CCK-A receptor. Thus, it can be assumed that normally CCK-8 is mainly utilized by the liver, which is altered during chronic hepatitis B resulting in elevated blood CCK-8 concentration. As a consequence, it enhances pancreatic secretion resulting in developing pancreatitis that is paralleled with inhibited gastric secretion and emerged atrophic gastritis.

Currently, the Russian market of phospholipase A2 enzyme preparations is represented by commercial preparations of foreign manufacturers: Nagase (Japan) and Maxapal (the Netherlands). However, the growing demand and the need to reduce the cost of production of phospholipase A2 require the development of new super-producers of phospholipase A2. In this connection, the aim of the work is to compare the expression of recombinant phospholipase A2 in Komagataella phaffii depending on the modification of the alpha-factor signaling peptide. The object of the study is the recipient yeast strain Komagataella phaffii X-33. The studies were conducted in accordance with generally accepted norms and approaches. Phospholipase A2 genes from Streptomyces violaceoruber were used for this worK. The target sequences were synthesized in the company "Eurogen" (Russia) and cloned as part of the TE vector pUC57. In the course of the work, the genetic constructs pPICZaA-Pla2 and PPICZmf4iA-Pla2 containing the Streptomyces violaceoruber phospholipase A2 gene were assembled under the native signal a-MF and its modification mf4i. The transformation of the yeast Komagataella phaffii X-33 with the obtained genetic constructs was also carried out. As a result of the conducted studies, it was shown that on average, there were no significant differences in the level of expression and specific activity of recombinant phospholipase A2 in methylotrophic yeast K. Phaffii X-33 when using the native a-MF secretion signal and its modified version mf4i. However, the use of the secretion factor mf4i allows for higher production of phospholipase A2 in individual clones. The obtained data indicate the prospects of using the secretion factor mf4i to create super-producers of enzymes based on yeast K. Phaffii X-33.

Macrophages (Mφ) play a key role in regulation of fibrogenesis, including proliferation of fibroblasts and myofibroblasts, differentiation of progenitor cells into myofibroblasts, as well as synthesis and secretion of the extracellular matrix, mainly collagen. The direction of the Mφ effects (stimulation or suppression) is determined by a number of factors, including the stage of the fibrotic process and the Mφ functional phenotype dependent on the signals of microenvironment. One of the feasible ways of the fibrogenesis regulating is the secretion of pro- or antifibrotic factors such as matrix metalloproteinases, inhibitors of metalloproteinases and some cytokines. However, existing data on ability to secrete these factors by various subpopulations of human Mφ are rare and controversial. The aim of this study was to characterize the ability of human M1, M2a, and M2c Mφ differentiating in the presence of GM-CSF to produce matrix metalloproteinases (MMP-9) and their tissue inhibitors (TIMP-1), as well as some cytokines and growth factors. As compared to M2 macrophages, the M1 macrophages polarized by lipopolysaccharide produced significantly more TNFα, IL-6 and IL-2 that have pro-inflammatory activity and are able to initiate a fibrotic process. In turn, M2a Mφ stimulated by IL-4 were characterized by a high level of VEGF production and, at the same time, low levels of TNFα and IL-6, which may determine the important role of these cells at the proliferative stage of fibrosis and stimulation of extracellular matrix deposition. Finally, M2c Mφ polarized by dexamethasone, exhibited the М2а-like cytokine profile, i.e., VEGF was actively produced against the background of low TNFα and IL-6 synthesis. Moreover, all three Mφ subpopulations did actively secrete MMP-9 and TIMP-1, without significant difference in production of these factors. However, M2c Mφ differed by a significantly higher MMP-9/TIMP-1 ratio index compared to M1 and M2a Mφ, and it is crucial at the rearrangement stage of the fibrotic process. Thus, the production of MMP-9 and TIMP-1, together with other pleiotropic cytokines and growth factors by various Mφ subtypes may reflect their role in regulation of fibrotic process at various stages.

Aim. To compare the influence of “lipophilic” atorvastatin and “hydrophilic” rosuvastatin on the parameters of cellular immunity in atherosclerosis patients.Material and methods. Totally, 35 participants included, mean age 62 [57;68] y.o., 18 males and 17 females, directed for follow-up to Myasnikov Cardiovascular Center with preliminary diagnosis coronary heart disease, atherosclerosis of coronary and carotid arteries, and with indications for intensified statin therapy. In 17 patients the dosage of atorvastatin was increased from 20 to 80 mg, in 18 — dosage of rosuvastatin from 10 to 40 mg. All patients at baseline and in 1 month, by the methods of direct immune fluorescence and cytofluometry underwent the measurement of content of monocytes and lymphocyte populations in peripheral blood, incl. regulatory and effectory subpopulations of the latter. Under the circumstances of cellular culture the influence studied, of atorvastatin and rosuvastatin on CD4+ T-lymphocytes populations, as the lipopolysaccharideinduced synthesis of cytokines by monocytes of donors blood.Results. At the background of atorvastatin, there was marked increase of relative content of circulating regulatory T-lymphocytes (Treg), increase of the relation of Treg/Thelper 17 (Th17) and changes of cellular immunity parameters. Statins did not influence subpopulations of blood monocytes. There was dose-dependent inhibition by statins of CD4+ T-lymphocytes proliferation: atorvastatin action was noted in 10 nM/L, rosuvastatin — in 10 times higher concentration. Introduction ofstatins, 10-100 nM/L, to the culture of monocytes did not influence neither spontaneous, nor endotoxin induced secretion of cytokines.Conclusion. In therapeutic dosages atorvastatin shows immune modulating activity presenting with an increase of relative content of regulatory T-lymphocytes subpopulations that might be determined by suppression of effectory cells proliferation.

It is known that a relationship exists between patient’s age and pathogenesis of malignant tumors associated with ability of tumor and its microenvironment to secrete cytokines that may play an important role in carcinogenesis. The aim of present study was a comparative analysis of cytokine production by peripheral blood cells and tumor samples when exposed to polyclonal activators (PA) in patients of different age groups with invasive ductal carcinoma. Peripheral blood samples of 54 women with invasive ductal carcinoma were under study. The patients were divided into the following groups: I, 40-60 years old; II, 61 year and older. Metastases into lymph nodes were revealed at the moment of study in 13 patients (42%) from group I. By contrast, only 3 of 23 (10%) patients with metastases were revealed in group II. Fisher’s ratio test showed significant differences between occurrence of metastases in lymph nodes, and patient age (p = 0.034). In both groups of patients, there was an increase of IL-6, IL-8, IL-10, IL-17, IL-18, IL-1ra, G-CSF, GM-CSF and VEGF concentrations in tumor supernates and its microenvironment, when compared with the production of cytokines by blood cells, thus indicating to high functional activity of tumor together with its microenvironment. Effect of PA led to increase of IL-2, IL-10, IL-17, IL-1β, IL-1ra, TNFα and IFNγ concentrations in blood cells supernatant which suggested their high ability to produce cytokines. In the group of patients who were 40-60 years old, the stimulatory index of PA (SIPA) for IFNγ production was several times higher than similar index in patients aged 61 years or more. Increase of SIPA for all cytokines in blood cells supernates, in comparison to SIPA of tumor supernates, could be explained by initially high levels of spontaneous cytokine production by the tumor and its microenvironment that are able to promote tumor growth. Therefore, effect of PA upon tumor tissue was not so expressed as its effect upon immunocompetent blood cells. Both tumor and microenvironment maintained high ability of IL-18 production, thus confirming its important role in carcinogenesis. In summary, these findings suggest that circulating immunocompetent cells may serve as an additional factor providing tumor progression, due to their high ability to secrete cytokines.

Numerous modern basic research done undeniable fact that neutrophilic granulocytes (NG) are key effector and regulatory circuits both innate and adaptive immunity, and play a crucial role in the pathogenesis of a wide range of diseases. NG have potent receptor repertoire, providing a connection between them, cells of the immune system, as well as communication with endothelial cells, epithelial and other tissues. NG inducing stimuli activate and promote the translocation of cytoplasmic granules and vesicles surface molecules on the cytoplasmic membrane the secretion of a large spectrum of pro-and anti-inf lammatory, immunoregulatory cytokines, colony, angiogenic factors and fibrogenic, TNF superfamily members, chemokines, regulatory protein, etc. Chromatin nuclei NG capable of restructuring under the influence of inducing stimuli, which is associated with the expression of multiple cytokine genes. NG receiving complex cytokine inf luence not only acquire new features, but also in various stages of activation and differentiation processes involved in intracellular intraphagosomalis degranulation and killing of implementing elimination microorganisms and extracellular neutrophil degranulation in the formation neutrophil extracellular traps (NET), while this dying through NETosis. Features NG phenotype and their functional properties, demonstrate the existence of subpopulations of NG with different capabilities: equipment of different receptor, the ability to restructure chromatin expressing cytokine genes and secrete cytokines to implement the contents of the granular system, produce reactive oxygen species, implement cytotoxicity form NET. In our opinion, there subpopulation NG: regulatory; suppressor; proinf lammatory — initiating an inf lammatory response; inf lammation with a positive potential microbicidal (antibacterial, antiviral, antifungal); inf lammatory cytotoxic potential of the negative — “aggressive”; anti-inf lammation regulating regression; antitumoral — TAN1; pro-tumoral — TAN2; hybrid, combining the characteristics of NG and dendritic cells. The absence of adequate response, or hyperactivation blockade NG functions leads to the development of low-intensity infectious and inf lammatory diseases, do not respond to conventional therapy of autoimmune diseases/chronic immune-dependent processes. Remodeling dysfunctions NG — the key to new immunotherapeutic strategies.

Chronic pancreatitis (CP) is one of the most widespread gastroenterological diseases, the pathogenic mechanisms of which are still under investigation. The authors gave a short overview of the main stages in the research of the relationships between gastric and pancreatic secretion, and the results of physiological studies, performed by I. P. Pavlov and his students. Being а strict follower of «nervousness» theory, the great physiologist and his associates made the incorrect conclusions about the presence of local nerve connections (peripheral reflex arc) between the duodenum and pancreas, not suggesting the presence of humoral interactions. The modern data have been presented that give the evidence that duodenal acidification results in the secondary deficiency of endogenous pancreatic enzymes, increased stimulation of pancreatic secretion, exacerbated symptoms of exocrine pancreatic insufficiency, increased secretory tension in pancreatic tissue, duct and tissue pressure, and increased risk of CP attack.However, a dot at the end of a statement «CP is an acid-dependent disease» is tentative. The proton pump inhibitors (PPIs) are used throughout for the treatment of acid-dependent diseases. However, neither European, nor Russian recommendations do include PPIs as a basic treatment for CP. The results of a prospective randomized controlled trial, which proved the effectiveness of CP treatment with the use of combination of analgesics and PPIs have been considered. However, the results of the only study are not enough to recommend the inclusion of PPI in the basic CP therapy. The analysis has been performed for the provisions of the European Guidelines for the Diagnosis and Treatment of Pancreatitis. These guidelines describe situations when PPIs should be included in the complex CP treatment. The data of evidence-based medicine have been presented, illustrating that suppression of gastric secretion can increase the effectiveness of enzyme preparations in comparison with the administration of pancreatin monotherapy. The advantages of pantoprazole among other of the PPIs’ representatives have been analyzed. A clinical case of CP treatment with a reasonable pantoprazole (Nolpaza) administration have been considered., Хронический панкреатит (ХП) — одно из наиболее распространенных гастроэнтерологических заболеваний, патогенетические механизмы возникновения которого продолжают исследовать. Описаны основные этапы изучения взаимосвязи желудочной и панкреатической секреции, результаты исследований И. П. Павлова и его учеников. Строго следуя принципам теории «нервизма», великий физиолог и его соратники сделали неправильные выводы о наличии локальных нервных связей (периферической рефлекторной дуги) между двенадцатиперстной кишкой и поджелудочной железой, не предполагая наличия гуморальных взаимодействий. Приведены современные данные, доказывающие, что ацидификация двенадцатиперстной кишки приводит ко вторичному дефициту эндогенных панкреатических ферментов, усилению стимуляции панкреатической секреции, усугублению симптомов экзокринной панкреатической недостаточности, повышению секреторного напряжения в ткани поджелудочной железы, протокового и тканевого давления и увеличению риска атаки ХП.Однако после утверждения «ХП — это кислотозависимое заболевание» можно поставить точку только предварительно. Для лечения кислотозависимых заболеваний в мире используют ингибиторы протонной помпы (ИПП). Однако ни в европейских, ни в российских рекомендациях ИПП не указаны в качестве базисного средства лечения ХП. Рассмотрены результаты проспективного рандомизированного контролируемого исследования, в котором доказана эффективность лечения ХП с использованием комбинации аналгетиков и ИПП. Результатов одного исследования недостаточно для рекомендации включения ИПП в базисную терапию ХП. Проанализированы положения европейского руководства по диагностике и лечению ХП, описывающие случаи обоснованного включения ИПП в комплексное лечение ХП. Приведены данные доказательной медицины, иллюстрирующие, что подавление желудочной секреции может повысить эффективность ферментных препаратов по сравнению с приемом одного панкреатина. Проанализированы преимущества пантопразола перед другими представителями группы ИПП. Рассмотрен клинический случай лечения ХП с обоснованным назначением пантопразола («Нольпазы»)., Хронічний панкреатит (ХП) — одне з найпоширеніших гастроентерологічних захворювань, патогенетичні механізми виникнення якого продовжують досліджувати. Описано основні етапи вивчення взаємозв’язку між шлунковою та панкреатичною секрецією, результати досліджень І. П. Павлова і його учнів. Суворо дотримуючись принципів теорії «нервізму», видатний фізіолог та його соратники зробили неправильні висновки щодо наявності локальних нервових зв’язків (периферичної рефлекторної дуги) між дванадцятипалої кишкою і підшлунковою залозою, не припускаючи наявність гуморальних взаємодій. Наведено сучасні дані, які доводять, що ацидифікация дванадцятипалої кишки призводить до вторинного дефіциту ендогенних панкреатичних ферментів, посилення стимуляції панкреатичної секреції, поглиблення симптомів екзокринної панкреатичної недостатності, підвищення секреторного напруження в тканині підшлункової залози, протокового і тканинного тиску та збільшення ризику атаки ХП.Однак після твердження «ХП — кислотозалежне захворювання» можна поставити крапку лише поперед­ньо. Для лікування кислотозалежних захворювань у світі використовують інгібітори протонної помпи (ІПП). Однак ні в європейських, ні в російських рекомендаціях ІПП не зазначені як базисний засіб лікування ХП. Розглянуто результати проспективного рандомізованого контрольованого дослідження, в якому доведено ефективність лікування ХП з використанням комбінації аналгетиків та ІПП. Результатів одного дослідження недостатньо для рекомендації включення ІПП у базисну терапію ХП. Проаналізовано положення європейських рекомендацій з діагностики та лікування ХП, які описують ситуації, коли ІПП слід включати в комплексне лікування ХП. Наведено дані доказової медицини, які ілюструють, що пригнічення шлункової секреції може підвищити ефективність ферментних препаратів порівняно з прийманням одного панкреатину. Проаналізовано переваги пантопразолу над іншими представниками групи ІПП. Розглянуто клінічний випадок лікування ХП з обґрунтованим призначенням пантопразолу («Нольпази»).

Большой физиологической важностью в организме поросят обладают тромбоциты, являющиеся важным компонентом системы поддержания гомеостаза. Их функциональные свойства в значительной степени определяют процессы гемоциркуляции в просвете капилляров, а следовательно, интенсивность метаболических процессов во всех тканях. Особую значимость в раннем онтогенезе поросят имеет фаза молочно-растительного питания. От уровня многих физиологических механизмов на ее протяжении зависят процессы развития и становления продуктивных качеств этих животных. Становится ясно, что большая функциональная значимость в этом плане у них принадлежит тромбоцитарному гемостазу, способному определять успешность процессов микроциркуляции и рост тканей, и в конечном счете, динамику хозяйственно значимых параметров поросят. В этой связи для дальнейшего развития науки и практики свиноводства необходимы дополнительные исследования различных аспектов функционирования тромбоцитарного гемостаза у поросят на протяжении фазы молочно-растительного питания. Данная информация может помочь в совершенствовании подходов к кормлению и лечению свиней, а следовательно, будет способствовать интенсификации свиноводства в целом. В исследовании выяснено, что у поросят за фазу молочно-растительного питания отмечается небольшое повышение функциональной активности тромбоцитарного гемостаза. Центральное место в этом занимают развивающиеся изменения рецепторных и пострецепторных процессов активации тромбоцитов. На физиологическом уровне они проявляются интенсификацией их адгезии, агрегации и процесса секреции. В исследовании выявлено нарастание в тромбоцитах поросят молочно-растительного питания холестерина, актина, миозина, АДФ и повышение степени секреции последнего. Данные изменения нужно расценивать как факторы, стимулирующие активность тромбоцитарного гемостаза у поросят на протяжении фазы молочно-растительного питания. Они, очевидно, способствуют стойкому сохранению у этих животных параметров гомеостаза и оптимального уровня микроциркуляции во всех тканях, адекватного имеющимся потребностям активно растущего организма в ходе смены питания с молочного на растительное., Of great physiological importance in the body of piglets are platelets, which are an important component of the system for maintaining homeostasis. Their functional properties largely determine the processes of blood circulation in the lumen of the capillaries, and, consequently, the intensity of metabolic processes in all tissues. Of particular importance in the early ontogeny of piglets is the phase of the milk-plant nutrition. The processes of development and formation of the productive qualities of these animals depend on the level of many physiological mechanisms throughout its length. It becomes clear that a large functional significance in this regard belongs to platelet hemostasis, which is able to determine the success of microcirculation processes and tissue growth, and ultimately, the dynamics of the economically significant parameters of piglets. In this regard, for the further development of the science and practice of pig breeding, additional studies are needed on various aspects of platelet hemostasis in piglets during the dairy-plant nutrition phase. This information can help to improve the approaches to feeding and treatment of pigs, and, therefore, will contribute to the intensification of pig breeding in general. The study found that in piglets during the dairy-plant nutrition phase a slight increase in the functional activity of platelet hemostasis was observed. Central to this are developing changes in receptor and post-receptor processes of platelet activation. At the physiological level, they are manifested by the intensification of their adhesion, aggregation and secretion process. The study revealed an increase in the platelets of piglets of the milk-plant cholesterol, actin, myosin, ADP and an increase in the secretion of the latter. These changes should be regarded as factors that stimulate the activity of platelet hemostasis in piglets during the milk-plant nutrition phase. They obviously contribute to the persistent preservation of the parameters of homeostasis in these animals and the optimal level of microcirculation in all tissues, which is adequate to the existing needs of an actively growing organism during a diet change from dairy to vegetable., №6 (2020)

The study presents results concerning effects of H3/H4 histamine receptors antagonist (Ciproxifan) upon in vitro chemokine secretion by dendritic cells (DC) and peripheral blood mononuclear cells (PBMC). We have been shown that in vitro inhibition of type H3/H4 histamine receptor affects in different ways production of major chemokines by PBMC and DC, including eotaxin, RANTES, MCP-1, MIP-1α, MIP-1β, GRO-α, and IL-8. E.g., RANTES and GRO-α secretion in DC cultures was reduced, respectively, by 20% and 40%. Meanwhile, MIP-1β secretion was virtually unchanged, whereas MCP-1 secretion exhibited a four-fold increase. In PBMC cultures, the H3/H4 receptor antagonist induced a significant increase in eotaxin, RANTES, MIP-1α, MIP-1β, GRO-α, and IL-8 secretion (p < 0.05).

Coxiella burnetii is an obligate intracellular gram-negative bacterial pathogen, an ethiological agent of Q-fever, a zoonotic disease, elapsing as an acute (mostly atypical pneumonia) or a chronic (mostly endocarditis) form. The host range is represented by wide range of mammal, avian and arthropod species, but the main source of human infection are farm animals. The main route of infection is aerosolic. In case of contact with organism pathogen binds with phagocytal monocytic-macrophagal cell line. C. burnetii promotes maturation of specific phagolysosome-like compartment in host cell, called coxiella-containing vacuole, within this vacuole pathogen becames metabolically activated and actively replicates. Coxiella persists as metabolically inactive spore-like form in environment. Internalisation of C. burnetii occurs using actin-mediated phagocytosis and zipper mechanism. After internalization of bacteria maturation of phagolysosome-like compartment and large coxiella-containing vacuole formation occure, and vacuole can occupy nearly the whole cytoplasm of the host cell. Survivance of infected cells is important for chronic infection with C. burnetii. C. burnetii elongate the viability of host cell by two ways: it actively inhibits apoptotic signal cascades and induce pro-survival factors. Exceptthat C. burnetii involves autophagic pathway during coxiella-containing vacuole formation, and induction of autophagy promotes pathogen replication. During infection C. burnetii translocates effector substrates from bacterial cytosole to euca ryotic host cell cytosole using type IV secretion system, where effectors modulate host cell proteins. Overall approximately 130 secreted effectors of type IV transport system, but function of most of them remains unknown to date. Specific sec reted proteins for variety of strains and isolates were identified, confirmed that certain pathotypes of C. burnetii can exist. Identification and characterization of novel virulence factors it is now possible through axenic media for C. burnetii cultivation and development of site-specific mutagenesis and other genetic technics, which is important for research of C. burnetii molecular pathogenesis.

Pathogenesis of serous discharge from the nipples of the breast is complex and includes the following components: hormonal imbalance as a backdrop, stimulating increased secretion in the ducts of serous content, resorption of the liquid component of a secret, resulting in local pathological effects of protein resorption, hormone content, autoimmune reactions ductless with subsequent development of proliferative processes in them. Under adverse conditions, infection occurs from an external or internal source of microorganisms. The presence of fluid in the ducts, mammary system ductal communication with the external environment, is an “open gate” for content ductal infection of the mammary glands. This creates the prerequisites for the development of proliferative processes in the first phase of the inflammatory origin. Application of 194 women with serous secretions from the nipples of the breast combined combinations of enzymes, glucocorticoids, progestogens, gonadoliberins and other agonists, suppress the proliferation of drugs that helped to put an end to abnormal secretion and prevent changes in the walls of the ducts and periductal breast tissue at different stages of disease.

Актуальность. Исследовали прямые эффекты гранулоцитмакрофагального колониестимулирующего фактора (GMCSF) человека на функциональную активность субпопуляций Tлимфоцитов. Методы. CD3 Тлимфоциты были выделены из крови здоровых доноров методом позитивной магнитной сепарации. Tклетки активировали частицами, конъюгированными с антителами (АТ) к молекулам CD3, СD28 и СD2 человека. Мембранную экспрессию CD3, CD4, СD45RA, СD197, CD25 и CD38 оценивали методом проточной цитофлюорометрии. Содержание интерферона Результаты. Установлено, что GMCSF в диапазоне концентраций 0,0110,0 нг/мл не оказывал существенного влияния на содержание CD25 клеток, среди активированных Тлимфоцитов. Вместе с тем, GMCSF в концентрации 0,1 и 1,0 нг/мл обладал способностью заметно увеличивать содержание CD38 клеток среди наивных СD45RA/СD197 Тклеток, а также среди СD45RA/СD197 Тклеток центральной памяти, не оказывая при этом существенного влияния на экспрессию CD38, выявляемую среди эффекторных СD45RA/СD197 и терминально дифференцированных СD45RA/СD197 эффекторных Тклеток. В относительно низкой концентрации (0,01 нг/мл) GMCSF заметно снижал Тклеточную продукцию INF, тогда как в высокой концентрации (10,0 нг/мл) усиливал продукцию IL2 и IL4, снижая при этом выработку IL10. Заключение. Полученные данные предполагают, что GMCSF способен поддерживать активацию относительно низкодифференцированных Тклеток, не оказывая при этом значимого влияния на активацию высоко дифференцированных Тклеток., Background. We investigated direct effects of granulocytemacrophage colonystimulating factor (GMCSF) on the functionality of Tlymphocyte subsets. Methods. CD3 T cells were isolated from the blood of healthy donors by positive magnetic separation. The isolated T cells were activated with particles conjugated with antibodies (Abs) to human CD3, CD28, and CD2 molecules. The membrane expression of CD3, CD4, СD45RA, СD197, CD25, and CD38 was evaluated by flow cytofluorometry. Contents of interferon (IFN ), interleukin2 (IL2), IL4, and IL10 in culture supernatants were determined by the enzyme immunoassay. Results. GMCSF at 0.0110.0 ng/ml had no significant effect on the content of CD25 cells among activated T lymphocytes. At the same time, GMCSF at 0.11.0 ng/ml was able to noticeably increase the content of CD38 cells among both naive CD45RA/CD197 T cells and central memory CD45RA/CD197 T cells without affecting the СD38 expression on effector CD45RA /CD197 and terminally differentiated CD45RA / CD197 effector T cells. At a relatively low concentration (0.01 ng/ml), GMCSF significantly decreased Tcell production of INF whereas at a high concentration (10.0 ng/ml), GMCSF detectably enhanced the secretion of IL2 and IL4 while lowering IL10 production. Conclusion. The findings suggest that GMCSF is capable of supporting the activation of relatively lowdifferentiated T cells without significantly affecting the activation of highly differentiated T cells., №4 (2019)

Эндогенные опиоидные пептиды играют важную роль в регуляции иммунных процессов. Цель работы - изучение влияния бета-эндорфина на уровни кортикостерона в плазме периферической крови мышей, секрецию IL-2, IL-4 и IFN-gamma мышиными спленоцитами, а также продукцию IL-1b и IL-10 перитонеальными макрофагами in vivo через 1 ч и через 6 ч после введения пептида. Методика. Исследования выполнены на беспородных мышах-самцах массой 22-25 г. Бета-эндорфин вводили однократно внутрибрюшинно в дозах 100-0,0005 мкг/кг. Через 1 или 6 ч после введения бета-эндорфина животных выводили из эксперимента, выделяли перитонеальные макрофаги и спленоциты для культивирования. Результаты. Установлено, что введение бета-эндорфина в дозах 100, 0,01, 0,0005 мкг/кг за 1 ч до выведения животных из эксперимента приводило к угнетению спонтанной продукции IL-1b макрофагами. Введение пептида за 6 ч в дозе 1 мкг/кг приводило к угнетению продукции IL-1b стимулированными макрофагами. b-эндорфин выражено снижал продукцию IL-10 во всех исследуемых дозах как при введении за 1ч, так и за 6 ч до выведения животных из эксперимента. Пептид усиливал Кон А - индуцированную продукцию IL-4 как через 1 ч, так и через 6 ч после инъекции. Продукция IL-2 спленоцитами угнеталась через 1 ч после введения пептида, и стимулировалась через 6 ч после введения. На продукцию IFN-g введение b-эндорфина влияния не оказывало. Концентрация кортикостерона в плазме крови интактных мышей под воздействием b-эндорфина не изменялась. Заключение. Бета-эндорфин, несмотря на быстрое ферментативное расщепление в организме, способен оказывать пролонгированные иммуномодулирующие эффекты, которые могут менять свою направленность с течением времени., Endogenous opioid peptides play an important role in regulation of immune processes. The work objective was to study the effect of beta-endorphin on corticosterone levels in murine peripheral blood plasma, secretion of IL-2, IL-4, and IFN-g by murine splenocytes, and production of IL-1b and IL-10 by peritoneal macrophages in vivo at 1 h and 6 h following administration of the peptide. Methods. Investigations were performed on mice weighing 22-25 g. Beta-endorphin (100-0,0005 mg/kg, i.p.) was administered once. One or six hours after the beta-endorphin administration, the animals were withdrawn from the experiment, peritoneal macrophages and splenocytes were isolated for culturing. Results. Administration of beta-endorphin at doses of 100, 0.01, and 0.0005 mg/kg 1 h before sacrificing animals led to suppression of macrophage spontaneous production of IL-1b. Administration of the peptide at a dose of 1 mg/kg within 6 hours prior to sacrificing resulted in inhibition of IL-1b production by stimulated macrophages. Βeta-endorphin at all doses reduced the IL-10 production both 1 h and 6 h before sacrificing the mice. The peptide enhanced Con A-induced production of IL-4 both 1 h and 6 h after injection. Production of IL-2 by splenocytes was inhibited 1 h after administration of the peptide and stimulated 6 h after the peptide administration. Administration of b-endorphin had no effect on the IFN-g production. b-endorphin did not change plasma corticosterone concentrations in intact mice. Conclusion. Beta-endorphin, despite its rapid enzymatic cleavage, is able to exert long-term immunomodulatory effects, which can change their direction over time., №1 (2018)

Diagnostics of thyroid cancer (TC) remains a challenging issue due to the high incidence of asymptomatic thyroid nodular pathologies and absence of non-invasive methods of their assessment. Thyroid tumors are classified as benign and malignant with incidence ration approximated as 9:1. Correct and timely differential diagnosis is the basis for correctly choosing a treatment policy and hence determines treatment results. Methods for molecular genetic analysis are being recently developed and introduced into clinical practice, enabling the diagnostic process to be optimized. Analysis of the intracellular and secreted (exosomal) fractions of small regulatory RNAs (microRNAs) is one of the most promising methods for the diagnosis of cancers, including TC. The stability of extracellular microRNA is determined by bonds to proteins, lipoproteins, or its encapsulation into the membrane microvesicles – exosomes. There is reason for suggesting that exosomes with the specific composition of microRNA are a result of the process of active and biologically important secretion while release of other microRNA forms accompanies apoptotic or necrotic cell death. This determines the special diagnostic value of the exosomal fraction of circulating microRNAs, which may reflect the presence and clinically relevant properties of a tumor. This paper discusses the state of the problem and presents methods and preliminary results of the studies conducted by the authors to develop a novel method for diagnosing and monitoring TC. Thus, level of plasma exosomal miR-21 was shown to distinguish patients with benign tumor and follicular CT, while miR-31 can help to distinguish patients with benign tumor and papillary TC. Moreover, reciprocal character of miR-21 and miR-181a concentration in plasma exosomes was detected by comparison of patient with papillary and follicular TC.

This article presents the statistical data and results of own research on microbiological study of the udder secret from cows with subclinical and clinically expressed mastitis in 20 farms (27 dairy farms) of dairy direction of the Vologda region. The decisive value in the frequency of seeding from the affected shares of the udder of cows is coccal microflora, the largest share of which is occupied by conditionally pathogenic and pathogenic staphylococci (30,2% and 23,2%, respectively). Significant variability (from 0 to 100,0) of indicators of particular microflora from the secretion of the udder of cows with mastitis indicates significant differences in the etiology of the disease on farms region. Pathogenic and opportunistic staphylococci containing enterotoxins of types A, C, D, B and E from the udder secret indicates the potential danger of such milk for both calves and people. The article is executed in accordance with the state order № 0578-2014-0030.

The results levels of insulin, testosterone and cortisol in patients with burn disease are presented. The analysis of the secretion hormones in groups with varying severity of the injury are provided. It was shown that in male burned patients was accompanied with prolonged decrease in testosterone secretion. Degree of inhibition testosterone secretion depends on the severity of burn injuries. Also there were revealed that burn disease was accompanied with prolonged increase in cortisol secretion. It was shown that increase IRI-activity on the 6 th and 14 th day of hospitalization, which may indicate emerging insulin resistance in this period disease. Severe burn injury were observed to hormonal failure, which may reduce the chances of survival of the injured.

Aim. To study the influence of biogenic stimulators Actovegine and Zifodine (deproteinized hemodrivates of calf blood) on reactive oxygen species formation by blood phagocytes in patients with chronic heart failure (CHF). Material and methods. Reactive oxygen species formation was registered by lucigenine-dependent chemiluminiscence on the luminomenter “Biotox-7” (Russia) in 20 patients with CHF of II-III functional classes by NYHA. The hemiluminiscences basal and stimulated by farbolic ether (RMA) were measured, as also inhibitory activity of Actovegine and Zifodine. Results. In CHF patients’ blood there was “spontaneous” secretion superoxide anion radicals, that is showing presence of preactivated phagocytes. In addition, a significant increase of oxygen radicals is shown as a response to RMA (1 mcM). Actovegine and Zifodine dose-dependently suppressed prominence of the response. While concomitantly applied Actovegine and Zifodine at the RMA background there was additive inhibitory effect found, which was not depended on consequence of additives. Conclusion. Actovegine and Zifodine decrease reactive oxygen species secretion by phagocytes, which are extracted from the blood of CHF patients. Addictiveness of such inhibitory effects of Actovegine and Zifodine shows absence of their identity by structure and of improper translation of this data into conclusions on effectiveness and safety, found in clinical practice while prescribing Actovegine and Zifodine.

Мonocytes in feto-placental circulation are exposed to factors secreted by placental tissue. These factors influence monocyte functions in pregnancy. In present study, an in vitro model (monocyte-like THP-1 cells) was used for assessing effects of soluble placental factors obtained from women with physiological pregnancies, or preeclampsia cases. The following effects of placental factors were revealed: increased secretion of VEGF by THP-1 cells along with decreased secretion of IL-6, IL-8 and MCP-1 under the influence of placental factors from the I. trimester of pregnancy in comparison with III. trimester. Secretion of IL-6 and MCP-1 by THP-1 cells was increased, and secretion of soluble TNFRII was decreased upon co-cultivation with soluble placental factors from the women with preeclampsia, as compared with placental products from physiological pregnancies. The work is supported by grants ГК № 02.740.11.0711 from Ministry of Education and Science, and НШ-3594.2010.7 grant from the President of Russian Federation.

Phenotypic and functional features of IFNα-induced dendritic cells (DCs) were studied in patients with chronic viral hepatitis B and C (HBV and HCV), and in cases with hepatitis-related liver cirrhosis (LC). It was shown that DCs are characterized by delayed differentiation/maturation which was more pronounced in HCV patients, as well as in all patients with LC, regardless of virus type. DCs from HBV patients were characterized by increased IFNγ secretion. Transformation of HBV-infection to LC is accompanied by a moderate decrease in IFNγ production, combined with a significantly increased IL-10 secretion. Irrespectively of fibrosis severity, the IFNα-induced DCs of HCV patients displayed active IL-10 synthesis. Moreover, ability of DCs to secrete IFNγ was significantly decreased only in cases of fibrosis-complicated HCV-infection. With respect to TNFα and IL-4 production levels, DCs of the patients were compatibe to normal donor cells, independently on the type of virus, or fibrosis severity. DCs from HBV- and HCV-patients were characterized by intact allostimulatory and Th1/Th2-stimulatory activities in MLC. At the same time, IFNα-induced DCs exhibited suppression of allostimulatory and increase in Th2-polarizing activity upon LC development, both in HBV and HCV patients.

Malignant diseases of blood system, associated with an eosinophilic syndrome, are accompanied by altered production of some key cytokines regulating proliferation, differentiation and activation events in the cells of eosinophilic lineage, as detected with ELISA approach. An imbalanced secretion of Th2 cytokines, as studied in lymphoproliferative diseases, may be a promoting factor of eosinophilia development in malignant disorders of blood system.

Influence of topical application of bacterial lipopolysaccharide (LPS), a Toll-like receptor 4 agonist, upon epithelization process was investigated, using a murine incisional skin wound model. It was shown that topical LPS application accelerates wound epithelization, and this process was associated with increased levels of epidermal growth factor secretion in wounded skin area.

Alpha-1-acid glycoprotein (orosomucoid) is a multifunctional acute phase reactant belonging to the family of lipocalines from plasma alpha-2 globulin fraction. In present study, we investigated dosedependent effects of orosomucoid upon secretion of IL-1â, IL-2, IL-3, IL-4 by mononuclear cells from venous blood of healthy volunteers. Mononuclear cells were separated by means of gradient centrifugation, followed by incubation for 24 hours with 250, 500, or 1000 mcg of orosomucoid per ml RPMI-1640 medium (resp., low, medium and high dose). The levels of cytokine production were assayed by ELISA technique. Orosomucoid-induced secretion of IL-1â and IL-4 was increased, whereas IL-3 secretion was inhibited. IL-2 production was suppressed at low doses of orosomucoid, and stimulated at medium and high doses. The effect of alpha-1-acid glycoprotein upon production of IL-2, IL-3 and IL-4 was dose-dependent. Hence, these data indicate that orosomucoid is capable of modifying IL-1â, IL-2, IL-3, and IL-4 secretion by blood mononuclear cells.

Modern literature and the results of our own studies on the role of neutrophils in the body of a healthy person and in a patient with diabetes mellitus type 1 (DM‑1) were analyzed. The article presents the latest data on the content, ultrastructure and functions (phagocytosis, netosis, secretion of bactericidal and immunoregulatory compounds) of blood neutrophils in various stages of DM‑1, starting from the preclinical period. Particular attention is paid to the involvement of neutrophils in various reactions of natural and adaptive immunity. There were discussed the new data on the neutrophil infiltration of the exocrine pancreas and islets of Langerhans, as well as the possibility of direct destructive effect of neutrophils on β-cells, which became the basis for the correction of existing hypotheses about the pathogenesis of DM‑1 in humans, i.e. for approval that not only T-cells of adaptive immunity, but also cellular elements of natural immunity, in particular neutrophils, play an important role in the mechanism of DM‑1 onset., Проведен анализ современной литературы и результатов собственных исследований, касающихся роли нейтрофилов в организме здорового человека и больного сахарным диабетом 1-го типа (СД‑1). Представлены последние данные о содержании, ультраструктуре и функциях (фагоцитоз, нетоз, секреция бактерицидных и иммунорегуляторных соединений) нейтрофилов крови на различных стадиях развития СД‑1, начиная с доклинического периода. Особое внимание уделено участию нейтрофилов в различных реакциях естественного и адаптивного иммунитета. Обсуждены новые данные об инфильтрации нейтрофилами экзокринной части поджелудочной железы и островков Лангерганса, а также возможность непосредственного деструктивного действия нейтрофилов на β-клетки, ставшие основанием для коррекции существующих гипотез о патогенезе СД‑1 у человека, то есть для утверждения, что в механизме возникновения СД‑1 существенную роль играют не только Т-клетки адаптивного иммунитета, но и клеточные элементы естественного иммунитета, в частности нейтрофилы., Проведено аналіз сучасної літератури і результатів власних досліджень, що стосуються ролі нейтрофілів в організмі здорової людини і хворого на цукровий діабет 1-го типу (ЦД‑1). Представлені останні дані про вміст, ультраструктуру і функції (фагоцитоз, нетоз, секреція бактерицидних і імунорегуляторних сполук) нейтрофілів крові на різних стадіях розвитку СД‑1, починаючи з доклінічного періоду. Особливу увагу приділено участі нейтрофілів у різних реакціях природного і адаптивного імунітету. Обговорено нові дані про інфільтрацію нейтрофілами екзокринної частини підшлункової залози і острівців Лангерганса, а також про можливість безпосередньої деструктивної дії нейтрофілів на β-клітини, що стали підставою для корекції існуючих гіпотез про патогенез ЦД‑1 у людини, тобто для ствердження, що в механізмі виникнення СД‑1 істотну роль відіграють не тільки Т-клітини адаптивного імунітету, але й клітинні елементи природного імунітету, зокрема нейтрофіли.

Cell-free nucleic acids (cfNA) may reach the urine through cell necrosis or apoptosis, active secretion of nucleic acids by healthy and tumor cells of the urinary tract, and transport of circulating nucleic acids (cir-NA) from the blood into primary urine. Even though urinary DNA and RNA are fragmented, they can be used to detect marker sequences. MicroRNAs are also of interest as diagnostic probes. The stability of cfNA in the urine is determined by their structure and packaging into supramolecular complexes and by nuclease activity in the urine. This review summarizes current data on the sources of urinary cfNA, their structural features, diagnostic potential and factors affecting their stability.

The paper analyzes the data of modern scientific literature on the new class of drugs that restore the effect of glucagon-like peptide-1 (GLP-1). According to modern concepts, the pathophysio-logical mechanisms of therapeutic action of GLP-1 analogues in patients with diabetes mellitus (DM) type 2 primarily due to their bene-ficial effects on β-cell dysfunction, whose role in the pathogenesis of DM type 2 until recently been underestimated. The main targets of GLP-1 are pancreatic islets, where it glucose-dependently stimulates production and secretion of insulin and somatostatin secretion and inhibits glucagon secretion. Furthermore, GLP-1 inhibits β-cell destruction and stimulates their proliferation, increasing their number. The article presents information on the main multicenter studies on analogues of human GLP-1, analyzes hypoglycemic effectiveness of incretins and others of their positive effects., В статье проанализированы данные современной научной литературы относительно нового класса препаратов, воссоздающих эффект глюкагоноподобного пептида-1 (ГПП-1). Согласно современным представлениям, патофизиологические механизмы лечебного действия аналогов ГПП-1 у пациентов с сахарным диабетом (СД) 2-го типа прежде всего обусловлены их благоприятным действием на дисфункцию β-клеток, роль которой в патогенезе СД 2-го типа до недавнего времени недооценивалась. Главной мишенью ГПП-1 являются панкреатические островки, где он глюкозозависимо стимулирует продукцию и секрецию инсулина и соматостатина и ингибирует секрецию глюкагона. Кроме того, ГПП-1 препятствует деструкции β-клеток и стимулирует их пролиферацию, увеличивая их количество. В статье приведены данные основных многоцентровых исследований аналогов человеческого ГПП-1, проанализированы сахароснижающая эффективность инкретинов и другие их положительные эффекты., У статті проаналізовано дані сучасної наукової літератури щодо нового класу препаратів, що відтворюють ефект глюкагоноподібного пептиду-1 (ГПП-1). Відповідно до сучасних уявлень, патофізіологічні механізми лікувальної дії аналогів ГПП-1 у пацієнтів із цукровим діабетом (ЦД) 2-го типу передусім обумовлені їх сприятливою дією на дисфункцію β-клітин, роль якої в патогенезі ЦД 2-го типу донедавна недооцінювалася. Головною мішенню ГПП-1 є панкреатичні острівці, де він глюкозозалежно стимулює продукцію та секрецію інсуліну і соматостатину та інгібує секрецію глюкагону. Крім того, ГПП-1 перешкоджає деструкції β-клітин і стимулює їх проліферацію, збільшуючи їх кількість. У статті наведено дані основних багатоцентрових досліджень аналогів людського ГПП-1, проаналізовано цукрознижувальну ефективність інкретинів та інші їх позитивні ефекти.