Kolesnik, Evgeniy A.

Kolesnik E.A., Derkho M.A., Rebezov M.B.

Relevance. Lysosomal cationic proteins (LCP) of granulocytic leukocytes: elastase, cathepsin G, proteinase-3, calgranulin, cathelicidins, defensins, lactoferrin, protegrins are active against viruses, bacteria, fungi, protozoa. The issues of physiological regulatory, immune and pathological effects of LCP and their derivatives – neutrophil (heterophil) extracellular traps (NETs) on pathogens, healthy cellular and tissue structures of the body are noted.Results. The increment of LCP granulocytes is realized by: 1. merocrine type — by degranulation; 2. exocytosis and false degranulation, that is, the process of decationization of lysosomes containing granules of cationic proteins with apocrine or holocrine type of secretion. Decationization implements exocytosis of LCP, extrusion of intact lysosomes from the cell with LCP, and diffusion of LCP through the lysosome membrane. Lysosome degranulation reactions with LCP form phagolysosomes and initiate phagocytosis, lysosome decationization reactions with LCP ensure the formation and functions of NETs. NETs is formed by non-lytic (non-lytic) and lytic (lyzed) pathways in septic and aseptic inflammation, with the ontogenetic development of immune links. NETs is stereotypically formed intravascular during aseptic inflammation, oxidative stress and in a physiological regime, when granulocytes are stimulated by products of oxidative metabolism. Using a cytochemical test with a highly sensitive acid-base bromophenol blue indicator, subcellular and cellular manifestations of the physiological age-related immune activity of cationic proteins accumulated in granulocyte lysosomes were studied on the avian model organism (Aves), and nonspecific adaptive reactions (NAR) of vertebrates in early postnatal ontogenesis were studied. The basis for the formation of NAR is the relationship of groups of leukocytes (lymphocytes, monocytes and granulocytes) with the dynamics of their lysosomal cationic proteins. The method for calculating the level of activity and potential capabilities of granulocytes in phagocytic reactions and in the formation of .NETs includes indices characterizing the directions and intensity of immune reactions of granulocytes, taking into account the processes: 1. degranulation of lysosomes with LCP — in the initiation of the cellular phagocytic link; 2. decationization of lysosomes with LCP — in the initiation of extracellular traps involved in the implementation of the humoral link of immunity.

Kolesnik E., Derho M., Rebezov M.

Abstract. The purpose is to characterize the development of the synthetic activity of polychromatophilic erythroblasts and erythrocytes, in particular the status of the hemoglobin-synthesized function of avian erythroid cells in early postembryonic ontogenesis. Methods. The experimental part of the study was carried out in the conditions poultry farm of “Chebarkul’skaya ptitsa” (Chebarkul district of the Chelyabinsk region, Russia). The whole blood of Hubbard ISA F15 broiler chickens of four age groups (n = 40) was studied: group I – 1-day-old chicks; II – 7-day-old chickens; III – 23-day-old broilers; IV – 42-day-old chickens. The scientific novelty. Functional morpho-densitometric parameters of chromatin of the nucleus and cytoplasm of erythroblasts and red blood cells of birds in postembryonic ontogenesis. Cytophysiological and epigenetic parameters of hemoglobin synthesis by erythroblasts and young erythrocytes are important in elucidating the regulation of the functions of blood cells in normal, adaptation and disorders. Criteria for the synthetic activity of erythroblasts and maturing erythrocytes of animals and humans are optical density as an indicator of the concentration of euchromatin and the metabolic dynamics of the cytoplasm, as well as area as an indicator of the distribution of euchromatin in the stroma of the nucleus and the size’s of the cytoplasm. Results. According to the results of calculation of spectral, morphometric and opticometric values of nuclear chromatin, cytoplasm and their indexed ratios for polychromatophilic erythroblasts and erythrocytes, hemoglobin synthesis in early postembryonic ontogenesis is characterized on a model organism of broiler chickens. According to the results of determining the geometric (n = 30) and optical (n = 300) parameters of euchromatin, heterochromatin of the nucleus and cytoplasm, their ratios and indices were calculated for polychromatophilic erythroblasts, polychromatophilic erythrocytes and mature erythrocytes in peripheral blood smears (n = 158) stained by Pappenheim. Cytochemical reactions reflecting the physiological and biochemical interrelations of erythroblasts and erythrocytes are the basis of a complex morpho-densitometric test of the activity level of hemoglobin synthesis by erythroid cells in early postnatal ontogenesis.

Kolesnik E.A., Derkho M.A., Rebezov M.B.

Relevance. An overview is presented of the patterns of appearance, physiological and clinical significance of shadows of cells, a comparative experimental cytomorphological analysis using the example of polychromatophilic erythroblasts (PolyErythro), shadows of erythroblast (ShadErythro) and undifferentiated shadows cells of in peripheral blood smears of chickens Gallus gallus L.Methods. The studied individuals were divided into four groups (n = 40) depending on the age of the chickens (Postembryonalis — P1, P7, P23, and P42): 1, 7, 23, and 42 days of postembryonic ontogenesis) of a factory herd. ShadErythro apoptotic changes, undifferentiated cell shadows were characterized. Were calculated and characterized the morphophysiological parameters of PolyErythro, ShadErythro and unidentified shadows of cells by high-resolution color microphotographs in blood smears of birds (Pappenheim stained); a total of 158 (n = 158) microphotographs were analyzed.Results. The structured ShadErythro were able in chromatinolized. Unidentified shadows of cells were in a state of cytolysis, fragmentation of the cytoplasm, karyopycnosis and karyorrhexis, chromatin fragmentation, and karyolysis were noted. Structured of ShadErythro and undifferentiated shadows of cells were light purple with a reddish tint. Significant difference in the area of ShadErythro (Sshadow, μm2) was recorded in the ascending area of PolyErythro (Scell, μm2), within 25% (p ≤ 0.001), an increase in the percentage of difference between the minimum and maximum values of the ShadErythro area (Pshad.-cell-differ.-min-max,%) of Pcell-differ.-min-max,% PolyErythro was 36% (p ≤ 0.001). The difference in the percentage difference between the minimum and maximum values of the area of the nucleus of erythroblasts (Pnucleus-differ.-min-max) and the shadows of the nucleus of erythroblasts (Pshad.-nucle.-differ.-min-max) is significantly less than the analogous values (noted above) for cells and shadows erythroblasts, for example, Pnucleus-differ.-min-max exceeded Pshad.-nucle.-differ.-min-max by only 21% (p ≤ 0.001). The percentage of difference between the minimum and maximum values of the nuclear-cytoplasmic ratio ShadErythro (PN/C Ratio-shad.-cell) significantly exceeded the analogous value of PolyErythro (PN/C Ratio) within 64% (p ≤ 0.001). At the same time, the nuclear-cytoplasmic ratio of PolyErythro (N/C Ratio) and ShadErythro (N/C Ratio-shad.-cell) had a relatively minimal difference. The ShadErythro nuclear-cytoplasmic ratio (N/C Ratio-shad.-cell) exceeded the PolyErythro nuclear-cytoplasmic ratio (N/C Ratio) by only 12% (p ≤ 0.001). ShadErythro is maximally represented in chickens aged P1 — 40% and up to 30% (p ≤ 0.05) in P23. In P7, the amount of ShadErythro decreased by 30% (p ≤ 0.05), in P42, a decrease in ShadErythro to 20% (p ≤ 0.05) was recorded. The described dynamics of ShadErythro is due to physiological adaptive processes in the body of broiler chickens in the early period of postembryonic ontogenesis. The data obtained by us from a comprehensive comparative morphophysiological analysis of forms of degeneration of blood cells will be useful in the clinical diagnosis of animal pathologies, experimental and theoretical studies of cellular adaptations of the functional blood system and the whole organism.

Kharisma V.D., Ansori A.N., Antonius Y., Rosadi I., Murtadlo A.A., Jakhmola V., Rebezov M., Maksimiuk N., Kolesnik E., Burkov P., Derkho M., Scherbakov P., Ullah M.E., Sucipto T.H., Purnobasuki H.

Context: Mangosteen (Garcinia mangostana L.) is used in traditional medicine as an antibacterial, antioxidant, and anti-inflammatory. Aims: To determine the molecular mechanism and potential of garciniaxanthone derivate compounds from G. mangostana as SARS-CoV-2 antiviral and prevent cytokine storm through in silico approach. Methods: Ligand and protein samples were obtained from databases such as PubChem and Protein Databank, then drug-likeness analysis using Lipinski, Ghose, Veber, Egan, and Muege rules on SwissADME server, prediction of antiviral probability through PASSOnline server. Furthermore, molecular docking simulation with PyRx v1.0 software (Scripps Research, USA) with an academic license, identification of interactions and chemical bond positions of ligands on the target by PoseView server, 3D visualization of PyMOLv.2.5.2 software (Schrödinger, Inc., USA) with an academic license, molecular dynamics simulation for molecular stability prediction by CABS-flex v2.0 server, target prediction of antiviral candidate compounds by SwissTargetPrediction server, pathway analysis through STRING v11.5 database, and toxicity by ProTox-II server were used. Results: Garciniaxanthone C from G. mangostana was found to be a drug-like molecule with low toxicity. This can be a candidate for SARS-Cov-2 antiviral through inhibitor activity on two viral enzymes consisting of Mpro and replicase with a binding affinity value that is more negative than other garciniaxanthone derivates and is stable. Garciniaxanthone C is predicted to bind and inhibit pro-inflammatory proteins that trigger cytokine storms, such as NFKB1 and PTGS2. Conclusions: Garciniaxanthone derivative compounds from G. mangostana may be candidates for SARS-CoV-2 antiviral and preventing cytokine storm through garciniaxanthone C activity.

Е.А. К., Н.В. М., Ю.В. К.

Введение. В статье рассмотрены особенности применения кейс-технологий в образовательном процессе и обоснована необходимость применения кейс-заданий при обучении медико-биологическим дисциплинам будущих учителей физической культуры в контексте формирования информационно-аналитической компетенции и представлений об организме человека как едином целом в неразрывной взаимосвязи с факторами окружающей среды. Материалы и методы. В качестве основных методов исследования выступили анализ литературы по проблемам применения методов активного обучения, в том числе кейс-технологий, формирования информационно-аналитической компетенции и подготовки учителей физической культуры. Диагностика проводилась по авторской методике. Результаты. Исследование уровня сформированности информационно-аналитической компетенции у студентов первого курса, обучающихся по профилям физкультурной направленности, показало, что у обучающихся компетенция сформирована на низком уровне. В качестве инструмента для решения проблемы предложено использовать кейс-задания в процессе обучения студентов дисциплинам медико-биологической направленности. Обсуждение. Высказано предположение, что кейс-задачи по медико-биологическим дисциплинам будут способствовать формированию информационно-аналитической компетенции в комплексном и самостоятельном освоении здоровьесберегающих технологий будущими учителями физической культуры на всех этапах обучения. Заключение. Сделан вывод о необходимости формирования у студентов профилей физкультурной направленности информационно-аналитической компетенции и применения физиологического холистического принципа, то есть представления об организме человека как единой системе в процессе обучения, выполнения кейс-заданий по медико-биологическим дисциплинам. Ключевые слова: кейс-технологии; информационно-аналитическая компетенция; физическая культура; медико-биологические дисциплины; холистический принцип. Основные положения: – проанализированы особенности применения кейс-технологий в образовательном процессе; – проведена диагностика уровня сформированности информационно-аналитической компетенции студентов-будущих учителей физической культуры; – предложено применять кейс-задачи при обучении студентов дисциплинам медико-биологического профиля для фор

Kolesnik E.A., Derkho M.A.

The work is devoted to the study of literature data (humane and veterinary medicine) and the practical analysis of artifacts formed elements and plasma in peripheral blood smears of birds in a model of broiler chickens Gallus gallus L. of early postnatal ontogenesis. The age of the studied clinically healthy chicks and young hens was: day 1, day 7, day 23 and day 42 (n = 40). We studied 158 (n = 158) high-resolution color micrographs of the fields of view, in blood smears stained according to Pappenheim. As a result, single artifacts of avian erythrocytes were identified: cytoplasmic vacuoles of various pattern character, scalloped «bitten» edges of cells. Artifacts of blood plasma were found: pericellular and adhesioned on the cell surface of colored coagulated granularity. In some cases, this granularity imitated the toxic forms of granulocytes and agranulocytes in the peripheral blood of birds. It is necessary to distinguish artifacts of cells and plasma in peripheral blood smears from adaptive changes and symptoms of infectious, invasive and non-infectious diseases.

Kolesnik E., Derkho M., Strizhikov V., Strizhikova S., Sereda T., Gizatullina F., Rebezov M.

The present research was carried out in an attempt to identify the problems associated with the morphofunctional analysis of vertebrate animal’s leukocyte blood cells. For this blood samples of four age groups chickens (Gallus gallus L.) were collected and analysis was carried out as per the recommendations of the International Council for Standardization in Hematology for the identifying morpho-physiological characteristics of leukocytes of peripheral blood of birds. Results of the current study were based on the sample of early postnatal ontogenesis based on the analysis of high-resolution color microphotographs taken by the method of light-optical microscopy. Results of the current study revealed that the cells have the well-designed of all granular leukocytes typical "eosinophilic" nucleus with two segments. Sometimes cells contain polysegmental nuclei that are eccentrically located. Further, in contrast to heterophils, the eosinophilic nucleus has well-expressed contours, formed by irregularly shaped chromatin blocks with an optically denser structure. There are also eosinophils with stick-nuclei. The smallest blood granulocytes of the birds are basophils. Basophils differ in structure slightly basophilic color cytoplasm with intensively basophilic granules of rounded or other forms of different size. The sizes of granules in basophils are smaller than the other types of granular leukocytes. Thus, based on the analysis of qualitative color microphotographs of white blood cells of chickens of neonatal ontogenesis performed by optical microscopy, the differential morpho-physiological markers of leukocytic cells of peripheral blood of birds were marked and characterized.

In accordance with the recommendations of The International Council for Standardization in Haematology (ICSH, https://icsh.org/), this article describes the morphophysiological characteristics of the precursors and mature erythroid cells in the early period of postnatal development of birds (Gallus gallus L.) including calculation of the surface area of these cells (S, μm2 , X±SEM). Depending on cell shape, cytoplasm color, and chromatin organization in nucleus, the following types are distinguished: basophilic erythroblasts (69.60±4.01 μm2 , p≤0.05), polychromatophilic erythroblasts (65.42±2.49 μm2 , p≤0.05), and oxyphilic erythroblasts (71.10±4.43 μm2). Formation of cell pool is characteristic for erythropoiesis in birds due to mitotic proliferation of basophilic erythroblasts. There are often proerythroblasts and polychromatophilic erythroblasts. The nucleus of a polychromatophilic proerythroblast contains a large number of histone proteins; therefore, it has an intensely basophilic color with a pronounced oxyphilic hue (proteinrelated oxyphilia). The accumulation of hemoglobin in the protoplasm of these cells contributes to the gradual transition of the basophilic staining of cytoplasm to the oxyphilic one which is typical for mature red blood cells (73.95±2.10 μm2 , p≤0.05). Cell shape and the structure of erythroblast nucleus approaches to these of mature red blood cells.

Kolesnik E.A., Derkho M.A., Lebedeva I.A.

М.А. Д., Е.А. К.

Целью работы была систематизация данных о соотношении морфологических и гормональных показателей периферической крови в раннем постнатальном онтогенезе цыплят-бройлеров в условиях промышленных технологий с использованием комплексных гемато-гормональных индексов. Из промышленного стада были сформированы четыре группы цыплят-бройлеров кросса Hubbard F15, по 10 цыплят в каждой, у которых в возрасте 1, 7, 23 и 42 суток определяли показатели клеточного состава, содержание АКТГ и кортизола в периферичской крови. Полученные данные сопоставляли с динамикой гемато-гормональных индексов, составленных на основе показателей количества эритроцитов (Э), гетерофилов (Г), лимфоцитов (Л) и содержания кортизола (К) в крови: эритроцитарно-лимфоцитарный (ЭЛИ, Э/Л), эритроцитарно-гетерофильный (ЭГИ, Э/Г), эритроцитарно-лимфоцитарно-кортизольный (ЭЛКИ, [(Э/Л)*К)]/100, эритроцитарно-гетерофильно-кортизольный (ЭГКИ, [(Э/Г)*К)]/100 и интегральный индекс эритроцитов-гетерофилов-лимфоцитов и кортизола (ИИЭГЛК, [((Э+Г)/Л)*К)]/100. Индекс ЭЛИ поcле достижения наибольшего снижения в 7 сут. (P<0,001 по отношению к 1 сут.), увеличивался к 42 сут. (P<0,001); ЭГИ существенно возрастал к 7 сут., к 23 сут. снижался (P<0,001) и к периоду 42 сут. вновь увеличивался (P<0,001). Для ЭЛКИ отмечено снижение в возрасте 1-7 сут. (P<0,001) с последующим снижением к 23 сут. (P<0,001) и небольшим возрастанием к 42 сут. Отмечен существенный рост ЭГКИ в период от 1 до 7 сут. (P<0,001). Индекс ИИЭГЛК характеризовался снижением в период от 1 до 7 сут. (P<0,001) и значительным приростом в период 7-23 сут. (P<0,001), коррелирующим со снижением сохранности. Заключили, что в периоды раннего постнатального онтогенеза формируется совокупный адаптационный процесс, повторяя в целом реакции в костном мозге, лимфоидных органах и периферической крови, которые составляют физиологическую основу развития общего адаптационного синдрома. При этом анаболический характер обмена веществ и неспецифических адаптационных реакций опосредует в раннем постнатальном онтогенезе цыплят-бройлеров формирование функциональной системы адаптационного гомеостаза. The aim of the work was to systematize data on the relationships of morphological and hormonal parameters of peripheral blood in the early postnatal ontogenesis of broiler chickens in industrial technology using complex hemato-hormonal indices. In order to characterize their age dynamics, four groups of broiler chickens Hubbard F15 were formed from the industrial herd, 10 chickens each. At the age of 1, 7, 23 and 42 days the parameters of the cellular composition, the content of ACTH and cortisol in the peripheral blood were determined. The obtained data were compared with the dynamics of hemato-hormonal indices, compiled on the basis of the numbers of erythrocytes (E), heterophiles (H), lymphocytes (L) and cortisol (C) in blood: erythrocyte-lymphocyte (ELI, E/L), erythrocyte-heterophilic (EHI, E/H), erythrocyte-lymphocytic-cortisol (ECI, [(E/L)*C]/100), erythrocyte-heterophilic-cortisol (EHCI, [(E/H)*C]/100) and the integral index of erythrocytes-heterophyll-lymphocytes and cortisol (EHLCI, [((E+H)/L)*C)]/100). The ELI index after reaching the greatest decrease at 7 d (P<0.001 in relation to 1 d) increased to 42 d (P<0.001), EHI increased significantly at 7 d, decreased at 23 d (P <0.001) and increased again in a period to 42 d (P<0.001), with a significant increase in EHCI in the period from 1 to 7 days (P<0.001). The index of EHLCI showed a significant decrease in the periods from 1 to 7 days (P<0.001), with a significant rise at the age 7-23 d (P<0.001) correlating with a decrease in survival. Concluded that in the periods of early postnatal ontogeny, a cumulative adaptation process is formed, repeating in general, the reactions in the bone marrow, lymphoid organs and peripheral blood that constitute the physiological basis for the development of the general adaptation syndrome. At the same time, the anabolic character of the metabolism and nonspecific adaptation reactions mediates the formation of a functional system of adaptive homeostasis in early postnatal ontogenesis of broiler chickens

Kolesnik E.A., Derkho M.A.

Kolesnik E.A., Derkho M.A.

Kolesnik E.A., Derkho M.A.

Kolesnik E.A., Derkho M.A.

Kolesnik E.A., Derkho M.A., Rebezov M.B.

Relevance. Lysosomal cationic proteins (LCP) of granulocytic leukocytes: elastase, cathepsin G, proteinase-3, calgranulin, cathelicidins, defensins, lactoferrin, protegrins are active against viruses, bacteria, fungi, protozoa. The issues of physiological regulatory, immune and pathological effects of LCP and their derivatives – neutrophil (heterophil) extracellular traps (NETs) on pathogens, healthy cellular and tissue structures of the body are noted.Results. The increment of LCP granulocytes is realized by: 1. merocrine type — by degranulation; 2. exocytosis and false degranulation, that is, the process of decationization of lysosomes containing granules of cationic proteins with apocrine or holocrine type of secretion. Decationization implements exocytosis of LCP, extrusion of intact lysosomes from the cell with LCP, and diffusion of LCP through the lysosome membrane. Lysosome degranulation reactions with LCP form phagolysosomes and initiate phagocytosis, lysosome decationization reactions with LCP ensure the formation and functions of NETs. NETs is formed by non-lytic (non-lytic) and lytic (lyzed) pathways in septic and aseptic inflammation, with the ontogenetic development of immune links. NETs is stereotypically formed intravascular during aseptic inflammation, oxidative stress and in a physiological regime, when granulocytes are stimulated by products of oxidative metabolism. Using a cytochemical test with a highly sensitive acid-base bromophenol blue indicator, subcellular and cellular manifestations of the physiological age-related immune activity of cationic proteins accumulated in granulocyte lysosomes were studied on the avian model organism (Aves), and nonspecific adaptive reactions (NAR) of vertebrates in early postnatal ontogenesis were studied. The basis for the formation of NAR is the relationship of groups of leukocytes (lymphocytes, monocytes and granulocytes) with the dynamics of their lysosomal cationic proteins. The method for calculating the level of activity and potential capabilities of granulocytes in phagocytic reactions and in the formation of .NETs includes indices characterizing the directions and intensity of immune reactions of granulocytes, taking into account the processes: 1. degranulation of lysosomes with LCP — in the initiation of the cellular phagocytic link; 2. decationization of lysosomes with LCP — in the initiation of extracellular traps involved in the implementation of the humoral link of immunity.

Lestari D., Murtini S., Ulupi N., Gunawan A., Khaerunnisa I., Anggraeni A., Sumantri C.

Background and Aim: The IPB-D2 chicken, a potential disease-resistant local Indonesian line, has been developed to improve poultry health and productivity for small-scale farmers. This study aimed to characterize the disease resistance traits of IPB-D2 chickens by analyzing key immunological parameters, including immunoglobulin Y (IgY) concentration, Newcastle disease (ND) antibody titers, and leukocyte profiles. Materials and Methods: A total of 100 IPB-D2 chickens were studied. Blood samples from 21-week-old chickens were collected for analysis of IgY concentrations using indirect enzyme-linked immunosorbent assay, ND antibody titers using the hemagglutination inhibition test, and leukocyte profiling using the Giemsa staining method. Correlations between parameters were determined using Pearson’s correlation analysis in RStudio. Results: The findings revealed that IPB-D2 chickens exhibited higher IgY concentrations, leukocyte counts, heterophil levels, and monocyte counts compared to their IPB-D1 progenitors. However, ND antibody titers in IPB-D2 chickens were lower than the protective threshold. Significant positive correlations were observed between leukocytes and lymphocytes, and negative correlations were identified between heterophils and lymphocytes. Conclusion: IPB-D2 chickens demonstrated a well-balanced immune system with enhanced non-specific (leukocyte and heterophil) and specific (IgY) immune responses. These traits highlight their potential as a disease-resistant poultry line. Future research should focus on molecular selection through transcriptomic analysis to identify candidate genes associated with disease resistance traits, enabling further genetic improvements. Keywords: IPB-D2 chicken, immunoglobulin Y concentration, ND antibody titer, leukocyte profile.

Bekshenova A.M., Alexandrova S.S.

Relevance. Contemporary poultry farming practices, often involving high stocking densities, can significantly alter the environmental conditions and dietary regimens experienced by chickens. These changes can have detrimental effects on the neurological and cardiovascular health of broiler chickens, ultimately impacting their meat yield. In response to these challenges, poultry producers are increasingly turning to pharmaceutical interventions. These medications aim to regulate cardiovascular function and metabolism, promote bone regeneration, and mitigate stress in birds. The purpose of the work is to assess the effect of “Butamax 200” and “Propranolol” on the productive qualities of broiler chickens.Results. The dynamics of live weight and growth rates during the rearing of broiler chickens indicate the effectiveness of the use of “Butaphosphane 200” and “Propranolol” preparations. The experimental groups differed in lower feed conversion compared to the control group by 23.11% and 28.3%, respectively. The efficiency index of broiler chicken rearing was significantly higher in the experimental groups. The EIPB of the 1st experimental group was 316 units, and the 2nd experimental group was 352 units, which exceeded the values of the control analogues by 114 and 150 units, respectively.

Parikesit A.A.

This chapter provides a comprehensive overview of the role of genomics in accelerating the drug development process. It highlights the significant advancements in large-scale population collections and whole genome sequencing approaches, which have significantly influenced multiple stages across the lifespan of a drug discovery program. The chapter emphasizes the increasing importance of genomic data in target identification, target prioritization, and tractability, as well as predicting outcomes from pharmacological perturbations. It also discusses integrating genomic technologies with other omics approaches, such as proteomics and metabolomics, to understand disease mechanisms better and identify potential therapeutic targets. The integration of bioinformatics is pivotal in harnessing the power of genomic data for drug discovery endeavors. This chapter delves into the computational approaches and analytical frameworks that enable extracting meaningful insights from vast genomic datasets. Furthermore, it examines the contributions of bioinformatics in the field of pharmacogenomics.

Burkov P.V., Derkho M.A., Rebezov M.B., Shcherbakov P.N., Derkho A.O.

Relevance. The article presents data on the toxicological assessment of the safety of the specific immunobiostimulant of antigen-directed action “Transfer Factor” in a form ready for intramuscular administration and obtained from the blood of hyperimmunized donors on laboratory animals in an acute experiment.Methods. The object of the study was the finished form of “Transfer Factor”, the acute toxicity of which was determined in models on laboratory animals (nonlinear mice, nonlinear rats).Results. It was found that after administration of the drug to mice intramuscularly (0.25 ml/head) and intraperitoneally (0.5 ml/head), to rats intramuscularly (2.5 ml/head) and intraperitoneally (2.5 ml/head), muscle tremor is observed, disappearing after 15–20 minutes. Intraperitoneal administration of the drug to mice and rats in dosages of 0.75 ml/head and 3.75 ml/head, respectively, determines the presence of tremor lasting 40 minutes. During the 14-day observation period, the body weight of mice and rats increases by 10.15-14.25 and 11.84-17.67%. Autopsy of animals does not reveal visible changes in the location of organs and fluid in the abdominal and pleural cavities. However, some pathological changes in color, consistency and size are found in the lungs, heart, spleen and liver with intramuscular administration of Transfer Factor to mice and rats at a dose of 0.25 ml/head and 2.5 ml/head, intraperitoneal administration to mice at a dose of 0.5 ml/head and 0.75 ml/head and to rats 2.5 ml/head and 3.75 ml/head. Based on the totality of changes in the vital signs of laboratory animals, body weight and macroscopic characteristics of internal organs according to GOST 12.1.007-76, the drug “Transfer Factor” can be classified as hazard class IV “Low-hazard substances”.

Astuti N.P., Manuaba I.B., Jawi I.M., Putra A.A., Wiradana P.A., Widhiantara I.G., Permatasari A.A., Ansori A.N., Kharisma V.D.

Traditional herbal remedies have an important role in human health. Empirically, Blumea balsamifera is often used as a traditional beverage to alleviate fever symptoms, lower cholesterol levels, and maintain body immunity. The purpose of this study was to discover the phytoconstituent profile that contributes to the anti-diabetic properties of B. balsamifera leaf extract (BBLE) using in silico approaches.LCMS/MS was used to identify the constituent profile of BBLE, and the ability of these compounds against diabetes-related proteins was analyzed computationally.Three proteins related to diabetes are NF-KB p65, GLP-1, and DPP-4. A total of 18 compounds were successfully identified through LCMS/MS, including 4 compounds known to be flavonoid derivatives and can be used as markers of BBLE. Pheophorbide A and 1,1-Cyclopentanediacetic acid were reported for the first time to inhibit the NF-KB p65, GLP-1, and DPP-4 proteins in docking simulation studies. Based on these findings, it can be confirmed that the bioactive compounds in BBLE show strong inhibitory potential against anti-diabetic proteins.

Pronina G.I., Bubunets E.V., Fedotova A.R., Zhelankin R.V.

Despite the great interest in the use of probiotics to strengthen the immune characteristics of animals, there is not enough work on this issue, so we set ourselves the goal of studying the physiological state and immune status of sterlet grown in the ultrasound, which was fed with the addition of the probiotic “Enzimsporin”. A control group and three experimental groups were selected for the experiment, individuals were selected according to the principle of analogues of 50 in each group. For 60 days, the sterlet of the experimental groups was fed with the addition of probiotic in dosages of 1, 1.5 and 2 g/kg. Based on the combination of high linear-weight indicators and survival, we evaluated the hematological characteristics of individuals in the group that received a probiotic dosage of 1 g/kg of feed. In this experimental group, the number of erythrocytes and leukocytes in the blood did not significantly differ. When using a probiotic in a fish leukogram, the proportion of immature myeloid cells, including rod-shaped neutrophils, decreases from 3.1% to 0.6%. At the same time, there were no significant differences in the proportion of mature segmented neutrophils. The use of probiotics significantly increases the amount of lysosomal cationic protein in neutrophils to a value of 1.94 units of blood compared to the control group, whose values were 1.05 units, which indicates a favorable state of cellular immunity and a high immune status of fish in this group as a whole.

Kolesnik E., Derho M., Rebezov M.

Abstract. The purpose is to characterize the development of the synthetic activity of polychromatophilic erythroblasts and erythrocytes, in particular the status of the hemoglobin-synthesized function of avian erythroid cells in early postembryonic ontogenesis. Methods. The experimental part of the study was carried out in the conditions poultry farm of “Chebarkul’skaya ptitsa” (Chebarkul district of the Chelyabinsk region, Russia). The whole blood of Hubbard ISA F15 broiler chickens of four age groups (n = 40) was studied: group I – 1-day-old chicks; II – 7-day-old chickens; III – 23-day-old broilers; IV – 42-day-old chickens. The scientific novelty. Functional morpho-densitometric parameters of chromatin of the nucleus and cytoplasm of erythroblasts and red blood cells of birds in postembryonic ontogenesis. Cytophysiological and epigenetic parameters of hemoglobin synthesis by erythroblasts and young erythrocytes are important in elucidating the regulation of the functions of blood cells in normal, adaptation and disorders. Criteria for the synthetic activity of erythroblasts and maturing erythrocytes of animals and humans are optical density as an indicator of the concentration of euchromatin and the metabolic dynamics of the cytoplasm, as well as area as an indicator of the distribution of euchromatin in the stroma of the nucleus and the size’s of the cytoplasm. Results. According to the results of calculation of spectral, morphometric and opticometric values of nuclear chromatin, cytoplasm and their indexed ratios for polychromatophilic erythroblasts and erythrocytes, hemoglobin synthesis in early postembryonic ontogenesis is characterized on a model organism of broiler chickens. According to the results of determining the geometric (n = 30) and optical (n = 300) parameters of euchromatin, heterochromatin of the nucleus and cytoplasm, their ratios and indices were calculated for polychromatophilic erythroblasts, polychromatophilic erythrocytes and mature erythrocytes in peripheral blood smears (n = 158) stained by Pappenheim. Cytochemical reactions reflecting the physiological and biochemical interrelations of erythroblasts and erythrocytes are the basis of a complex morpho-densitometric test of the activity level of hemoglobin synthesis by erythroid cells in early postnatal ontogenesis.

Kolesnik E.A., Derkho M.A., Rebezov M.B.

Relevance. An overview is presented of the patterns of appearance, physiological and clinical significance of shadows of cells, a comparative experimental cytomorphological analysis using the example of polychromatophilic erythroblasts (PolyErythro), shadows of erythroblast (ShadErythro) and undifferentiated shadows cells of in peripheral blood smears of chickens Gallus gallus L.Methods. The studied individuals were divided into four groups (n = 40) depending on the age of the chickens (Postembryonalis — P1, P7, P23, and P42): 1, 7, 23, and 42 days of postembryonic ontogenesis) of a factory herd. ShadErythro apoptotic changes, undifferentiated cell shadows were characterized. Were calculated and characterized the morphophysiological parameters of PolyErythro, ShadErythro and unidentified shadows of cells by high-resolution color microphotographs in blood smears of birds (Pappenheim stained); a total of 158 (n = 158) microphotographs were analyzed.Results. The structured ShadErythro were able in chromatinolized. Unidentified shadows of cells were in a state of cytolysis, fragmentation of the cytoplasm, karyopycnosis and karyorrhexis, chromatin fragmentation, and karyolysis were noted. Structured of ShadErythro and undifferentiated shadows of cells were light purple with a reddish tint. Significant difference in the area of ShadErythro (Sshadow, μm2) was recorded in the ascending area of PolyErythro (Scell, μm2), within 25% (p ≤ 0.001), an increase in the percentage of difference between the minimum and maximum values of the ShadErythro area (Pshad.-cell-differ.-min-max,%) of Pcell-differ.-min-max,% PolyErythro was 36% (p ≤ 0.001). The difference in the percentage difference between the minimum and maximum values of the area of the nucleus of erythroblasts (Pnucleus-differ.-min-max) and the shadows of the nucleus of erythroblasts (Pshad.-nucle.-differ.-min-max) is significantly less than the analogous values (noted above) for cells and shadows erythroblasts, for example, Pnucleus-differ.-min-max exceeded Pshad.-nucle.-differ.-min-max by only 21% (p ≤ 0.001). The percentage of difference between the minimum and maximum values of the nuclear-cytoplasmic ratio ShadErythro (PN/C Ratio-shad.-cell) significantly exceeded the analogous value of PolyErythro (PN/C Ratio) within 64% (p ≤ 0.001). At the same time, the nuclear-cytoplasmic ratio of PolyErythro (N/C Ratio) and ShadErythro (N/C Ratio-shad.-cell) had a relatively minimal difference. The ShadErythro nuclear-cytoplasmic ratio (N/C Ratio-shad.-cell) exceeded the PolyErythro nuclear-cytoplasmic ratio (N/C Ratio) by only 12% (p ≤ 0.001). ShadErythro is maximally represented in chickens aged P1 — 40% and up to 30% (p ≤ 0.05) in P23. In P7, the amount of ShadErythro decreased by 30% (p ≤ 0.05), in P42, a decrease in ShadErythro to 20% (p ≤ 0.05) was recorded. The described dynamics of ShadErythro is due to physiological adaptive processes in the body of broiler chickens in the early period of postembryonic ontogenesis. The data obtained by us from a comprehensive comparative morphophysiological analysis of forms of degeneration of blood cells will be useful in the clinical diagnosis of animal pathologies, experimental and theoretical studies of cellular adaptations of the functional blood system and the whole organism.

Vdovin V.M., Suzopov E.V., Shakhmatov I.I., Bobrov I.P., Orekhov D.A., Dolgatov A.Y., Momot A.P.

Background. In our previously published studies, we demonstrated a high hemostatic activity of a low dose of exogenous fibrin monomer during its systemic administration in a model of dosed liver injury with preliminary inhibition of platelet aggregation. However, the analysis of platelet involvement in the mechanisms of local fibrin formation has not been analyzed. The aim of the study. To conduct a comparative analysis of the cellular composition of venous and wound blood, as well as blood in the wound vessels to assess the contribution of platelets to the hemostatic effect of exogenously administered fibrin monomers in dosed liver injury under conditions of pharmacologically determined thrombocytopathy. Methods. In a model of dosed liver injury in rabbits after inhibition of platelet aggregation by  acetylsalicylic acid in combination with clopidogrel, the effect of the administration of fibrin monomer was evaluated in comparison with the use of tranexamic acid. We studied the number of platelets in venous and wound blood smears, as well as in the contents of wound vessels. Results. It has been established that with the systemic administration of exogenous fibrin monomer, the number of platelets in wound blood smears decreases by 17.2 % in comparison with free circulating venous blood. Platelets in wound blood form aggregates and are in an activated state. In the wound vessels, the number of these cells was maximum (150 per lower field) compared with the number of platelets in the placebo and tranexamic acid groups (55 and 84 per lower field, respectively). Also in the wound blood, erythrocytes with altered forms (echinocytes, schistocytes, stomatocytes and ovalocytes) were found. Conclusion. Systemic administration of exogenous fibrin monomer affects the redistribution of platelets between the systemic circulation, wound vessels and wound blood, determining its hemostatic effect and local wound fibrin formation in dosed liver injury. The presence of receptor-mediated platelets recruitment due to fibrin monomer in the wound vessels with the  participation of damaged erythrocytes is assumed.

Gorelik O.V., Harlap S.Y., Struin A.A., Belookov A.A., Belookova O.V., Chuhutin E.V.

   The main costs of livestock production, including poultry farming, are related with the feed. Therefore it is very relevant to increase digestibility of feed, apply compound feeds from cheap raw materials produced domestically without reducing of nutritional value, and fully provide the poultry with substances necessary for their normal life. It is possible to achieve with the application of new feed additives, including biologically active substances. As a result of the research it was found that all broiler chickens showed high parameters of body condition during the boilers growing. The chickens from the II experimental group, which received the additive “Ares” in amount of 0.6 g/kg of feed, showed the highest live weight gain. Lower values were observed among the chickens of I and III experimental groups. However, in terms of live weight gain at the age of 36 days, these chickens exceeded the normal value parameters of the cross and the results obtained in the control group. The increase in the average daily weight gains in the first and third groups was observed throughout the entire period of fattening, and a slight decrease was observed in the second group in the period from 24th to 31th day with a sharp surge of weight gain in the last period from 31th to 36th day.

Derkho M.A., Sajfutdinova L.N., Strizhikov V.K., Strizhikova S.V., Ponomaryova T.A.

The influence of hen layer density on the variability of the number of red blood cells, heterophiles and lymphocytes in the blood, the secretory activity of adrenal glands, estimated by the level of corti-costerone and cortisol, as well as the presence of interrelations between hormones and blood cells by calculating complex indices, were studied. Chickens, as the research object, were kept in cages, under conditions of standard layer density and increased by 1.5 and 2.0 times. We found that chickens adapt to an increase in layer density by one and a half times, pro-vided that egg production decreases to 33.33%; two times exceed of the regulatory requirements for laying does not correspond to the adaptive abilities of birds. Depending on the level of layer density excess (stress factor) in chicken blood, the concentration of corticosterone and cortisol increases, determining a decrease in the number of lymphocytes and an in-crease in heterophiles against the background of the preservation of red blood cells, reflecting the “energy price” of adaptation. Corticosterone af-fects the relationship of red blood cells with lymphocytes and heterophiles, determining the variability of the values of the indices reflecting the ratio of red blood cells and lymphocytes (ISEL), red blood cells and hetero-philes (ISEG), red blood cells, lymphocytes and corticosterone (ISELC), red blood cells, heterophiles and corticosterone (ISEGC) and the integral index of red blood cells-heterophiles-lymphocytes and corticosterone (IIEGLC).

Saken A.K., Fatkullin R.R., Mustafin M.K., Daribaeva S.A.

Abstract The direct consequences of a complex of structural changes in the form of intensification, industrialization and geographic conditioning of heavy metals that can be utilized through soil destruction often lead to environmental problems and cyclically affect the food chain. In this regard, one of the most important areas of research in the field of poultry farming with the identification of chemical waste is the protection of environmental objects, namely the soil as an object characterized by the accumulative properties of heavy metals, which entail a number of consequences. The normal vital activity of a living organism requires an optimal amount of minerals with trace elements and heavy metals, which creates the need to monitor this cyclic system. The paper presents the results of a study of heavy metals from soil samples, samples of plant food, as well as poultry dung and the effect of their excess or deficiency on the biochemical parameters of the body of chickens. Pursuing the relevance of the content of heavy metals in objects eliminated by the body in the form of excrement, containing substances permeable to the entire body, today it is undergoing intensive study, since plant zones are an integral part of the food chain, accompanied by immuno-biochemical significance for the healthy state and development of birds.

Kolesnik E., Derkho M., Strizhikov V., Strizhikova S., Sereda T., Gizatullina F., Rebezov M.

The present research was carried out in an attempt to identify the problems associated with the morphofunctional analysis of vertebrate animal’s leukocyte blood cells. For this blood samples of four age groups chickens (Gallus gallus L.) were collected and analysis was carried out as per the recommendations of the International Council for Standardization in Hematology for the identifying morpho-physiological characteristics of leukocytes of peripheral blood of birds. Results of the current study were based on the sample of early postnatal ontogenesis based on the analysis of high-resolution color microphotographs taken by the method of light-optical microscopy. Results of the current study revealed that the cells have the well-designed of all granular leukocytes typical "eosinophilic" nucleus with two segments. Sometimes cells contain polysegmental nuclei that are eccentrically located. Further, in contrast to heterophils, the eosinophilic nucleus has well-expressed contours, formed by irregularly shaped chromatin blocks with an optically denser structure. There are also eosinophils with stick-nuclei. The smallest blood granulocytes of the birds are basophils. Basophils differ in structure slightly basophilic color cytoplasm with intensively basophilic granules of rounded or other forms of different size. The sizes of granules in basophils are smaller than the other types of granular leukocytes. Thus, based on the analysis of qualitative color microphotographs of white blood cells of chickens of neonatal ontogenesis performed by optical microscopy, the differential morpho-physiological markers of leukocytic cells of peripheral blood of birds were marked and characterized.

Renganathan B., Moore A.S., Yeo W., Petruncio A., Ackerman D., Wiegel A., Pasolli H.A., Xu C.S., Hess H.F., Serpinskaya A.S., Zhang H.F., Lippincott-Schwartz J., Gelfand V.I.

AbstractVimentin intermediate filaments (VIFs) form complex, tight-packed networks; due to this density, traditional ensemble labeling and imaging approaches cannot accurately discern single filament behavior. To address this, we introduce a sparse vimentin-SunTag labeling strategy to unambiguously visualize individual filament dynamics. This technique confirmed known long-range dynein and kinesin transport of peripheral VIFs and uncovered extensive bidirectional VIF motion within the perinuclear vimentin network, a region we had thought too densely bundled to permit such motility. To examine the nanoscale organization of perinuclear vimentin, we acquired high-resolution electron microscopy volumes of a vitreously frozen cell and reconstructed VIFs and microtubules within a ∼50 µm3window. Of 583 VIFs identified, most were integrated into long, semi-coherent bundles that fluctuated in width and filament packing density. Unexpectedly, VIFs displayed minimal local co-alignment with microtubules, save for sporadic cross-over sites that we predict facilitate cytoskeletal crosstalk. Overall, this work demonstrates single VIF dynamics and organization in the cellular milieu for the first timeSummarySingle-particle tracking demonstrates that individual filaments in bundles of vimentin intermediate filaments are transported in the cytoplasm by motor proteins along microtubules. Furthermore, using 3D FIB-SEM the authors showed that vimentin filament bundles are loosely packed and co-aligned with microtubules.

Barron A.J., Agrawal S., Lesperance D.N., Doucette J., Calle S., Broderick N.A.

Summary Microbial invasions underlie host-microbe interactions resulting in pathogenesis and probiotic colonization. In this study, we explore the effects of the microbiome on microbial invasion in Drosophila melanogaster. We demonstrate that gut microbes Lactiplantibacillus plantarum and Acetobacter tropicalis improve survival and lead to a reduction in microbial burden during infection. Using a microbial interaction assay, we report that L. plantarum inhibits the growth of invasive bacteria, while A. tropicalis reduces this inhibition. We further show that inhibition by L. plantarum is linked to its ability to acidify its environment via lactic acid production by lactate dehydrogenase, while A. tropicalis diminishes the inhibition by quenching acids. We propose that acid from the microbiome is a gatekeeper to microbial invasions, as only microbes capable of tolerating acidic environments can colonize the host. The methods and findings described herein will add to the growing breadth of tools to study microbe-microbe interactions in broad contexts.

Lima-Gomes P.D., do Nascimento M.T., Nadaes N.R., de Campos S.G., Tavares Haido R.M., Danelli M.D., Pinto-da-Silva L.H., Saraiva E.M.

Aspergillosis is a common fungal disease in avian species, causing high mortality in young chicks in agricultural farms and yards. It is caused by fungi belonging to the genus Aspergillus. Aspergillosis occurs by inhalation of fungal conidia, and in chickens, effective infection control relies on a rapid and large influx of heterophils to the lungs. Heterophils, upon different stimuli, release to the extracellular milieu their chromatin associated with several proteins that ensnare and kill different pathogens similarly to neutrophil extracellular traps. Here, we showed that Aspergillus fumigatus conidia and the peptidogalactomannan (PGM), isolated from the fungus cell wall, induce the release of DNA extracellular traps (DETs) in chicks' blood and lung heterophils. We demonstrated that reactive oxygen species, elastase and peptidyl arginine deiminase (PAD) were involved in DETs extrusion, the occurrence of DETs in the lungs of A. fumigatus-exposed chicks in vivo, and its role in chick survival. These results may contribute to developing more efficient tools for the therapeutic and diagnosis of aspergillosis.

Kolesnik E.A., Derkho M.A., Rebezov M.B.

Relevance. An overview is presented of the patterns of appearance, physiological and clinical significance of shadows of cells, a comparative experimental cytomorphological analysis using the example of polychromatophilic erythroblasts (PolyErythro), shadows of erythroblast (ShadErythro) and undifferentiated shadows cells of in peripheral blood smears of chickens Gallus gallus L.Methods. The studied individuals were divided into four groups (n = 40) depending on the age of the chickens (Postembryonalis — P1, P7, P23, and P42): 1, 7, 23, and 42 days of postembryonic ontogenesis) of a factory herd. ShadErythro apoptotic changes, undifferentiated cell shadows were characterized. Were calculated and characterized the morphophysiological parameters of PolyErythro, ShadErythro and unidentified shadows of cells by high-resolution color microphotographs in blood smears of birds (Pappenheim stained); a total of 158 (n = 158) microphotographs were analyzed.Results. The structured ShadErythro were able in chromatinolized. Unidentified shadows of cells were in a state of cytolysis, fragmentation of the cytoplasm, karyopycnosis and karyorrhexis, chromatin fragmentation, and karyolysis were noted. Structured of ShadErythro and undifferentiated shadows of cells were light purple with a reddish tint. Significant difference in the area of ShadErythro (Sshadow, μm2) was recorded in the ascending area of PolyErythro (Scell, μm2), within 25% (p ≤ 0.001), an increase in the percentage of difference between the minimum and maximum values of the ShadErythro area (Pshad.-cell-differ.-min-max,%) of Pcell-differ.-min-max,% PolyErythro was 36% (p ≤ 0.001). The difference in the percentage difference between the minimum and maximum values of the area of the nucleus of erythroblasts (Pnucleus-differ.-min-max) and the shadows of the nucleus of erythroblasts (Pshad.-nucle.-differ.-min-max) is significantly less than the analogous values (noted above) for cells and shadows erythroblasts, for example, Pnucleus-differ.-min-max exceeded Pshad.-nucle.-differ.-min-max by only 21% (p ≤ 0.001). The percentage of difference between the minimum and maximum values of the nuclear-cytoplasmic ratio ShadErythro (PN/C Ratio-shad.-cell) significantly exceeded the analogous value of PolyErythro (PN/C Ratio) within 64% (p ≤ 0.001). At the same time, the nuclear-cytoplasmic ratio of PolyErythro (N/C Ratio) and ShadErythro (N/C Ratio-shad.-cell) had a relatively minimal difference. The ShadErythro nuclear-cytoplasmic ratio (N/C Ratio-shad.-cell) exceeded the PolyErythro nuclear-cytoplasmic ratio (N/C Ratio) by only 12% (p ≤ 0.001). ShadErythro is maximally represented in chickens aged P1 — 40% and up to 30% (p ≤ 0.05) in P23. In P7, the amount of ShadErythro decreased by 30% (p ≤ 0.05), in P42, a decrease in ShadErythro to 20% (p ≤ 0.05) was recorded. The described dynamics of ShadErythro is due to physiological adaptive processes in the body of broiler chickens in the early period of postembryonic ontogenesis. The data obtained by us from a comprehensive comparative morphophysiological analysis of forms of degeneration of blood cells will be useful in the clinical diagnosis of animal pathologies, experimental and theoretical studies of cellular adaptations of the functional blood system and the whole organism.

Rzhepakovsky I.V., Timchenko L.D., Piskov S.I., Avanesyan S.S., Sizonenko M.N., Shakhbanov M.S., Nagdalian A.A., Rebezov M.B.

Relevance. The chicken embryo model is ideal for studying the development of the heart, which is the first functioning organ in embryogenesis. Particular attention is drawn to the embryonic period, when the embryonic heart is both functioning and forming, and small deviations in this process can lead to developmental anomalies. The current lack of results of accurate measurements of the morphology of the heart of a chicken embryo requires high-quality visualization, the complexity of which is determined by dynamic changes in the morphology of the heart, small size and complex spatial details of the organ. Threedimensional imaging methods are indispensable tools in this aspect, among which X-ray microtomography has special advantages.Methods. Embryo scanning (4–8 days, embryonic stages HH22–HH34) was performed using a highresolution X-ray microtomograph Skyscan 1176. A 1% solution of phosphoric-tungstic acid was used as a contrast. Post-processing, reconstruction, visualization of 3D-images, morphometry and X-ray density assessment were carried out using software packages DataViewer (1.5.6.2), CTvox (3.3.0r1403), CT-analyser (1.18.4.0).Results. The results obtained consist of three components: 1) general three-dimensional X-ray microtomographic visualization of chicken embryos in the early stages of embryogenesis with the designation of internal organs, allowing to determine the correct location of the heart; 2) microtomography of the heart with the designation of the main structures; 3) morphometric parameters of the embryo, heart and their X-ray density in units of the Hounsfield scale (HU). The methodological approach used has shown high efficiency. With an increase in the linear size of the chicken embryo and internal organs, including the heart, a high level of both general X-ray contrast and diffraction remains. At all the studied stages of the embryonic period of embryogenesis (4–8 days; HH22–HH34), the main visual and quantitative characteristics of cardiac morphogenesis were revealed.

Kharisma V.D., Ansori A.N., Antonius Y., Rosadi I., Murtadlo A.A., Jakhmola V., Rebezov M., Maksimiuk N., Kolesnik E., Burkov P., Derkho M., Scherbakov P., Ullah M.E., Sucipto T.H., Purnobasuki H.

Context: Mangosteen (Garcinia mangostana L.) is used in traditional medicine as an antibacterial, antioxidant, and anti-inflammatory. Aims: To determine the molecular mechanism and potential of garciniaxanthone derivate compounds from G. mangostana as SARS-CoV-2 antiviral and prevent cytokine storm through in silico approach. Methods: Ligand and protein samples were obtained from databases such as PubChem and Protein Databank, then drug-likeness analysis using Lipinski, Ghose, Veber, Egan, and Muege rules on SwissADME server, prediction of antiviral probability through PASSOnline server. Furthermore, molecular docking simulation with PyRx v1.0 software (Scripps Research, USA) with an academic license, identification of interactions and chemical bond positions of ligands on the target by PoseView server, 3D visualization of PyMOLv.2.5.2 software (Schrödinger, Inc., USA) with an academic license, molecular dynamics simulation for molecular stability prediction by CABS-flex v2.0 server, target prediction of antiviral candidate compounds by SwissTargetPrediction server, pathway analysis through STRING v11.5 database, and toxicity by ProTox-II server were used. Results: Garciniaxanthone C from G. mangostana was found to be a drug-like molecule with low toxicity. This can be a candidate for SARS-Cov-2 antiviral through inhibitor activity on two viral enzymes consisting of Mpro and replicase with a binding affinity value that is more negative than other garciniaxanthone derivates and is stable. Garciniaxanthone C is predicted to bind and inhibit pro-inflammatory proteins that trigger cytokine storms, such as NFKB1 and PTGS2. Conclusions: Garciniaxanthone derivative compounds from G. mangostana may be candidates for SARS-CoV-2 antiviral and preventing cytokine storm through garciniaxanthone C activity.

Wu H., Zhu X., Wu Z., Li P., Chen Y., Ye Y., Wang J., Zhou E., Yang Z.

Fumonisin B1 (FB1), a worldwide contaminating mycotoxin produced by Fusarium, poses a great threat to the poultry industry. It was reported that extracellular traps could be induced by FB1 efficiently in chickens. However, the relevance of autophagy and glycolysis in FB1-triggered heterophil extracellular trap (HET) formation is unclear. In this study, immunostaining revealed that FB1-induced HETs structures were composed of DNA coated with histones H3, and elastase, and that heterophils underwent LC3B-related autophagosome formation assembly driven by FB1. Western blotting showed that FB1 downregulated the phosphorylated phosphoinositide 3-kinase3-kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin complex 1 (mTORC1) axis and raised the AMP-activated kinase α (AMPKα) activation protein. Furthermore, rapamycin- and 3-Methyladenine (3-MA)-treatments modulated FB1-triggered HET formation according to the pharmacological analysis. Further studies on energy metabolism showed that glucose/lactate transport and glycolysis inhibitors abated FB1-induced HETs. These results showed that FB1-induced HET formation might interact with the autophagy process and relied on glucose/monocarboxylic acid transporter 1 (MCT1) and glycolysis, reflecting chicken's early innate immune responses against FB1 intake.

Takahata S., Murakami Y.

DNA is stored in the nucleus of a cell in a folded state; however, only the necessary genetic information is extracted from the required group of genes. The key to extracting genetic information is chromatin ambivalence. Depending on the chromosomal region, chromatin is characterized into low-density “euchromatin” and high-density “heterochromatin”, with various factors being involved in its regulation. Here, we focus on chromatin regulation and gene expression by the yeast FACT complex, which functions in both euchromatin and heterochromatin. FACT is known as a histone H2A/H2B chaperone and was initially reported as an elongation factor associated with RNA polymerase II. In budding yeast, FACT activates promoter chromatin by interacting with the transcriptional activators SBF/MBF via the regulation of G1/S cell cycle genes. In fission yeast, FACT plays an important role in the formation of higher-order chromatin structures and transcriptional repression by binding to Swi6, an HP1 family protein, at heterochromatin. This FACT property, which refers to the alternate chromatin-regulation depending on the binding partner, is an interesting phenomenon. Further analysis of nucleosome regulation within heterochromatin is expected in future studies.

Zhang Z., Zhang R., Xiao K., Sun X.

G-quadruplex (G4) structures are critical epigenetic regulatory elements, which usually form in guanine-rich regions in DNA. However, predicting the formation of G4 structures within living cells remains a challenge. Here, we present an ultra-robust machine learning method, G4Beacon, which utilizes the Gradient-Boosting Decision Tree (GBDT) algorithm, coupled with the ATAC-seq data and the surrounding sequences of in vitro G4s, to accurately predict the formation ability of these in vitro G4s in different cell types. As a result, our model achieved excellent performance even when the test set was extremely skewed. Besides this, G4Beacon can also identify the in vivo G4s of other cell lines precisely with the model built on a special cell line, regardless of the experimental techniques or platforms. Altogether, G4Beacon is an accurate, reliable, and easy-to-use method for the prediction of in vivo G4s of various cell lines.

Chen Y., Ye Y., Wu H., Wu Z., Li P., Fu Y., Sun Y., Wang X., Wang J., Yang Z., Zhou E.

Citrinin (CTN), a secondary fungal metabolite produced by several Aspergillus, Penicillium, and Monascus genera species, is a toxin with a wide range of biological activities. Neutrophil extracellular traps represent a novel potential mechanism of the neutrophil response to foreign matters, and chicken heterophils can release similar heterophil extracellular traps (HETs). In this study, we aimed to investigate the effect of CTN on HET formation. Density gradient centrifugation was used to isolate chicken peripheral blood heterophils, and then immunofluorescence was used to observe the effects of CTN on HET formation. The mechanisms of HET formation were analyzed using pharmacological inhibitors and quantification of extracellular DNA, and the production of reactive oxygen species was detected with a fluorescent probe. Our results revealed that CTN (50-400 μM) had no cytotoxic effect on heterophils. CTN exposure induced the release of HETs composed of chromatin decorated with histones and elastase, and CTN-triggered HETs were dose- and time-dependent to some extent. Furthermore, CTN increased ROS production and activated p38 and ERK1/2 signaling pathways in heterophils. However, inhibition of the p38 signaling pathway, ERK1/2 signaling pathway, and NADPH oxidase pathway did not block HET formation induced by CTN. Inhibition of peptidyl arginine deiminase 4 (PAD4) enzyme and P2×1 receptor decreased HET formation after CTN stimulation, suggesting that HET formation exposed to CTN was mediated by PAD4 and P2×1 receptor. In conclusion, these findings may suggest a canonical mechanism relevant to the innate immunity caused by mycotoxins in chickens.

Gasparotto M., Lee Y., Palazzi A., Vacca M., Filippini F.

Aberrant mitochondrial phenotypes are common to many central nervous system (CNS) disorders, including neurodegenerative and neurodevelopmental diseases. Mitochondrial function and homeostasis depend on proper control of several biological processes such as chromatin remodeling and transcriptional control, post-transcriptional events, vesicle and organelle subcellular trafficking, fusion, and morphogenesis. Mutation or impaired regulation of major players that orchestrate such processes can disrupt cellular and mitochondrial dynamics, contributing to neurological disorders. The first part of this review provides an overview of a functional relationship between chromatin players and mitochondria. Specifically, we relied on specific monogenic CNS disorders which share features with mitochondrial diseases. On the other hand, subcellular trafficking is coordinated directly or indirectly through evolutionarily conserved domains and proteins that regulate the dynamics of membrane compartments and organelles, including mitochondria. Among these “building blocks”, longin domains and small GTPases are involved in autophagy and mitophagy, cell reshaping, and organelle fusion. Impairments in those processes significantly impact CNS as well and are discussed in the second part of the review. Hopefully, in filling the functional gap between the nucleus and cytoplasmic organelles new routes for therapy could be disclosed.

Pastore A., Badocco D., Cappellin L., Pastore P.

The dichromatism of Bromophenol blue (BPB) was investigated by varying its concentration in the absence and presence of surfactant. A model of the indicator behavior was carried out, justifying the experimental shapes of the sigmoidal profiles of the hue (H) coordinate. The model applied to the solution was compared with the performance of colorimetric sensor arrays (CSAs) with increasing BPB concentrations. The H shape and the prediction errors of the CSAs were very similar to those predicted. The experimental results enable the changing of the slope of the calibration profiles, at will, by varying only the BPB concentration.

Wu Z., Zhu X., Li P., Wang X., Sun Y., Fu Y., Wang J., Yang Z., Zhou E.

Fumonisin B1 (FB1) is a common mycotoxin contamination in agricultural commodities being considered as a significant risk to human and livestock health, while the mechanism of FB1 immunotoxicity are less understood, especially in chicken. Given that extracellular traps as a novel defense mechanism of leukocytes play an important role against foreign matters, in this study we aimed to investigate the effects of FB1 on chicken heterophil extracellular traps (HETs) formation. Our result showed that FB1 induced HETs release in chicken heterophils observed via immunostaining, and it was concentration-dependent during 10 to 40 μM. Moreover, in 40 μM FB1-exposed chicken heterophils, reactive oxygen species (ROS) level was increased, while catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activity and glutathione (GSH) content were decreased. Simultaneously, FB1 (40 μM) activated ERK and p38 MAPK signaling pathways via increasing the phosphorylation level of ERK and p38 proteins. However, pretreatment of SB202190, U0126, and diphenyleneiodonium chloride (DPI) did not change FB1-triggered ROS production and HETs formation, suggesting FB1-induced HETs was a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, p38, and extracellular regulated protein kinases (ERK) signaling pathways-independent process. Inhibition of peptidyl arginine deiminase 4 (PAD4) enzyme and P2 × 1 receptor showed their vital role in 40 μM FB1-triggered HETs. This study reported for the first time that 40 μM FB1 induced the release of HETs in heterophils, and it was related to ROS production, PAD4, and P2 × 1, but was independent of NADPH oxidase, p38 and ERK signaling pathways, which might provide a whole novel perspective of perceiving and understanding the role of FB1 in immunotoxicity.

Yap K.N., Zhang Y.

Erythrocyte enucleation is thought to have evolved in mammals to support their energetic cost of high metabolic activities. However, birds face similar selection pressure yet possess nucleated erythrocytes. Current hypotheses on the mammalian erythrocyte enucleation claim that the absence of cell organelles allows erythrocytes to 1) pack more hemoglobin into the cells to increase oxygen carrying capacity and 2) decrease erythrocyte size for increased surface area-to-volume ratio, and improved ability to traverse small capillaries. In this article, we first empirically tested current hypotheses using both conventional and phylogenetically informed analysis comparing literature values of mean cell hemoglobin concentration (MCHC) and mean cell volume (MCV) between 181 avian and 194 mammalian species. We found no difference in MCHC levels between birds and mammals using both conventional and phylogenetically corrected analysis. MCV was higher in birds than mammals according to conventional analysis, but the difference was lost when we controlled for phylogeny. These results suggested that avian and mammalian erythrocytes may employ different strategies to solve a common problem. To further investigate existing hypotheses or develop new hypothesis, we need to understand the functions of various organelles in avian erythrocytes. Consequently, we covered potential physiological functions of various cell organelles in avian erythrocytes based on current knowledge, while making explicit comparisons with their mammalian counterparts. Finally, we proposed by taking an integrative and comparative approach, using tools from molecular biology to evolutionary biology, would allow us to better understand the fundamental physiological functions of various components of avian and mammalian erythrocytes.

Schreier S., Budchart P., Borwornpinyo S., Arpornwirat W., Triampo W.

Background: The circulating rare cell population is diverse and rich in diagnostic information. Its characterization and clinical exploitation by cell-based liquid biopsy is an ongoing research task. Bone marrow is one of the major contributors to the peripheral blood rare cell population and, consequently, determines individual rare cell profiles thus depending on bone marrow health status. Bone marrow damage has been associated with aggressive or late-stage systemic diseases and egress of various bone marrow cells into the blood circulation. The association of quantity and heterogeneity of circulating erythroblast with bone marrow damage is of particular interest. Methods: Circulating CD71high/CD45-/Hoechsthigh blast cells from healthy, noncancer- and cancer-afflicted donors were enriched by CD45 depletion and analyzed by immunofluorescence microscopy. Results: A new finding of aberrant and mitotic circulating erythroid-like cells that appear similar across blood donors afflicted with various systemic pathologies is reported. Further presented is a classification of said erythroblast-like cells in nine subcategories according to morphological differences between phenotypically similar cells. Conclusion: Aberrant and mitotic bone marrow-derived rare circulating erythroid-like cells can be detected in the blood of afflicted individuals but not in healthy donors, suggesting the cause of bone marrow damage.