Browsing by Author "Onishchenko, Anatolii"

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A possible contribution of carrageenan to diabetes mellitus type 2 development
(ХНМУ, 2019-04) Alam, J.; Tkachenko, Anton; Onishchenko, Anatolii
A study of erythrocyte membranes in carrageenaninduced gastroenterocolitis by method of fluorescent probes
(HVM Bioflux, 2018) Tkachenko, Anton; Marakushyn, Dmytro; Rezunenko, Yurii; Onishchenko, Anatolii; Nakonechna, Oksana; Posokhov, Yevgen
Objective. The aim of our research was to investigate erythrocyte membranes of rats with carrageenan-induced gastroenterocolitis using fluorescent probes (ortho-hydroxy derivatives of 2,5-diaryloxazole).Materials and methods. Fluorescence of O1O (2 - (2’-OH-phenyl)-5-phenyl-1,3-oxazole) and PH7 (2-(2’-OH-phenyl)-phenanthro[9,10]-1,3-oxazole) probes in suspensions of erythrocytes of rats with chronic carrageenan-induced gastroenterocolitis was determined and compared to the control group consisted of intact animals.Results. It has been found that the hydration of erythrocyte membranes increases in rats with carrageenan-induced gastroenterocolitis. The disease is associated with the changes in the polar regions of the membranes, while no changes in the hydrophobic areas of the erythrocyte membranes are observed. Conclusion. The detected changes allow us to make a conclusion that the fluidity of the erythrocyte membranes of the rats with chronic carrageenan-induced gastroenterocolitis increases.
Antimicrobial activity and cytotoxicity study of cerium oxide nanoparticles with two different sizes
(2022-11-06) Yefimova, Svetlana; Klochkov, Vladimir; Kavok, Nataliya; Tkachenko, Anton; Onishchenko, Anatolii; Chumachenko, Tatyana; Чумаченко, Тетяна Олександрівна; Dizge, Nadir; Özdemir, Sadin; Gonca, Serpil; Ocakoglu, Kasim
The control over bacterial diseases requires the development of novel antibacterial agents. The use of antibacterial nanomedicines is one of the strategies to tackle antibiotic resistance. The study was designed to assess the antimicrobial activity of cerium oxide (CeO2) nanoparticles (NP) of two different sizes (CeO2 NP1 [1–2 nm] and CeO2 NP2 [10–12 nm]) and their cytotoxicity towards eukaryotic cells. The antimicrobial activity, effects of nanoparticles on DNA cleavage, microbial cell viability, and biofilm formation inhibition were analyzed. The impact of cerium oxide nanoparticles on eryptosis of erythrocytes was estimated using annexin V staining by flow cytometry. The newly synthesized CeO2 NP1 and CeO2 NP2 displayed moderate antimicrobial activities. CeO2 NP1 and CeO2 NP2 exhibited single-strand DNA cleavage ability. CeO2 NPs were found to show 100% microbial cell viability inhibition at a concentration of 500 mg/L. In addition, CeO2 NP1 and CeO2 NP2 inhibited the biofilm formation of S. aureus and P. aeruginosa. Larger cerium oxide nanoparticles were found to be less toxic against erythrocytes compared with the smaller ones. CeO2 nanoparticles demonstrate moderate antimicrobial activity and low cytotoxicity towards erythrocytes, which make them promising antibacterial agents.
Assessment of HSP70 and HSP90α expression in nasal epithelial cells of patients with chronic rhinosinusitis with nasal polyps
(portobiomedicaljournal, 2019) Kharchenko, Elina; Skliaruk, Diana; Onishchenko, Anatolii; Tkachenko, Anton
Changes in cell membranes of white blood cells treated with a common food additive E407a
(2021) Tkachenko, Anton; Onishchenko, Anatolii; Posokhov, Yevgen; Roshal, Alexander; Myasoedov, Valeriy; Nakonechna, Oksana
Objectives: To estimate the state of phospholipid bilayer of rats WBCs exposed to a common food additive E407a, which is used as a thickener and emulsifier, during 4 h using a fluorescent probe − ortho-hydroxy derivative of 2,5-diaryl-1,3-oxazole. Materials and methods: Steady-state fluorescence spectroscopy: a study by the environment-sensitive fluorescent probe − 2-(2′-hydroxy-phenyl)-5-phenyl-1,3-oxazole (probe O1O). Results: Changes are detected in the spectra of the fluorescent probe bound to rat WBCs treated with the solutions of E407a of various concentrations in comparison with the corresponding spectra of the probe incubated with the untreated leukocytes. The decrease in polarity and protondonor ability is observed in the lipid membranes of leukocytes in the region, where the probe locates. Conclusions: Our findings suggest a higher dehydration of leukocyte membranes of rats treated with a common food additive E407a at high concentrations and, thus, indicate that exposure to high doses of E407a leads to the increase in the lipid order (i.e. to decrease in fluidity) of the membranes of rat WBCs.
Changes in S100 protein expression in the cerebellum of rats orally exposed to energy drinks
(The Official Publication of The Malaysian Society for Biochemistry and Molecular Biology, 2019-12-20) Ткаченко, Антон Сергійович; Губіна-Вакулік, Галина Иванівна; Каук, Оксана Іванівна; Оніщенко, Анатолій Ігорович; Шеховцова, Есміра Вадимівна; Tkachenko, Anton; Gubina-Vakulyck, Galina; Kauk, Oksana; Onishchenko, Anatolii; Shekhovtsova, Esmira; Nakonechna, Oksana
The aim of our study was of assess the rate of S100 protein expression in the cerebellum of rats orally exposed to caffeinated energy drinks (CED) during 2 and 4 weeks. Twenty WAG rats were enrolled in the experiment. They were subdivided into three groups. The animals from group 1 and group 2 (n=7 in each) were orally exposed to a caffeinated energy beverage “Black” at a dose of 12 ml per kg of body weight during two and four weeks, respectively. The control group consisted of 6 intact rats obtained drinking water instead of a caffeine-containing energy drink. Cerebellar S100 expression was evaluated immunohistochemically. Against the background CED oral administration during two and four weeks, the density of granule cell layer becomes lower indicating neuronal loss. S100 protein was upregulated in group 1 in the entire cerebellar cortex. Thus, CED oral exposure resulted in the development of cerebellar astrogliosis. Four-week-long CED consumption resulted in S100 downregulation in the molecular and granular layers. Our findings demonstrate the damage to the cerebellum in rats exposed to CEDs with overexpression of S100, followed by a decrease in its expression.
Comparative analysis of HSP70 and HSP90alpha expression in nasal epithelial cells of patients with chronic rhinosinusitis with nasal polyps
(Тернопільський національний медичний університет імені І. Я. Горбачевського МОЗ України, 2019) Tkachenko, Anton; Onishchenko, Anatolii; Gubina-Vakulyck, Galina; Nakonechna, Oksana; Gorbach, Tetiana; Stetsenko, S.
Cytotoxicity of Hybrid Noble Metal-Polymer Composites
(2022-10-11) Tkachenko, Anton; Virych, Pavlo; Myasoedov, Valeriy; Prokopiuk, Volodymyr; Onishchenko, Anatolii; Butov, Dmytro; Kuziv, Yuliia; Yeshchenko, Oleg; Zhong, Sican; Nie, Guochao; Kutsevol, Nataliya
The aim of the present research was to assess the cytotoxicity of gold and silver nanoparticles synthesized into dextran-graft-polyacrylamide (D-PAA) polymer nanocarrier, which were used as a basis for further preparation of multicomponent nanocomposites revealed high efficacy for antitumor therapy. The evaluation of the influence of Me-polymer systems on the viability and metabolic activity of fibroblasts and eryptosis elucidating the mechanisms of the proeryptotic effects has been done in the current research. The nanocomposites investigated in this study did not reduce the survival of fibroblasts even at the highest used concentration. Our findings suggest that hybrid Ag/D-PAA composite activated eryptosis via ROS- and Ca2+-mediated pathways at the low concentration, in contrast to other studied materials. Thus, the cytotoxicity of Ag/D-PAA composite against erythrocytes was more pronounced compared with D-PAA and hybrid Au/polymer composite. Eryptosis is a more sensitive tool for assessing the biocompatibility of nanomaterials compared with fibroblast viability assays.
Effects of E407a on the viability, metabolic and functional activity of dermal fibroblasts
(2021) Tkachenko, Anton; Prokopiuk, Volodymyr; Onishchenko, Anatolii; Shevchenko, Maria
Aim: To evaluate the effects of semi-refined carrageenan (E407a) on the viability, metabolic and functional activity of skin fibroblasts in vitro. Material and methods: Various concentrations of E407a (0 – 10 mg/ml) were incubated for 24 h with skin fibroblast cultures isolated from rat embryos. The viability of cells exposed to semi-refined carrageenan was assessed by the neutral red uptake assay. MTT test was selected to estimate the metabolic activity of cells. In addition, scratch assay was used to determine the functional activity of dermal fibroblasts. Results: Direct exposure to semi-refined carrageenan did not affect the viability of rat dermal fibroblasts, evidenced by the results of MTT and neutral red uptake assays. However, experimental data of MTT and scratch assays revealed that high concentrations of this food additive increased the metabolic activity of fibroblasts and diminished the motility of these cells, suggesting that E407a is cytotoxic at high concentrations. Conclusion: Food-grade semi-refined carrageenan is cytotoxic towards skin fibroblasts at high concentrations.
Effects of semi-refined carrageenan (food additive E407a) on cell membranes of leukocytes assessed in vivo and in vitro
(2021) Tkachenko, Anton; Onishchenko, Anatolii; Roshal, Alexander; Nakonechna, Oksana; Chumachenko, Tetyana; Чумаченко, Тетяна Олександрівна; Чумаченко, Татьяна Александровна; Yevgen, Posokhov
Effects of semi-refined carrageenan (food additive E407a) oncell membranes of leukocytes assessed in vivo and in vitro
(2021) Tkachenko, Anton; Onishchenko, Anatolii; Roshal, Alexander; Nakonechna, Oksana; Chumachenko, Tetyana; Posokhov, Yevgen
Aim: To assess the impact of semi-refined carrageenan (E407a) on hydrophobic regions of phosphololipid bilayer in cell membranes of leukocytes collected from rats orally administered this food additive and white blood cells incubated with E407a. Methods: Fluorescent probes (ortho-hydroxy derivatives of 2,5-diaryl-1,3-oxazole) were used to estimate the state of lipid bilayer in leukocytes obtained from rats orally exposed to the food additive E407a and in white blood cells incubated with E407a. Results: No noticeable changes in the physico-chemical properties were observed in the lipid membranes of leukocytes in the region where the probes locate in response to oral intake of semi-refined carrageenan. Incubation of leukocytes with E407a solutions resulted in a decrease in polarity and proton-donor ability of leukocytes in the area of carbonyl groups of phospholipids and in the area of hydrocarbon chains of phospholipids near the polar region of the bilayer. Conclusion: Membrane fluidity abnormalities found in cells exposed to E407a are similar to those observed in patients with IBD suggesting that contribution of carrageenan to IBD development may be partially explained by leukocyte membrane modifications.
Epithelial-to-mesenchymal transition and some parameters of extracellular matrix remodeling in chronic rhinosinusitis with nasal polyps
(HVM Bioflux, 2018-09) Onishchenko, Anatolii; Lupyr, Andrii; Tkachenko, Anton; Gorbach, Tatyana; Nakonechna, Oksana; Gubina-Vakulyck, Galina
Objective. The purpose of our study was to evaluate vimentin expression in nasal polyp tissue, concentrations of monocyte chemoattractant protein-1 (MCP-1) and matrix metalloproteinase-9 (MMP-9) in blood serum of patients with chronic rhinosinusitis with nasal polyps (CRSwNP).Materials and methods. Levels of MMP-9 and MCP-1 were determined in blood serum of twenty patients with CRSwNP using the commercially produced ELISA kits. Immunohistochemical vimentin staining using specimens of nasal polyp tissue was performed. The results were compared to the control group consisted of twenty relatively healthy subjects.Results. Vimentin was found to be strongly expressed in both the lamina propria and epithelial cells in nasal polyp tissue of patients with CRSwNP. Blood serum concentrations of MMP-9 and MCP-1 were higher in CRSwNP compared to the control individuals. MCP-1/MMP-9 ratio was higher as well. Conclusion. The development of nasal polyps is accompanied by an increase in the amount of vimentin-positive cells in the epithelial layer, which indicates the activation of endothelial-to-mesenchymal transition (EMT). CRSwNP is associated with the imbalance between profibrotic and antifibrotic factors.
Experimental Evaluation of Food-Grade Semi-Refined Carrageenan Toxicity
(2021) Pogozhykh, Denys; Posokhov, Yevgen; Myasoedov, Valeriy; Gubina-Vakulyck, Galina; Chumachenko, Tetyana; Knigavko, Oleksandr; Polikarpova, Hanna; Kalashnyk-Vakulenko, Yuliia; Sharashydze, Ketino; Nakonechna, Oksana; Prokopyuk, Volodymyr; Onishchenko, Anatolii; Tkachenko, Anton
The safety of food additives E407 and E407a has raised concerns in the scientific community. Thus, this study aims to assess the local and systemic toxic effects of the common food additive E407a in rats orally exposed to it for two weeks. Complex evaluations of the effects of semi-refined carrageenan (E407a) on rats upon oral exposure were performed. Local effects of E407a on the intestine were analyzed using routine histological stains and CD68 immunostaining. Furthermore, circulating levels of inflammatory markers were assessed. A fluorescent probe O1O (2- (2′-OH-phenyl)-5-phenyl-1,3-oxazole) was used for evaluating the state of leukocyte cell membranes. Cell death modes of leukocytes were analyzed by flow cytometry using Annexin V and 7-aminoactinomycin D staining. Oral administration of the common food additive E407a was found to be associated with altered small and large intestinal morphology, infiltration of the lamina propria in the small intestine with macrophages (CD68+ cells), high systemic levels of inflammation markers, and changes in the lipid order of the phospholipid bilayer in the cell membranes of leukocytes, alongside the activation of their apoptosis. Our findings suggest that oral exposure to E407a through rats results in the development of intestinal inflammation.
Experimental Evaluation of Food-Grade Semi-Refined Carrageenan Toxicity
(2021-10-16) Pogozhykh, Denys; Posokhov, Yevgen; Myasoedov, Valeriy; Gubina-Vakulyck, Galina; Chumachenko, Tetyana; Knigavko, Oleksandr; Polikarpova, Hanna; Kalashnyk-Vakulenko, Yuliia; Sharashydze, Ketino; Nakonechna, Oksana; Prokopyuk, Volodymyr; Onishchenko, Anatolii; Tkachenko, Anton
Abstract: The safety of food additives E407 and E407a has raised concerns in the scientific community. Thus, this study aims to assess the local and systemic toxic effects of the common food additive E407a in rats orally exposed to it for two weeks. Complex evaluations of the effects of semi-refined carrageenan (E407a) on rats upon oral exposure were performed. Local effects of E407a on the intestine were analyzed using routine histological stains and CD68 immunostaining. Furthermore, circulating levels of inflammatory markers were assessed. A fluorescent probe O1O (2- (20-OH-phenyl)-5-phenyl- 1,3-oxazole) was used for evaluating the state of leukocyte cell membranes. Cell death modes of leukocytes were analyzed by flow cytometry using Annexin V and 7-aminoactinomycin D staining. Oral administration of the common food additive E407a was found to be associated with altered small and large intestinal morphology, infiltration of the lamina propria in the small intestine with macrophages (CD68+ cells), high systemic levels of inflammation markers, and changes in the lipid order of the phospholipid bilayer in the cell membranes of leukocytes, alongside the activation of their apoptosis. Our findings suggest that oral exposure to E407a through rats results in the development of intestinal inflammation.
Experimental Evaluation of Food-Grade Semi-Refined Carrageenan Toxicity
(2021) Pogozhykh, Denys; Posokhov, Yevgen; Myasoedov, Valeriy; Gubina-Vakulyck, Galina; Knigavko, Oleksandr; Polikarpova, Hanna; Kalashnyk-Vakulenko, Yuliia; Sharashydze, Ketino; Nakonechna, Oksana; Prokopyuk, Volodymyr; Onishchenko, Anatolii; Tkachenko, Anton; Chumachenko, Tatyana; Чумаченко, Тетяна Олександрівна; Чумаченко, Татьяна Александровна
Fascin is upregulated in nasal mucosa in chronic rhinosinusitis with nasal polyps
(Kyiv, Palladin Institute of Biochemistry National Academy of Sciences of Ukraine, 2019-03) Tkachenko, Anton; Onishchenko, Anatolii
Gadolinium Orthovanadate GdVO4:Eu3+ Nanoparticles Ameliorate Carrageenan-Induced Intestinal Inflammation
(2021) Tkachenko, Anton; Pogozhykh, Denys; Onishchenko, Anatolii; Myasoedov, Valeriy; Podrigalo, Leonid; Klochkov, Vladimir; Chumachenko, Tetyana; Prokopyuk, Volodymyr; Yefimova, Svetlana; Gubina-Vakulyck, Galina; Kavok, Nataliya; Butov, Dmytro; Andrieiev, Andrii; Polikarpova, Hanna; Nakonechna, Oksana
HSP90α overexpression in small intestinal mucosa and high blood serum levels of HSP70 and 8-isoprostane in carrageenan-induced intestinal inflammation
(Bioflux, 2019) Tkachenko, Anton; Onishchenko, Anatolii; Gorbach, Tetiana; Nakonechna, Oksana; Shekhovtsova, E.; Gubina-Vakulyck, Galina
Aims. The aim of our research was to evaluate HSP90α expression in small intestinal tissue, levels of HSP70 and 8-isoprostane in blood serum of rats with chronic carrageenan-induced enteritis. Methods. We determined immunohistochemically HSP90α expression in small intestine of ten rats with intestinal inflammation caused by the four-month-long oral administration of k-carrageenan-containing 1% processed Eucheuma seaweed solution and compared it with the small intestinal HSP90α expression in ten healthy animals. Levels of 8-isoprostane and HSP70 were determined in blood serum of animals from both groups using commercially available ELISA kits. Results. HSP90α overexpression was observed in small intestine of rats with carrageenan-induced enteritis. The disease was also associated with the elevated circulating levels of 8-isoprostane, indicating the development of oxidative stress. The level of HSP70 in blood serum was also found to be higher in animals with carrageenan-induced intestinal inflammation compared with control rats. Conclusion. Our findings suggest that HSP90α and HSP70 play an important role in the pathogenesis of chronic carrageenan-induced enteritis.
Human chemerin induces eryptosis at concentrations exceeding circulating levels
(Tech Science Press, 2024) Tkachenko, Maryna; Onishchenko, Anatolii; Tryfonyuk, Liliya; Butov, Dmytro; Kot, Kateryna; Novikova, Viktoriia; Fan, Liwei; Prokopiuk, Volodymyr; Kot, Yurii; Anton Tkachenko
Introduction: Human chemerin is an adipokine that regulates chemotaxis, inflammation, and glucose metabolism. In addition, accumulating evidence suggests that chemerin promotes apoptosis, autophagy, and pyroptosis. However, there are no data on its impact on eryptosis. The current study aimed to analyze the effects of human active Glu21-Ser157 chemerin on eryptosis in vitro. Materials and Methods: Human chemerin 0-2-10-50 µg/mL was incubated for 24 h with human erythrocytes (hematocrit 0.4%) obtained from eight healthy individuals. Flow cytometry-based determination of phospholipid scrambling, reactive oxygen species (ROS) production, and intracellular Ca2+ levels was performed. To supplement data on ROS and Ca2+ signaling in chemerin-mediated eryptosis, incubation in the presence or absence of antioxidants vitamin C and N-acetylcysteine and Ca2+-binding agent EGTA was carried out, respectively. Confocal microscopy-based techniques were used to detect reactive nitrogen species (RNS) generation, involvement of caspase-3 and caspase-8, as well as the state of lipid order in cell membranes of erythrocytes exposed to human Glu21-Ser157 chemerin. Results: Our observations suggest that human Glu21-Ser157 chemerin had no impact on eryptosis parameters at 2 µg/mL. However, chemerin stimulated phosphatidylserine externalization, ROS production, and Ca2+ accumulation at higher concentrations suggesting activation of eryptosis. Ca2+ uptake turned out to be at least partly required for chemerin-mediated eryptosis. Chemerin-mediated erythrotoxicity was additionally mediated by RNS, caspase-3, and caspase-8. Moreover, Glu21-Ser157 chemerin promoted reduction in the liquid-ordered phase of cell membranes in erythrocytes. Conclusions: The present study first discloses that human chemerin can induce eryptosis via Ca2+-dependent mechanisms at concentrations noticeably exceeding circulating levels. Thus, chemerin-induced eryptosis can hardly contribute to eryptosis-mediated anemia in diseases associated with enhanced levels of chemerin in blood.
Human HMGB1 does not induce eryptosis in vitro
(2022) Tkachenko, Maryna; Onishchenko, Anatolii; Butov, Dmytro; Butova, Tetyana; Tkachenko, Anton
Aim: To study the ability of human high mobility group box protein 1 (HMGB1) to induce eryptosis in vitro. Material and methods: Blood collected from six healthy volunteers was incubated with HMGB1 (0-0.2-1-5 ng per ml). Eryptosis of red blood cells was assessed by Annexin V staining and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) staining by flow cytometry. The forward scatter (FSC) fluorescence was used to evaluate the morphology of red blood cells. Results: Exposure of erythrocytes to HMGB1 did not affect the morphology of erythrocytes, evidenced by no changes in the percentage of cells with small volume, i.e. shrunken cells, and erythrocytes with large volume, i.e. enlarged cells. HMGB1 had no impact on phosphatidylserine externalization, which is confirmed by the absence of statistically significant changes in the amount of phosphatidylserine-displaying cells and the mean fluorescence intensity (MFI) values of Annexin V-FITC in cells exposed to different concentrations of HMGB1. Furthermore, H2DCFDA staining revealed that the HMGB1 did not induce oxidative stress. Conclusion: HMGB1 does not promote eryptosis of human erythrocytes at concentrations of up to 5 ng/ml.