Browsing by Author "Posokhov, Yevgen"

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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.
Assessment of apoptosis in chronic carrageenan-induced gastroenterocolitis by fluorescent probes
(2017) Корниенко, Евгений Михайлович; Посохов, Евгений Александрович; Ткаченко, Антон Сергеевич; Korniyenko, Ye.; Posokhov, Yevgen; 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.
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.
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
Influence of Carrageenan (E 407) on the Membrane of Enterocytes Investigated by Fluorescent Probes Research Institute of Chemistry (Kharkiv)
(2013) Посохов, Евгений Александрович; Ткаченко, Антон Сергеевич; Корниенко, Евгений Михайлович; Posokhov, Yevgen; Tkachenko, Anton; Korniyenko, Ye.
The influence on enterocytes membrane structure of added to the diet of rats food-additive carrageenan (E 407) was studied using fluorescent probes (derivatives of ortho-hydroxy oxazole). It was found that carrageenan changed hydration membrane of enterocytes in experimental animals. It was shown that carrageenan (E 407) caused changes in the polar regions of biological membranes and at the same time, the consumption of this food-additive didn’t lead to any changes in the hydrophobic parts of the membrane of enterocytes. Nature of detected changes allows to make a conclusion that enterocyte membranes in rats with chronic carrageenan-induced gastroenterocolitis have increased microviscosity.
The study of phospholipid bilayer of cell membranes in leukocytes incubated with high concentrations of the food additive E407a
(2021) Tkachenko, Anton; Onishchenko, Anatolii; Roshal, Alexander; Posokhov, Yevgen
Aim: The aim of the study was to assess the impact of semi-refined carrageenan (E407a) on the hydrophobic region of phospholipid bilayer in cell membranes of leukocytes exposed to the solution with high concentrations of this food additive. Material and methods: Fluorescent probe (2-phenylphenanthro[ 9,10-d]-1,3-oxazole) was used to investigate the influence of E407a on the state of lipid bilayer in leukocytes extracted from rats and treated with a 5% solution of the food additive E407a during 4 hours. Results: The shapes of the probe fluorescence spectra did not differ in leukocytes of rats treated with the 5% solution of semi-refined carrageenan and white blood cells of control samples. Such findings suggest that exposure to the E407a solution causes no changes in the proton-donor ability of the media in the lipid membranes of leukocytes in the region where the probe locates. Conclusion: Exposure of white blood cell suspensions to the semirefined carrageenan solution does not affect the membrane hydration of the hydrophobic region of leukocyte phospholipid bilayer.
The fluorescent probe method in investigation of the state of erythrocyte membranes in white rats at exposure to chemical environmental factors
(Чорноморський національний університет ім. Петра Могили (м. Миколаїв), 2018) Наконечна, Оксана Анатоліївна; Безродна, Анастасія Ігорівна; Корнієнко, Євген Михайлович; Ткачова, Т.М.; Єфімова, С.Л.; Посохов, Євгеній Олександрович; Максимова, Ірина Геннадіївна; Nakonechna, Oksana; Bezrodna, A.; Kornienko, E.; Tkacheva, T.; Efimova, S.; Posokhov, Yevgen; Maksymova, I.
The purpose of the presented work was to study the possibility of using the fluorescent probes method to diagnose the harmful effects of chemical factors on the example of polyethylene glycol on the organism of white rats by evaluating the state of erythrocyte membranes. Material and methods. We used the following fluorescent probes in the studies: ortho-hydroxy derivatives of 2.5-diaryl-1.3-oxazole. Results and discussion. In the case of erythrocytes of rats, which are toxic to PEG-400, there is a marked decrease in the fluorescence intensity of all the probes used. The discussed decrease in the fluorescence intensity of the probes indicates a decrease in the number of molecules of each of the probes associated with erythrocyte membranes per one hour of incubation, which indicates a decrease in the rate of binding of probes to membranes. Such a decrease in the binding rate can be explained by the formation of an additional protective membrane around each lipid membrane. Conclusion. The established fact of formation of an additional coat of polyethylene glycol molecules on the surface of erythrocyte membranes can be treated as a stable standardized indicator for the method of fluorescent probes, which may indicate the absence of damaging effect on the membranes at the object of study, which requires confirmation in further studies.