Category Archives: Uncategorized
Metallothioneins contribution to the response of bivalve mollusk to xenobiotics
V. V. Khoma1, L. L. Gnatyshyna1,2, V. V. Martyniuk1,
T. R. Mackiv1,2, N. Y. Mishchuk1, O. B. Stoliar1
1Ternopil Volodymyr Hnatiuk National Pedagogical University, Ukraine;
2I.Ya. Horbachevsky Ternopil National Medical University, Ukraine;
*e-mail: Oksana.Stolyar@tnpu.edu.ua
Received: 17 April 2020; Accepted: 25 June 2020
Estimation of cellular thiols metallothioneins (MTs) sensitivity to continuous pressure of environmental chemical ‘cocktail’ of xenobiotics needs investigation in correct model experiments. The aim of this study was to elucidate MT s contribution into bivalve mollusk response to co-exposure to xenobiotics and elevated temperature. We treated the mussels Unio tumidus Philipson, 1788 (Unionidae) with drugs diclofenac (Dc, 2 nM), nifedipine (Nf, 2 nM) or with organophosphonate herbicide glyphosate (Gl, formulation Roundup MAX, 79 nM) separately at 18°C and in combination at 18°C (DcNfGl) and 25°C (DcNfGl+T) during 14 days. MTs were isolated from digestive gland by size-exclusion chromatography. The concentration of MTs in the tissue was assessed according to metals (Zn, Cu, Cd) in the eluted peak of MTs (MT-Me) and thiols (MT-SH) content. Tissue redox status was assessed using lactate/pyruvate ratio. The assay of cells viability was based on the lysosomes ability of hemocytes to concentrate the Neutral Red (NR) dye. It was found, that MT-SH content in the digestive gland was increased under all exposures. Treatment with Dc increased the level of MT-Me, whereas treatment with Gl decreased it and increased lactate/pyruvate ratio. Nf decreased this ratio by elevating pyruvate level and increased lysosomal membrane stability in hemocytes. At co-exposure to xenobiotics and elevated temperature the number of hemocytes with nuclear abnormalities was increased indicating the exceeding of organisms’ adaptive limits. Multivariate statistical analyses showed negative correlations in pairs MT-SH/MT-Me and MT-SH/pyruvate and distinguished Gl and DcNfGl+T exposed groups from other groups.
Effects of ethylthiosulfanylate and chromium (VI) on the state of pro/antioxidant system in rat liver
B. І. Kotyk1, R. Ya. Iskra1, O. M. Slivinska1, N. M. Liubas1,
A. Z. Pylypets1, V. I. Lubenets2, V. I. Pryimych3
1Institute of Animal Biology, NAAS of Ukraine, Lviv;
2Lviv Polytechnic National University, Ukraine;
3Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine;
e-mail: kicyniabo@gmail.com
Received: 04 April 2020; Accepted: 25 June 2020
Ethylthiosulfanylate is alkyl ester of thiosulfoacid and belongs to the class of thiosulfonate compounds. Structurally, thiosulfonates are synthetic analogues of natural phytoncides. It is known that, natural organic sulfur-containing compounds are characterized by antioxidant and detoxification properties against heavy metals toxicity. Therefore, the purpose of the study was to investigate the influence of ethylthiosulfanylate, as a synthetic analogue of natural phytoncides, on the state of the pro/antioxidant system in the liver of laboratory rats exposed to Cr(VI). It was found that ethylthiosulfanylate exposure at a dose 100 mg/kg body weight daily for 14 days led to a decrease in the intensity of increasing of the lipid hydroperoxides (LHP) content in the rat liver caused by Cr(VI) action. In addition, ethylthiosulfanylate pretreatment prevented depletion of reduced glutathione (GSH) pool under the action of potassium dichromate oxidative stress and performed the accumulation of cellular GSH in rat liver.
The oxidative status in patients with chronic kidney disease
V. S. Vasylchenko1,2*, L. V. Korol1, O. B. Kuchmenko2, N. M. Stepanova1
1State Institution “Institute of Nephrology of the National Academy
of Medical Sciences of Ukraine”, Kyiv;
2National University of Kyiv-Mohyla Academy, Ukraine;
*e-mail: vasylchenkovita@gmail.com
Received: 26 March 2020; Accepted: 25 June 2020
An excess of free radicals accompanies the development of renal pathologies and causes numerous concomitant complications and syndromes. The most common of these are cardiometabolic syndromes in patients with chronic kidney disease. Therefore, the purpose of the study was to determine the activity of paraoxonase-1 and myeloperoxidase, which are associated with indicators of high-density lipoproteins content and oxidative stress in the blood of patients with the chronic stage of kidney disease. The activity of the enzymes, thiobarbiturate-active products concentration and transferrin, ceruloplasmin, thiol compounds content were determined in the blood of patients with chronic kidney disease. The oxidative status was shown to be changed. Thus, myeloperoxidase activity, the content of oxidized proteins and the concentration of thiobarbiturate-positive components were increased, while the activity of the antioxidant enzyme paraoxonase-1, the content of transferrin, ceruloplasmin and thiol compounds were decreased. The ratio of myeloperoxidase/paraoxonase-1 activities was progressively increased up to 9-fold, indicating the presence of cardiovascular complications in patients. The data obtained allowed to extend the range of indicators for monitoring the development of cardiometabolic disorders in the progression of chronic kidney disease.
DNA loop domain rearrangements in blast transformed human lymphocytes and lymphoid leukaemic Jurkat T cells
K. Afanasieva1, V. Olefirenko1, A. Martyniak1,
L. Lukash2, A. Sivolob1*
1Taras Shevchenko National University of Kyiv, Ukraine;
2Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: sivolob@univ.kiev.ua
Received: 06 April 2020; Accepted: 25 June 2020
Chromatin loops are important elements of both chromatin higher-order structure and transcription regulation system. Our previous works have shown that several features of the loop domain organization could be investigated by single cell gel electrophoresis (the comet assay) using the kinetic approach. In this study we applied this technique to study DNA loop domain organization in lymphoid cells: human lymphocytes, lymphoblasts cultivated during 24 h and 44 h, and T cells of Jurkat cell line. Two features of the loop domain organization were found to depend on the cell functional state. First, DNA fraction in the loops of large sizes (more than ~200 kb) was essentially increased in proliferating (de-differentiated) cells in comparison with terminally differentiated lymphocytes. Second, the linear density of the loops not larger than ~200 kb was decreased in transcriptionally active cells and was increased upon their inactivation.
ERN1 dependent regulation of TMED10, MYL9, SPOCK1, CUL4A and CUL4B genes expression at glucose and glutamine deprivations in U87 glioma cells
O. H. Minchenko*, O. S. Hnatiuk, D. O. Tsymbal, Y. M. Viletska,
S. V. Danilovskyi, O. V. Halkin, I. V. Kryvdiuk, O. V. Rudnytska
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: ominchenko@yahoo.com
Received: 05 April 2020; Accepted: 25 June 2020
It was shown previously that inhibition of ERN1 (endoplasmic reticulum to nucleus signaling 1) pathway, a central mediator of the unfolded protein response, leads to suppression of tumor growth through down-regulation of key pro-proliferative and up-regulation of tumor suppressor factors and modifies the sensitivity of these genes to glucose and glutamine deprivation. However, the executive mechanisms of ERN1 mediated control of glioma cell proliferation are not yet known. The goal of this study was to estimate the effect of glucose and glutamine deprivations on expression of cancer related genes in glioma U87 cells at ERN1 signaling inhibition for evaluation of their possible significance in ERN1 mediated control of glioma cell proliferation. We have studied the effect of glucose and glutamine deprivations on the expression level of cancer related genes encoding TMED10 (transmembrane p24 trafficking protein 10), MYL9 (myosin, light chain 9, regulatory), SPOCK1 (sparc/osteonectin, cwcv and kazal-like domains proteoglycan 1), CUL4A (cullin 4A), and CUL4B in U87 glioma control cells and cells with ERN1 knockdown. It was shown that at glucose deprivation the expression level of MYL9, SPOCK1 and CUL4B genes was significantly up-regulated in control glioma cells. ERN1 knockdown modified the sensitivity to glucose deprivation of all studied genes except TMED10 gene. At glutamine deprivation the expression of MYL9, CUL4A and CUL4B genes was shown to be up-regulated in control glioma cells. The sensitivity of MYL9, TMED10 and CUL4B gene expression to glutamine deprivation in glioma cells with ERN1 knockdown was significantly modified, while CUL4A and SPOCK1 gene expression did not respond to ERN1 inhibition. The present study demonstrates that glucose and glutamine deprivation affected the expression of the most studied genes in a specific manner and that inhibition of ERN1 signaling preferentially modified their expression at glucose and glutamine deprivation.
Cryoprotective agents affect amino acids incorporation into total proteins in cells of lymphoid organs and liver of experimental animals
A. K. Gulevskyy*, Yu. S. Akhatova, A. Yu. Nikolchenko
Institute for Problems of Cryobiology and Cryomedicine,
National Academy of Sciences of Ukraine, Kharkiv;
*e-mail: profgulevskyy@gmail.com
Received: 07 July 2019; Accepted: 25 June 2020
The effect of penetrating (glycerol, DMSO) and poorly penetrating (PEG-400) cryoprotective agents on labeled amino acids incorporation into de novo synthesized proteins in the cells of mice thymus, lymph nodes, spleen and cell-free rat liver extract was studied. Cryoprotective agents within the range of concentrations which provide a cryoprotective effect were found to inhibit significantly protein synthesis in cell-free systems under investigation. The most effective inhibition was exerted by polymeric cryoprotective agent PEG-400. Cryoprotective agents more efficiently inhibited protein synthesis at a cell level as compared with that in cell-free system that was likely associated with their effect on amino acids transport system. An inhibitory effect of cryoprotective agents on the protein synthesizing apparatus of cells was determined to be Mg2+-dependent and reversible.
L-carnitine administration effects on AMPK, APPL1 and PPARγ genes expression in the liver and serum adiponectin levels and HOMA-IR in type 2 diabetes rat model induced by STZ and nicotinamide
B. Shahouzehi1,2, H. Fallah3, Y. Masoumi-Ardakani4*
1Student Research Committee, Kerman University of Medical Sciences, Kerman, Iran;
2Cardiovascular Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
3Department of Biochemistry, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;
4Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
*e-mail: ymab125@gmail.com
Received: 18 January 2020; Accepted: 25 June 2020
Diabetes is a chronic disease and a public health problem globally. L-Carnitine is synthesized in the liver, promotes fatty acids oxidation and currently is used as a supplement against weight gain. Carnitine level is found to be reduced in diabetic patients and to be beneficial as a supplement at diabetes, but the mechanisms of this effect is not fully understood. Therefore, we evaluated the oral L-carnitine supplementation on expression of AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor gamma (PPARγ), adaptor protein APPL1 genes in the liver and insulin and adiponectin levels in the serum of diabetic rats. Rats were randomly divided into three groups (n = 8) as follow: group 1 – control without any treatment, group 2 – diabetic control rats which received STZ (45 mg/kg) and nicotinamide (200 mg/kg) by i.p. injection, group 3 – diabetic rats which received 600 mg/kg/day carnitine orally for 35 days. It was found that L-carnitine supplementation reduced the level of fasting glucose compared to that in control and diabetic groups (P = 0.001, P = 0.0001 respectively) and increased adiponectin level compared to diabetic nontreated rats (P = 0.0001). Homeostasis model assessment of insulin resistance (HOMA-IR) was significantly increased in the diabetic group and reduced in the group that received L-carnitine. These promising beneficial effect of L-carnitine on the type 2 diabetes in rats’ model was shown to be conducted through the up-regulation of AMPK, PPARγ and APPL1 genes expression in the liver and elevation of serum adiponectin level.
Selected 5-amino-1-aryl-1H-1,2,3-triazole scaffolds as promising antiproliferative agents
N. Pokhodylo1*, O. Shyyka1, N. Finiuk2, R. Stoika2
1Ivan Franko National University of Lviv, Ukraine;
2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
*e-mail: pokhodylo@gmail.com; stoika@cellbiol.lviv.ua
Received: 09 January 2020; Accepted: 25 June 2020
Development of a new effective drugs with low side effects and definite chemical characteristics needs indentification of bioactive scaffolds for further structural optimization. New synthesized derivatives of 4-hetaryl-5-amino-1-aryl-1H-1,2,3-triazoles and 3H-[1,2,3]triazolo[4,5-b]pyridines were tested for anticancer activity using 60 human tumor cell lines within 9 cancer types. The selective influence of (5-amino-1H-1,2,3-triazol-4-yl)quinazolin-4(3H)-ones: 2-(5-amino-1-(4-chlorophenyl)-1H-1,2,3-triazol-4-yl)quinazolin-4(3H)-one and 2-(5-amino-1-phenyl-1H-1,2,3-triazol-4-yl)-6-bromoquinazolin-4(3H)-one on ovarian cancer OVCAR-4 cells with growth percentage (GP) = -4.08 and 6.63%, respectively, was found. The derivative 5,7-diamino-3-(3-(trifluoromethyl)phenyl)-3H-[1,2,3]triazolo[4,5-b]pyridine-6-carbonitrile possessed high activity towards lung cancer EKVX cells (GP = 29.14%). The compounds were shown to be less toxic than doxorubicin towards non-tumor human embryonic kidney cells of HEK293 line. Thus, the results of our study confirm the anticancer potential of compounds based on 5-amino-1-aryl-1H-1,2,3-triazoles scaffolds and their fused polycyclic derivatives.
Kinetics of interaction between polyreactive immunoglobulins and antigen
S. A. Bobrovnik1*, O. V. Ogloblya2, M. O. Demchenko1, S. V. Komisarenko1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: s-bobrov@ukr.net
Received: 28 January 2020; Accepted: 25 June 2020
A number of experimental kinetics curves of mice polyreactive immunoglobulins (PRIGs) binding to ovalbumin immobilized on immunologic plates were obtained at different temperatures. Analysis of these curves allowed us to conclude that the model of PRIGs interaction with antigens proposed by us earlier and consisted on PRIGs activation (i.e. exposition of hydrophobic patches on PRIGs surface) and either sequential binding to antigen or inactivation was is in a good agreement with the experimental data obtained in this study. We have designed a method of the rate constants evaluation from experimental binding curves. It was found that the rate constant of the activated PRIGs binding to immobilized antigen did not depend on temperature. The rate constant of PRIGs activation occurred to be depend on temperature more strongly than the rate constant of PRIGs inactivation. We have concluded from the acquired dependences that at 37°С the number of activated PRIGs was 15 times higher than that at 0°С.
ATP-sensitive potassium transport in rat brain mitochondria is highly sensitive to mK(ATP) channels openers: a light scattering study
O. V. Akopova*, L. I. Kolchinskaya, V. I. Nosar,
A. N. Smirnov, L. V. Bratus
Bogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: ov_akopova@ukr.net
Received: 17 January 2020; Accepted: 25 June 2020
The aspects of ATP-sensitive K+ transport regulation by mitochondrial K+,ATP-sensitive (mKATP) channels openers are important for understanding the properties of these channels. The effect of KATP channels openers (KCOs) diazoxide and pinacidil on ATP-sensitive K+ transport in isolated brain mitochondria was studied in the absence and the presence of MgATP using light scattering technique. Without MgATP we observed high sensitivity of ATP-sensitive K+ transport to both drugs with full activation at ≤ 0.5 µM. ATP-sensitive K+ transport was specifically blocked by ATP in the presence of Mg2+. Neither Mg2+ nor ATP affected Vmax of ATP-sensitive K+ transport activated by KCOs, but MgATP shifted the activation curve to micromolar scale. The blockage of ATP-sensitive K+ transport by KATP channels blockers glibenclamide and 5-hydroxydecanoate in the absence and the presence of MgATP proved the sensitivity of ATP-sensitive K+ transport to the blockers of mKATP channel. Full activation of mKATP channel by diazoxide and pinacidil on sub-micromolar scale in the absence of MgATP was shown. The sensitivity of ATP-sensitive K+ transport to the known modulators of mKATP channel (diazoxide, pinacidil, glibenclamide, 5-HD and MgATP) proved the identity of ATP-sensitive K+ transport with mKATP channel activity. Based on our studies, we hypothesized that mKATP channel might comprise high affinity sites for KCOs binding screened by MgATP. The results of this work reveal novel not described earlier aspects of the regulation of ATP-sensitive K+ transport by mKATP channels openers, important for understanding of mKATP channel properties.