Category Archives: Uncategorized
Proteins oxidative modification and antioxidant enzymes activity in the liver mitochondria of rats under laser irradiation and administration of ω-3 polyunsaturated fatty acids
O. V. Ketsa*, M. O. Zelinska, M. M. Marchenko
Fedkovich Chernovtsy National University, Chernovtsy, Ukraine;
*e-mail: o.ketsa@chnu.edu.ua
Received: 12 August 2021; Accepted: 21 January 2022
The effect of laser irradiation of rats combined with omega-3 polyunsaturated fatty acids (ω-3 PUFA) administration on the proteins oxidative modification and superoxide dismutase and catalase activity in the mitochondrial fraction of the liver was investigated. Animals were irradiated with a 650 nm laser diode in the abdomen daily for 4 min at the distance of 10 cm from the skin surface. ω-3 PUFA were administered per os at a daily dose of 120 mg/kg of the body weight. Fatty acids in the fish oil were identified by gas chromatography. Animals were divided into five groups (12 animals in each group): I – intact rats (control); II – rats exposed to the daily laser irradiation for 7 or 14 days ; III – rats that received ω-3 PUFA two hours after irradiation; IV – rats that received ω-3 PUFA two hours before irradiation; V – rats that received ω-3 PUFA for 7 days before irradiation. The mitochondrial fraction of rat liver was obtained by differential centrifugation. The increase in the content of protein carbonyl derivatives and a decrease of protein thiol groups in the liver mitochondrial fraction were detected after seven-day laser irradiation of rats. As the duration of irradiation increases, superoxide dismutase and catalase activity was decreased, indicating a depletion of mitochondrial antioxidant reserves. No antioxidant effect was observed when ω-3 PUFA were administrated after laser irradiation and a slight antioxidant effect was shown when ω-3 PUFA were administrated two hours before irradiation. Preliminary seven-day administration ω-3 PUFA before laser irradiation was the most effective, as it reduced the level of protein carbonyl derivatives and O2•--generation, increased proteins SH-groups content and antioxidant enzymes activity.
Systemic inflammation biomarkers in 6-OHDA- and LPS-induced Parkinson’s disease in rats
Zh. Oliynyk*, M. Rudyk, V. Svyatetska, T. Dovbynchuk, G. Tolstanova, L. Skivka
ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: ojankin@yahoo.com
Received: 14 December 2021; Accepted: 21 January 2022
Hematological and immunological markers of systemic inflammation were studied in 6-hydroxydopamine (6-OHDA)- and lipopolysaccharide (LPS)-induced models of Parkinson’s disease (PD). Experiments were carried out on adult male Wistar rats: 1 – intact animals; 2 – sham-operated animals and 3 – 6-OHDA- and LPS-lesioned animals. PD development was confirmed by the results of behavioral testing (apomorphine test, open field test) and immunohistochemical detection of the loss of dopaminergic neurons. Hematological indices (complete blood count and differential leukocyte count (DLC)) were examined using hematological analyser. Immunological indices included phenotypic (CD206 and CD80/86) and metabolic (oxidative metabolism and phagocytic activity) characteristics of circulating monocytes (Mo) and granulocytes (Gr), which were determined by flow cytometry, as well as plasma levels of C-reactive protein, which were determined by ELISA. LPS-induced PD was associated with neutrophilia, 1.9 times increased neutrophil-to-lymphocyte ratio, 3 times increased platelet-to-lymphocyte ratio, and 3 times increased systemic immune inflammation index as compared to intact animals. Functional profile of circulating phagocytes from LPS-lesioned animals was characterized by the pro-inflammtory metabolic shift, as was indicated by 5 times increased oxidative metabolism indices and up-regulated CD80/86 expression along with decreased phagocytic activity and CD206 expression. 6-OHDA-lesioned rats demonstrated decreased DLC indices as compared to intact and sham-operated rats. Functional profile of circulating phagocytes in this model was characterized by anti-inflammatory shift. The results obtained from this study demonstrated that stereotaxic LPS-induced PD is appropriate rodent model for the study of systemic inflammation which is inherent for the disease pathophysiology.
Rhabdomyolysis attenuates activity of semicarbazide sensitive amine oxidase as the marker of nephropathy in diabetic rats
O. Hudkova*, I. Krysiuk, L. Drobot, N. Latyshko
Department of Cell Signaling, Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: ogudkova@biohem.kiev.ua
Received: 22 December 2021; Accepted: 21 January 2022
Amine oxidases are involved in the progression of many diseases and their complications, including renal failure, due to the generation of the three toxic metabolites (H2O2, aldehydes, and ammonia) in the course of biogenic amines oxidative deamination. The participation of the first two products in kidney pathogenesis was confirmed, whereas the role of ammonia as a potential inducer of the nitrozative stress is not yet understood. The aim of the present study was to test how further intensification of oxidative stress would affect diabetes-mediated metabolic changes. For this purpose, a rat model of glycerol-induced rhabdomyolysis, as a source of powerful oxidative stress due to the release of labile Fe3+ from ruptured myocytes, on the background of streptozotocin-induced diabetes was used. The experimental animal groups were as follows: group 1 – ‘Control’, group 2 – ‘Diabetes’, group 3 – ‘Diabetes + rhabdomyolysis’. A multifold increase in semicarbazide sensitive amine oxidase (SSAO) activity in the kidney and blood, free radicals (FR), MetHb and 3-nitrotyrosine (3-NT) levels in the blood, as well as the emergence of HbNO in plasma and dinitrosyl iron complexes (DNICs) in the liver of animals in group 2 as compared to control were revealed. An additional increase in FR, HbNO levels in the blood, and DNICs in the liver of animals in the diabetes + rhabdomyolysis group as compared to the diabetes group, which correlated with the appearance of a large amount of Fe3+ in the blood of group 3 animals, was detected. Unexpectedly, we observed the positive regulatory effects in animals of the diabetes + rhabdomyolysis group, in particular, a decreased SSAO activity in the kidney and 3-NT level in plasma, as well as the normalization of activity of pro- and antioxidant enzymes in the blood and liver compared to animals of diabetes group. These consequences mediated by rhabdomyolysis may be the result of NO exclusion from the circulation due to the excessive formation of NO stable complexes in the blood and liver. The data obtained allow us to consider SSAO activity as a marker of renal failure in diabetes mellitus. In addition, we suggest a significant role of nitrosative stress in the development of pathology, and, therefore, recommend NO-traps in the complex treatment of diabetic complications.
Glucose deprivation-induced glycogen degradation and viability are altered in peripheral blood mononuclear cells of type 2 diabetes patients
K. S. Praveen Kumar1, P. Kamarthy2, S. Balakrishna1*
1Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education, Kolar, India;
2Department of General Medicine, Sri Devaraj Urs Medical College, Tamaka, Kolar, India;
*e-mail: sharath@sduu.ac.in
Received: 07 September 2021; Accepted: 21 January 2022
The glycogen pathway plays an important role in glucose homeostasis. Impairment of the glycogen pathway has been linked to diabetes mellitus. The aim of the study is to compare the levels of glucose deprivation-induced glycogen degradation and cell viability in peripheral blood mononuclear cells from type 2 diabetes mellitus patients and healthy controls. This was a case-control study comprising 45 T2DM patients and 45 healthy controls. PBMCs were prepared from peripheral blood by density gradient centrifugation. Glycogen levels were measured by the periodic acid-schiff (PAS) staining method. Glycogen degradation was measured as percent change in PAS-stained cells before and after glucose deprivation. PBMC viability was measured by trypan-blue assay. The levels of glucose deprivation-induced glycogen degradation were 55.4% (IQR: 50.6–61.3) in the T2DM group and 70.5% (IQR: 63.9–72.2) in the healthy control group. The difference between the two groups was statistically significant (P = 0.001). The levels of glucose deprivation-induced cell viability were 70.9% (IQR: 66.3–77.1) in the T2DM group and 87.8% (IQR: 83.7–90.7) in the healthy control group. The difference between the two groups was statistically significant (P = 0.001). Together these results indicate that the glucose deprivation-induced glycogen degradation and viability are reduced in PBMCs of T2DM patients.
Pyridoxine, folate, and cobalamin and the condition of the innate and acquired immunity
M. I. Kinash, O. R. Boyarchuk
Ivan Horbachevsky Ternopil National Medical University, Ukraine;
e-mail: kinash@tdmu.edu.ua, boyarchuk@tdmu.edu.ua
Received: 23 September 2021; Accepted: 21 January 2022
The immune response to the pathogen depends on the state and coordinated activity of innate and acquired immunity. To date, the positive role of many micronutrients (vitamins and minerals) in maintaining homeostasis of the human immune system has been proven. The aim of the review was to analyze the main manifestations of vitamins B6, B9, B12 influence on the state of innate and adaptive immunity. These vitamins play an important role in the synthesis of DNA and RNA, cytokines and immunoglobulins, thus maintaining the proper state of the immune system. The functioning of the main links of specific and nonspecific immunity at the normal vitamin status and at pyridoxine, folate and cobalamin deficiency is compared.
Breaking the genetic code – a new revolutionary stage in the development of molecular biology: 1968 Nobel Prize laureates M. W. Nirenberg, H. G. Khorana, R. W. Holley
O. P. Matyshevska*, V. M. Danilova, S. V. Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: matysh@yahoo.com
Received: 28 October 2021; Accepted: 12 November 2021
This review presents the life stories of M. Nirenberg, H. Khorana, and R. Holley, winners of the 1968 Nobel Prize in Physiology or Medicine, the history of the discoveries made by these scientists, and the methodological approaches used in their works. Owing to the M. Nirenberg and H. Khorana research, the nucleotide compositions of all mRNA triplet codons were decoded. H. Khorana was the first scientist to experimentally prove the direct link between the nucleotide sequence of DNA and the amino acid sequence of the synthesized protein and to obtain a synthetic gene. R. Holley was the first to completely decode the sequence of transport RNA, determine its secondary structure and role in protein synthesis on the ribosome. The Nobel Prize awarded to the scientists was a recognition of their contribution in understanding the mechanisms of coding and reading genetic information and marked a breakthrough moment in the development of molecular biology.
Differential impact of the temperature stress and soil drought on lipoxygenase activity in winter rye plants
L. M. Babenko, K. O. Romanenko*, I. V. Kosakivska
M.G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: katerynaromanenko4@gmail.com
Received: 5 July 2021; Accepted: 12 November 2021
Lipoxygenase cascade is a source of physiologically active compounds, the presence of which is considered not only as a sign of damage but also as a trigger of adaptive responses to stress. The aim of the study was to determine the effects of short-term (2 h) heat (40°C) and cold (4°C) temperature stress and moderate soil drought on lipoxygenase (LOX) activity in 14-day-old winter rye (Secale cereale L. ‘Boguslavka’) plants. The shoots were found to have both membrane-bound 9-LOX1 and 9-LOX2 and soluble 13-LOX activity, the roots – membrane-bound 9-LOX activity. After heat stress, the activity of 9-LOX1 and 9-LOX2 in the shoots increased by 3 and 2 times, respectively, the activity of 9-LOX in the roots – by 2 times, and 13-LOX activity in the shoots decreased by 1.5 times. After the cold stress, the activity of 9-LOX1 and 9-LOX2 in the shoots raised by almost 1.5 times, the activity of 9-LOX in the roots – by 1.2 times. Moderate soil drought caused enhancement in the activity of both membrane-bound isoforms of 9-LOX in the shoots by 1.5–2 times and in the roots – by 3 times. The established fluctuations indicate that molecular forms of LOX with different localization are differentially involved in the winter rye response to temperature stress and moderate soil drought.
Prediction of pEC50(M) and molecular docking study for the selective inhibition of arachidonate 5-lipoxygenase
N. R. Das1, P. G. R. Achary2*
1Department of CSIT, Siksha ‘O’ Anusandhan deemed to be University, Bhubaneswar, Odisha, India;
2Department of Chemistry, Faculty of Engineering and Technology (ITER), Siksha ‘O’ Anusandhan deemed to be University, Bhubaneswar, Odisha, India;
*e-mail: pgrachary@soa.ac.in
Received: 28 April 2021; Accepted: 12 November 2021
Arachidonate 5-lipoxygenase (ALOX5) is considered a prime target for drug discovery in the area of liver fibrosis, rheumatoid arthritis, atherosclerosis, cancer and asthma. To date, the lead rate in the discovery of drugs that inhibit ALOX5 for the treatment of the above diseases is not satisfactory. So, the development of powerful and effective ALOX5-targeted drugs is desired. In this regard, Quantitative Structure-Activity Relationship (QSAR) and molecular docking can have a major role in screening and designing drugs. In this work, 3D-QSAR models were proposed, which were built using the techniques like Multiple Linear Regression (MLR), and Partial Least Squares (PLS) for the pEC50(M) taking a diverse dataset of 112 molecules. The technique of the ‘Index of Ideality of Correlation (IIC)’ was also investigated to generate an optimal descriptor derived from the SMILES molecular structure. The effect of the number and nature of descriptors on the model were analyzed. The models can be helpful in providing better directions for the development of novel drug targets for 5-lipoxygenase. A significant improvement in the stability of the model was observed by the incorporation of the optimal descriptor. The molecular docking results showed that the ALOX5 receptor was well inhibited by the 112 ligands showing the least binding energy (-10.8 Kcal/mol). In order to validate the binding mode of the ligands docked with AutoDock Vina software, the top-scored compounds were re-docked using DockThor online docking server. The results obtained from docking suggest that the ligands with IDs 18, 20, 24, 30 and 44 are some of the potential inhibitors for ALOX5.
Indicators of mineral metabolism and dental status of young rats born from female with methionine-induced hyperhomocysteinemia
O. Kutelmakh1*, R. Lesyk2,3, Yu. Chumakova4,
S. Bondar1, V. Dronenko1, L. Shkilniak1
1Vinnytsia National Pirogov Memorial Medical University, Vinnytsya, Ukraine;
2Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
3University of Information Technology and Management in Rzeszow, Rzeszow, Poland;
4International Humanitarian University, Odessa, Ukraine;
*e-mail: ahzelo1968vin@i.ua
Received: 12 July 2021; Accepted: 12 November 2021
We assume that the violation of homocysteine (Hcy) metabolism and vitamin D level in mothers’ utero causes various abnormalities in offspring, including hard-woven teeth and periodontium impairments. The aim of this study was to evaluate the biochemical parameters of blood and alveolar bone homogenates and assess dental status of offspring born from female rats under long-term methionine hyperhomocysteinemia (HHcy). Clinical, biochemical and morphometric studies were performed on 30 female rats and 281 infantile and early juvenile rats. Serum Hcy levels were determined by competitive immunoassay with electrochemiluminescent detection, 25-hydroxyvitamin D (25OHD) level was estimated by ELISA. No violation of Hcy serum level was found in rats born from females with long-term methionine-induced hyperhomocysteinaemia. Hyperhomocysteinaemic diet caused by long-term use of methionine reduced the level of total and ionized calcium and phosphates in the blood of pregnant females and their offspring. Elevated levels of homocysteine and decreased vitamin D levels in pregnant females caused a delay in the complete physiological eruption of permanent molars in their offspring. Biochemical changes in the blood, jawbones and the presence of early carious processes in the teeth of young rats, in our opinion, are the result of metabolic disorders in utero caused by hyperhomocysteinaemia and vitamin D deficiency in their mothers.