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The Nobel Prize in Chemistry 2018

The Nobel Prize in Physiology or Medicine 2018

The FEBS3+ Meeting – XIth Parnas Conference – Young Scientists Forum ‘Biochemistry and Molecular Biology for Innovative Medicine’

(September 3-5, 2018, Kyiv, Ukraine)

Founder of molecular immunology in Ukraine, well-known political and public figure

On the 75th birthday of Academician of the NAS of Ukraine S. V. Komisarenko

S. O. Kosterin, V. M. Danilova

July 9 marks the 75th birthday of Serhiy Vasylyovych Komisarenko – the well-known scientist-biochemist, founder of the scientific school in molecular immunology, diplomat, state and public figure, laureate of the State Prize of Ukraine in the Field of Science and Technology (1979), Honored Science and Technology Worker of Ukraine (2008), Winner of the O.V. Palladin Prize (2003) and I.I. Mechnikov Prize (2012) of the NAS of Ukraine, Academician-Secretary of the Division of Biochemistry, Physiology and Molecular Biology of the NAS of Ukraine, director of the O.V. Palladin Institute of Biochemistry of the NAS of Ukraine, Doctor of sciences (biology), Professor, Member of the NAS and NAMS of Ukraine.

Purification procedure and assay for the activity of lysyl oxidase

O. O. Gudkova, N. V. Latyshko, O. V. Zaitseva, S. G. Shandrenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

The goal of the present study was to extract and purify lysyl oxidase from rodent’s tissues by a fast, simple, effective and inexpensive method and to develop a sensitive, time-saving lysyl oxidase specific activity assay for routine in vitro experiments. Lysyl oxidase was purified by elaborated purification procedure which relies on negative adsorption principle, that is, an effective decrease in the concentration of ballast components by the polar hydrophilic adsorbent and increasing the concentration of the protein of interest. Peroxide-coupled lysyl oxidase activity quantification methods based on luminol chemiluminescence in the presence of horseradish peroxidase as a catalyst and fluorescent detection using folic acid and Cu(II)  with 1,5-diaminopentane as the substrate, were designed. Lysyl oxidase was partially purified from urea extracts of rodent’s tissues. Used purification procedure ensures the fast release of 93% of ballast proteins as shown by polyacrylamide gel electrophoresis. Lysyl oxidase specific activity after purification was 10-22-fold higher than that of the original extract. The molecular mass of murine lysyl oxidase from lung and heart was estimated to be ~32 kDa.
We elaborated two sensitive methods for lysyl oxidase activity quantification and fast inexpensive procedure for partial enzyme purification useful in bulky in vitro experiments.

The indices of nitrogen (II) oxide system in experimental hepatopulmonary syndrome

I. Ya. Krynytska, M. I. Marushchak

I. Horbachevsky Ternopil State Medical University, Ternopil, Ukraine;

Hepatopulmonary syndrome (HPS) is a pulmonary complication of liver disease characterized by arterial hypoxemia. Altered nitrogen (II) oxide (NO) production has also been implicated in the pathogenesis of the HPS. The present study was designed to evaluate the indices of NO system in the blood serum and lung tissue of animals with different models of hepatopulmonary syndrome. The total NOS activity was performed by monitoring the rate of conversion of L-arginine into citrulline. The total contents of NO metabolites was assessed by evaluation of their amount, which included nitrite ions that were previously presented in the sample (NO2) and also nitrate ions reducted to nitrites (NO3). We found a significant increase in total NOS activity in the lung tissue of Rats of both experimental groups  as compared to control animals, but it was greater in the rats on the 28th day after the common bile duct ligation. The total concentration of NO2 + NO3 in the lung tissue of the rats in the experimental group N 1 also significantly increased (5.8 times) and in the rats of the experimental group with carbon – 4.5 times (P < 0.001) vs the control group. Thus, in rats with different models of hepatopulmonary syndrome the activation of nitroxydergic process by a significant increase in nitrogen (II) oxide metabolites contents and total NO synthases activity has been established. Herewith a more pronounced intensification of nitroxydergic processes was observed in rats on the 28th day after the common bile duct ligation.

The effect of N-stearoylethanolamine on adipocytes free cholesterol content and phospholipid composition in rats with obesity-induced insulin resistance

O. S. Dziuba, Ie. A. Hudz, H. V. Kosiakova, T. M. Horid’ko, V. M. Klimashevsky, N. M. Hula

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

Obesity induces molecular changes that promote associated disorders, such as insulin resistance (IR) and type 2 diabetes. Low insulin sensitivity occurs primarily due to defects in the pathway of insulin action in target tissues, and there is a hypothesis that IR may originate in adipose tissue and is followed by dyslipidemia. In this study using methods of thin-layer and gas-liquid chromatography we investigated free cholesterol content and phospholipid composition of adipocytes of obesity-induced IR rats and its changes induced by the N-stearoylethanolamine (NSE) administration. The results we obtained demonstrated that free cholesterol content significantly increased in adipocytes of IR rats compared to control. The analysis of phospholipid composition indicated a reduction of phosphatidylcholine and the total content of phosphatidylinositol with phosphatidylserine, whereas the content of lysophosphatidylcholine, sphingomyelin and phosphatidylethanolamine increased in IR group compared to control. NSE administration caused a statistically significant decrease in total cholesterol level and had a considerable effect on normalization of individual phospholipids content. As far as NSE administration caused a statistically significant decrease in free cholesterol level and had a considerable effect on normalization of individual phospholipids content of adipocytes, we can consider NSE as a prospective compound worthy more complex investigation of its action under the pathological conditions.

Evaluation of metallothioneins, oxidative stress and signs of cytotoxicity in young obese women

H. I. Falfushynska, O. I. Horyn, V. V. Khoma, G. V. Tereshchuk,
D. V. Osadchuk, N. I. Rusnak, O. B. Stoliar

Ternopil Volodymyr Hnatiuk National Pedagogical University, Ukraine;

Obesity is rapidly increasing all over the world and pretends to be the global medical and social problem. Thus, the understanding of early signs of obesity and suitable biomarkers is urgently needed for developing an adequate strategy of the obesity prevention and a decrease in its growth rate. The parameters of the lipids’ metabolism and oxidative stress, metallothioneins and signs of cytotoxicity have been investigated in blood samples of young obese women (O-group, 32 < Body Mass Index (BMI) < 37). With regard to persons of O-group they had higher catalase activity (by 435%), level of reactive oxygen species (by 129%), level of oxidised glutathione (by 55%), lipid peroxidation (by 26%) and protein carbonyls (by 345%) in the blood, when compared with control. The obesity was accompanied by an increase in concentration of metallothioneins which have a partial tread effect on radical processes and reduce manifestations of oxidative damage to biomolecules in obese patients. The obese women had the signs of cytotoxicity as higher lactate dehydrogenase activity (by 387%) and DNA fragmentation (by 42%). The principal component analysis revealed the set of biological traits which describes the obesity progress and it included metallothioneins, parameters of oxidative stress, cytotoxicity, BMI and a concentration of low density lipoproteins and total cholesterol. The BMI was in a good correlation with parameters of the lipid metabolism, oxidative injury and cytotoxicity (r > |0.73|, P < 0.001).

Overall hemostasis potential of the blood plasma and its relation to some molecular markers of the hemostasis system in patients with chronic renal disease of stage VD

B. G. Storozhuk1, L. V. Pyrogova2, T. M. Chernyshenko2, O. P. Kostiuchenko2,
I. M. Kolesnikova2, T. M. Platonova2, O. B. Storozhuk1, L. O. Storozhuk1,
G. K. Bereznitsky2, P. Yu. Tsap2, O. O. Masenko2,
E. M. Makogonenko2, E. V. Lugovskoy2

1Pyrogov National Medical University of Vinnytsa, Ukraine;
2Palladin Instiute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;

The values of the coagulation, overall and fibrinolysis potentials were estimated by the method of the global potential of Blomback M., as well as the values of concentrations of molecular markers of the hemostasis system: soluble fibrin (sf), D-dimer, fibrinogen (Fg) and protein C (88 patients, 52 of them men, 36 women). It was shown that hemostasis system activity in women plasma is higher than that in men plasma. The division of patients into 3 groups, depending on the concentration of sf: less than normal – sf ≤ 3, about norm – 3 < sf < 4 and more than norm – sf > 4 μg/ml, allowed establishing the growth of the parameters of both the hemostatic potential and concentrations of molecular markers in accordance with concentration of sf in the groups of patients. Paerson’s correlation analysis of the relationship between the parameters of the hemostasis potential and concentrations of molecular markers revealed an increase in the correlation relationship to the strong and very strong between the parameters of coagulation, fibrinolysis and protein C systems with an increase in the concentration of soluble fibrin in plasma of patients.

Action of methyl jasmonate and salt stress on antioxidant system of Arabidopsis plants defective in jasmonate signaling genes

Т. О. Yastreb1, Yu. E. Kolupaev1,2, N. V. Shvidenko1, A. P. Dmitriev3

1Dokuchaev Kharkiv National Agrarian University, Ukraine;
2Karazin Kharkiv National University, Ukraine;
3Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv;

Role of jasmonate signaling in the regulation of stress-protective systems in Arabidopsis under salt stress remains insufficiently studied. For its clarification, comparative studies with mutants lacking various protein components of jasmonate signaling are advisable. In this connection, effects of methyl jasmonate (MJ, 50 μM) and salt stress (NaCl, 150 mM) on functioning of antioxidant and osmoprotective systems of wild-type Arabidopsis plants (Col-0) and ones defective in jasmonate signaling, namely coi1 (mutant for gene coding the protein COI1, which participates in removal of repressor proteins of transcription factors of jasmonate signaling) and jin1 (mutant defective in gene encoding the transcription factor JIN1/MYC2, one of the key in jasmonate signaling), were investigated. Salt stress inhibited growth of plants of all three genotypes. Treatment with MJ before salt stress positively influenced only the growth of wild-type plants. In contrast to mutants coi1 and jin1, Col-0 plants treated with MJ, under conditions of salt stress, kept close to the control values of water and total chlorophylls content, and the content of carotenoids increased. The coi1 plants under normal conditions differed from wild-type plants and jin1 mutants by reduced activity of guaiacol peroxidase and catalase and increased proline content. Treatment with MJ did not affect the activity of antioxidant enzymes and proline content in both mutants defective in jasmonate signaling. Under salt stress, the activity of superoxide dismutase, catalase and guaiacol peroxidase, as well as the content of proline and anthocyanins, in wild-type plants treated with MJ, were significantly higher than in control plants. The role of jasmonate-dependent protective systems in resistance of Arabidopsis plants to salt stress is discussed.