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The Nobel laureates’ contributions to the study of carbohydrate metabolism and its regulation. A. Harden, H. Euler-Chelpin, C. F. Cori, G. T. Cori, E. Sutherland, L. F. Leloir, H. Krebs, F. Lipmann, P. Mitchell

R. P. Vynogradova, V. M. Danilova, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kiyv;
e-mail: valdan@biochem.kiev.ua

Received: 13 November 2019; Accepted: 29 November 2019

Carbohydrate metabolism is a complex and multi-stage process. Many scientists (biochemists, physiologists, chemists) worked on deciphering this process, but only some of them were awarded the Nobel Prize. Thus, in the early XXth century, the work of A. Garden and H. Euler-Chelpin with yeast cells revealed that the conversion of carbohydrates (sugars) into end products occurs in living cells in several steps with the involvement of enzymes and that this conversion requires the presence of phosphoric acid residue. These studies were the beginning of exploring the chemical reactions (the reactions of intermediate metabolism) that are fundamentals for the vital functions of cells. In 1932, Hans Krebs discovered the ornithine cycle, a sequence of chemical reactions, in which the end product of nitrogen metabolism, urea, is formed in the liver of animals. The apogee of his research was the discovery of tri- and dicarboxylic acid cycle, which combines the oxidation of almost all organic compounds in living organisms. Fritz Lipmann, who in 1945 discovered coenzyme A and identified its role in the activation of organic compounds, furthered the works of H. Krebs. At the time, it became clear how inactive acetic acid and other organic acids are activated in living organisms to be oxidized in the tricarboxylic acid cycle. The great work was done by the spouses Gerty and Carl Cori,  and Bernardo Houssay, as well as their students and followers, in particular, Luis Leloir to clarify the mechanism of conversion (synthesis and breakdown) of glycogen in the liver and muscles. The peak of studying the carbohydrate metabolism was the research of Earl Sutherland who in 1958 revealed the regulation of the activity of enzymes involved in the conversion of chemical compounds (the example – phosphorylase), with the participation of the adenylate cyclase enzyme and c-AMP. The discovery of c-AMR established one of the fundamental principles of almost all vital processes. And the culmination of research on carbohydrate metabolism was the unrivaled works of Peter Mitchell, who studied the course of biochemical reactions in cells relative to certain intracellular “landmarks” and who formulated the chemiosmotic theory of oxidative phosphorylation that underlies bioenergetics.

Free radical oxidation in liver mitochondria of tumor-bearing rats and its correction by essential lipophilic nutrients

O. V. Ketsa, M. M. Marchenko

Fedkovich Chernivtsy National University, Ukraine;
e-mail: o.ketsa@chnu.edu.ua

Received: 21 May 2019; Accepted: 29 November 2019

The role of free radical oxidation in the increase of mitochondrial membranes permeability in organs which are not involved in oncogenesis and the development of the methods for preventing mitochondria dysfunction remain topical problems. In this work, the interconnection of lipid peroxidation (LPO) in liver mitochondrial fraction with the processes of mitochondrial swelling and cytochrome с release to the cytosol under separate and combined administration of ω-3 polyunsaturated fatty acids (PUFAs) and retinol acetate (vitamin A acetate) to rats with transplanted Guerin’s carcinoma was studied.  During the intensive tumor growth (14 days) the increase of superoxide radical generation and the content of primary (triene conjugates, TC), secondary (ketodienes and coupled trienes, CD+CT) and terminal (Schiff bases) lipid peroxidation products­ in the mitochondrial fraction of tumor-bearing rats was detected, which contributed to the mitochondrial swelling and cytochrome с release to the cytosol. Separate administration of ω-3 PUFAs to tumor-bearing rats decreased both free radical processes in mitochondrial fraction and mitochondrial swelling. Separate administration of retinol acetate in a high dose (3000 IU/kg of body weight) intensified free radical processes in the mitochondrial fraction of tumor-bearing rats, while administration of retinol acetate in a physiological dose (30 IU/kg of body weight) did not lead to changes compared to tumor-bearing rats that did not receive the drug. The prooxidant effects of retinoid were partially eliminated in the case of combined administration with ω-3 PUFA.

Cytotoxic activity of the cluster rhenium compound with β-alanine ligands

K. V. Polokhina1, D. E. Kytova1, A. V. Shtemenko1, N. I. Shtemenko1,2

1Ukrainian State University of Chemical Technology, Dnipro, Ukraine;
2Dnipro University of Technology, Ukraine;
e-mail: n.shtemenko@i.ua

Received: 25 February 2019; Accepted: 29 November 2019

Earlier we have shown that cluster rhenium compounds not only inhibited tumor growth in vivo but also supported the antioxidant state of experimental animals. Further investigation of new dirhenium(III) and cluster rhenium compounds in human leukemic cells is of great importance. The aim of the recent work was to investigate the cytotoxic activity of the new cluster rhenium compound with β-alanine ligands [Re2Cl6(C3H7NO2)2]·1.5H2O (I) in the solutions and nanoliposomes alone and together with cisplatin in Jurkat cells. It was shown that I in solution had cytotoxicity close to cisplatin (LC50 = 2.06·10-6 M). The administration of the rhenium-platinum system with І showed  increased cytotoxic activity, especially high when both components of the system were in the mixed liposomes together (LC50 = 4.93·10-10 M). The new dirhenium dicarboxylate complex with zwitterionic amino acid ligands possesses an appreciable cytotoxic and proapoptotic activity against leukemic cells, especially in combination with cisplatin, guiding the search for novel active rhenium compounds and development of improved regimens for combined chemotherapy based on combination of rhenium-platinum compounds.

Indexes of citrulline metabolism in rat liver under the toxic injury against the background of alimentary protein deficiency

H. P. Kopylchuk, I. M. Nykolaichuk, I. S. Lylyk

Yuriy Fedkovych Chernivtsi National University, Ukraine;
Institute of Biology, Chemistry and Bioresources, Chernivtsi, Ukraine;
e-mail: g.kopilchuk@chnu.edu.ua

Received: 29 May 2019; Accepted: 29 November 2019

It is known that citrulline is converted into arginine in the series of metabolic transformations. Results of our previous studies showed that acetaminophen-induced toxic injury on the background of the alimentary deprivation of protein is accompanied by a decrease in arginine level in rat hepatocytes, but citrulline liver metabolism at these conditions remains incompletely clear. In this work, the content of citrulline in the rat liver mitochondrial and cytosolic fractions and the activity of citrulline-degrading enzymes – argininosuccinate synthase and argininosuccinate lyase were investigated. It was found that in the mitochondrial fraction a maximal reduction of the citrulline levels occurred after administration of acetaminophen toxic doses regardless of the protein amount in the ration, while in the cytosolic fraction the alimentary protein deficiency was a key factor in decreasing the activity of argininosuccinate synthase and arginino-succinate lyase. The data obtained indicated the disturbances of the urea cycle functioning and explained the   decrease of L-arginine level in hepatocytes in conditions of acetaminophen-induced toxic injury against the background alimentary protein deficiency.

Biochemical indicators of green photosynthetic bacteria Chlorobium limicola response to Cu(2+) action

T. B. Sehin1, S. O. Hnatush1, O. D. Maslovska1,
A. A. Halushka1, Y. H. Zaritska2

1Ivan Franko National University of Lviv, Ukraine;
2State Scientific-Research Control Institute of Veterinary
Medicinal Products and Feed Additives, Lviv, Ukraine;
e-mail: SeginT@ukr.net

Received: 11 March 2019; Accepted: 29 November 2019

Photolithotrophic sulfur bacteria are involved in  biota functioning and have  a biotechnological potential for bioremediation of contaminated environment, but the mechanisms of xenobiotics, in particular of heavy metal ions damaging action and the pathways of photolithotrophic bacteria adaptation  under these conditions have not been established. In this work, the biochemical indicators of green photosynthetic bacteria Chlorobium limicola response to Cu ions were studied. C. limicola cells were incubated during one hour in buffer containing copper (II) sulfate in 0.05–0.5 mM concentrations and grown for 8 days in GSB medium. The content of Cu2+ in cells was estimated by atomic absorption spectroscopy. The activity of enzymes of antioxidant defense, photosynthetic pigments and glutathione content, indexes of lipids unsaturation and membrane viscosity as markers of  membrane fluidity were estimated. It was shown that the response of green photosynthetic bacteria C. limicola to Cu2+ action varied  depending on cations concentration. Under the influence of metal salt at 0.05 mM concentration, the activity of antioxidant enzymes, GSH/GSSG ratio, the content of photosynthetic pigments and membrane fluidity indexes were higher as compared with control. Under the increase of copper (II) sulfate concentration to 0.25 mM, the activity of antioxidant  enzymes was lower compared to the response of the cells under the influence of 0.05 mM copper (II) and the GSSG content was increased.  Under the influence of 0.5 mM copper (II) the indexes of membrane fluidity did not differ from the control, but superoxide dismutase and peroxidase activity inhibition and  the further decrease of GSH/GSSG ratio were observed followed by the highest Cu2+ cations accumulation in cells and significant decrease of  bacteria biomass growth.

Production and physicochemical characterization of xanthan gum by native lactose consuming isolates of Xanthomonas citri subsp. citri

R. Moravej1, S. M. Alavi2, M. Azin3, A. H. Salmanian2

1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran;
2Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran;
3Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran;
e-mail: mealavi@nigeb.ac.ir

Received: 30 September 2019; Accepted: 29 November 2019

Xanthan is a biopolymer produced by Xanthomonas bacteria which is widely used in many industries such as food and oil. In this work, three Xanthomonas strains (X. citri/NIGEB-88, X. citri/NIGEB-386 and X. citri/NIGEB-K37) were used to evaluate their industrial potential to produce xanthan gum in whey medium. Bacteria growth rate, viscosity, biomass, dry weigh of produced xanthan and β-galactosidase activity were studied during the fermentation process and the presence of β-galactosidase genes was assessed by PCR technique. Strain NIGEB-386 had the best ability to utilize lactose in the whey medium. The highest amount of xanthan production and viscosity were 22.7 g/l and 2066.6 mPa·s, respectively. The presence of six β-galactosidase genes in strains NIGEB-386 and NIGEB-K37 was confirmed. The pyruvate and acetyl contents in xanthan gum were 2.1 and 0.29 %, respectively. Fourier-transform infrared spectroscopy analysis determined the position of the functional groups in the structure of the fermentation product. In whey medium, the performance of both NIGEB-386 and NIGEB-K37 strains were better than the X. campestris. The findings showed that Xanthomonas citri/NIGEB-386 is suitable for industrial production of xanthan using cheese whey as a low-cost medium.

Phenol-oxidizing activity and fatty acid profile of Brevibacillus centrosporus F14 strain

Т. V. Gudzenko, O. V. Voliuvach, O. G. Gorshkova,
А. М. Ostapchuk, V. O. Ivanytsia

I.I. Mechnykov Оdessa National University, Ukraine;
e-mail: tgudzenko@ukr.net

Received: 19 June 2019; Accepted: 29 November 2019 2019

Biotechnological treatment of enterprises wastewater, which contains toxic phenolic compounds, is the most acceptable method today. Therefore, the search for new non-pathogenic biochemically active microorganisms – destructors of phenol and their identification is relevant. The aim of this study was to determine phenol-oxidizing activity and the fatty acid profile of Brevibacillus centrosporus F14 – destructor of phenol, isolated from pharmaceutical plant wastewater. Fatty acids analysis of the microorganism culture was carried out using automatic system of microorganisms identification MIDI Sherlock based on gas chromatograph Agilent 7890. The phenol-oxidizing activity of bacteria was determined by photometric method accor­ding to the degree of phenol extraction from the water. It was experimentally confirmed that Brevibacillus sp. F14 strain had a phenol-destructive activity. When phenol-containing water was treated with Brevibacillus sp. F14 cells, phenol concentration in the water decreased from 200.0 ± 12.0 mg/l to 6.8 ± 0.8 mg/l during 15 days of exposure. Chromatographic analysis showed that saturated fatty acids i-14:0 (14.9%), i-15:0 (14.8%), a-15:0 (34.9%), i-16:0 (11.10%) dominated in the strain’s fatty acids profile. The total content of the branched saturated fatty acids i-14:0, i-15:0, a-15: 0, i-16: 0, i-17: 0, a-17:0, ∑(a-17:1/i-17:1) was 85.0% and that of saturated and unsaturated fatty acids with the normal structure – 7% of the total fatty acids pool. The distinction of Brevibacillus centrosporus F14 strain’s  fatty acid profile from other bacteria of Brevibacillus genus was  the presence of more i-14:0 (14.85%), i-16:0 (11.10%) fatty acids and 16: 1 w7c alcohol (7.71%), as well as the  minor content of fatty acids markers 16:0 N alcohol (0.60%), 18:3 w6c (0.4%) and ∑(a-17:1/i-17:1) (1.94%). According to the phenotypic characteristics and fatty acid composition, the tested phenol-oxidizing strain belongs to Brevibacillus centrosporus species. The index of Brevibacillus centrosporus F14 strain’s fatty acid profile similarity with the library was 0.645.

Indexes of nitric oxide system in experimental antiphospholipid syndrome

O. Z. Yaremchuk, K. A. Posokhova, І. P. Kuzmak,
M. I. Kulitska, I. М. Klishch, M. M. Korda

I. Horbachevsky Ternopil National Medical University, Ukraine;
e-mail: yaremchuk@tdmu.edu.ua

Received: 11 November 2019; Accepted: 21 January 2020

Antiphospholipid syndrome (APS) is an autoimmune disease characterized by the presence of antibo­dies to negatively charged membrane phospholipids (aPL).  Endothelial dysfunction is one of the most dangerous­ APS manifestations followed by thrombosis, placental insufficiency and often foetal death due to circulatory disorders in placenta blood vessels. It is established that synthesis and bioavailability of nitric oxide (NO) in the endothelium are impaired at APS, but the role of NO system in pregnancy failure at this pathology remains ambiguous. The aim of this research was to estimate the indexes of the nitric oxide system in animals with an experimental antiphospholipid syndrome before pregnancy and on the 18th day of pregnancy, without­ treatment and under treatment with nitric oxide synthesis modulators (L-arginine and aminoguanidine). In the blood serum and liver of the BALB/c mice with experimental APS, the content of eNOS and iNOS by ELISA and the level of NO2 and NO3 with the use of Gris reagent were determined before pregnancy and on the 18th day of pregnancy. The data obtained indicate the relative inefficient NO production by eNOS and NO hyperproduction by iNOS in the blood serum and liver of mice in the pathogenesis of experimental APS. Thus, in mice with APS before pregnancy and on the 18th day of the pregnancy, the eNOS content and NO2 level were decreased while the iNOS content and NO3 level were increased compared to the indexes in the control animal group. L-arginine administration to the animals with APS at the follow-up periods resulted in an increased eNOS content and NO2, NO3 levels in blood serum and liver with the simultaneous decrease in iNOS content in the liver as compared to indexes in untreated mice with APS. The combined use of L-arginine and selective iNOS inhibitor aminoguanidine caused a significant increase in eNOS content and a decrease in iNOS content followed by normalization of NO2 and NO3 levels in blood and liver of mice with  experimental APS before pregnancy and on the 18th day of pregnancy compared to untreated mice with APS.

Methotrexate effect on biochemical indices of psoriasis patients depends on MTHFR gene polymorphism

O. M. Fedota1, L. V. Roschenyuk2,3, T. V. Tyzhnenko1,
N. G. Puzik1,3, V. M. Vorontsov1, P. P. Ryzhko1

1V.N. Karazin Kharkiv National University, Ukraine;
2Kharkiv Regional Clinical Skin and Venereal Diseases Dispensary №1, Ukraine;
3Kharkiv National Medical University, Ukraine;
e-mail: tyzhnenko@ukr.net

Received: 13 June 2019; Accepted: 29 November 2019

Methotrexate (MTX) is the immunosuppressive anti-inflammatory drug and the antagonist of the enzyme dihydrofolate reductase. Pharmacogenomic studies and clinical evidences suggest that altered response to MTX  in patients with different diseases is associated with polymorphisms of genes that regulate folate metabolism. The purpose of the article was to analyze the methotrexate effect on the biochemical indices of psoriasis patients depending on methylenetetrahydrofolate reductase gene (MTHFR) polymorphisms. Effects of two single-nucleotide polymorphisms, C677T and A1298C, were studied. An increase of alanine aminotransferase and aspartate aminotransferase activity above the normal level in the patients with both MTHFR gene polymorphisms after methotrexate intake was observed. In patients with CC, TT, CT genotypes for C677T polymorphism and AA genotype for A1298C polymorphism of MTHFR gene, significant differen­ces in alpha-amylase activity before and after treatment with methotrexate were detected. Analysis of the biochemical indices of  patients with arthropathic and vulgaris psoriasis showed that the positive effect of MTX treatment could be associated with wild-type alleles in both  polymorphisms of MTHFR gene, while the ineffectiveness of methotrexate was associated with the dihеterozygous genotype. The largest number of smokers was found within the CTAA genotype group (37.5%), while no smokers were observed within TTAA patients and most of CCAA patients. The data obtained testify the utility of the individual approach to the psoriasis patients therapy taking into account genetic background.

Profiling of metabolic biomarkers in the serum of prostate cancer patients

F. Ali1, S. Akram1, S. Niaz1,2, N. Wajid1

1Institute of Molecular Biology and Biotechnology (IMBB) & Centre for Research In Molecular Medicine (CRIMM), The University of Lahore, Raiwind Road Lahore, Pakistan;
2Social Security Hospital Multan Chungi, Multan Road, Lahore;
e-mail: Fatima.ali@imbb.uol.edu.pk; fatemei.ali@gmail.com

Received: 26 July 2019; Accepted: 29 November 2019

Prostate cancer (PCa) is the major cause of the death of men population globally. Multiple factors are involved in the initiation and progression of PCa. This study aimed to evaluate different metabolic parameters in the serum of PCa patients. Males of 50 years and above age with the recent diagnosis of PCa (digital rectal examination, and elevated serum prostate-specific antigen (PSA) level) were included in the study. Glucose and serum electrolytes level, lactate dehydrogenase activity, parameters of lipid metabolism and liver and kidney functioning were measured on a fully automated analyzer using standard reagent kits. Oxidative stress was evaluated by measuring MDA, CAT, GSH, and SOD in serum. Detection of C-reactive protein (CRP), insulin-like growth factor (IGF-1) and vascular endothelial growth factor (VEGF) was performed by immunoassay. It was shown that serum glucose and HDL levels were lower while total cholesterol, LDL and triglyceride levels were significantly higher in PCa group than in the control group. PCa patients had an elevated level of liver and kidney functional markers. Comparison of the oxidative stress markers in patient and control groups showed significant difference. It was detected that serum levels of CRP, IGF-1 and VEGF were significantly higher in PCa group, compared the control to group (P < 0.05). Low level of glucose and dyslipidemia indices in prostate cancer patients indicated metabolic changes and demonstrated the importance of multiple parameters analysis (free PSA, dyslipidemia, VEGF, IGF-1, CRP, and oxidative stress markers) for early PCa diagnostics.