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Scientific and educational activity of the Palladin Institute of Biochemistry among students

V. I. Nazarenko, T. O. Borisova

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

Received: 19 May 2021; Accepted: 07 July 2021

The paper article is dedicated to scientific and educational activity of Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine among youth students, which is implemented­ via its scholars­’ work in certain programs. The authors consider in chronological order the forms and methods of scientific and scientific-educational activity of the Institute in cooperation with the Municipal Out-of-School Educational Institution “Kyiv Junior Acade­my of Sciences of Student Youth” for ten years, since its founding. This activity’s root dates back to the time of Oleksandr Volodymyrovych Palladin’s work, which is currently performed together with the Youth Section of the Ukrainian Biochemical Society. In particular, the emphasis is on the structures of the “University of Young Biochemists”: Lectures “Advanced Frontiers of Biology”, “Laboratory-practical activities” and seminars. The authors consider in detail the role of the OV Palladin Memorial museum in outreach activity and disseminating biochemical knowledge, achievements, and contribution of Ukrainian scientists to the world mainstream of life sciences. Its work plan includes implementing some scientific and educational activities in conjunction with out-of-school educational institutions. The Palladin Institute’s scholars heavily contribute to the local and international scien­tific and educational programs. The article also teels about the life success of students who performed research work under the guidance and assistance of scientists of the Palladin Institute of Biochemistry at the contests of youth scientific creativity and provides examples of participation of students of the University of Young Biochemists­ in the youth scientific conferences and symposia.

The discovery of genetic control of enzyme and virus synthesis: 1965 Nobel Prize Laureates André Lwoff, François Jacob, Jacques Monod

O. P. Matyshevska, V. M. Danilova, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kiyv;
e-mail: matysh@yahoo.com

Received: 19 April 2021; Accepted: 07 July 2021

The middle of the 20th century was marked by a number of significant events in molecular biology, among which the groundbreaking discovery of the double helix of DNA, which could self-replicate and thus perform the main life function; the isolation of enzymes for DNA synthesis, and DNA synthesis outside the cell, to name but a few. However, the question of how the information transmission from DNA to proteins is regulated remained open. The concept of the mechanism of regulation of prokaryotic gene activity developed by three French scientists (André Lwoff, François Jacob, Jacques Monod; Nobel Prize 1965), which was a logical outcome of the research in genetics and biochemistry over the previous decades, is recognized to be one of the remarkable achievements in molecular biology. This article describes the essence of the discovery of Lwoff, Jacob and Mono that is the identification of two different groups of genes – structural and functional – and the role that these genes perform in the transmission of genetic information.

Identification of cortactin molecular forms in human urine and their possible diagnostic value

M. Starykovych1, S. Souchelnytskyi2, O. Fayura3, O. Abrahamovych3,
M. Abrahamovych3, N. Lukavetskyy4, R. Stoika1, Y. Kit1*

1Department of Regulation of Cell Proliferation and Apoptosis, Institute of Cell Biology, National Academy of Scoences of Ukraine;
2College of Medicine, QU Health, Qatar University, Doha, Qatar;
3Department of Internal Medicine № 1, Danylo Halytsky Lviv National Medical University, Ukraine;
4Lviv State Oncology Regional Treatment and Diagnostic Center, Lviv, Ukraine;
*e-mail: kit@cellbiol.lviv.ua

Received: 14 September 2020; Accepted: 07 July 2021

The protein composition of human urine reflects changes in the biochemical and physiological status of an individual and has an essential diagnostic value. Using precipitation/extraction methods we isolated a protein with Mr ~100 kDa in a human urine. MALDI TOF/TOF mass spectrometry identified this protein as human Src protein kinase substrate cortactin (UniProtKB/Swiss-Prot: Q14247). Screening of urine samples using Western blotting with specific anti-human cortactin antibodies revealed   different proteins immunologically related to cortactin in healthy humans and patients with liver cirrhosis and lung cancer diseases.  These data  suggest that the level of cortacins isoform in urine might serve as a potential marker for testing acute and systemic diseases.

Biochemical and tensometric analysis of C(60) fullerenes protective effect on the development of skeletal muscle fatigue

D. M. Nozdrenko1, K. I. Bogutska1, I. V. Pampuha1,
O. O. Gonchar2, O. M. Abramchuk3, Yu. I. Prylutskyy1*

1Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: prylut@ukr.net;
2Bogomolets Institute of Physiology, National Academy of Sciences of Ukraine, Kyiv;
3Lesya Ukrainka Volyn National University, Lutsk, Ukraine

Received: 19 May 2021; Accepted: 07 July 2021

TThe protective effect of water-soluble C60 fullerenes on the development of slow and rapid fatigue of rat skeletal muscles was analyzed. It was found that the reduction of muscle contraction force (muscle soleus) by 50% of the initial values is almost twice as slow as stimulation with a frequency of 1 Hz (slow muscle fatigue) than with 2 Hz (rapid muscle fatigue) stimulation after intramuscular injection of C60  fullerenes (dose 0.5 mg/kg). There is a clear tendency to decrease the values of biochemical parameters of the blood of animals with the therapeutic effect of water-soluble C60 fullerenes by approximately 45-60% and 35-40% with the development of slow and rapid muscle fatigue, respectively. Thus, C60 fullerenes, as powerful antioxidants, are able to efficiently affect the prooxidant-antioxidant homeostasis of muscle tissue and thus help maintain its normal physiological state.

Personalised diet improve intestine microbiota and metabolism of obese rats

V. V. Bati1*, T. V. Meleshko1, O.V. Pallah1,
I. P. Zayachuk2, N. V. Boyko1

1RDE Centre of Molecular Microbiology and Mucosal Immunology, Uzhhorod National University, Ukraine;
2Department of Physiology and Pathophysiology, Uzhhorod National University, Ukraine;
*e-mail: victoria.bati@uzhnu.edu.ua

Received: 25 April 2020; Accepted: 07 July 2021

Recent research on human microbiome provide opportunities to develop functional foods of new generation that can regulate intestinal microbiota and the biochemical status of the individual. The aim of the study was to determine the effect of individually designed nutrition on the intestinal microbiota and metabolic parameters of rats. Outbred laboratory rats with obesity  were randomly divided into 9 groups (= 12) depending on the type of food ingredients taken orally for three months. The ratio of the intestinal commensal microorganisms main groups, as well as the lipid profile and the content of glucose, urea, calcium in the serum of animals were determined. It was shown that cholesterol level in the serum was reduced in experimental groups after consumption of lactobacilli suspension, blueberry juice, fermented milk drink based on lactobacilli, fermented milk drink with blueberry juice, sauerkraut. In most cases, the gut microbiome of experimental animals was characterized by a consistently high level of lacto and other beneficial bacteria and decreased amount of opportunistic microorganisms at the end of the experiment compared with animals in the control group. Based on the obtained data, we first proposed the principles of creating functional products by synergistically combining components of edible plants that act as prebiotics and microorganisms that act as probiotics for personalized use, targeted correction of intestinal microbiome and prevention of noncommunicable diseases.

Protective effect of Atriplex halimus extract against benzene-induced haematotoxicity in rats

K. Zeghib1*, D. A. Boutlelis2, S. Menai3, M. Debouba4

1Department of chemistry, Faculty of exact sciences, University of El-Oued, El-Oued, Algeria;
2Department of Biology, Faculty of natural sciences and life, University of El-Oued, El-Oued, Algeria;
3The mother-child hospital (Bachir Bennacer) of El-Oued, El-Oued, Algeria;
4Higher Institute of Applied Biology of Medenine, University of Gabès, Tunisia;
*e-mail: zeghib-khaoula@univ-eloued.dz

Received: 24 December 2020; Accepted: 07 July 2021

Benzen (BZ) is a ubiquitous environmental pollutant with a toxic effect mainly aimed at the hematopoietic­ and immune systems. Atriplex halimus L. (Amaranthaceae) is a Mediterranean halophytic shrub traditionally used in North Africa as medicinal plant  for several therapeutic uses. The present study aimed to estimate the preventive and curative effects of Atriplex halimus L. (Ah) aqueous extract against BZ-induced hematotoxici­ty in rats. Analysis of the extract by the method of LC-MS revealed the presence of 7 vitamins, among which vitamin  C content was the highest. Adult rats were divided into five groups as follow: Group 1 received water (control); Group 2 received orally Ah aqueous extract (200 mg/kg) 3 days/week  for 15 weeks; Group 3 received BZ (100 mg/kg b.w) daily in drinking water for 15 weeks; Group 4 received concomitantly BZ (100 mg/kg) and  preventive treatment with Ah (200 mg/kg) for 15 weeks (AhP+BZ); Group 5 first received BZ (100 mg/kg) for 11 weeks and then curative treatment with Ah extract (300 mg/kg) daily for 30 days (BZ+AhC). It was shown that sub-chronic exposure to benzene induced leukopenia, lymphocytopenia, granulocytopenia and massive degeneration of the bone marrow tissue. The level of GSH and activity of GST and CAT were significantly lowered and the level of MDA was increased in the blood and bone marrow in rats of BZ-intoxicated group compared to the control rats. Administration of Ah extract recovered the bone marrow structure, dramatically decreased MDA content and increased GSH and CAT activity and GST level in the blood and bone marrow as compared with the indices in BZ-treated group. These observations demonstrate that curative and, to a lesser extent, preventive treatment with Atriplex halimus extract have therapeutic potential against hematotoxicity induced by benzene.

Expression of antioxidant enzymes genes in the liver and cardiac tissues of rats under L-carnitine administration and high-intensity interval exercise training

B. Shahouzehi1,2, Y. Masoumi-Ardakani3, S. Aminizadeh3, H. Nasri2*

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;
3Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran;
*e-mail: dr_hrnasri@yahoo.com
 
Received: 29 September 2020; Accepted: 07 July 2021

Reactive oxygen and nitrogen species are produced in the body both in normal and pathological processes and can alter cell redox and affect cell functions. Exercise training is able to modulate oxidant/antioxidants balance. In this study, we aimed to evaluate expression of antioxidant enzymes genes in the liver and cardiac tissues of rats that performed high-intensity interval training (HIIT) and received L-carnitine (LCAR). Thirty-two male Wistar rats were were randomly assigned into 4 groups (n = 8) as follows: 1. Untreated control; 2. The group that received LCAR (200 mg/kg/day i.p.); 3. The group that performed HIIT on a readmill (5 days/week for 4 weeks); 4. The group  that received LCAR and performed HIIT. At the end of the study, liver and cardiac tissues were excised and used to quantify glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT) and NF-κB genes expression by real-time PCR. It was found that both in LCAR and  HIIT groups GPX, SOD and NF-κB (P < 0.01) expression in cardiac and liver tissues was  significantly increased compared to the indices in the control group. In LCAR-HIIT group SOD and NF-κB expression in the liver was significantly increased compared to the group that received LCAR only (P = 0.046).  Our results showed that LCAR supplementation is useful to improve oxidative status in cardiac and liver tissues of rat during exercise training.

Effect of IRAK1/4 inhibitor on IL-1β, IL-6, INF-γ and TNF-α expression in breast cancer cells of several lines

M. Rezaei1, B. Shahouzehi2,4, S. Rahemi1,3, H. Fallah1*, M. Salarkarimi1

1Department of Clinical Biochemistry, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;
2Cardiovascular Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
3Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
4Student Research Committee, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;
*e-mail: hf59ma@gmail.com

Received: 28 July 2020; Accepted: 07 July 2021

Recent studies have shown that inflammation mediated via interleukin-1 receptor-associated kinases (IRAKs) is associated with cancer cells drug resistance. We aimed to evaluate the expression of inflammatory cytokines as the potential mechanism involved in the development of cancer cells resistance to conventional chemotherapy drugs. Breast cancer cells of BT549, BT20 and MB468 lines were treated with IRAK 1/4 inhibitor alone or in combination with chemotherapeutic agents methotrexate and topotecan. Expression of IL-1β, IL-6, TNF-α, and IFN-γ genes was quantified by real-time PCR. It was found that IRAK1/4 inhibitor suppressed IL-1β expression in BT549 cells at most and had minimal effect on IL-6 expression in MB468 cells. For the first time we showed that concomitant use of IRAK1/4 inhibitor with topotecan and methotrexate reduced IL-1β, IFN γ, TNF-α and IL-6 expression in BT-20, BT-549, MB-468 cell lines compared to the controls. It is suggested that specific IRAK inhibitors in combination with conventional chemotherapy can be used in cancer treatment to increase drug sensitivity and decrease the risk of tumor recurrence.

miR-329-containing exosomes derived from breast tumor cells suppress VEGF and KDM1A expression in endothelial cells

N. Maleki1,2,3*, F. Karami1, S. Heyati2, M. HadiZadeh3, Gh. Parnian4*

1Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Islamic Azad University-Tehran North Branch, Tehran, Iran;
*e-mail: dr.nargesmaleki@yahoo.com;
2Gynecology and reproductive biology Department, Kowsar poly-clinic, Tehran, Iran;
3Cancer Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
4Appletree Medical group, 275 Dundad W (Grange), Toronto, Ontario, Canada;
*e-mail: ghazalehparnian1@gmail.com

Received: 03 February 2021; Accepted: 07 July 2021

The exosomal transfer of miRNAs from tumor cells is considered to modulate VEGF expression and angiogenesis in endothelial cells. The aim of our investigation was to focus exclusively on the ability of specific exosomal miR329 to regulate angiogenesis within breast tumor. All experiments were done on MCF-7 and HUVEC cell lines. Exosomes were derived from MCF-7 cells both untreated and treated with tamoxifen that is an effecrive suppressor of hormone receptor-positive breast cancer. The level of miR32 and its targeted genes VEGF and lysine (K)-specific demethylase 1A (KDM1A) expression was estimated with q-RT-PCR. The PKH26 red fluorescent labeling kit was used to label the isolated exosomes and monitor their uptake. It was shown that the relative amount of miR-329 in exosomes was twice as large as in breast cancer  cells. Fluorescence microscopy imaging presented that exosomes from  MCF-7 cells were able to penetrate into endothelial cells and concentrate in the cytoplasm. It was observed that exosomes derived from untreated breast cancer cells induced KDM1A and VEGF gene expressions whereas exosomes from tamoxifen-treated cancer cells induced time-dependent decrease of KDM1A and VEGF expression in endothelial cells. It is assumed that the transfer of miR-329 containing exosomes from tamoxifen treated breast cancer cells to the endothelial cells could repress angiogenic molecular signaling pathway and be used as a supplementary strategy in breast cancer treatment.

Тhiacalix[4]arene phosphonate C-800 as a novel fluorescent probe for zinc in living cells

V. I. Yavorovska1, R. D. Labyntseva1*, O. V. Bevza1, A. Y. Pugach1,
A. B. Drapailo2, S. O. Cherenok2, V. I. Kalchenko2, S. O. Kosterin1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: raisa.lab@gmail.com

Received: 07 April 2021; Accepted: 07 July 2021

Zn ions are significant for maintaining the proper human organism functioning, thus monitoring­ the zinc content in living cells and the development of sensitive tracking systems and sensors for Zn is particularly important. The purpose of the work was to study the properties of synthetic thiacalix[4]arene C-800 (5,11,17,23-tetrakis[(hydroxy-ethoxyphosphonyl)methyl])-25,26,27,28-tetrahydroxythiacalix[4]arene) as a fluo­rescent sensor for zinc ions in living cells. Our studies demonstrated that thiacalix[4]arene C-800 containing­   four hydroxy-ethoxyphosphonylmethyl groups on the upper rim exhibited fluorescent properties at 340 nm excitation wavelength. Fluorescence intensity of thiacalix[4]arene C-800 was increased significantly in the presence of Zn cations, while cations of other metals, such as Mg2+, Ca2+, Cd2+, and Pb2+ did not affect it. Computer modeling demonstrated that two Zn cations interact with the oxygen atoms of four hydroxy-ethoxyphosphonylmethyl groups. It was shown that thiacalix[4]arene C-800 quickly penetrated rat myometrial cells that led to an increased intracellular fluorescence level. The addition of Zn2+ to cells, stained with thiacalix[4]arene C-800, was followed an even greater increase of intracellular fluorescent signal intensity. No effect of thiacalix[4]arene C-800 on reactive oxygen species production in myometrial cells was detected as well as on cells viability in the range of its 50-250 μM concentrations. Thus, thiacalix[4]arene C-800 can potentially be used as a selective fluorescent probe for the detection of Zn2+ in living cells.