Category Archives: Uncategorized
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.
Chlorine-binding structures: role and organization in different proteins
R. Yu. Marunych*, O. O. Hrabovskyi, G. K. Bereznytskyj,
L. V. Pyrogova, G. K. Gogolinskaya, Ye. M. Makogonenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: rostmarbiotech@gmail.com
Received: 29 September 2020; Accepted: 07 July 2021
The review focuses on chloride-binding structures in the proteins of bacteria, plants, viruses and animals. The structure and amino acid composition of the chloride-binding site and its role in the functioning of structural, regulatory, transport, receptor, channel proteins, transcription factors and enzymes are considered. Data on the important role of chloride-binding structures and chloride anions in the polymerization of fibrin are presented.
A legend in his own lifetime: double Nobel prize winner Linus Pauling
T. V. Danylova1*, S. V. Komisarenko2
1National University of Life and Environmental Sciences of Ukraine, Kyiv;
*e-mail: danilova_tv@ukr.net;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail:svk@biochem.kiev.ua
Received: 19 April 2021; Accepted: 17 May 2021
Linus Pauling – a prominent American chemist, biochemist, chemical engineer, peace activist, author, and educator – was one of the scientists-humanists whose life echoed the key milestones of the 20th century and who could be compared with Leonardo da Vinci in terms of the breadth of interests and creative achievements. He is one of the four scientists to have won two Nobel Prizes (the Nobel Prize in Chemistry 1954; the Nobel Peace Prize 1962) and the only one to have been awarded two unshared Nobel Prizes. As a result of his long-lasting research, Pauling formulated a theory of structure and function of proteins; studied the effect of oxygen saturation of hemoglobin on its magnetic properties, laid the foundations for structural analysis of protein molecules, made a contribution to the study of antibodies. Believing that people had to develop a new type of thinking for the sake of the survival of humanity, Linus Pauling spoke out strongly against nuclear testing, and the contemporary world is indebted to Pauling for his courage and moral leadership in reminding us about the dangers of nuclear war. The versatile innovative activities of the brilliant scientist and humanist Linus Pauling, his unconventional personality, and the huge scientific heritage have left a deep mark in the history of humankind.
Discovery of cell apoptosis regulation genes: Sydney Brenner, John E. Sulston and H. Robert Horvitz (The Nobel Prize in Physiology or Medicine 2002)
M. V. Grigorieva*, V. M. Danilova, S. V. Komisarenko
Palladin Biochemistry Institute, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: mvgrigorieva@biochem.kiev.ua
Received: 24 Nobember 2020; Accepted: 23 April 2021
The Nobel Prize in Physiology or Medicine 2002 was awarded to Sydney Brenner, Howard Robert Horvitz and John Edward Sulston for their discoveries concerning “genetic regulation of organ development and programmed cell death”. The scientists studied cell division and differentiation in the nematode Caenorhabditis elegans from the fertilized egg to the adult organism. As a result of their studies, key genes regulating organ development and programmed cell death (apoptosis) were identified, and corresponding genes were shown to exist in higher species, including humans. These discoveries shed light on the pathogenesis of many diseases and were important for further medical research.
Application of petri nets methodology to determine biophysicochemical parameters of mitochondria functioning
H. V. Danylovych*, A. Yu. Chunikhin, Yu. V. Danylovych, S. O. Kosterin
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: danylovych@biochem.kiev.ua
Received: 01 Nobember 2020; Accepted: 17 May 2021
With the use of Petri net methodology a mathematical simulation model able to predict simultaneous changes in biophysicochemical parameters of mitochondria functioning was developed. The model allowed to interconnect in time the changes in mitochondria hydrodynamic diameter, electronic transport chain functioning, endogenous fluorescence of adenine nucleotides, DCF fluorescence signal of ROS production and NaN3 effects. It was shown that the calculated values of the studied biophysicalchemical parameters correspond to those obtained experimentally. The model permit to link mitochondrial functional changes and their structural representation and to optimize significantly experimental procedures.
NO-synthase activity and nitric oxide content in lymphoid cells of thymus and spleen of rats under conditions of diet-induced obesity
L. Kot*, V. Konopelnyuk, K. Dvorshchenko, V. Vereschaka
ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: kot_lora@ukr.net
Received: 10 August 2020; Accepted: 17 May 2021
Metabolic disorders under conditions of obesity are known to be accompanied by systemic inflammation and immune system imbalance while nitric oxide (NO) system is one of the main key links in the immune responses regulation. The aim of the research was to evaluate NO-synthase (NOS) activity and NO content in the cells of thymus and spleen of rats with experimental obesity induced by 14 weeks of high-calorie diet. NO content in cell suspensions was determined using Griess reaction, NOS activity was estimated by NADPH++Н+ specific cleavage. It was shown that under conditions of experimental obesity NO content in cells of thymus and spleen as well as NOS activity in the spleen were decreased with simultaneous increase in both NO content and NOS activity in the serum. The obtained data testify the changes in lymphoid cells functional activity and in organism general adaptive capacity under conditions of obesity.
Sex dependent differences in oxidative stress in the heart of rats with type 2 diabetes
N. I. Gorbenko1*, O. Yu. Borikov2, O. V. Ivanova1, T. V. Kiprych1,
E. V. Taran1, T. I. Gopciy2, Т. S. Litvinova1
1SI “V. Danilevsky Institute for Endocrine Pathology Problems of the NAMS of Ukraine”, Kharkiv;
2V.N. Karazin Kharkiv National University, Kharkov, Ukraine;.
*е-mail: Gorbenkonat58@ukr.net
Received: 17 September 2020; Accepted: 17 May 2021
Type 2 diabetes mellitus is known to double mortality from cardiovascular diseases (CVD), in which oxidative stress plays an important role. It is suggested that the impact of diabetes on CVD risk may vary depending on gender. The aim of the study was to assess oxidative stress parameters in the heart of 12 weeks old male and female Wistar rats with type 2 diabetes mellitus (T2DM) induced by high-calorie diet followed by intraperitoneal streptozotocin injections. The level of advanced oxidation protein products, superoxide dismutase, glutathione reductase and glutathione peroxidase activity in the isolated heart mitochondria and NADPH-oxidase and xanthine oxidase activity in the post-mitochondrial supernatant fraction were determined. It was shown that T2DM induced more pronounced oxidative stress confirmed by the increased level of advanced oxidation protein products in the heart mitochondria of males than females. The data obtained indicate that the main reason of oxidative stress in the heart of diabetic males is the activation of non-mitochondrial sources of reactive oxygen species. While in the heart of diabetic female rats it is the decrease in antioxidant enzymes activity in mitochondria. These results justify the necessity of gender-specific therapy for the prevention and management of diabetic CVD.