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In silico identification and biochemical validation of plausible molecular targets of 4-thiazolidinone derivative Les-3833 as a potential anticancer agent
L. Kоbylinska1*, D. Khylyuk2, I. Subtelna2,
M. Kitsera3, R. Lesyk2
1Department of Biochemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
2Department of Pharmaceutical, Organic and Bioorganic Chemistry, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
3Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, Ukraine;
*e-mail: Kobylinska_Lesya@meduniv.lviv.ua; lesya8@gmail.com
Received: 16 January 2021; Accepted: 23 April 2021
Synthetic 4-thiazolidinone derivatives have a broad range of pharmacologic activities. Thus, 4-thiazolidinones are being investigated to create new molecules and develop active pharmaceutical substances for anticancer treatment. In our previous study, we investigated the pyrazoline-thiazolidinone-isatin conjugates, and determined that Les-3833 was the most active compound and might act through inhibition of PARP-, MAPK-, JNK-, Bcl-2-, CDK1/cyclin B, and/or the caspase family. The aim of this research was to perform molecular docking studies to enable the construction of a pharmacophore model for the Les-3833 compound and investigate probable biological targets. Pharmacophore modeling software packages performed molecular docking studies of probable biological targets and enabled the construction of a pharmacophore model. Docking models of Les-3833 with 11 enzymes involved in apoptotic mechanisms were studied. Based on the pharmacophore modeling results for all 11 enzymes, Les-3833 is predicted to be most active in Chk‑1, caspase-6, and caspase-8. Immunoblot analysis proved that the application of Les-3833 led to inhibition of Ser345 phosphorylation, which is induced by etoposide, the most important modification responsible for Chk‑1 activity. Taken together with the results of the docking studies, several mechanisms for the expression of antitumor activity by 4-thiazolidinones are suggested, and such multi-affinity is a characteristic feature of all these derivatives. The docking analysis confirmed the affinity of test compound Les-3833 for a topoisomerase II inhibitor and a high possibility of inhibitory interaction with Chk-1, caspase-6, and caspase-8.
Inflammation is the common mechanism of diseases (CMD) in COVID-19 disease during pregnancy and in gestational diabetes mellitus
Sandor G. Vari
Cedars-Sinai Medical Center, International Research and Innovation in Medicine Program, Los Angeles, California, United States
The Regional Cooperation for Health, Science and Technology (RECOOP HST) Consortium, led by Cedars-Sinai Medical Center was formed in 2006, was transformed into an Association in 2012 and includes 17 universities and academic organizations from eight countries: seven in Central and Eastern Europe (Croatia, Czech Republic, Hungary, Poland, Romania, Slovakia, Ukraine) and the United States. RECOOP builds multinational, multidisciplinary collaborations, and assists as well as coordinates the research activities of the sixteen research groups that are the Cedars-Sinai Medical Center – RECOOP Research Centers (CRRCs). https://www.cedars-sinai.org/research/administration/recoop.html.
Implementations of RECOOP’s strategic goals enable diverse talents geared towards integration of new knowledge derived from multiple specialties to investigate Common Mechanism of Diseases (CMD). While some may consider RECOOP’s CMD research strategy unorthodox, recent and timely scientific evidence shows that inflammation is the triggering event in the change of vascularization and it is the common mechanism of these two diseases: COVID-19 Disease during pregnancy and gestational diabetes mellitus (GDM).
Binding of the SARS-CoV-2 virus to the ACE2 receptor and its entrance into endothelial cells plays a role in vascular thrombosis but has a lesser effect placental endothelial dysfunction. The latter is induced by inflammation and exacerbated by proinflammatory cytokines, resulting in ischemic events and creating an upward spiral of an inflammatory reaction in pregnant women, accompanied by similar conditions in the placenta that will ultimately affect fetal development. In mild or moderate COVID-19 disease, changes in placental vascularization and blood flow have similarities to comorbidities in pregnancy such as GDM. However, during severe or critical stages of COVID-19 Disease, the changes could be harsher than those observed in GDM.
In COVID-19 Disease and GDM the immune status of pregnant women and consequently the newborn is altered due to inflammation and characterized by changes in levels of C-reactive protein (CRP), immunoglobulins (IgG, IgM, IgA) and proinflammatory cytokines that are detectable in maternal and umbilical cord blood and in mother milk.
To examine changes and monitor placental angiogenesis it is necessary to measure Vascular Endothelial Growth Factor (VEGF), Placental Growth Factor (PLGF), and Umbilical Cord Blood Sclerostin (UCBS) in maternal and umbilical cord blood serum. The angiogenic activity of sclerostin must be validated with the well-known marker VEGF, which is a proven indicator for changes in vascularization. The morphology of the vascular tree and blood flow in the placenta could be evaluated with three-dimensional power Doppler. The proinflammatory and ischemic effects in the placenta should be quantified with histopathology and immunohistochemistry. Changes in blood flow in the placenta and the morphology of the vascular tree in COVID-19 Disease during pregnancy may have similarities to those observed in GDM.
In summary, to improve maternal and fetal outcomes it is imperative to formulate better strategies for managing pregnancies during COVID-19 Disease and comorbidities like GDM. VEGF, PLGF and UCBS could be predictors of placental weight, birth weight, and fetal outcomes. In addition, further studies are needed to investigate the effects, if any, of proinflammatory and anti-inflammatory cytokines on postnatal development.
The discovery of the mechanisms of biological synthesis of nucleic acids: 1959 Nobel laureates S. Ochoa and A. Kornberg
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: 12 September 2020; Accepted: 17 December 2020
Alongside the chemical and physical research of nucleic acids in the 1940s-50s, the mechanisms of their biosynthesis were investigated. Thus, in 1959, Severo Ochoa and Arthur Kornberg were awarded the Nobel Prize in Physiology or Medicine for the discovery of the mechanisms of biological synthesis of RNA and DNA. The experiments performed by Ochoa and Kornberg are considered today the cornerstone of genetic engineering, as they first demonstrated the possibility of synthesizing RNA and DNA outside the living cell, and also as the enzymes they discovered were among the first tools of this technology.
Prospects of genome editing using CRISPR/CAS or how to master genetic scissors. Nobel Prize in Chemistry 2020
S. V. Komisarenko, S. I. Romaniuk
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: svk@biochem.kiev.ua
The Nobel Prize in Chemistry 2020 was awarded to two researchers in the field of molecular biology – the French Emmanuelle Charpentier, who now heads the Max Planck Unit for the Science of Pathogens in Berlin, and the American Jennifer Doudna from the University of California, Berkeley – for the ‘development of genome editing method’. A press release of the Nobel Committee states that the winners discovered one of the most powerful genetic technology tools – CRISPR/Cas9, or so-called ‘genetic scissors’. This method has contributed to many important results of basic research. In particular, plant researchers have managed to create crops resistant to mold, pests and drought. As for medicine, clinical trials of new cancer treatment techniques are underway, and a dream of curing hereditary diseases is about to become a reality. Genetic scissors have brought the life sciences to a new stage of development and greatly contributed to the benefit of mankind.
Application of fluorescein copolymer to improve the efficiency of counter-immunoelectrophoresis for diagnostics of animal infectious diseases
D. D. Ostapiv1, N. V. Kuz’mina1, М. R. Kozak1, Shan Hu2,
V. V. Vlizlo3*, I. Ya. Kotsiumbas3, S. M. Varvarenko4,
V. Ya. Samaryk4, N. G. Nosova4, M. V. Yakoviv4
1Institute of Animal Biology, NAAS of Ukraine, Lviv, Ukraine;
2Qingdao Agricultural University, Qingdao, China;
3State Scientific-Research Control Institute of Veterinary Medicinal Products and Feed Additives, Lviv, Ukraine;
4Lviv Polytechnic National University, Lviv, Ukraine;
*e-mail: vasyl.vlizlo@scivp.lviv.ua
Received: 12 May 2020; Accepted: 17 December 2020
The counter-immunoelectrophoresis (CIE) is a highly specific, simple and rapid method, which does not require expensive equipment, input materials and long-term staff training as compared with other express methods of viral infections diagnostic. Despite the CIE high informativeness, the results in practice can be doubtful at times. Therefore, the purpose of the study was to improve CIE method by strengthening the antigen-antibody interaction with the use of synthesized amphiphilic fluorescein-containing copolymer. It was demonstrated that the use of the fluorescein copolymer -antigen complex enhanced the interaction with serum/plasma antibodies of rabbits immunized against hemorrhagic disease and increased protein content in the area of precipitation. This effect was more pronounced when more hydrophobic copolymer containing 5.87% fluorescein was introduced into the complex with antigen. The ability of the amphiphilic fluorescein copolymer to enhance the antigen-antibody interaction and the ability to visualize the fluorescent precipitation zone may increase CIE effectiveness for express diagnostic of infectious diseases.
Quercetin and histamine effects on free radical reactions in rat erythrocytes
N. P. Harasym*, M. Y. Booklyv, A. R. Zyn, S. M. Mandzynets,
A. O. Bezkorovainy, D. I. Sanahursky
Ivan Franko National University of Lviv, Faculty of Biology, Departments of Biophysics and Bioinformatics, Ukraine;
*e-mail: garasymnataly@gmail.com
Received: 12 June 2020; Accepted: 17 December 2020
The effects of quercetin and histamine separately or in combination on the free radical state of rat erythrocytes were estimated in vitro. Quercetin (0.1; 0.5; 3.0; 5.0 mM) or histamine (0.01; 10.0 μM) were added to whole blood separately or in combination. The content of hydroperoxides, TBA-active products and carbonyl groups of proteins in erythrocytes after hemolysis was determined. The greatest influence of quercetin and histamine on erythrocytes state indicators was revealed under their combined action, when the level of TBA-active products and the content of carbonyl groups of proteins were found to be increased substantially.
Comparative characteristic of lung cancer stem-like cells generated in vitro under different culture conditions
O. V. Skachkova1*, O. I. Gorbach1, M. V. Inomistova1,
L. V. Garmanchuk2, N. M. Khranovska1
1National Cancer Institute, Kyiv, Ukraine;
2ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: oksanaskachkova@ukr.net
Received: 20 June 2020; Accepted: 17 December 2020
Cancer stem cells (CSCs) play an important role in resistance to cancer treatment and recurrence developing. The aim of this study was to obtain cell culture of MOR line non-small cell lung cancer cells enriched in CSCs and to investigate its functional and molecular-genetic properties. Tumor spheroids (TS) of MOR cell line were generated in vitro under normal adhesive (0.2% carboxymethyl cellulose, CMC) or low-adhesive (2% agarose) culture conditions. Lateral population of TS was evaluated by flow cytometry with the use of R-123 fluorescent dye, the index of R-123 exclusion was also assessed. Expression of CD44, ALDHA1, CD133, Sox2 and Nanog mRNA was determined with RT-qPCR. It was found that regardless of the culture conditions tumor spheroids form a lateral population characterized by an increased dye exclusion index. Expression levels of CD44, ALDHA1, CD133, Sox2 and Nanog mRNA in TS cells obtained under low-adhesive (2% agarose) conditions were significantly higher than in monolayer cells and cells obtained using 0.2% CMC. Thus, the proposed method of culturing in low-adhesive conditions allowed to enrich significantly tumor spheroids of MOR line in cells with CSC properties.
MicroRNAs hsa-mir-34a and hsa-mir-124 as biomarkers for predicting and monitoring the lithium treatment in bipolar disorder: in silico analysis
Orcun Avsar
Hitit University, Department of Molecular Biology and Genetics, Corum, Turkey;
e-mail: orcunavsar.gen@gmail.com
Received: 14 July 2020; Accepted: 17 December 2020
Lithium is known to be efficient in treatment for mental disorders in people with bipolar disorder. The aim of the study was to identify bipolar disorder-specific MicroRNAs (miRNAs) that are associated with genes targeted by lithium treatment and lead to variance in the clinical response. miRNAs which are experimentally validated and shown to have differential expression pattern in bipolar disorder were selected from online public databases miRTarbase and HMDD v3.2. Target prediction was carried out for each miRNA, and experimentally validated miRNA-target mRNA pairs were obtained and analyzed by miRTarbase, HMDD v3.2, TargetScan, and DIANA databases. miRNA-target genes that are associated with lithium was determined by the use of DrugBank. According to the in silico analysis, it has been found that IMPA1 gene was targeted by hsa-mir-34a and GRIA3 gene was targeted by hsa-mir-124. The present study demonstrated that IMPA1, GRIA3, hsa-mir-34a, hsa-mir-124 may have the potential to be used as molecular biomarkers for estimating and monitoring the response to lithium treatment in bipolar disorder.
Organo-specific accumulation of phenolic compounds in a buckwheat seedlings under aluminium-acid stress
O. E. Smirnov, A. M. Kosyan, Yu. V. Pryimak,
O. I. Kosyk, N. Yu. Taran
ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: plantaphys@gmail.com
Received: 19 September 2020; Accepted: 17 December 2020
Toxic effect of aluminum contamination is one of the causes of valuable crops yield loss all over the world. It is considered that plants’ phenolic compounds play a key role in aluminium detoxification by chelation of aluminium ions in the aboveground part of aluminium-accumulating plants. However, recent evidence shows the chelating ligands involvement in both the internal and external aluminium detoxification in plants. The aim of the study was to determine the total phenolic compounds, flavonoids, anthocyanins accumulation and the activity of phenylalanine ammonia-lyase (PAL) as the key enzyme in phenolic compounds synthesis in seedlings of common (Fagopyrum esculentum Moench.) and tartary (Fagopyrum tataricum (L.) Gaertn.) buckwheat in response to the chronic aluminium-acid stress. It was recorded that addition of 50 μM Al2(SO4)3·18H2O to the nutrient medium led to the accumulation of phenolic compounds in all organs of both studied species on the tenth day of the plant exposure to stress. Species-specific and organ-specific accumulation of certain classes of phenylpropanoids was recorded. On the tenth day of stress, PAL activity was increased in the leaf tissues of both buckwheat species, but was decreased in common buckwheat root tissues and no statistically significant changes were observed in tartary buckwheat root tissues. Species and organ specificity of phenylpropanoids accumulation in the studied species is considered to be an adaptive reaction under conditions of aluminum stress.
Male and female rats differ in homeostatic shifts during pre-slaughter fear stress
S. S. Grabovskyi1, R. R. Panchuk2, N. R. Skorokhyd2, R. S. Stoika2*
1Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies, Lviv, Ukraine;
2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
*e-mail: stoika.rostyslav@gmail.com
Received: 19 June 2020; Accepted: 17 December 2020
In this investigation, the level of сortisol in blood plasma and splenocytes apoptosis in male and female rats in response to fear stress created by animals’ waiting for different terms (20, 40, 60 min) before the slaughter were evaluated. A significant and dependent on the stress state duration increase in the level of cortisol in the blood plasma of both male and female rats compared with the rats of control groups was found, which was more pronounced in females compared to males. The number of Annexin V-positive (apoptotic) cells was determined by FACS analysis. It was shown that the content of apoptotic splenocytes during the stress before slaughter was higher in females compared to males. The prolongation of the stress period was accompanied by an increase in the content of apoptotic splenocytes in males and its decrease in females. The potential hormone-dependent attenuation of the mechanisms of adaptation to stress before slaughter in female rats is discussed.