Category Archives: Uncategorized

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.

Effect of N-acetyl cysteine on oxidative stress and Bax and Bcl2 expression in the kidney tissue of rats exposed to lead

M. Gholami1, A. B. Harchegani2, S. Saeedian3,
M. Owrang4, M. R. Parvizi1*

1Department of Physiology, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran;
2Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
3Medical Genomic Research Center, Tehran Medicine Sciences Islamic Azad University, Tehran, Iran;
4Faculty of Medicine, Sari Branch, Islamic Azad University, Sari, Iran;
*e-mail: mparvizi@alumnus.tums.ac.ir

Received: 30 May 2020; Accepted: 17 December 2021

This study aimed to consider the lead-induced oxidative damage of the kidney of male rats and the role of antioxidant N-acetylcysteine (NAC) in preserving cells against Pb toxicity. Rats were randomly divided into five groups including G1 (control), G2 (single 70 mg/kg dose of Pb), G3 (continuous daily 2 mg/kg dosing of Pb for 4 weeks), G4 (single dose of Pb + 50 mg/kg NAC), and G5 (continuous daily dosing of Pb + 50 mg/kg NAC). The level of malonic dialdehyde (MDA) and total antioxidant capacity were measured spectrophotometrically.The level of Pb in  serum and kidney tissue was measured by atomic absorption spectroscopy. Expression of Bax and Bcl2 genes was estimated using RT-PCR.  It was shown that single and continuous exposure to Pb caused a considerable increase of Pb content in serum and kidney tissue of rats in G2 and G3 groups compared to other groups. NAC treatment significantly improved TAC values and decreased MDA values in the serum of rats exposed to Pb. Single and continuous Pb dosing caused a 3.9- and 13.1-fold increase in Bax expression and 1.5-fold and 2.1-fold decrease in Bcl2 expression in a kidney tissue respectively. The current study revealed that single and  especially continuous Pb exposure  was strongly associated with Pb accumulation, antioxidant depletion, oxidative stress and kidney cells apoptosis. NAC can help protect kidney tissue against Pb by elevating antioxidant capacity, mitigating oxidative stress and normalizing Bax and Bcl2 genes expression.

Leptin and curcumin affect renal ischemia-reperfusion injury via modulation of P65 and Bax genes expression

M. M. Ragy1, M. M. Ramzy2*

1Physiology Department, Faculty of Medicine, Minia University, Misr-Aswan Road, Egypt;
2Biochemistry Department, Faculty of Medicine, Minia University, Misr-Aswan Road, Egypt;
*e-mail: maggiemaher24@gmail.com

Received: 01 June 2020; Accepted: 17 December 2020

Ischemia and reperfusion are natural steps during kidney transplantation, and ischemia-reperfusion injury is a critical condition in which physicians must preserve organ function and control cell damage. As leptin is thought to play an important role in the regulation of the immune system and inflammation and curcumin is a potent anti-fibrotic agent, both agents are promising to have therapeutic impact on renal damage. The present study was designed to evaluate the effects of leptin and curcumin on renal ischemia-reperfusion injury. Forty adult male albino rats were divided into four groups: control; ischemia-reperfusion (I/R), leptin-treated (leptin was injected intraperitoneally at a dose 100 μg/kg for 3 days prior to ischemia) and curcumin-treated (curcumin was given orally at a dose of 50 mg/kg/day for 5 days before ischemia). All rats were sacrificed 24 hours after reperfusion. Serum urea and creatinine, renal malondialdehyde and total antioxidant capacity were measured. Renal TNF-α was assayed by ELISA and P65 and Bax mRNA expression were determined using RT-PCR. Our results demonstrated a significant increase in P65 and Bax mRNA expression after renal ischemia-reperfusion injury compared to control group. Both leptin and curcumin prevented oxidative damage of the renal tissues as they lowered MDA and nitric oxide levels, increased antioxidant capacity and decreased TNF-α level. It was shown that protective leptin and curcumin effect against kidney IR-induced oxidative injury was associated with a down-regulation of P65 and Bax expression. These results show that ischemia-reperfusion leads to renal damage and also they reveal that both leptin and curcumin have protective implications which may be promising agents for avoiding various adverse effects.