Category Archives: Uncategorized
Double Nobel prize winner: Frederick Sanger – the father of genomics
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: 02 February 2021; Accepted: 23 April 2021
This paper aims to outline briefly the main stages of Frederick Sanger’s scientific activity – the only person to have won two Nobel Prizes in Chemistry (1958, 1980). His work on the structure of proteins, especially that of insulin, and the determination of base sequences in nucleic acids made an immense impact on the development of biochemistry and especially on the development of a new scientific field – molecular biology. His methods for determining the primary structure of proteins and nucleic acids helped biochemists and molecular biologists to determine the structure of many proteins and nucleic acids and laid the basis for genetic engineering.
Inclusion of enterosorbents in anti-inflammatory therapy improve treatment effectiveness in copd patients during exacerbations
S. V. Kucher*, O. V. Lototska
I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: kuchersv@tdmu.edu.ua
Received: 29 January 2021; Accepted: 23 April 2021
Inflammation plays a key role in the development of chronic obstructive pulmonary disease (COPD), and manifests as increased levels of inflammation markers such as cytokines. The purpose of this study was to examine the effect of enterosorbents on IL-1β, TNF-α and IL-10 levels in COPD patients with exacerbations, while taking into account their ages. Patients were divided into two age groups: age group 1 (40-59 years) and age group 2 (>60 years). Levels of IL-1β, TNF-α and IL-10 were determined by enzyme-linked immunosorbent assay. Levels of pro-inflammatory and anti-inflammatory cytokines were increased in both age groups of COPD patients during exacerbations compared to the healthy control group. Pro-inflammatory cytokines (IL-1β, TNF-α) increased more in age group 2. Basic therapy (BT) had positive impacts. Addition to BT of the enterosorbents Enterosgel and Carboline decreased serum levels of IL-1β, TNF-α and IL-10 in both age groups more than after BT alone. Greater changes were observed in age group 2. We found that addition of the enterosorbents Carboline or Enterosgel to basic therapy of COPD patients during exacerbations decreased levels of IL-1β, TNF-α and IL-10 in patients of both age groups (40-59 years and >60 years).
Clinical and immunological peculiarities of parasitic infections in children with digestive system disorders in Western Ukraine
K. T. Hlushko*, H. A. Pavlyshyn, K. V. Kozak
Department of Pediatrics No 2, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: glushko_kt@tdmu.edu.ua
Received: 29 January 2021; Accepted: 23 April 2021
Diseases of the digestive system such as gastroduodenal disorders (GDD) and hepatobiliary diseases (HBD) are fairly common among children. At the same time, intestinal parasites are widespread gastrointestinal tract infectious agents. We examined 108 children with GDD (n = 54) and HBD (n = 54) (mean age 11.8 ± 4.3 years) who were treated in a children’s hospital. The following were performed: blood test for specific Ig to Ascaris lumbricoides, Toxocara canis; feces analysis for helminth eggs, Giardia cysts; and pinworm test. The serum level of interleukin-4 (IL-4) was measured in 97 of 108 children. Overall parasitic co-infections were found in 60.2% of children: giardiasis in 30.6%, toxocariasis in 17.6%, ascariasis seropositivity in 13.9% and enterobiasis in 8.3% of cases. Parasitic infections (PIs) occurred in 72.2% of the HBD group and in 48.2% of the GDD group (P = 0.01). Among the PIs only ascariasis was found more often in the HBD group (22.2%) compared with the GDD group (5.6%) (P = 0.01). The mean age of children who were positive for PI (PI+) was 9.9 ± 4.6 years; while in the PI negative (PI–) group the mean age was 12.8 ± 3.3 years (P < 0.001). In GDD the IL-4 level in PI+ group was 9.3 ± 0.9 pg/ml in the versus 6.9 ± 1.8 pg/ml in the PI‒ group (P = 0.02). Children with HBD and PI had higher IL-4 levels (14.2 ± 18.8 pg/ml) compared to those without PI (7.5 ± 2.4 pg/ml) (P = 0.03). The differing impact on the course and immune response in children depending on co-infection with PIs indicates the need for additional testing for these infections.
Thromboelastographic study of fibrin clot and molecular basis of maximum clot firmness
D. S. Korolova1*, Y. M. Stohnii1, V. I. Gryshchuk1, S. I. Zhuk2,
I. V. Us2, T. M. Chernyshenko1, O. P. Kostiuchenko1, K. P. Klymenko1,
O. M. Platonov1,3, O. I. Ivashchenko3, V. O. Chernyshenko1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine;
3ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: d.korolova@gmail.com
Received: 29 January 2021; Accepted: 23 April 2021
Maximum clot firmness (MCF) is the main parameter of thromboelastography (TEG) reflecting the stability of a clot. In this work, we looked for markers that can influence the enhancement of MCF and detected molecular markers and blood clotting parameters that can be involved in such mechanisms. Blood samples of pregnant women with placental disorders were collected in the Kyiv Perinatal Center. TEG was performed on whole blood in EXTEM and INTEM tests. APTT, INR, fibrinogen concentration and platelet aggregation were measured using traditional laboratory approaches. D-dimer was detected in sandwich ELISA using monoclonal antibodies III-3B and II-4D. The relative cross-linking activity of factor XIIIa was measured by the direct quantification of the cross-linked γ-chain of fibrin using Western-Blotting with monoclonal antibody II-4D. D-dimer and fibrinogen concentrations, clotting time in the APTT test, INR and rate of platelet aggregation did not correlate with the MCF. However, we found positive correlations of MCF with factor XIIIa activity: 0.51 and 0.87 for EXTEM and INTEM, respectively. These data indicate that for normal and slightly increased fibrinogen concentrations, fibrin clot firmness will depend mostly on the activity of factor XIIIa. Thus the direct determination of factor XIIIa activity in blood plasma of patients can be relevant for predicting the risk of intravascular coagulation. Evaluation of the content and activity of individual clotting factors or other components of the coagulation system can be useful additions to the TEG diagnostics and should not be neglected.
Oxidative stress suppression contributes to antiseizure action of axitinib and rapamycin in pentylenetetrazol-induced kindling
O. B. Poshyvak1*, O. R. Pinyazhko1,2, L. S. Godlevsky3
1Pharmacology Department, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
2Department of Civilization Diseases and Regenerative Medicine, WSIiZ, Rzeszow, Poland;
3Department of Biophysics, Informatics and Medical Devices, Odesa National Medical University, Odesa, Ukraine;
*e-mail: olesya.poshyvak@gmail.com
Received: 29 January 2021; Accepted: 23 April 2021
Rapamycin and axitinib block different kinases in signaling pathways such as PI3K-Akt-mTOR and BDNF-TrkB, respectively. Both have antiseizure and antioxidative actions, which justify studying the combined effects of these drugs upon seizures and oxidative stress in the chronic model of epilepsy. The investigation aimed to look for the combined effect of rapamycin and axitinib upon pentylenetetrazol (PTZ)-kindled seizures and oxidative stress. Experiments were performed on 300 two- to four-month-old Wistar male rats, which had been kindled daily with PTZ (35.0 mg/kg, i.p.). Malondialdehyde (MDA) level, superoxide dismutase (SOD) activity, and glutathione (GSH) level were determined in brain tissues of kindled rats before and after the treatment. The analysis of antiseizure and antioxidative actions was performed using ED50 of rapamycin and axitinib, with their combined administration using graded dosages of ED50 of each drug. The median effective dose (ED50) for rapamycin and axitinib was 0.93 and 4.97 mg/kg, respectively. ED50 of rapamycin when combined with axitinib (2.0 mg/kg) was 0.60 mg/kg, which was reduced by 35.6% when compared with the ED50 administered alone (P < 0.05). The MDA level increased from 152.9±24.8 to 388.3±49.2 nmol/mg of protein (P < 0.05), while SOD activity reduced from 11.14±2.33 to 3.54±1.08 IU/mg of protein (P < 0.05) in brain tissues of the kindled rats. Combined treatment with rapamycin (0.56 mg/kg, i.p.) and axitinib (2.0 mg/kg, i.p.) resulted in a significant rise in SOD activity (11.09±1.86 IU/mg) and GSH level (7.32±1.34 µg/mg) when compared with the kindled rats (P < 0.05). Combined axitinib and rapamycin therapy have an antiepileptic and antioxidative effect on PTZ-kindled seizures.
Effect of a novel thiazole derivative and its complex with a polymeric carrier on stability of DNA in human breast cancer cells
N. S. Finiuk1,2, O. Yu. Klyuchivska1, I. I. Ivasechko1,
N. E. Mitina3, Yu. V. Ostapiuk2, M. D. Obushak2,
O. S. Zaichenko3, A. M. Babsky2, R. S. Stoika1,2*
1Institute of Cell Biology, NAS of Ukraine, Lviv, Ukraine;
2Ivan Franko National University of Lviv, Lviv, Ukraine;
3Lviv Polytechnic National University, Lviv, Ukraine;
*e-mail: stoika.rostyslav@gmail.com
Received: 26 January 2021; Accepted: 2021
Thiazole derivatives are perspective antitumor compounds characterized by a broad range of bioactivity, while polymeric carriers are widely used to enhance the efficiency of biological action of drugs, improve their biocompatibility and water solubility. Previously, we identified that the thiazole-based derivative BF1 (N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide) possessed differential toxicity towards targeted tumor cell lines. The aim of the present work was to investigate the action in vitro of BF1 and its complex with the polymeric carrier (PC) poly(PEGMA-co-DMM) (BF1-РС complex) towards human breast adenocarcinoma cells of the MDA-MB-231 and MCF-7 lines. DNA comet analysis, diphenylamine DNA fragmentation assay, gel retardation assay of plasmid DNA, DNA intercalation assay using methyl green dye and fluorescent microscopy were used to study the effects of BF1 on DNA stability in breast cancer cells. The ІС50 of cytotoxic action towards MDA-MB-231 cells was 26.5 ± 2.9 µМ for BF1, while the ІС50 for the BF1-PC complex was 6.9 ± 0.4 µМ, and the PC demonstrated low toxicity (ІС50 ˃ 50 µМ). The BF1-PC complex possessed higher toxicity towards MCF-7 cells than free BF1, with ІС50 of 9.6 ± 0.8 µМ and 15.8 ± 0.9 µМ, respectively. BF1 and BF1-PC induced an increase in the number of damaged cells of the MDA-MB-231 line with blebbing of plasma membrane, condensed chromatin and/or fragmented nucleus and micronuclei formation. Both BF1 and the BF1-PC complex induced single-strand breaks in DNA and its fragmentation in treated MDA-MB-231 cells. The studied compounds were not bound to plasmid DNA and did not intercalate into DNA molecules.
Locally delivered lovastatin-containing chitosan nanoparticles promote bone regeneration in rats
O. O. Shevchuk1*, Ya. V. Panasiuk2, M. M. Korda3
1Department of Pharmacology and Clinical Pharmacology, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
2Department of Functional and Laboratory Diagnostics, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
3Department of Medical Biochemistry, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: shevchukoo@tdmu.edu.ua
Received: 24 February 2021; Accepted: 23 April 2021
Hypolipidemic statins can stimulate osteoregeneration. However, such effects are observed only after administration methods that are unacceptable for patients (prolonged infusions or huge oral doses). The aim of our research was to compare the osteoregeneration effects of lovastatin administered alone as a common pharmaceutical formulation and as lovastatin-containing chitosan nanoparticles (LCCN) in a drill-hole model of bone damage in rats. White inbred rats were randomly divided into four groups: group 1 – intact rats; group 2 – rats with bone defect without treatment (control group); group 3 – rats with bone defect, which received common pharmaceutical formulation of lovastatin at doses of 0.1, 1.0 and 5.0 mg/kg; group 4 – rats, which received 0.1 mg/kg lovastatin in the form of lovastatin-containing chitosan nanoparticles (LCCN). A dental drill of 2.0 mm in diameter was used to form the tibial bone defect. Rats were sacrificed 3, 7, 14 and 28 days after bone defect formation. Calcium (Ca), phosphorus (P) and sialic acid concentrations, alkaline and acidic phosphatase activities, mineralization index, and collagenolytic activity were measured in blood serum. Computed tomography (CT) and histological study were used to estimate the regenerative processes in the bone. It was found that therapeutic doses of lovastatin (0.1 and 1.0 mg/kg) are ineffective for bone defect healing. Only high doses of lovastatin (5.0 mg/kg) promote osteoregeneration. LCCN were more efficient compared to lovastatin alone, as confirmed by CT examination of bone defects and significant changes of Ca, P, and sialic acid concentrations, alkaline and acidic phosphatase activities, mineralization index, and collagenolytic activity. Lovastatin-containing chitosan nanoparticles effectively enhance fracture healing in used preclinical model. This finding suggests the possibility that a similar approach may be effective in hastening fracture repair in humans.