Category Archives: Uncategorized
The impact of vitamin D(3) on bone remodeling in different types of experimental pathology
A. O. Mazanova*, O. O. Makarova, A. V. Khomenko, V. M. Vasylevska,
O. Yu. Lototska, I. O. Shymanskyi, M. M. Veliky
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: ann.mazanova@gmail.com
Received: 17 June 2022; Revised: 28 July 2022;
Accepted: 29 September 2022; Available on-line: 06 October 2022
Osteoporosis is a progressive systemic skeletal disease characterized by a decrease in bone density, impairment of its microarchitectonics, and an increased risk of fractures that occur under minimal or no mechanical stress. One of the main causes of osteoporosis is vitamin D deficiency, which leads to disruption of normal bone remodeling. The aim of our study was to analyze the features of the process of bone tissue remodeling by measuring the key biochemical markers of bone formation/resorption in primary and secondary osteoporosis, as well as to investigate the potential corrective effect of vitamin D3 supplementation. The work was conducted on rats with different osteoporosis models: alimentary, dysfunctional and secondary osteoporosis associated with diabetes mellitus. We used ELISA to measure 25(OH)D content in blood serum. Blood serum and bone tissue calcium, and alkaline phosphatase activity were determined with bioassay kits. The content of inorganic phosphate in blood serum and ash was assayed by the Dyce method. It was shown that all the studied pathological conditions were accompanied by vitamin D deficiency, which led to impaired absorption of calcium in the intestine and reabsorption of inorganic phosphates by the kidneys, reducing, as a result, their concentration in the blood serum. Hypocalcemia and hypophosphatemia contributed to the disruption of normal bone remodeling, excessive activation of alkaline phosphatase, and a decrease in the content of calcium and phosphate in bone tissue. Thus, sufficient vitamin D bioavailability was confirmed to be critical for effective bone remodeling in primary and secondary osteoporosis.
Unraveling the mystery of nitric oxide: Nobel prize winners Robert Furchgott, Louis Ignarro, and Ferid Murad
T. V. Danylova1*, S. V. Komisarenko2
1Institute for Social and Political Psychology, National Academy of Educational 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: 21 February 2022; Accepted: 01 July 2022
In the 21st century, none of the scientists denies the determining role of the cardiovascular system and its central organ, the heart. The ongoing attempts to design new medications, elaborate effective trainings, heart transplant programs testify that humanity does not abandon attempts to improve and prolong human life, especially given the fact that the world’s biggest killer is ischemic heart disease. The most significant achievements in this field receive the highest rating in the scientific community – the Nobel Prize. In 1998, the Nobel Prize in Physiology or Medicine was awarded jointly to Robert F. Furchgott, Louis J. Ignarro and Ferid Murad “for their discoveries concerning nitric oxide as a signaling molecule in cardiovascular system”. Their discovery triggered an international boom in research on nitric oxide. The paper aims to outline briefly the main stages of the scientific activity of R.F. Furchgott, L.J. Ignarro and F. Murad.
Analysis of chitosan molecular weight profile by electrophoresis in a porosity step gradient polyacrylamide gel
M. D. Lootsik1*, N. O. Manko1, R. O. Bilyy2,
M. M. Lutsyk (Jr.)2, R. S. Stoika1
1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Danylo Halytsky National Medical University, Lviv, Ukraine;
*e-mail: lootsikmaxim@gmail.com
Received: 12 April 2022; Accepted: 01 July 2022
Chitosan is biocompatible and biodegradable natural biopolymer widely applied in various fields of biology, medicine, and pharmacy, however, its effects significantly depend on the degree of polymerization (DP) and the degree of deacetylation (DDA) of polymer chains. Evaluation of the chitosan chain diversity by DP requires the use of a highly expensive method of high-performance size exclusion chromatography. The aim of our study was to determine the molecular weight profile of chitosan specimens by the use of electrophoresis in a porosity step gradient polyacrylamide gel and to evaluate the efficacy of this method in monitoring the purification of chitosan fragments and its derivatives. Two types of step gradient porosity gels were used: 1) gels of layers with acrylamide concentration 2.5, 3.5, 5.0, 10.0, 15.0, 20.0 % w/v for native chitosan or its high molecular fragments; 2) gels of layers with acrylamide concentration 2.5, 5.0, 10.0, 15.0, 20.0, 25.0 % w/v for low molecular chitosan fragments. The main amount of molecules from the chitosan pool was localized in the type 1 gel in the region of 550-40 kDa and distributed among three bands, which in different samples differed significantly in percentage. Electrophoresis of chitosan fragments fractionated by gel permeation chromatography provided a clear separation of medium molecular weight fragments (50–400 kDa) in type 1 gel and of low molecular weight fragments (3–40 kDa) in type 2 gel. Thus the method of chitosan electrophoresis in a step-gradient porosity of polyacrylamide gel was developed which permits to characterize the molecular weight profile of chitosan specimens polymer chains and is effective in monitoring the isolation of chitosan fragments by gel penetration chromatography of molecular weights from 3 to 400 kDa.
Application of gold nanoparticles to determine spermine in the presence of other polyamines
Yu. V. Yanish1, M. P. Prylutskyi1*, S. P. Zaletok1, Yu. P. Mukha2
1R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, National Academy of Sciences of Ukraine, Kyiv;
2O. O. Chuiko Institute of Surface Chemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: prilutskiy.maxim@gmail.com
Received: 19 October 2021; Accepted: 01 July 2022
The development of methods for the detection of polyamines in biological fluids is essential to improve early diagnosis and treatment of patients with prostate cancer. One of the promising areas is the use of noble metal nanoparticles. According to the literature data, there is no methodological approach have been developed to reliably distinguish spermine from other polyamines, in particular, from their acetylated forms and related compounds present in biological fluids. The paper presents the results of spectrophotometric determination of spermine both alone and in the presence of putrescine, spermidine or urea in the urine using gold nanoparticles. The results of the experiments proved that the developed method is suitable for the selective determination of spermine. It was shown that the presence of spermidine, putrescine, acetylated forms of polyamines or carbamide does not affect the results of the analysis.
Changes in the expression of TRPV4 and TRPM8 channels in the colon of rats with 6-OHDA-induced Parkinson’s disease
V. О. Stetska1, T. V. Dovbynchuk1, N. V. Dziubenko2,
A. V. Zholos1, G. M. Tolstanova2*
1ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Ukraine;
2Institute of High Technologies, Taras Shevchenko National University of Kyiv, Ukraine;
*e-mail: ganna.tolstanova@knu.ua
Received: 01 June 2022; Accepted: 01 July 2022
Parkinson’s disease (PD) is neurodegenerative disease, which is accompanied by degeneration of dopaminergic neurons in subtantia nigra. Non-motor symptoms, in particular, disorders of the gastrointestinal (GI) tract are observed in 20-80% of patients some 15-20 years before clinically diagnosed PD and are not a least important feature of PD pathogenesis. The transient receptor potential (TRP) channels are expressed throughout the GI tract, where they play an important role in taste, thermoregulation, pain, mucosal function and homeostasis, control of interstitial motility etc. The aim of this study was to investigate the contribution of TRPV4 and TRPM8 channels in the GI motor function in the colon of rats with PD, incduced by injection of the 12 μg 6-hydroxydopamine (6-OHDA). The studies were performed on the 4th week and the 7th month after PD induction The rats were randomly divided into: I group – the sham-lesioned rats, 4 μl 0.9% NaCl, autopsy 4 weeks after injection (n = 5); II group – the 6-OHDA-PD rats, 4 μl 12 μg of 6-OHDA, autopsy 4 weeks after injection (n = 5); III group – the sham-lesioned rats, 4 μl 0.9% NaCl, autopsy 7 months after injection (n = 4); IV group – the 6-OHDA-PD rats, 4 μl 12 μg of 6-OHDA, autopsy 7 months after injection (n = 5). We evaluated the body weight of rats, GI transit time, the cecum weight index and immunohistochemical identification of tyrosine hydroxylase (TH) -positive cells, and TRPV4, TRPM8 expression in rat’s colon. We showed that on the 7th month of the experiment, the GI transit time doubles over time; the cecum weight index of 6-OHDA rats increased by 57%; the number of TH-positive cells in colon rats decreased 2-fold, while TRPM8 ion channels were downregulated in PD rats and TRPV4 ion channels were upregulated in the colon of rats with 6-OHDA-PD. It was concluded that TRPV4 and TRPM8 ion channels may be considered pharmacological targets in the progression of PD pathology.
Thrombomodulin and von willebrand factor as markers of endothelial dysfunction in patients with chronic kidney disease
I. S. Mykhaloiko*, R. I. Yatsyshyn, N. V. Cherniuk, M. Ja. Humeniuk
Ivano-Frankivsk National Medical University, Ivano-Frankivsk, Ukraine;
*e-mail: iralisn@gmail.com
Received: 23 December 2021; Accepted: 01 July 2022
The aim of research was to study the levels of thrombomodulin (TM) and von Willebrand factor (VWF) in the serum and urine of patients with chronic kidney disease (CKD)as diagnostic markers of endothelial dysfunction. The study involved 140 patients with CKD. The clinical diagnosis was determined based on standard methods of patients examination according to the kidney diseases classification and protocols of CKD patients management. The concentrations of TM and VWF in serum and urine were quantified by ELISA. A generalized endothelial dysfunction in the vessels of the whole body, including the kidneys and high concentration of TM and FVF in the serum and urine of patients with a diabetic nephropathy have been found. The concentration of TM and VWF in the serum of patients with a chronic glomerulonephritis was at the same level as in the serum of healthy individuals, while those in urine significantly exceeded the control values, indicating endothelial damage in the glomeruli of the kidneys due to exposure to pro-inflammatory cytokines. In our opinion, the studied markers will contribute to the timely diagnosis of endothelial dysfunction in patients with CKD and to the development of criteria for prescribing antiplatelet agents in glomerular kidney disease.
Tumor biomarkers CEA, CA19.9, CA15.3 and AFP levels in the serum of patients with COVID-19
Abubaker H. Ali1*, Abdullah H. Yaqub2, Ihssin A. Faraj2
1Department of Biotechnology, Faculty of Science, Sebha University, Libya;
2Department of Zoology, Faculty of Science, Sebha University, Libya;
*e-mail: Abu.Ali@sebhau.edu.ly
Received: 11 December 2021; Accepted: 01 July 2022
Early diagnosis is very important to reduce morbidity and mortality in COVID-19 infected patients. The aim of this study was to detect of tumor antigens CEA, CA19.9, CA15.3, and AFP and to compare their levels in the serum of 69 COVID-19 patients and 69 healthy individuals who did not have COVID-19. The levels of CEA, CA19.9, CA15.3, and AFP in the serum were measured using ELISA. The levels of the tumor biomarkers in the serum of COVID-19 patients were found to be 7.74 ± 4.65 ng/ml for CEA, 29.33 ± 16.35 U/ml for CA19.9, 23.24 ± 13.48 U/ml for CA15.3 and 7.46 ± 5.57 ng/ml for AFP, while in the serum of healthy control patients 9.73 ± 43.58 ng/ml for CEA, 20.66 ± 11.1 for CA19.9, 19.64 ± 10.99 U/ml for CA15.3, and 3.83 ± 9.20 ng/ml for AFP, indicating no differences in the levels of the studied tumor biomarkers in the two experimental groups. It is concluded that tumor biomarkers CEA, CA19.9, CA15.3, and AFP cannot be used as effective screening tools for patients with COVID-19.
Comparison of adjuvant properties of chitosan during oral and subcutaneous immunization of mice with BSA
M. R. Kozak1*, I. M. Petruh1, V. V. Vlizlo2
1Institute of Animal Biology NAAS, Lviv, Ukraine;
2Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine;
*e-mail: mariyarkozak@gmail.com
Received: 15 December 2021; Accepted: 01 July 2022
Vaccination is the best method to prevent the spread of infectious diseases, its disadvantages are side effects. Potentially safe DNA, RNA or protein molecules possess antigenic properties, but are low-immunogenic and therefore require conjugation with an adjuvant. The aim of the research was to evaluate Chitosan (CS) potency as an adjuvant and compare its effectiveness depending on the route of drug administration. The experiments were carried out on 3 groups of BALB/c mice. Mice of the first group were injected subcutaneously with 20 µl of a mixture of CS (3.3 mg/kg) and BSA (1.7 mg/kg). The mixture of CS and BSA at the same doses and volume was administered orally to mice of the second experimental group. The third group – control – unvaccinated mice. Anti-BSA antibody levels were measured by ELISA. Aspartate aminotransferase, alanine aminotransferase activity and cholesterol, creatinine and urea levels were determined in the serum. It was found that both subcutaneous and mucosal immunizations provided a 2-fold increase in anti-BSA antibody titers against the background of maintaining all biochemical blood parameters at the level of the physiological norm. However, AST activity in the serum of oral-immunized mice was elevated as compared to subcutaneous-immunized mice. Serum cholesterol level in the group of subcutaneously immunized mice and creatinine and urea levels in both experimental groups were reduced compared to the control. It is concluded that oral immunization with CS is the optimal route for antigen-specific IgG antibody response induction.
Calix[4]arene chalcone amide C-1011 elicits differential effects on the viability of 4T1 mouse breast adenocarcinoma cells with different levels of adaptor protein Ruk/CIN85 expression
L. G. Babich1*, S. G. Shlykov1, O. A. Yesypenko2, A. O. Bavelska-Somak1,
A. G. Zahoruiko1, I. R. Horak1, L. B. Drobot1, 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: babich@biochem.kiev.ua
Received: 07 February 2021; Accepted: 01 July 2022
According to our earlier data, calix[4]arene chalcone amides modulate Ca ions exchange in the myometrium mitochondria and the level of inner membrane polarization that can potentially affect cell survival. To test this hypothesis, we studied the effect of calix[4]arene with 4 chalcone amide groups on mitochondria membrane polarization and viability of 4T1 mouse breast adenocarcinoma cells, a surrogate model of human triple-negative breast cancer, and on its highly malignant subline overexpressing the adaptor protein Ruk/CIN85. Mitochondria membrane potential was measured by flow cytometry, and cell viability was assessed using Trypan blue dye exclusion. It was shown that mitochondrial membranes of control (Mock) cells had a higher polarization level (67.80 ± 8.82 r.u., n = 5) compared to 4T1 cells with up-regulation of Ruk/CIN85 (RukUp cells) (25.42 ± 2.58 r.u., n = 4). Upon incubation of cells with 1 μM calix[4]arene C-1011, the CCCP-sensitive component of mitochondrial membranes polarization decreased (by almost 50%) in 4T1 Mock cells and did not change in RukUp cells compared with the control. It was demonstrated that 1 μM calix[4]arene C-1011 suppressed the viability of 4T1 Mock cells by 45%, but did not affect RukUp cells considerably. It was suggested that calix[4]arene chalcone amide С-1011 decreased mouse breast adenocarcinoma 4T1 cell viability at least by affecting mitochondrial membrane polarization.The data obtained indicate the prospects of further studies of calix[4]arene chalcone amide as a potential anticancer drug candidate.