Studia Biologica

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ISSN / EISSN : 1996-4536 / 2311-0783
Published by: Ivan Franko National University of Lviv (10.30970)
Total articles ≅ 632
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I. V. Shydlovskyy, Y. V. Tsaryk
Studia Biologica, Volume 15, pp 73-88;

Introduction. Information on historical changes in the fauna of geographical regions and biota habitats is important for understanding the processes that take place in ecosystems. The results of such an analysis in combination with modern research can indicate not only the global trends in changes of animal associations and the status of particular populations of species, but also reveal the reasons that caused the situation. Materials and discussion. The first lists of bird species in the 18th – early 19th centuries showed the nesting of 12 species of waders. However, during the 19th century 37 species of waders were found, 12 of which were nesting. 13 wader species were permanent migratory and 8 – rare migratory species. In the first half of the 20th century, there were 27 species of waders on the territory of Prykarpattia, and 25 in Volyn, among which there is a larger list of migratory and vagrant species in the foothills of the Carpathians. In the second half of the 20th century, 39 species of waders were described within the western regions of Ukraine, of which 17 were nesting, and four of them were recorded during the last decade. Conclusions. There are 42 species of waders (39 of them are recent) registered in the western part of Ukraine. Among them 17 species are nesting, 15 – migratory and seven – vagrant species. During the 20th–21st centuries, 23 species of waders, including only 7 nesting species, have not changed their status of stay. However, it has changed for 17 species: two species have disappeared; four – have become vagrant; the Ruff Calidris pugnax and the Marsh Sandpiper Tringa stagnatilis have changed their status from nesting to migratory. The Northern Lapwing Vanellus vanellus, the Green Sandpiper Tringa ochropus and the Wood Sandpiper T. glareola that were common, somewhere numerous bird, – have become few in numbers; and the Great Snipe Gallinago media and the Black-tailed Gotwit Limosa limosa – have become rare. The Jack Snipe Lymnocryptes minimus has become migratory. The nesting of sandpipers, with a southern area of distribution, in the western regions of Ukraine proves the fact of the global warming. However, the drop of the water level, drying out of wet meadows and swamps and, as a consequence, plant succession, have terminated the nesting of such species as the Ruff and the Marsh Sandpiper; the Black-tailed, the Great Snipe and the Wood Sandpiper become rare; the Northern Lapwing and the Green Sandpiper have become much less numerous.
N. V. Hrushanyk, Y. I. Fedorko, , Institute of Cell Biology NAS of Ukraine
Studia Biologica, Volume 15, pp 41-50;

Background. Improper folding and accumulation of a-synuclein aggregates are among the causes of Parkinson’s disease. The most important factor influencing the process of α-synuclein aggregation is the level of this protein in neurons which depends on the balance between its synthesis, degradation and secretion. Under certain conditions, when α-synuclein is synthesized at a high level, monomers of this protein can aggregate on the lipid membrane, which leads to the formation of amyloids, fibrils and protofibrils unable to perform their physiological functions. Since it is virtually impossible to study the properties of α-synuclein in vivo, researchers are actively using model biological systems (single-celled microorganisms, human cell lines, animal models etc.). The aim of this study was to construct a recombinant strain of Saccharomyces cerevisiae with controlled expression of human α-synuclein to study the regulation and properties of this protein and for screening for new low molecular weight chemi­cal compounds which can induce α-synuclein aggregation and/or degradation. Materials and methods. A recombinant strain of S. cerevisiae with controlled expression of α-synuclein conjugated to a green fluorescent protein was isolated. Western blotting with specific anti-α-synuclein antibodies was used to detect recombinant α-synuclein in yeast cells. Intracellular localization of heterologous chimeric green fluorescent protein conjugated to α-synuclein was also examined by fluorescence microscopy. Results. To construct a recombinant strain of S. cerevisiae, the coding sequence of the human wild-type α-synuclein gene was expressed under the regulated promoter of the ScMET25 gene. Analysis of the effect of different concentrations of exogenous methionine as a factor regulating the expression of the ScMET25 promoter on the content of recombinant protein showed that the expression of the human α-synuclein gene in S. cerevisiae is repressed in the presence of methionine at a concentration of 10 mg/L and higher. During long-term cultivation of yeast cells, this effect decreased due to the depletion of methionine in the growth medium. As a result, recombinant protein synthesis was restored, and α-synuclein content in such cells approached that of cells grown in a medium with a low concentration of (5 mg/L), or without methionine. It was also found that overproduction of recombinant α-synuclein in S. cerevisiae cells had virtually no effect on culture growth, indicating the absence or a very weak toxic effect of human α-synuclein on yeast physiology. Conclusions. The obtained data indicate a concentration-dependent effect of methionine on the level of recombinant α-synuclein synthesis in S. cerevisiae yeast cells. Such controlled expression of the studied protein can be used to screen for compounds capable of promoting dose-dependent aggregation or degradation of α-synuclein in yeast cells and potentially in human cells as well.
A. I. Polishchuk,
Studia Biologica, Volume 15, pp 51-60;

Background. Various modes of transport are among the main sources of environmental pollution with heavy metals and other pollutants. Bryophytes are known to accumulate heavy metals; however, metabolic changes in mosses under conditions of metal accumulation have not been extensively studied. The aim of this article was to analyze the accumulation of heavy metals, the process of lipid peroxidation (LPO) and the activity of enzymes of the antioxidant system in Didymodon rigidulus Hedw. moss collected in urban areas with an intense load created by road and railway transport. Materials and Methods. Gametophyte shoots were collected at three sampling sites in the city of Lviv (Ukraine). Site 1 was selected in a park zone, which was considered a control one; sites 2 and 3 were selected in areas with heavy road and rail traffic, respectively. Concentrations of chromium (Cr), nickel (Ni), lead (Pb), and zinc (Zn) in the moss material were determined by atomic absorption spectrophotometry. The levels of LPO products, namely lipid hydroperoxides and thiobarbituric acid reactive substances (TBARS), as well as superoxide dismutase (SOD) and catalase activities, were determined by standard methods. The results were processed using the methods of variation statistics. Results. Moss D. rigidulus growing in the park area (site 1) accumulated metals in the following order of decreasing concentration: Zn> Cr> Ni> Pb. Gametophyte shoots of D. rigidulus collected at site 2 accumulated higher levels of Pb, Zn and Ni (by 2.27, 1.78 and 1.45 times, respectively), and at site 3, higher levels of Pb and Zn (by 1.8 and 1.67 times, respectively) compared to gametophytes collected in the park zone. In the moss samples from these sites, no significant differences in the Cr content were found as compared to the control. Concentration of lipid hydroperoxides in the moss sampled at sites 2 and 3 was 4.26 and 3.75 times higher, respectively, compared to the control, and TBARS production was more intense in plant material from site 2 than from the control site. SOD and catalase activities were considerably increased in D. rigidulus moss from site 2 compared with those from the control area; however, the activity of both enzymes in the moss samples from site 3 did not significantly exceed the control levels. Conclusions. Both road and rail traffic loads contribute to the accumulation of heavy metals, especially Zn and Pb, in D. rigidulus moss growing in the surrounding areas. Under such conditions, LPO process is stimulated, which is more pronounced in moss growing in area with heavy road traffic. The increased activity of antioxidant enzymes (SOD and catalase) in moss growing in this area can play an important role in protecting bryophyte cells against metal-induced oxidative stress under conditions of intense metal accumulation.
T. Shupova, Institute for Evolutionary Ecology NAS of Ukraine, A. Chaplygina
Studia Biologica, Volume 15, pp 61-72;

Background. In the 21st century, landscape transformation processes are underway in large cities, which affects the stability of wildlife habitats. Habitat transformations often reduce species richness due to a decrease in the population sizes of some species, and therefore, small in number and rare species are eliminated from bird communities. Rare species can have unique consortive relationships, which makes them particularly important for the long-term ecosystem functioning. A study of the avifauna of forest parks makes it possible to develop an algorithm for the coexistence of human and birds. Methods. The number and distribution of birds were determined by route counting. The total length of the fixed route was 5.7 km in Kyiv and 3.5 km in Kharkiv. On each route, observations were carried out annually with three repetitions during the nesting period when the birds are most attached to their habitats (end of April–May–June). The average data for the total study period (2013–2017) were calculated for each city. For the average number, the standard deviation was calculated. An analysis of the faunogenetic structure of avifauna was carried out according to the method developed by V.P. Belik. A faunogenetic complex is a group of animal species associated by a common origin with ecosystems of a certain landscape-geographical zone. We also classified bird species into ecological groups according to the patterns of microhabitat choice. To compare the α-diversity of bird in the forest-park zones of cities, a number of commonly accepted indices that express the correlation between the number and density of species were calculated: 1) Berger–Parker dominance index: DBP = Nimax / N; 2) Shannon diversity index: H´ = -∑(Pi × LnPi); 3) Pielou evenness index: E = H´ / LnS; where: Nі– the number of each species; Nimax – the maximum value of Nі; N = ∑Ni – the total number of all species (pairs/km); Pi = Ni / N – the ratio of each species; S – total number of the species. Results. The study presents a comparative analysis of diversity and faunogenetic structure of avifauna in the forest park zones of Kyiv and Kharkiv, inhabited by 71 breeding species of birds that belong to 10 orders. In the eastern region, the proportion of birds of the boreal and the European forest-steppe complex decreases, but the share of the desert-mountain complex increases. The fauna of the European nemoral complex dominates (32.8 % in Kyiv and 40.4 % in Kharkiv). The basis of the communities are dendrophils: 83.6 % (n = 67) in Kyiv and 82.7 % (n = 52) in Kharkiv. The dominant species in all forest parks are the great tit (Parus major) and chaffinch (Fringilla coelebs). Conclusions. The differences in the faunogenetic structure of bird communities are due to the proximity of model forest parks on the territory of Kyiv to the forest natural geographical zone, and on the territory of Kharkiv to the steppe, which leads in the eastern region to a decrease in the proportion of birds of the boreal and the European forest-steppe complex while the proportion of desert-mountain complex increases. Dendrophils predominate significantly, and the share of sclerophils and limnophils in total is less than 20 % of the bird community in the forest parks of each city. As a consequence of the fragmentation of the Kiev forest park zone, the diversity of nesting birds communities in the forest-park zone of Kyiv is slightly lower than of Kharkiv, and the pressure of the dominant species is more significant.
Studia Biologica, Volume 15, pp 29-40;

Introduction. Genetic engineering in plants is of great importance for agriculture, biotechnology and medicine, and nanomaterials are widely used for genetic engineering. The aim of present study was to evaluate the potential of poly(2-dimethylamino)ethyl methacrylate (DMAEMA)-based comb-like polymers as gene delivery systems in moss Ceratodon purpureus (Hedw.) Brid. protoplasts and determine the level of phytotoxicity of these polymers. Materials and Methods. In order to confirm the formation of complex of poly-DMAEMA carrier with plasmid DNA pSF3, gel retardation assay was used. The PEG-mediated transformation protocol was adapted to transform the protoplasts of C. purpureus moss with poly-DMAEMA carriers. Light microscopy was used to study a toxicity of polymers for moss protoplasts. The level of the polymers toxicity was estimated as IC50 value. Results and Discussion. The formation of pDNA complex with DMAEMA-based carriers took place at 0.03% concentration of the polymers BGA-21, BGA-22(2ph), BG-24, BG-25, BG-26 or 0.1% concentration of the BGA-22 polymer. Poly-DMAEMA carriers were able to deliver plasmid DNA pSF3 into protoplasts of C. purpureus moss. Three stable transformants of C. purpureus were obtained at using BGA-22 polymer, 2 clones – at using BGA-21 carrier, and 1 clone – at using BGA-22(2ph), BG-24, BG-25, BG-26 polymers. The poly-DMAEMA carriers at the working 0.0025% dose were relatively non-toxic for protoplasts of C. purpureus moss. 83.1-88.4% of viable protoplasts of C. purpureus moss were detected after treatment with studied carriers at 0.0025% dose. A survival ratio of protoplasts reached 66.7-72.9% under the effect of these polymers at 0.025% dose, which is 10 times higher than their working concentration. The IC50 value of poly-DMAEMA carriers was in the range of 0.113-0.164%, that was approximately 10 times higher than that of the PEG-6000 used for gene delivery in plants. Conclusion. Novel synthetic poly-DMAEMA carriers delivered the gene of interest into moss C. purpureus protoplasts and possessed a low phytotoxicity. Thus, these carriers can be useful for gene delivery into plant cells.
Studia Biologica, Volume 15, pp 17-28;

Background. Previously, we have detected the antitumor and anti-inflammatory activities of pyrrole-derived protein kinase inhibitors - MI-1 (1-(4-Cl-benzyl)-3-Cl-4-(CF3-phenylamino) -1H-pyrrole-2,5-dione 1) and D1 (5-amino-4-(1,3-benzothiazole-2-yl)-1-(3-methoxyphenyl)-1,2-dihydro-3H-pyrrole-3-one) using rat colon cancer model. Therefore, pyrrole derivatives was aimed at detecting the anti-inflammatory effect on the model of ulcerative colitis caused by acetic acid in rats. Materials and Methods. Prednisolone was used as a reference anti-inflammatory drug of glucocorticoid nature. It was administered intraperitoneally at a dose of 0.7 mg/kg. The compounds were administered in 2 h after the first administration of acetic acid. Total protein was estimated quantitatively, as described by Lowry et al., 1951. Content of the malonic dialdehyde, protein carbonyl groups, and the activity of antioxidant enzymes as indicators of colon mucosa redox status were measured spectrophotometrically. Statistical analysis of the results was performed using MS Excel-2013. Results and Discussion. In case of chronic colitis, the number of carbonyl groups and lipid peroxidation products in the colonic mucosa are increased, indicating the development of oxidative stress. The injection of pyrrole derivatives separately contributes to the approaching these indicators to normal. Adding prednisolone does not have this effect. Colitis has been shown to have a decrease in superoxide dismutase activity, which is a typical phenomenon for chronic inflammation and may indicate depletion of the enzyme. In case of colitis, alanine aminotransferase activity and the content of direct bilirubin are increased, which indicates a liver injury and are systemic manifestations of inflammation of the colon. Pyrrole derivatives help to reduce the liver injury, which indicates the restoration of normal alanine aminotransferase activity and direct bilirubin content. Conclusion. It has been found that at chronic colitis pyrrole derivatives reduce the manifestations of inflammation, contribute to the normal structure of the mucous membrane (comparative to prednisolone as a standard anti-inflammatory drug). It suggests their anti-inflammatory effectiveness, while an increase in total bilirubin under exposition to pyrrole derivatives may be a sign of the adverse effects on the rat’s liver.
O. I. Karmash, , N. V. Yefimenko,
Studia Biologica, Volume 15, pp 3-16;

Background. Diabetes mellitus is a chronic endocrine-metabolic disease caused by an absolute or relative insulin deficiency. During diabetes, there are perfect conditions for the development of oxidative stress: the content of substrates for oxidation increases, the content of natural antioxidants decreases and the activity of antioxidant systems is suppressed. It is known that photobiomodulation therapy produce antioxidant and antihyperglycemic effects. Here we investigated its influence on blood system functioning. Materials and Methods. The study was performed on male Wistar rats. Experimental diabetes mellitus was induced by the intraperitoneal injection of streptozotocin. Leukocyte formula was calculated using blood smears stained by Romanowsky–Giemsa. Catalase activity was determined spectrophotometrically. Affinity of hemoglobin to oxygen was evaluated by spectrophotometric method in Ivanov’s modification by drawing oxygenation curves. The protoporphyrin content in whole blood was measured by analyzing its fluorescence spectra. The content of NO2-, total and inducible NO synthase activity was determined spectrophotometrically. Results. Under the action of photobiomodulation therapy on healthy animals, there was a shift of oxygenation curves to the left and a decrease of P50, whereas under irradiation of rats with diabetes, there was a shift of oxygenation curves to the right and increase in P50 compared to indices in nonirradiated animals. During diabetes, there was a decrease in protoporphyrin content compared to control, but there was a tendency to increase under photobiomodulation. Photobiomodulation therapy of rats with diabetes increased catalase activity in erythrocyte hemolysates. We revealed significant changes in leukocyte formula under photobiomodulation. The total NO synthase activity in leukocytes of rats with diabetes was higher compared to healthy animals, but decreased under the action of photobiomodulation. We found an increase in inducible NO synthase activity in leukocytes of rats with diabetes and in leukocytes of irradiated healthy animals. An increase in NO2- content in leukocytes of rats with diabetes was detected. Under photobiomodulation, NO2- content was significantly lower in rats with diabetes. Conclusion. Photobiomodulation therapy produces a corrective action on blood system during diabetes, in particular, it improves oxygen release from hemoglobin and prevents hypoxia. Simultaneously with the increase in tissue oxygen saturation, a decrease in NO synthase activity and nitrite content along with an increase in catalase activity prevents the development of oxidative stress.
D. Yu. Hertsyk, , L. V. Kaprelyants, , Ukraine Odesa National Academy Of Food Technologies
Studia Biologica, Volume 15, pp 37-48;

Background. Diabetes mellitus is accompanied by oxidative-nitrative stress, which is caused both by an increased generation of the Reactive Oxygen Species and the Reactive Nitrogen Species under the conditions of this pathology and by the disorders of the antioxidant defense system, especially its enzymatic part. This leads alterations in the morpho-functional state of cells, organs, and the whole organism. Free radicals are involved in the destruction of pancreatic cells in type 1 diabetes mellitus, which leads to even greater inhibition of insulin secretion, worsening of the course of the disease and occurrence of diabetic complications. The complications of diabetes mellitus include often diagnosed cardiovascular diseases. Therefore, the use of treatments characterized not only by hypoglycemic properties for normalization of blood sugar level in diabetes mellitus, but also by antioxidant properties for normalization of oxidative/antioxidant balance of the organism in the studied pathology can be promising. Scientists pay a great attention to the study of substances with such properties, especially natural phenolic compounds of grape, which are also characterized by immunomodulatory properties. Materials and Methods. The study was performed on cardiac tissues of control rats, control animals treated with wine polyphenolic complex concentrate, animals with streptozotocin-induced diabetes mellitus and animals with experimental diabetes mellitus treated with a concentrate of wine polyphenolic complex. The activity of catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase were detected to examine the corrective effect of the concentrate of red wine natural polyphenolic comp­lex on the state of the enzymatic part of the antioxidant defense system. Results. The results have shown the normalization of activity of catalase, superoxide dismutase, and changes in the activity of enzymes of glutathione cycle after oral administration of polyphenolic complex concentrate for 14 days to rats with streptozo­tocin-induced diabetes mellitus. Conclusions. The results confirm a hypothesis about the antioxidant effect of the studied concentrate and the ability of natural polyphenolic complexes to serve as the basis for new drugs for treatment of diabetes-induced disorders.
M. D. Lootsik,
Studia Biologica, Volume 15, pp 49-60;

Introduction. An acquired resistance to experimental tumors was detected in animals that recovered from a primary transplanted tumor due to treatment or spontaneously, and demonstrated intolerance to a renewal tumor inoculation. This phenomenon is much less frequently observed, although it is of great scientific interest and medical significance. Here, we have addressed the expression of the resistance phenomenon in a model tumor in mice – Nemeth–Kelner lymphoma (NK/Ly). The aim of our study was to elaborate a reproducible method for induction of resistance to transplantation of lymphoma NK/Ly in mice and to investigate the mechanisms of its development. Methods and Results. Three schemes for induction of resistance were tested. The first one included treatment of tumor-bearing mice with vinblastine and, thereafter, reconvalescent animals were checked for the development of resistance expressed as a complete suppression of tumor growth after re-inoculation of tumor cells. Mice were inoculated intraperitoneally with NK/Ly ascitic cells and then subjected to 2–4 intraperitoneal injections of vinblastine at a dose of 1mg/g of body weight. The recovered mice were re-inoculated with tumor cells and the absence of tumor growth was considered as resistance development. The disadvantage of this approach is that less than 5% of mice achieve a long lasting recovery due to the treatment. The second scheme included the immunization of mice with intraperitoneal injection of the minimal number of viable tumor cells that do not cause tumor growth, but initiate the immune response. However, this approach was not effective, since there was no reliable number of cells correspon­ding to these demands. The minimal number of 15×103 injected cells per mouse caused a retarded but still progressive tumor growth. In the third scheme, the immunization of mice was conducted by the intraperitoneal injections of NK/Ly cells permeabilized with saponin. It should be noted that treatment with saponin leads to cell death with a minimal damage to cell morphology. The scheme of immunization with permeabilized NK/Ly cells appeared to be simple and effective. It provided a reproducible resistance to transplanted tumor and might be used as a model in studies of the mechanisms of this phenomenon. Cytological investigation of tumor and immunocompetent cells in ascites of control and of tumor-resistant mice was conducted. As revealed, the number of lymphocytes in ascites of tumor-resistant mice was about 4 times higher than such amount in the control (non-resistant) mice. A destruction of tumor cells by the adherent mono-nuclears was observed. Conclusions. The method of induction of resistance to transplantation of experimental tumor NK/Ly by immunization of mice with tumor cells permeabilized with saponin is described. The intraperitoneal inoculation of tumor cells to the tumor-resistant mice caused the marked increase of the mononuclear leukocytes population in the peritoneal fluid, which showed a harmful effect upon tumor cells. Thus, the induction of resistance to transplantation of NK/Ly lymphoma in mice might be provided mainly via the mechanisms of cell immunity, in particular, by the appearance of cytotoxic lymphocytes specific to distinct tumor cells.
Studia Biologica, Volume 15, pp 25-34;

Background. Diabetes mellitus is a chronic endocrine metabolic disease with absolute or relative insufficiency of insulin, accompanied by impaired metabolism. Endogenous bioamine agmatine may become a basis of new antidiabetic drugs, as it is capable to induce the release of some peptide hormones, in particular insulin, and can regulate NO synthesis. Natural polyphenols are potential multifunctional agents that also can reduce the risk of diabetes and diabetic complications. The aim of the study was to evaluate the effect of agmatine and red wine concentrate, enriched with polyphenolic compounds, on NO-synthase activity and the content of NO stable metabolites under experimental diabetes mellitus. Materials and Methods. The experiments were conducted on white Wistar male rats. Diabetes was induced by intra-abdominal injection of streptozotocin. From the 14th day after the induction of diabetes, agmatine was injected intramuscularly or red wine concentrate, enriched with polyphenolic compounds was administrated orally to animals for 14 days. Rats were decapitated under ether anesthesia on the 28th day of the experiment. In the brain of rats, the activity of constitutive (Ca2+-dependent) and inducible (Ca2+-independent) isoforms of NO-synthase and the content of nitrite and nitrate anions were determined. Results and Discussion. The activities of constitutive and inducible isoforms of NO-synthase were increased in the brain of diabetic rats. The administration of both agmatine and red wine concentrate, enriched with polyphenolic compounds, caused the reduction of the activities of NO-synthase isoforms. In the case of diabetes, the administration of agmatine contributes to the increase of nitrite and nitrate content in brain cells compared to diabetes. The administration of red wine concentrate, enriched with polyphenolic compounds, also promotes nitrite levels but does not affect the nitrate level. Conclusion. We found that the red wine concentrate, enriched with polyphenolic compounds, has a stronger effect on the activity of Ca2+-dependent and Ca2+-independent isoforms of NO-synthase, as well as the content of nitrites and nitrates in brain of rats with experimental diabetes mellitus, compared to the effect of agmatine.
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