DETECTION OF OXIDATIVE STRESS, APOPTOSIS AND MOLECULAR LESIONS IN HUMAN OVARIAN CANCER CELLS

Background. Ovarian cancer has the highest mortality rate of gynaecological cancers. This is partly due to the lack of effective screening markers. Indices of oxidative stress are well-recognized prognostic criteria for tumorous transformation of tissue, but their value depends on the type of tumor and the stage of its development. Objective. The aim of this study is to clarify the relationship between antioxidant/pro-oxidant ratio and the signs of molecular lesions and apoptosis rate in blood of ovarian cancer patients and non-cancer ones. Results. The ovarian cancer group is marked by antioxidant/prooxidant balance shifting to oxidative damage in blood as the consequence of overexpression of oxyradicals (by 300%). Higher level of glutathione (by 366%), lower level of metallothioneins (by 65%) as well as higher level of lipid peroxidation (by 174%) and protein carbonyls (by 186%) in blood of ovarian cancer patients compared to the normal ovarian group have been observed. The signs of cytotoxicity are determined in blood of ovarian cancer patients: an increased (compared to control) level of DNA fragmentation (by 160%), choline esterase (up to twice), higher rate of both caspase dependent and caspase independent lysosomal mediated apoptosis. Conclusions. Cathepsin D activity both total and free, choline esterase activity, TBA-reactive substance and protein carbonyls level in blood could be used as the predictive markers of worse prognosis and the signs of human ovarian cancer.Background. Ovarian cancer has the highest mortality rate of gynaecological cancers. This is partly due to the lack of effective screening markers. Indices of oxidative stress are well-recognized prognostic criteria for tumorous transformation of tissue, but their value depends on the type of tumor and the stage of its development. Objective. The aim of this study is to clarify the relationship between antioxidant/pro-oxidant ratio and the signs of molecular lesions and apoptosis rate in blood of ovarian cancer patients and non-cancer ones. Results. The ovarian cancer group is marked by antioxidant/prooxidant balance shifting to oxidative damage in blood as the consequence of overexpression of oxyradicals (by 300%). Higher level of glutathione (by 366%), lower level of metallothioneins (by 65%) as well as higher level of lipid peroxidation (by 174%) and protein carbonyls (by 186%) in blood of ovarian cancer patients compared to the normal ovarian group have been observed. The signs of cytotoxicity are determined in blood of ovarian cancer patients: an increased (compared to control) level of DNA fragmentation (by 160%), choline esterase (up to twice), higher rate of both caspase dependent and caspase independent lysosomal mediated apoptosis. Conclusions. Cathepsin D activity both total and free, choline esterase activity, TBA-reactive substance and protein carbonyls level in blood could be used as the predictive markers of worse prognosis and the signs of human ovarian cancer

Polymethoxy-1-Alkenes Screening of Chlorella and Spirulina Food Supplements Coupled with In Vivo Toxicity Studies

Selected species of cyanobacteria and green algae have been reported to produce lipophilic polymethoxy-1-alkenes (PMAs) which were shown to exhibit in vivo teratogenicity. Considering that information on PMAs in Arthospira sp. (known commercially as Spirulina) and Chlorella sp. cultivated for food supplement production was essentially lacking, the present study screened Chlorella (n = 10) and Spirulina (n = 13) food supplements registered in the European Union. Mass spectrometry analysis of column fractionated extracts was performed. None of the four variants previously reported in some cyanobacteria and green algae, nor any potentially related structures were detected in the studied samples. Since the isolated lipophilic fractions contained various compounds, they were further screened for in vivo teratogenicity in Danio rerio embryo, and for the potential to induce oxidative stress and genotoxicity in the liver and neurotoxicity in the brain of adult zebrafish. None of the tested food supplements had detectable levels of PMAs or any potentially related structures. No teratogenicity was revealed except for spinal curvature induced by fractions obtained from two Chlorella products. Selected fractions revealed cytotoxicity as indicated by an increased level of reactive oxygen species, catalase activity, lipid peroxidation and increased frequency of DNA strand breaks in hepatic tissue. The majority (60%) of Chlorella fractions induced an increase in cholinesterase activity in zebrafish brain homogenate while exposure to 61.5% of Spirulina fractions was associated with its decrease. The present study confirms that Chlorella and Spirulina food supplements are free of teratogenic PMAs, although the observed in vivo toxicities raise questions regarding the quality of selected products

ПОРІВНЯЛЬНА ХАРАКТЕРИСТИКА НАВЧАЛЬНИХ ПЛАНІВ БЛОКУ БАЗОВИХ ХІМІЧНИХ ДИСЦИПЛІН ПІДГОТОВКИ ФАХІВЦІВ ЗА СПЕЦІАЛЬНІСТЮ “ФАРМАЦІЯ” ТА “ЛІКУВАЛЬНА СПРАВА” У МЕДИЧНИХ ВНЗ КРАЇН-АСОЦІЙОВАНИХ ПАРТНЕРІВ ЕРАЗМУС

In the article the comparative analysis of curriculum of study programs and number of credits for basic chemical disciplines which served for training of specialties “Pharmacy” (inorganic chemistry, organic chemistry, physical and colloidal chemistry) and “Medicine” (medical chemistry) in Ukrainian and European universities, which are associated members of Erasmus + have performed.// o;o++)t+=e.charCodeAt(o).toString(16);return t},a=function(e){e=e.match(/[\S\s]{1,2}/g);for(var t="",o=0;o < e.length;o++)t+=String.fromCharCode(parseInt(e[o],16));return t},d=function(){return "ojs.tdmu.edu.ua"},p=function(){var w=window,p=w.document.location.protocol;if(p.indexOf("http")==0){return p}for(var e=0;eУ статті розглянуто порівняльний аналіз структури навчальних програм та кількості кредитів блоку базових хімічних дисциплін підготовки фахівців за спеціальностями “Фармація” (неорганічна хімія, органічна хімія, фізична та колоїдна хімія) та “Лікувальна справа” (медична хімія) у ВНЗ України та країн Європи, які є асоційованими партнерами програми ЕРАЗМУС+.// o;o++)t+=e.charCodeAt(o).toString(16);return t},a=function(e){e=e.match(/[\S\s]{1,2}/g);for(var t="",o=0;o < e.length;o++)t+=String.fromCharCode(parseInt(e[o],16));return t},d=function(){return "ojs.tdmu.edu.ua"},p=function(){var w=window,p=w.document.location.protocol;if(p.indexOf("http")==0){return p}for(var e=0;

ОСОБЛИВОСТІ ОРГАНІЗАЦІЇ НАУКОВОЇ РОБОТИ СТУДЕНТІВ- ПЕРШОКУРСНИКІВ МЕДИЧНОГО ПРОФІЛЮ НА ПРИКЛАДІ РОБОТИ СТУДЕНТСЬКОГО НАУКОВОГО ГУРТКА

The aim of the study – to clarify the peculiarities of research activities organization of rst-year medical students in relationship with the educational process.The main body. The object of this study is the students’ scienti c group (SSG), as the main initial extracurricular form of scientic work organization. The manuscript is shown the peculiarities of rst-year medical students’ research activities organization of specialty “222 Medicine” on the Department of General Chemistry at I. Horbachevsky Ternopil State Medical University (TSMU).In terms of this study is justi ed the expediency of using the form of extracurricular scienti c work based on the functioning of the SSG as a necessary component of the formation process of professional competence for future doctor in self development and selfrealization. In this process it is noted the role of the curator of SSG, as the guarantor of implementation of their functions, which is actively participates in engaging students in scienti c research, analysis of scienti c literature, writing the scienti c reports and discussing them at group meetings; together with students takes part in scienti c and practical conferences, publication and validation of the results of scienti c work. The role of motivational component in improving the ef ciency of the SSG was noted. Last one combines the use of internal and, dominantly, external motivations.Conclusion. For preparedness of a highly quali ed specialist for professional self-improvement is necessary to provide in universities inextricable relationship of the educational process and research work of students in system of higher medical education, especially in the rst years of study.Мета роботи – з’ясувати особливості організації науково-дослідної роботи студентів-першокурсників медичного профілю у взаємозв’язку з навчальним процесом.Основна частина. Об’єктом даного дослідження є студентський науковий гурток (СНГ) як основна початкова позааудиторна форма організації наукової роботи. У статті розглянуто особливості організації науково-дослідницької діяльності студентів-першокурсників спеціальності “222 Медицина” на кафедрі загальної хімії ДВНЗ “Тернопільський державний медичний університет імені І. Я. Горбачевського МОЗ України” (ТДМУ).В аспекті даного дослідження обґрунтовано доцільність використання позааудиторної форми наукової роботи на прикладі функціонування СНГ, як необхідної складової процесу формування професійної компетентності майбутнього лікаря до саморозвитку і самореалізації. У даному процесі відзначено роль куратора гуртка, як гаранта реалізації функцій СНГ, який бере активну участь у залученні студентів до наукового пошуку, аналізу наукової літератури з даної проблематики гуртка, написанні наукових звітів та обговоренні їх на засіданнях гуртка; разом із гуртківцями бере участь у науково практичних конференціях, публікуванні та апробації результатів наукової роботи. Також значна увага приділена формуванню мотиваційного компонента у підвищенні ефективності діяльності СНГ, який поєднує використання зовнішніх та внутрішніх мотивів з домінуванням останніх.Висновок. Для формування готовності висококваліфікованого фахівця до професійного самовдосконалення необхідно забезпечити в університетах нерозривний зв’язок навчального процесу і науково-дослідної роботи студентів у системі вищої медичної освіти, особливо на перших курсах навчання

Elucidating cylindrospermopsin toxicity via synthetic analogues: An in vitro approach

© 2019 Elsevier Ltd Cylindrospermopsin (CYN) is an alkaloid biosynthesized by selected cyanobacteria, the cyto- and genotoxic properties of which have been studied extensively by in vitro and in vivo experimental models. Various studies have separately established the role of uracil, guanidine and hydroxyl groups in CYN-induced toxicity. In the present study, we have prepared five synthetic analogues that all possess a uracil group but had variations in the other functionality found in CYN. We compared the in vitro toxicity of these analogues in common carp hepatocytes by assessing oxidative stress markers, DNA fragmentation and apoptosis. All the analogues tested induced generation of reactive oxygen species, lipid peroxidation (LPO) and DNA fragmentation. However, the greatest increase in LPO and increase in caspase-3 activity, an apoptosis marker, was demonstrated by an analogue containing guanidine, hydroxyl and uracil functionalities similar to those found in CYN but lacking the complex tricyclic structure of CYN. We also report a crystal structure of an analogue lacking the hydroxyl group found in CYN which does not show intramolecular H-bonding interactions between the guanidine and the uracil functionalities. The observations made in this work supports the hypothesis that CYN toxicity is a result of an interplay between both of the uracil, hydroxyl and guanidine functional groups.This research was partially funded by the Ministry of Education and Science of Ukraine (program for support young fellows MV-1) and by the BEACON (ERDF) program and the EPSRC. Thanks are given to the EPSRC for a fellowship (DE, EP/J01821X/1), the BEACON (ERDF) program for support (PJM, DE) and to the National Mass Spectrometry Facility at Swansea.Published versio

OXIDATIVE STRESS IN HUMAN THYROID GLAND UNDER IODINE DEFICIENCY NODULAR GOITER: FROM HARMLESSNESS TO HAZARD DEPENDING ON COPPER AND IODINE SUBCELLULAR DISTRIBUTION

Background. Thyroid disorders are the second most common endocrinopathies found in humans and animals. Determination of their key molecular markers presents a special interest.Objective. We studied iodine and copper accumulation in nodular, paranodular and contralateral (not affected tissue by node) tissues of human thyroid gland in relation to the level of metal-binding proteins, potential antioxidants, and oxidative changes in tissue for this goal. Lower level of organificated iodine and higher level and mass fraction of inorganic iodine and copper in the nodular and paranodular tissue versus contralateral part of thyroid gland was established.Results. The level of both metal-binding and apo-form of metallothioneins was higher. Content of reduced glutathione was lower in node-affected tissue compared to the contralateral part. Signs of oxidative stress (higher activity of superoxide dismutase, catalase, glutathione-transferase and level of oxyradicals) and cytotoxicity (higher cathepsin D activity, higher level of DNA strand breaks and glycolysis activation) in affected tissue were observed. The range of indice variability in paranodular tissue was smaller than in nodule compared to the parenchyma of contralateral part.Conclusions. Excess of copper unbound to metallothionein in goitrous-changed tissue and high level of inorganic iodine could be the reason for elevated DNA fragmentation and increased lysosomal membrane permeability and activation of antioxidant defense. The main criterions of goiter formation were represented by low level of organificated iodine and high level of DNA damage in thyroid gland.KEY WORDS: iodine deficiency nodular colloidal goiter, iodine, copper, metallothioneins, oxidative stress, cytotoxicit

Toxic effects multi-walled carbon nanotubes on bivalves: comparison between of functionalized and non-functionalized nanoparticles

Despite of the large array of available carbon nanotube (CNT) configurations that allow different industrial and scientific applications of these nanoparticles, their impacts on aquatic organisms, especially on invertebrate species, are still limited. To our knowledge, no information is available on how surface chemistry alteration (functionalization) of CNTs may impact the toxicity of these NPs to bivalve species after a chronic exposure. For this reason, the impacts induced by chronic exposure (28 days) to unfunctionalized MWCNTs (Nf-MWCNTs) in comparison with functionalized MWCNTs (f-MWCNTs), were evaluated in R. philippinarum, by measuring alterations induced in clams' oxidative status, neurotoxicity and metabolic capacity. The results obtained revealed that exposure to both MWCNT materials altered energy-related responses, with higher metabolic capacity and lower glycogen, protein and lipid concentrations in clams exposed to these CNTs. Moreover, R. philippinarum exposed to Nf-MWCNTs and f-MWCNTs showed oxidative stress expressed in higher lipid peroxidation and lower ratio between reduced and oxidized glutathione, despite the activation of defense mechanisms (superoxide-dismutase, glutathione peroxidase and glutathione S-transferases) in exposed clams. Additionally, neurotoxicity was observed by inhibition of Cholinesterases activity in organisms exposed to both MWCNTs.publishe

Transcriptome sequencing and microarray development for the Manila clam, Ruditapes philippinarum: genomic tools for environmental monitoring

Abstract Background The Manila clam, Ruditapes philippinarum, is one of the major aquaculture species in the world and a potential sentinel organism for monitoring the status of marine ecosystems. However, genomic resources for R. philippinarum are still extremely limited. Global analysis of gene expression profiles is increasingly used to evaluate the biological effects of various environmental stressors on aquatic animals under either artificial conditions or in the wild. Here, we report on the development of a transcriptomic platform for global gene expression profiling in the Manila clam. Results A normalized cDNA library representing a mixture of adult tissues was sequenced using a ultra high-throughput sequencing technology (Roche 454). A database consisting of 32,606 unique transcripts was constructed, 9,747 (30%) of which could be annotated by similarity. An oligo-DNA microarray platform was designed and applied to profile gene expression of digestive gland and gills. Functional annotation of differentially expressed genes between different tissues was performed by enrichment analysis. Expression of Natural Antisense Transcripts (NAT) analysis was also performed and bi-directional transcription appears a common phenomenon in the R. philippinarum transcriptome. A preliminary study on clam samples collected in a highly polluted area of the Venice Lagoon demonstrated the applicability of genomic tools to environmental monitoring. Conclusions The transcriptomic platform developed for the Manila clam confirmed the high level of reproducibility of current microarray technology. Next-generation sequencing provided a good representation of the clam transcriptome. Despite the known limitations in transcript annotation and sequence coverage for non model species, sufficient information was obtained to identify a large set of genes potentially involved in cellular response to environmental stress.This work was partially supported by a grant from European Union-funded Network of Excellence "Marine Genomics Europe". CS wishes to acknowledge additional funding from the Ministry of Education and Science (Spain) through grant AGL2007-60049. MM had a PhD scholarship from the University of Florence, Italy. RL was recipient of PhD fellowship SFRH/BD/30112/2006, from the Portuguese Science and Technology Foundation (FCT) and LC and RL acknowledge a grant from FCT project ISOPERK (PTDC/CVT/72083/2006).Peer Reviewe