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1.
Mesenchymal stem cells differentially affect the invasion of distinct glioblastoma cell lines
Barbara Breznik, Helena Motaln, Miloš Vittori, Ana Rotter, Tamara Lah Turnšek, 2017, original scientific article

Abstract: Glioblastoma multiforme are an aggressive form of brain tumors that are characterized by distinct invasion of single glioblastoma cells, which infiltrate the brain parenchyma. This appears to be stimulated by the communication between cancer and stromal cells. Mesenchymal stem cells (MSCs) are part of the glioblastoma microenvironment, and their ‘cross-talk’ with glioblastoma cells is still poorly understood. Here, we examined the effects of bone marrow-derived MSCs on two different established glioblastoma cell lines U87 and U373. We focused on mutual effects of direct MSC/glioblastoma contact on cellular invasion in three-dimensional invasion assays in vitro and in a zebrafish embryo model in vivo. This is the first demonstration of glioblastoma cell-type-specific responses to MSCs in direct glioblastoma co-cultures, where MSCs inhibited the invasion of U87 cells and enhanced the invasion of U373. Inversely, direct cross-talk between MSCs and both of glioblastoma cell lines enhanced MSC motility. MSC-enhanced invasion of U373 cells was assisted by overexpression of proteases cathepsin B, calpain1, uPA/uPAR, MMP-2, -9 and -14, and increased activities of some of these proteases, as determined by the effects of their selective inhibitors on invasion. In contrast, these proteases had no effect on U87 cell invasion under MSC co-culturing. Finally, we identified differentially expressed genes, in U87 and U373 cells that could explain different response of these cell lines to MSCs. In conclusion, we demonstrated that MSC/glioblastoma cross-talk is different in the two glioblastoma cell phenotypes, which contributes to tumor heterogeneity.
Keywords: glioblastoma multiforme, proteases, mesenchymal stem cells, tumor heterogeneity, zebrafish model
Published in DiRROS: 24.07.2024; Views: 19; Downloads: 9
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2.
Non-indigenous species in the Mediterranean Sea : turning from pest to source by developing the 8Rs model, a new paradigm in pollution mitigation
Ana Rotter, Katja Klun, Janja Francé, Patricija Mozetič, Martina Orlando-Bonaca, 2020, original scientific article

Abstract: For mitigation of the effects of pollution, the media, policy makers and, in turn, the scientific community and industry each provide contributions through development of a sense of urgency, and with guidelines and solutions. For non-indigenous species (NIS) that can frequently have negative impacts on the native biota, this is often conveyed in an emotive way to the general public, who are typically keen to help and to get personally involved. This might be through organization of cleaning campaigns, influence on the media, or collaboration with scientists, to inform them of the local presence and abundance of NIS. Alternatively, they might proactively develop technological solutions themselves. To assess the current state of affairs, we reviewed the presence and effects of NIS in the Mediterranean Sea. As so often, any well-planned and successful activity is directly linked to financing, or a lack thereof, and this leads to sometimes untargeted and sporadic measures that are developed within a project or over a limited timeframe, without any sustainability measures. Therefore, we also assessed the activities and strategies that have been financed in this area of NIS mitigation. Based on this review of the presence and impact of NIS, and previous and ongoing activities, we propose a new paradigm to mitigate such pollution: the 8Rs model (i.e., Recognize, Reduce, Replace, Reuse, Recycle, Recover/Restore, Remove, and Regulate). This model extends from the more traditional 3Rs model (i.e., Reduce, Reuse, and Recycle) that is often used and promoted for innovative waste management strategies. Importantly, the 8Rs model can be applied sequentially, for either prevention of NIS introduction, or preparation of mitigation measures. The 8Rs model was constructed based on Mediterranean NIS, although we believe it can be applied to other sources of pollution and other geographic areas. Importantly, the 8Rs model represents a general framework to organize and categorize future pollution mitigation strategies. This approach is essential for development of any action plan to influence the administrative and financial decision makers who essentially enable these activities, and therefore who have important roles in the guarantee of sustainability of these actions, and the creation of innovative societies.
Keywords: non-indigenous species, pollution mitigation, pollution mitigation, 8Rs model, quadruple helix, Mediterranean Sea, citizen science, communication, sustainability
Published in DiRROS: 22.07.2024; Views: 28; Downloads: 8
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3.
Biogeography of Long-Jawed Spiders Reveals Multiple Colonization of the Caribbean
Klemen Čandek, Ingi Agnarsson, Greta Binford, Matjaž Kuntner, 2021, original scientific article

Abstract: Dispersal ability can affect levels of gene flow thereby shaping species distributions and richness patterns. The intermediate dispersal model of biogeography (IDM) predicts that in island systems, species diversity of those lineages with an intermediate dispersal potential is the highest. Here, we tested this prediction on long-jawed spiders (Tetragnatha) of the Caribbean archipelago using phylogenies from a total of 318 individuals delineated into 54 putative species. Our results support a Tetragnatha monophyly (within our sampling) but reject the monophyly of the Caribbean lineages, where we found low endemism yet high diversity. The reconstructed biogeographic history detects a potential early overwater colonization of the Caribbean, refuting an ancient vicariant origin of the Caribbean Tetragnatha as well as the GAARlandia land-bridge scenario. Instead, the results imply multiple colonization events to and from the Caribbean from the mid-Eocene to late-Miocene. Among arachnids, Tetragnatha uniquely comprises both excellently and poorly dispersing species. A direct test of the IDM would require consideration of three categories of dispersers; however, long-jawed spiders do not fit one of these three a priori definitions, but rather represent a more complex combination of attributes. A taxon such as Tetragnatha, one that readily undergoes evolutionary changes in dispersal propensity, can be referred to as a ‘dynamic disperser’.
Keywords: Tetragnatha, dynamic disperser, intermediate dispersal model of biogeography, GAARlandia
Published in DiRROS: 19.07.2024; Views: 53; Downloads: 14
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4.
On the potential of ensemble forecasting for the prediction of meteotsunamis in the Balearic Islands : sensitivity to atmospheric model parameterizations
Baptiste Mourre, A. Santana, A. Buils, L. Gautreau, Matjaž Ličer, Agusti Jansá, B. Casas, B. Amengual, Joaquín Tintoré, 2021, original scientific article

Abstract: This study investigates the potential of ensemble forecasting using full realistic high-resolution nested atmosphere–ocean models for the prediction of meteotsunamis in Ciutadella (Menorca, Spain). The sensitivity of model results to the parameterizations of the atmospheric model is assessed considering the ten most significant recent meteotsunami events for which observations are available. Different schemes adapted to high-resolution Weather Research and Forecasting model simulations were used for the representation of cumulus, microphysics, planetary boundary layer and longwave and shortwave radiations. Results indicate a large spread of the ensemble simulations in terms of the final magnitude of the meteotsunamis. While the modeling system is shown to be able to realistically trigger tsunamigenic atmospheric disturbances in more than 90% of the situations, the small-scale characteristics of these disturbances are significantly modified with the change of parameterizations, leading to significant differences in the magnitude of the simulated sea-level response. No preferred set of parameterizations can be identified that leads to either the largest or the most realistic magnitudes in the majority of situations. Instead, the performance of a given set of parameterizations is found to change with the meteotsunami event under consideration. Importantly, the observed magnitude of the extreme sea-level oscillations lies within the range of a nine-member ensemble in 70% of the cases. This ensemble approach would then allow to generate a realistic range of possibilities in the majority of events, thus improving the realism of meteotsunami predictions compared to single deterministic forecasts.
Keywords: meteotsunamis prediction, atmosphere, ocean modeling, ensemble forecasting, atmospheric model parameterizations
Published in DiRROS: 19.07.2024; Views: 66; Downloads: 34
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5.
Hepatocellular carcinoma (HepG2/C3A) cell-based 3D model for genotoxicity testing of chemicals
Martina Štampar, Helle Frandsen, Adelina Rogowska-Wrzesinska, Krzysztof Wrzesinski, Metka Filipič, Bojana Žegura, 2021, original scientific article

Abstract: The major weakness of the current in vitro genotoxicity test systems is the inability of the indicator cells to express metabolic enzymes needed for the activation and detoxification of genotoxic compounds, which consequently can lead to misleading results. Thus, there is a significant emphasis on developing hepatic cell models, including advanced in vitro three-dimensional (3D) cell-based systems, which better imitate in vivo cell behaviour and offer more accurate and predictive data for human exposures. In this study, we developed an approach for genotoxicity testing with 21-day old spheroids formed from human hepatocellular carcinoma cells (HepG2/C3A) using the dynamic clinostat bioreactor system (CelVivo BAM/bioreactor) under controlled conditions. The spheroids were exposed to indirect-acting genotoxic compounds, polycyclic aromatic hydrocarbon [PAH; benzo(a) pyrene B(a)P], and heterocyclic aromatic amine [PhIP]) at non-cytotoxic concentrations for 24 and 96 h. The results showed that both environmental pollutants B(a)P and PhIP significantly increased the level of DNA strand breaks assessed by the comet assay. Further, the mRNA level of selected genes encoding metabolic enzymes from phase I and II, and DNA damage responsive genes was determined (qPCR). The 21-day old spheroids showed higher basal expression of genes encoding metabolic enzymes compared to monolayer culture. In spheroids, B(a)P or PhIP induced compound-specific up-regulation of genes implicated in their metabolism, and deregulation of genes implicated in DNA damage and immediate-early response. The study demonstrated that this model utilizing HepG2/C3A spheroids grown under dynamic clinostat conditions represents a very sensitive and promising in vitro model for genotoxicity and environmental studies and can thus significantly contribute to a more reliable assessment of genotoxic activities of pure chemicals, and complex environmental samples even at very low for environmental exposure relevant concentrations.
Keywords: in vitro 3D cell model, 21-day old spheroids, cytotoxic, genotoxic, gene expression
Published in DiRROS: 19.07.2024; Views: 52; Downloads: 38
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6.
Water quality characteristics and source analysis of pollutants in the Maotiao river basin (SW China)
Yinjiu Li, Qiuhua Li, Shulin Jiao, Chen Liu, Liuying Yang, Guojia Huang, Si Zhou, Mengshu Han, Anton Brancelj, 2022, original scientific article

Abstract: Rivers are an important mediator between human activities and the natural environment. They provide multiple functions, including irrigation, transportation, food supply, recreation, and water supply. Therefore, evaluations of water quality and pollution sources are of great significance for ecological restoration and management of rivers. In this study, the improved “vušekriterijumska optimizacija i kompromisno rješenje” (VIKOR in Serbian; in English: Multicriteria Optimization and Compromise Solution), and a geodetector were used to analyze the water quality characteristics and pollution sources of the Maotiao River Basin (Gizhou province, SW China). The results showed that the water quality of the Maotiao River Basin deteriorated significantly during the summer drought period, as was evident in the reservoirs and lakes. It improved in the wet season (i.e., during the summer period) due to runoff dilution. Water quality decreased along the river’s course, from upstream to downstream sections. The results of the geographic detector analysis showed that agricultural areas were the primary factor affecting the spatial distribution of water quality in the river basin. In July, August, and November 2020, the influence of agricultural land was 0.72, 0.60, or 0.80, respectively, and the interactions among urban, industrial, agricultural, and forested areas explained 99.2%, 83.2%, or 99.9% of the spatial differentiation of water quality, respectively. Due to the influence of spatial scale, settlements have a small influence on the spatial distribution of water quality. Their impact factors were 0.38, −0.24, and −0.05, respectively. Notably, the negative relationship of water quality and forested areas reflects that topography, types of landscapes, and soil thickness have considerable influences on the Maotiao River Basin’s water quality. Based on the findings, we infer that good farmland water conservancy projects and comprehensive management of different types of landscapes, such as forests, agriculture, and urban area and water bodies, are of great significance for improving water quality.
Keywords: water quality, statistical data, VIKOR model, CRITIC model
Published in DiRROS: 16.07.2024; Views: 73; Downloads: 35
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7.
HepG2 spheroids as a biosensor-like cell-based system for (geno)toxicity assessment
Martina Štampar, Sonja Žabkar, Metka Filipič, Bojana Žegura, 2022, original scientific article

Abstract: 3D spheroids developed from HepG2 cells were used as a biosensor-like system for the detection of (geno)toxic effects induced by chemicals. Benzo(a)pyrene (B(a)P) and amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) with well-known mechanisms of action were used for system validation. HepG2 spheroids grown for 3 days were exposed to BaP and PhIP for 24 and 72 h. The growth and viability of spheroids were monitored by planimetry and Live/Dead staining of cells. Multi-parametric flow cytometric analysis was applied for simultaneous detection of specific end-effects including cell cycle analysis (Hoechst staining), cell proliferation (KI67 marker), and DNA double-strand breaks (ℽH2AX) induced by genotoxic compounds. Depending on the exposure concentration/time, BaP reduced spheroid growth, affected cell proliferation by arresting cells in S and G2 phase and induced DNA double-strand breaks (DSB). Simultaneous staining of ℽH2AX formation and cell cycle analysis revealed that after BaP (10 μM; 24 h) exposure 60% of cells in G0/G1 phase had DNA DSB, while after 72 h only 20% of cells contained DSB indicating efficient repair of DNA lesions. PhIP did not influence the spheroid size whereas accumulation of cells in the G2 phase occurred after both treatment times. The evaluation of DNA damage revealed that at 200 μM PhIP 50% of cells in G0/G1 phase had DNA DSB, which after 72-h exposure dropped to 40%, showing lower repair capacity of PhIP-induced DSB compared to BaP-induced. The developed approach using simultaneous detection of several parameters provides mechanistic data and thus contributes to more reliable genotoxicity assessment of chemicals as a high-content screening tool.
Keywords: in vitro 3D cell model, HepG2, flow cytometry, cell cycle, proliferation, DNA strand, breaks
Published in DiRROS: 16.07.2024; Views: 58; Downloads: 36
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8.
Combined toxic effects of BPA and its two analogues BPAP and BPC in a 3D HepG2 cell model
Martina Štampar, Tim Ravnjak, Ana-Marija Domijan, Bojana Žegura, 2023, original scientific article

Abstract: Bisphenol A (BPA) is one of the most commonly used substances in the manufacture ofvarious everyday products. Growing concerns about its hazardous properties, including endocrinedisruption and genotoxicity, have led to its gradual replacement by presumably safer analogues inmanufacturing plastics. The widespread use of BPA and, more recently, its analogues has increasedtheir residues in the environment. However, our knowledge of their toxicological profiles is limitedand their combined effects are unknown. In the present study, we investigated the toxic effectscaused by single bisphenols and by the combined exposure of BPA and its two analogues, BPAP andBPC, after short (24-h) and prolonged (96-h) exposure in HepG2 spheroids. The results showed thatBPA did not reduce cell viability in HepG2 spheroids after 24-h exposure. In contrast, BPAP andBPC affected cell viability in HepG2 spheroids. Both binary mixtures (BPA/BPAP and BPA/BPC)decreased cell viability in a dose-dependent manner, but the significant difference was only observedfor the combination of BPA/BPC (both at 40μM). After 96-h exposure, none of the BPs studiedaffected cell viability in HepG2 spheroids. Only the combination of BPA/BPAP decreased cellviability in a dose-dependent manner that was significant for the combination of 4μM BPA and 4μMBPAP. None of the BPs and their binary mixtures studied affected the surface area and growth ofspheroids as measured by planimetry. In addition, all BPs and their binary mixtures studied triggeredoxidative stress, as measured by the production of reactive oxygen species and malondialdehyde,at both exposure times. Overall, the results suggest that it is important to study the effects of BPsas single compounds. It is even more important to study the effects of combined exposures, as thecombined effects may differ from those induced by single compounds.
Keywords: BP analogues, hepatic in vitro 3D cell model, combined exposure, viability, oxidative stress, toxicology
Published in DiRROS: 12.07.2024; Views: 66; Downloads: 42
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10.
Uporabnost nacionalnih podatkov laserskega skeniranja in cikličnega aerofotografiranja pri zaznavanju gozdnih vrzeli
Anže Martin Pintar, Mitja Skudnik, 2024, original scientific article

Abstract: Gozdna tla zastirajo drevesne krošnje in gozdna vrzel je vsaka večja ali manjša površina tal, ki je ne prekrivajo krošnje zgornjega sloja dreves. Velikost vrzeli vpliva na to, katera drevesna vrsta se bo pomladila, pa tudi na vertikalno in horizontalno strukturo gozda. Na Pohorju (Pahernikova gozdna posest, za katero so značilni gozdovi, v katerih se na majhnih površinah pojavljajo drevesa različnih premerov in starosti) smo analizirali možnosti samodejnega zaznavanja vrzeli na podlagi podatkov Laserskega skeniranja Slovenije (LSS) in Cikličnega aerofotografiranja Slovenije (CAS). Uporabili smo digitalna modela krošenj (DMK), pridobljena iz obeh virov podatkov. Skupna natančnost zaznavanja tudi najmanjših vrzeli (manjših od 50 m2 ) na podlagi podatkov LSS je znašala 94,4 %, na podlagi podatkov CAS pa 83,1 %. Na podlagi podatkov LSS je mogoče zaznati večje število in površino vrzeli, prav tako so vrzeli, pridobljene na podlagi podatkov LSS, bolj razčlenjene in podolgovate. Zaznavanje vrzeli s podatki CAS in LSS je medsebojno bolj primerljivo pri najmanjših površinah vrzeli 100 m2 , natančnost zaznave takšnih vrzeli je 85,4 oziroma 94,4 %. Rezultati kažejo, da je ob odsotnosti podatkov LSS uporaba podatkov CAS za določanje vrzeli smiselna na velikopovršinski ravni in pri zaznavanju vrzeli, večjih od 100 m2 .
Keywords: raznomerni gozdovi, raznodobni gozdovi, digitalni model krošenj, gozdna vrzel, daljinsko zaznavanje, Lasersko skeniranje Slovenije, LSS, Ciklično aerofotografiranje Slovenije, CAS
Published in DiRROS: 08.07.2024; Views: 90; Downloads: 34
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