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1271 - 1280 / 2000
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1271.
The role of heparan sulfate and neuropilin 2 in VEGFA signaling in human endothelial tip cells and non-tip cells during angiogenesis in vitro
Marchien G. Dallinga, Yasmin I. Habani, Alinda W. M. Schimmel, Geesje M. Dallinga-Thie, Cornelis J. F. van Noorden, Ingeborg Klaassen, Reinier O. Schlingemann, 2021, original scientific article

Abstract: During angiogenesis, vascular endothelial growth factor A (VEGFA) regulates endothelial cell (EC) survival, tip cell formation, and stalk cell proliferation via VEGF receptor 2 (VEGFR2). VEGFR2 can interact with VEGFR2 co-receptors such as heparan sulfate proteoglycans (HSPGs) and neuropilin 2 (NRP2), but the exact roles of these co-receptors, or of sulfatase 2 (SULF2), an enzyme that removes sulfate groups from HSPGs and inhibits HSPG-mediated uptake of very low density lipoprotein (VLDL), in angiogenesis and tip cell biology are unknown. In the present study, we investigated whether the modulation of binding of VEGFA to VEGFR2 by knockdown of SULF2 or NRP2 affects sprouting angiogenesis, tip cell formation, proliferation of non-tip cells, and EC survival, or uptake of VLDL. To this end, we employed VEGFA splice variant 121, which lacks an HSPG binding domain, and VEGFA splice variant 165, which does have this domain, in in vitro models of angiogenic tip cells and vascular sprouting. We conclude that VEGFA165 and VEGFA121 have similar inducing effects on tip cells and sprouting in vitro, and that the binding of VEGFA165 to HSPGs in the extracellular matrix does not seem to play a role, as knockdown of SULF2 did not alter these effects. Co-binding of NRP2 appears to regulate VEGFA–VEGFR2-induced sprout initiation, but not tip cell formation. Finally, as the addition of VLDL increased sprout formation but not tip cell formation, and as VLDL uptake was limited to non-tip cells, our findings suggest that VLDL plays a role in sprout formation by providing biomass for stalk cell proliferation.
Keywords: endothelial cells, angiogenesis, VEGFA, tip cells, SULF2, NRP2, HSPG
Published in DiRROS: 19.07.2024; Views: 249; Downloads: 196
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1272.
Multi-platform, high-resolution study of a complex coastal system : the TOSCA experiment in the Gulf of Trieste
Stefano Querin, Simone Cosoli, Riccardo Gerin, Célia Laurent, Vlado Malačič, Neva Pristov, Pierre-Marie Poulain, 2021, original scientific article

Abstract: Abstract Although small in size, the Gulf of Trieste (GoT), a marginal coastal basin in the northern Adriatic Sea, is characterized by very complex dynamics and strong variability of its oceanographic conditions. In April–May 2012, a persistent, large-scale anticyclonic eddy was observed in the GoT. This event was captured by both High Frequency Radar (HFR) and Lagrangian drifter observations collected within the European MED TOSCA (Tracking Oil Spill and Coastal Awareness) project. The complexity of the system and the variety of forcing factors constitute major challenges from a numerical modeling perspective when it comes to simulating the observed features. In this study, we implemented a high-resolution hydrodynamic model in an attempt to reproduce and analyze the observed basin-wide eddy structure and determine its drivers. We adopted the Massachusetts Institute of Technology General Circulation Model (MITgcm), tailored for the GoT, nested into a large-scale simulation of the Adriatic Sea and driven by a tidal model, measured river freshwater discharge data and surface atmospheric forcing. Numerical results were qualitatively and quantitatively evaluated against HFR surface current maps, Lagrangian drifter trajectories and thermohaline data, showing good skills in reproducing the general circulation, but failing in accurately tracking the drifters. Model sensitivity to different forcing factors (wind, river and tides) was also assessed.
Keywords: Adriatic Sea, surface circulation, HF coastal radars, Lagrangian drifters, wind-driven currents, ocean circulation models
Published in DiRROS: 19.07.2024; Views: 299; Downloads: 211
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1273.
Detection of four new tomato viruses in Serbia using post hoc high-throughput sequencing analysis of samples from a large-scale field survey
Ana Vučurović, Denis Kutnjak, Nataša Mehle, Ivana Stanković, Anja Pecman, Aleksandra Bulajić, Branka Krstić, Maja Ravnikar, 2021, original scientific article

Abstract: Tomato production worldwide is affected by numerous plant virus species. The early and accurate detection of viruses is a critical step for disease control. However, the simultaneous detection of the most known tomato viruses can be difficult because of the high number and diversity of tomato-infecting viruses. Here, we have identified four new viruses in Serbia by applying target-independent small RNA high-throughput sequencing (HTS). HTS was applied on pools of samples and separate samples, in total comprising 30 tomato samples that exhibited (severe) virus-like symptoms and were collected in Serbia during three annual surveys (2011 to 2013). These samples had previously tested negative for the presence of 16 tomato viruses using targeted detection methods. Three divergent complete genome sequences of Physostegia chlorotic mottled virus were obtained from different localities, indicating for the first time that this virus is widespread in Serbia and might represent an emergent viral pathogen of tomato. The tomato torrado virus was detected at one locality with devastating yield losses. The southern tomato virus was detected at two localities, and the spinach latent virus was detected at one locality. In addition, we detected the presence of one already-known virus in Serbia, the tomato spotted wilt orthotospovirus. All the HTS results were subsequently confirmed by targeted detection methods. In this study, the successful application of post hoc HTS testing of a limited number of pooled samples resulted in the discovery of new viruses. Thus, our results encourage the use of HTS in research and diagnostic laboratories, including laboratories that have limited resources to resolve disease etiology.
Keywords: viruses, tomato, detection, identification
Published in DiRROS: 19.07.2024; Views: 335; Downloads: 262
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1274.
Inactivation of pepper mild mottle virus in water by cold atmospheric plasma
Arijana Filipić, David Dobnik, Magda Tušek-Žnidarič, Bojana Žegura, Alja Štern, Gregor Primc, Miran Mozetič, Maja Ravnikar, Jana Žel, Ion Gutiérrez-Aguirre, 2021, original scientific article

Abstract: Water scarcity is one of the greatest threats for human survival and quality of life, and this is increasingly contributing to the risk of human, animal and plant infections due to waterborne viruses. Viruses are transmitted through polluted water, where they can survive and cause infections even at low concentrations. Plant viruses from the genus Tobamovirus are highly mechanically transmissible, and cause considerable damage to important crops, such as tomato. The release of infective tobamoviruses into environmental waters has been reported, with the consequent risk for arid regions, where these waters are used for irrigation. Virus inactivation in water is thus very important and cold atmospheric plasma (CAP) is emerging in this field as an efficient, safe, and sustainable alternative to classic waterborne virus inactivation methods. In the present study we evaluated CAP-mediated inactivation of pepper mild mottle virus (PMMoV) in water samples. PMMoV is a very resilient water-transmissible tobamovirus that can survive transit through the human digestive tract. The efficiency of PMMoV inactivation was characterized for infectivity and virion integrity, and at the genome level, using test plant infectivity assays, transmission electron microscopy, and molecular methods, respectively. Additionally, the safety of CAP treatment was determined by testing the cytotoxic and genotoxic properties of CAP-treated water on the HepG2 cell line. 5-min treatment with CAP was sufficient to inactivate PMMoV without introducing any cytotoxic or genotoxic effects in the in-vitro cell model system. These data on inactivation of such stable waterborne virus, PMMoV, will encourage further examination of CAP as an alternative for treatment of potable and irrigation waters, and even for other water sources, with emphasis on inactivation of various viruses including enteric viruses.
Keywords: enteric viruses, pepper mild mottle virus, virus inactivation, water decontamination, cold atmospheric plasma
Published in DiRROS: 19.07.2024; Views: 286; Downloads: 185
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1275.
Cytotoxic and genotoxic effects of cyanobacterial and algal extracts-microcystin and retinoic acid content
Michal Bittner, Alja Štern, Marie Smutna, Klara Hilscherova, Bojana Žegura, 2021, original scientific article

Abstract: In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04–2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2–2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs.
Keywords: cyanobacteria, algae, extracts, complex mixtures, genotoxicity, cytotoxicity, retinoic acids, microcystins, cyanotoxins, chemical analysis
Published in DiRROS: 19.07.2024; Views: 298; Downloads: 208
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1276.
First restoration experiment for Gongolaria barbata in Slovenian coastal waters : What can go wrong?
Martina Orlando-Bonaca, Valentina Pitacco, Petra Slavinec, Milijan Šiško, Tihomir Makovec, Annalisa Falace, 2021, original scientific article

Abstract: The global decline of brown algal forests along rocky coasts is causing an exceptional biodiversity loss. Regardless of conservation efforts, different techniques have been developed for large-scale restoration strategies in the Mediterranean Sea. In this study we tested ex situ pilot restoration of Gongolaria barbata (=Treptacantha barbata) for the first time in Slovenian coastal waters. Healthy apical fronds of the species were collected and the development of recruits on clay tiles was followed under laboratory conditions for 20 days. Despite the experimental difficulties experienced, especially due to the lack of antibiotics to prevent the growth of the biofilm, G. barbata recruits were outplanted in the sea on two concrete plates with 48 tiles each, protected by purpose-built cages to avoid grazing by herbivorous fish. The high survival rate of juveniles after four months in the field (89% of the tiles on the plate that was constantly protected) suggests that outplanting G. barbata is an operable approach for restoration efforts in the northern Adriatic Sea. Our first experiment in Slovenian coastal waters provides new information for the optimization of the best practices during the laboratory cultivation and addresses the early steps of restoration and introduction of young thalli in the natural environment.
Published in DiRROS: 19.07.2024; Views: 276; Downloads: 191
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1277.
Molecular systematics of the long-snouted deep water dogfish (Centrophoridae, Deania) with implications for identification, taxonomy, and conservation
Sergio Stefanni, Diana Catarino, Pedro A. Ribeiro, Mafalda Freitas, Gui Menezes, Francis Neat, David Stanković, 2021, original scientific article

Abstract: According to the most recent taxonomical revision, the deep-sea dogfish genus Deania encompasses four species. Three of them, D. calcea, D. profundorum, and D. hystricosa, occur in the North Atlantic. Whilst D. profundorum can be identified by the presence of a subcaudal keel, the other two species are not easily visually distinguished. Uncertainties over identification raises concerns over stock units and whether management plans are adequate. In this study we compared onboard visual identification of Deania specimens, with morphological inspection of skin denticles under stereo microscope and with independent molecular taxonomical assignment using two molecular markers. Particular emphasis was paid to specimens identified as D. calcea and D. hystricosa in the NE Atlantic where these species potentially occur sympatrically and may be easily confused. In the past the species have been discriminated on the basis of the size of skin denticles (skin roughness), but our study showed that the crown length of skin denticles covaries with size (and sex), irrespective of species, and therefore this is not a reliable morphological character and should not be used to discriminate between the two species. Phylogenetic analyses did not indicate that D. hystricosa to be a distinct lineage from D. calcea. Interestingly, however four individuals (specimens from: UK, Azores Is., Madeira Is. and Seine seamount) formed a well-defined sub-clade nested within the D. calcea clade, possibly a signature of a past vicariance event or a result of coalescent stochasticity.
Keywords: deep-sea sharks, fishery by-catch, phylogeny, Atlantic Ocean, deep-sea dogfish, skin denticles
Published in DiRROS: 19.07.2024; Views: 283; Downloads: 164
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1278.
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: 268; Downloads: 188
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1279.
Metagenomic characterization of parental and production CHO cell lines for detection of adventitious viruses
Katarina Bačnik, Denis Kutnjak, Barbara Jerič Kokelj, Nika Tuta, Tan Lončar, Matjaž Vogelsang, Maja Ravnikar, 2021, original scientific article

Abstract: Viral contamination is a major concern for biological products. Therefore, virus testing of raw materials and cells is essential for the safety of the final product. We used high-throughput sequencing to detect viral-like sequences in selected CHO cell lines. Our aim was to test various approaches of sample preparation, to establish a pipeline for metagenomic analysis and to characterize standard viral metagenome of production and parental CHO cell lines. The comparison of the metagenomics composition of the differently prepared samples showed that among four tested approaches sequencing of ribosomal RNA depleted total RNA is the most promising approach. The metagenomics investigation of one production and three parental CHO cell lines of diverse origin did not indicate the presence of adventitious viral agents in the investigated samples. The study revealed an expected background of virus-like nucleic acids in the samples, which originate from remains of expression vectors, endogenized viral elements and residuals of bacteriophages.
Published in DiRROS: 19.07.2024; Views: 482; Downloads: 170
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1280.
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: 242; Downloads: 223
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