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1.
In-depth comparison of adeno-associated virus containing fractions after CsCl ultracentrifugation gradient separation
Mojca Janc, Kaja Zevnik, Ana Dolinar, Tjaša Jakomin, Maja Štalekar, Katarina Bačnik, Denis Kutnjak, Magda Tušek-Žnidarič, Lorena Zentilin, Dmitri G. Fedorov, David Dobnik, 2024, izvirni znanstveni članek

Povzetek: Recombinant adeno-associated viruses (rAAVs) play a pivotal role in the treatment of genetic diseases. However, current production and purification processes yield AAV-based preparations that often contain unwanted empty, partially filled or damaged viral particles and impurities, including residual host cell DNA and proteins, plasmid DNA, and viral aggregates. To precisely understand the composition of AAV preparations, we systematically compared four different single-stranded AAV (ssAAV) and self-complementary (scAAV) fractions extracted from the CsCl ultracentrifugation gradient using established methods (transduction efficiency, analytical ultracentrifugation (AUC), quantitative and digital droplet PCR (qPCR and ddPCR), transmission electron microscopy (TEM) and enzyme-linked immunosorbent assay (ELISA)) alongside newer techniques (multiplex ddPCR, multi-angle light-scattering coupled to size-exclusion chromatography (SEC-MALS), multi-angle dynamic light scattering (MADLS), and high-throughput sequencing (HTS)). Suboptimal particle separation within the fractions resulted in unexpectedly similar infectivity levels. No single technique could simultaneously provide comprehensive insights in the presence of both bioactive particles and contaminants. Notably, multiplex ddPCR revealed distinct vector genome fragmentation patterns, differing between ssAAV and scAAV. This highlights the urgent need for innovative analytical and production approaches to optimize AAV vector production and enhance therapeutic outcomes.
Ključne besede: recombinant adeno-associated viruses (rAAVs), CsCl ultracentrifugation gradient, analytical methods, digital droplet PCR (ddPCR), transmission electron microscopy (TEM), analytical ultracentrifugation (AUC), size-exclusion chromatography coupled with multi-angle light scattering (SEC-MALS), Illumina sequencing, virology
Objavljeno v DiRROS: 07.08.2024; Ogledov: 49; Prenosov: 40
.pdf Celotno besedilo (8,47 MB)
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2.
Coastal high-frequency radars in the Mediterranean : Applications in support of science priorities and societal needs
Emma Reyes, Eva Aguiar, Michele Bendoni, Maristella Berta, Carlo Brandini, Alejandro Cáceres-Euse, Fulvio Capodici, Vanessa Cardin, Daniela Cianelli, Giuseppe Ciraolo, Matjaž Ličer, 2022, pregledni znanstveni članek

Povzetek: The Mediterranean Sea is a prominent climate-change hot spot, with many socioeconomically vital coastal areas being the most vulnerable targets for maritime safety, diverse met-ocean hazards and marine pollution. Providing an unprecedented spatial and temporal resolution at wide coastal areas, high-frequency radars (HFRs) have been steadily gaining recognition as an effective land-based remote sensing technology for continuous monitoring of the surface circulation, increasingly waves and occasionally winds. HFR measurements have boosted the thorough scientific knowledge of coastal processes, also fostering a broad range of applications, which has promoted their integration in coastal ocean observing systems worldwide, with more than half of the European sites located in the Mediterranean coastal areas. In this work, we present a review of existing HFR data multidisciplinary science-based applications in the Mediterranean Sea, primarily focused on meeting end-user and science-driven requirements, addressing regional challenges in three main topics: (i) maritime safety, (ii) extreme hazards and (iii) environmental transport process. Additionally, the HFR observing and monitoring regional capabilities in the Mediterranean coastal areas required to underpin the underlying science and the further development of applications are also analyzed. The outcome of this assessment has allowed us to provide a set of recommendations for future improvement prospects to maximize the contribution to extending science-based HFR products into societally relevant downstream services to support blue growth in the Mediterranean coastal areas, helping to meet the UN's Decade of Ocean Science for Sustainable Development and the EU's Green Deal goals.
Ključne besede: coastal monitoring, Mediterranean Sea, multi-platform observing systems, oceanography
Objavljeno v DiRROS: 05.08.2024; Ogledov: 70; Prenosov: 80
.pdf Celotno besedilo (12,59 MB)
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3.
Coastal high-frequency radars in the Mediterranean : Status of operations and a framework for future development
Pablo Lorente, Eva Aguiar, Michele Bendoni, Maristella Berta, Carlo Brandini, Alejandro Cáceres-Euse, Fulvio Capodici, Daniela Cianelli, Giuseppe Ciraolo, Lorenzo Corgnati, Matjaž Ličer, 2022, pregledni znanstveni članek

Povzetek: Due to the semi-enclosed nature of the Mediterranean Sea, natural disasters and anthropogenic activities impose stronger pressures on its coastal ecosystems than in any other sea of the world. With the aim of responding adequately to science priorities and societal challenges, littoral waters must be effectively monitored with high-frequency radar (HFR) systems. This land-based remote sensing technology can provide, in near-real time, fine-resolution maps of the surface circulation over broad coastal areas, along with reliable directional wave and wind information. The main goal of this work is to showcase the current status of the Mediterranean HFR network and the future roadmap for orchestrated actions. Ongoing collaborative efforts and recent progress of this regional alliance are not only described but also connected with other European initiatives and global frameworks, highlighting the advantages of this cost-effective instrument for the multi-parameter monitoring of the sea state. Coordinated endeavors between HFR operators from different multi-disciplinary institutions are mandatory to reach a mature stage at both national and regional levels, striving to do the following: (i) harmonize deployment and maintenance practices; (ii) standardize data, metadata, and quality control procedures; (iii) centralize data management, visualization, and access platforms; and (iv) develop practical applications of societal benefit that can be used for strategic planning and informed decision-making in the Mediterranean marine environment. Such fit-for-purpose applications can serve for search and rescue operations, safe vessel navigation, tracking of marine pollutants, the monitoring of extreme events, the investigation of transport processes, and the connectivity between offshore waters and coastal ecosystems. Finally, future prospects within the Mediterranean framework are discussed along with a wealth of socioeconomic, technical, and scientific challenges to be faced during the implementation of this integrated HFR regional network.
Ključne besede: coastal regions, Mediterranean Sea, multi-platform observing systems, oceanography
Objavljeno v DiRROS: 05.08.2024; Ogledov: 57; Prenosov: 87
.pdf Celotno besedilo (2,10 MB)
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4.
DiNAR: revealing hidden patterns of plant signalling dynamics using Diferential Network Analysis in R
Maja Zagorščak, Andrej Blejec, Živa Ramšak, Marko Petek, Tjaša Stare, Kristina Gruden, 2018, izvirni znanstveni članek

Povzetek: Background Progress in high-throughput molecular methods accompanied by more complex experimental designs demands novel data visualisation solutions. To specifically answer the question which parts of the specifical biological system are responding in particular perturbation, integrative approach in which experimental data are superimposed on a prior knowledge network is shown to be advantageous. Results We have developed DiNAR, Differential Network Analysis in R, a user-friendly application with dynamic visualisation that integrates multiple condition high-throughput data and extensive biological prior knowledge. Implemented differential network approach and embedded network analysis allow users to analyse condition-specific responses in the context of topology of interest (e.g. immune signalling network) and extract knowledge concerning patterns of signalling dynamics (i.e. rewiring in network structure between two or more biological conditions). We validated the usability of software on the Arabidopsis thaliana and Solanum tuberosum datasets, but it is set to handle any biological instances. Conclusions DiNAR facilitates detection of network-rewiring events, gene prioritisation for future experimental design and allows capturing dynamics of complex biological system. The fully cross-platform Shiny App is hosted and freely available at https://nib-si.shinyapps.io/DiNAR. The most recent version of the source code is available at https://github.com/NIB-SI/DiNAR/ with a DOI 10.5281/zenodo.1230523 of the archived version in Zenodo.
Ključne besede: biological networks, clustering, gene expression, time series, dynamic network analysis, dynamic data visualisation, web application, multi-conditional datasets, background knowledge
Objavljeno v DiRROS: 24.07.2024; Ogledov: 141; Prenosov: 126
.pdf Celotno besedilo (1,63 MB)
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5.
Temperature effects explain continental scale distribution of cyanobacterial toxins
Evanthia Mantzouki, Tina Eleršek, 2018, izvirni znanstveni članek

Povzetek: nsight into how environmental change determines the production and distribution of cyanobacterial toxins is necessary for risk assessment. Management guidelines currently focus on hepatotoxins (microcystins). Increasing attention is given to other classes, such as neurotoxins (e.g., anatoxin-a) and cytotoxins (e.g., cylindrospermopsin) due to their potency. Most studies examine the relationship between individual toxin variants and environmental factors, such as nutrients, temperature and light. In summer 2015, we collected samples across Europe to investigate the effect of nutrient and temperature gradients on the variability of toxin production at a continental scale. Direct and indirect effects of temperature were the main drivers of the spatial distribution in the toxins produced by the cyanobacterial community, the toxin concentrations and toxin quota. Generalized linear models showed that a Toxin Diversity Index (TDI) increased with latitude, while it decreased with water stability. Increases in TDI were explained through a significant increase in toxin variants such as MC-YR, anatoxin and cylindrospermopsin, accompanied by a decreasing presence of MC-LR. While global warming continues, the direct and indirect effects of increased lake temperatures will drive changes in the distribution of cyanobacterial toxins in Europe, potentially promoting selection of a few highly toxic species or strains.
Ključne besede: microcystin, anatoxin, cylindrospermopsin, temperature, direct effects, indirect effects, spatial distribution, European Multi Lake Survey
Objavljeno v DiRROS: 24.07.2024; Ogledov: 110; Prenosov: 86
.pdf Celotno besedilo (16,03 MB)
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6.
Worldwide tests of generic attractants, a promising tool for early detection of non-native cerambycid species
Alain Roques, Lili Ren, Davide Rassati, Juan Shi, Evgueni Akulov, Alenka Žunič Kosi, 2023, izvirni znanstveni članek

Povzetek: A large proportion of the insects which have invaded new regions and countries are emerging species, being found for the first time outside their native range. Being able to detect such species upon arrival at ports of entry before they establish in non-native countries is an urgent challenge. The deployment of traps baited with broad-spectrum semiochemical lures at ports-of-entry and other high-risk sites could be one such early detection tool. Rapid progress in the identification of semiochemicals for cerambycid beetles during the last 15 years has revealed that aggregation-sex pheromones and sex pheromones are often conserved at global levels for genera, tribes or subfamilies of the Cerambycidae. This possibly allows the development of generic attractants which attract multiple species simultaneously, especially when such pheromones are combined into blends. Here, we present the results of a worldwide field trial programme conducted during 2018–2021, using traps baited with a standardised 8-pheromone blend, usually complemented with plant volatiles. A total of 1308 traps were deployed at 302 sites covering simultaneously or sequentially 13 European countries, 10 Chinese provinces and some regions of the USA, Canada, Australia, Russia (Siberia) and the Caribbean (Martinique). We intended to test the following hypotheses: 1) if a species is regularly trapped in significant numbers by the blend on a continent, it increases the probability that it can be detected when it arrives in other countries/continents and 2) if the blend exerts an effective, generic attraction to multiple species, it is likely that previously unknown and unexpected species can be captured due to the high degree of conservation of pheromone structures within related taxa. A total of 78,321 longhorned beetles were trapped, representing 376 species from eight subfamilies, with 84 species captured in numbers greater than 50 individuals. Captures comprised 60 tribes, with 10 tribes including more than nine species trapped on different continents. Some invasive species were captured in both the native and invaded continents. This demonstrates the potential of multipheromone lures as effective tools for the detection of ‘unexpected’ cerambycid invaders, accidentally translocated outside their native ranges. Adding new pheromones with analogous well-conserved motifs is discussed, as well as the limitations of using such blends, especially for some cerambycid taxa which may be more attracted by the trap colour or other characteristics rather than to the chemical blend.
Ključne besede: pheromone trapping, invasion, early detection, Holarctic, multi-pheromone blend
Objavljeno v DiRROS: 12.07.2024; Ogledov: 109; Prenosov: 121
.pdf Celotno besedilo (4,79 MB)
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7.
An investigation into corrosion around voids at the steel-concrete interface
Miha Hren, Tadeja Kosec, Andraž Legat, 2024, izvirni znanstveni članek

Povzetek: In this study, the influence of voids on corrosion process at the steel-concrete interface was investigated, as the exact influence of these features on corrosion processes under various environmental conditions is not entirely understood. Coupled multi electrode arrays and electrical resistance probes were implemented to monitor the evolution of corrosion under cyclic exposure to chlorides and carbonation. MicroCT was used to determine the location and volume of corrosion damage. It was found that, in most cases, corrosion damage initiated outside the voids. During initiation and the early propagation phase, the steel beneath the voids rarely participated in the redox reaction. In following phases, various kinds of corrosion evolution were observed. Specific corrosion mechanisms were proposed and discussed to explain these corrosion processes.
Ključne besede: microtomography, corrosion, coupled multi-electrode array, steel-concrete interface
Objavljeno v DiRROS: 06.06.2024; Ogledov: 193; Prenosov: 255
.pdf Celotno besedilo (23,73 MB)
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8.
Multi-scale and multi-modal imaging study of mantle xenoliths and petrological implications
Marco Venier, Luca Ziberna, Lucia Mancini, Alexander P. Kao, Federico Bernardini, Giacomo Roncoroni, Sula Milani, Nasrrddine Youbi, Yondon Majigsuren, Angelo De Min, Davide Lenaz, 2024, izvirni znanstveni članek

Povzetek: The accurate textural characterization of mantle xenoliths is one of the fundamental steps to understanding the main processes occurring in the upper mantle, such as sub-solidus recrystallization, magmatic crystallization, and metasomatism. Texture, composition, and mineralogy reflect the temperature, pressure, stress conditions, melting, and/or contamination events undergone before and during the entrapment in the host magma. For these reasons, characterizing the three-dimensional (3D) texture of silicate, oxide, sulfide, and glass phases has great importance in the study of mantle xenoliths. We performed a multi-scale and multi-modal 3D textural analysis based on X-ray computed microtomography (µ-CT) data of three mantle xenoliths from different geodynamic settings (i.e., mobile belt zone, pericraton, oceanic hotspot). The samples were selected to represent different, variably complex internal structures composed of grains of different phases, fractures, voids, and fluid inclusions of different sizes. We used an approach structured in increasing steps of spatial and contrast resolution, starting with in-house X-ray µ-CT imaging (spatial resolution from 30 µm down to 6.25 µm) and moving to high-resolution synchrotron X-ray µ-CT at the micrometer scale. We performed a 3D characterization of mantle xenoliths, comparing the results with the analysis of conventional 2D images (thin sections) obtained by optical microscopy and simulating the random sectioning of several thin sections to estimate the probability of correct modal classification. The 3D models allow the extraction of textural information that cannot be quantified solely from thin sections: spinel layering, distribution of silicic glass, and related vesicles. Moreover, high-density volumes identified as sulfides were detected in two xenoliths, showing no relation with the spinel layering in one case and a preferential concentration along fractures in the other. Given the variety of textures and mineral assemblages of mantle xenoliths worldwide, the results are used to suggest experimental and analytical protocols for the characterization of these materials.
Ključne besede: petrology, mantle xenoliths, X-ray microtomography, multi-scale image analysis
Objavljeno v DiRROS: 23.05.2024; Ogledov: 171; Prenosov: 37
.pdf Celotno besedilo (1,64 MB)
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9.
A cross-benchmark examination of feature-based algorithm selector generalization in single-objective numerical optimization
Gjorgjina Cenikj, Gašper Petelin, Tome Eftimov, 2024, izvirni znanstveni članek

Povzetek: The task of selecting the best optimization algorithm for a particular problem is known as algorithm selection (AS). This involves training a model using landscape characteristics to predict algorithm performance, but a key challenge remains: making AS models generalize effectively to new, untrained benchmark suites. This study assesses AS models’ generalizability in single-objective numerical optimization across diverse benchmark suites. Using Exploratory Landscape Analysis (ELA) and transformer-based (TransOpt) features, the research investigates their individual and combined effectiveness in AS across four benchmarks: BBOB, AFFINE, RANDOM, and ZIGZAG. AS models perform differently based on benchmark suite similarities in algorithm performance distributions and single-best solvers. When suites align, these models underperform against a baseline predicting mean algorithm performance; yet, they outperform the baseline when suites differ in performance distributions and solvers. The AS models trained using the ELA landscape features are better than the models trained using the TransOpt features on the BBOB and AFFINE benchmark suites, while the opposite is true for the RANDOM benchmark suite. Ultimately, the study reveals challenges in accurately capturing algorithm performance through problem landscape features (ELA or TransOpt), impacting AS model applicability.
Ključne besede: algorithm selection, multi-target regression, generalization, benchmarking
Objavljeno v DiRROS: 21.05.2024; Ogledov: 277; Prenosov: 383
.pdf Celotno besedilo (2,49 MB)
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10.
Invariants of multi-linkoids
Boštjan Gabrovšek, Neslihan Gügümcü, 2023, izvirni znanstveni članek

Povzetek: In this paper, we extend the definition of a knotoid to multilinkoids that consist of a finite number of knot and knotoid components. We study invariants of multi-linkoids, such as the Kauffman bracket polynomial, ordered bracket polynomial, the Kauffman skein module, and the $T$-invariant in relation with generalized $\Theta$-graphs.
Ključne besede: knotoid, multi-linkoid, spatial graph, Kauffman bracket polynomial, Kauffman bracket skein module, theta-curve, theta-graph
Objavljeno v DiRROS: 15.03.2024; Ogledov: 267; Prenosov: 146
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