31. Postmortem chondrocyte viability in porcine articular cartilage : influence of time, temperature, and burial under winter conditionsMarko Cvetko, Tanja Knific, Robert Frangež, Helena Motaln, Boris Rogelj, Armin Alibegović, Mitja Gombač, 2024, original scientific article Abstract: The aim of the present study was to investigate the effects of time, temperature, and burial in a natural environment on the viability of chondrocytes in porcine femoral condyles using confocal laser scanning microscopy. Hind trotters from 10 pigs were buried or left unburied. Samples were collected daily and stained with a combination of vital dyes (calcein-AM and ethidium homodimer-1). The chondrocytes showed an intense staining corresponding to their vitality. In the first 3 days, viability decreased slowly and showed no statistical difference between buried and unburied samples. After the first 3 days, it decreased rapidly, with the viability of the buried samples being 66% on day 4, decreasing to 25% on day 8 and to 16% on day 10, while in the unburied samples it decreased to 43% on day 4, 13% on day 8 and 5% on day 10. Our results indicate a time, temperature, and burial dependent decrease in chondrocyte viability and suggest the use of chondrocyte viability as a marker for estimating PMI in both the natural environment and in animals, as well as its potential use in humans.
Published in DiRROS: 08.01.2025; Views: 32; Downloads: 11
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32. Transcriptomic and proteomic profiling of bi-partite and tri-partite murine iPSC-derived neurospheroids under steady-state and inflammatory conditionJulia Di Stefano, Laura Garcia-Pupo, Helena Motaln, Boris Rogelj, Peter Ponsaerts, 2024, original scientific article Abstract: Induced-pluripotent stem cell (iPSC)-derived neurospheroid (NSPH) models are an emerging in vitro toolkit to study the influence of inflammatory triggers on neurodegeneration and repair in a 3D neural environment. In contrast to their human counterpart, the absence of murine iPSC-derived NSPHs for profound characterisation and validation studies is a major experimental research gap, even though they offer the only possibility to truly compare or validate in vitro NSPH responses with in vivo brain responses. To contribute to these developments, we here describe the generation and characterisation of 5-week-old CX3CR1eGFP+/- CCR2RFP+/- murine (m)iPSC-derived bi-partite (neurons + astrocytes) and tri-partite (neurons + astrocytes + microglia) NSPH models that can be subjected to cellular activation following pro-inflammatory stimulation. First, cytokine analysis demonstrates that both bi-partite and tri-partite NSPHs can be triggered to release IL6 and CXCL10 following three days of stimulation with, respectively, TNFα + IL1β + IFNγ and LPS + IFNγ. Additionally, immunocytochemical analysis for G3BP1 and PABPC1 revealed the development of stress granules in both bi-partite and tri-partite NSPHs after 3 days of stimulation. To further investigate the observed signs of inflammatory response and cellular stress, we performed an untargeted transcriptomic and proteomic analysis of bi- and tri-partite NSPHs under steady-state and inflammatory conditions. Here, using the combined differential gene and protein expression profiles between unstimulated and stimulated NSPHs, Ingenuity Pathway Analysis (IPA) confirms the activation of canonical pathways associated with inflammation and cellular stress in both bi-partite and tri-partite NSPHs. Moreover, our multi-omics analysis suggests a higher level of downstream inflammatory responses, impairment of homeostatic and developmental processes, as well as activation of cell death processes in stimulated tri-partite NSPHs compared to bi-partite NSPHs. Concluding, these results emphasise the advantages of including microglia in NSPH research to study inflammation-induced neurodegeneration in a 3D neural environment.
Keywords: murine induced-pluripotent cells, neurospheroids, inflammatory response, stress granules, transcriptomics, proteomics
Published in DiRROS: 08.01.2025; Views: 31; Downloads: 10
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33. Oka-1 manifolds: new examples and propertiesFranc Forstnerič, Finnur Lárusson, 2025, original scientific article Abstract: In this paper we investigate Oka-1 manifolds and Oka-1 maps, a class of complex manifolds and holomorphic maps recently introduced by Alarcón and Forstnerič. Oka-1 manifolds are characterised by the property that holomorphic maps from any open Riemann surface to the manifold satisfy the Runge approximation and Weierstrass interpolation conditions, while Oka-1 maps enjoy similar properties for liftings of maps from open Riemann surfaces in the absence of topological obstructions. We also formulate and study the algebraic version of the Oka-1 condition, called aOka-1. We show that it is a birational invariant for compact algebraic manifolds and holds for all rational manifolds. This gives a Runge approximation theorem for maps from compact Riemann surfaces to uniformly rational projective manifolds. Finally, we study a class of complex manifolds with an approximation property for holomorphic sprays of discs. This class lies between the smaller class of Oka manifolds and the bigger class of Oka-1 manifolds and has interesting functorial properties.
Keywords: Riemann surfaces, complex manifolds, algebraic manifolds, Oka-1 manifolds, Oka manifolds, algebraically elliptic manifold, Runge approximation
Published in DiRROS: 08.01.2025; Views: 32; Downloads: 12
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35. Backward dynamics of non-expanding maps in Gromov hyperbolic metric spacesLeandro Arosio, Matteo Fiacchi, Lorenzo Guerini, Anders Karlsson, 2024, original scientific article Abstract: We study the interplay between the backward dynamics of a non-expanding self-map $f$ of a proper geodesic Gromov hyperbolic metric space $X$ and the boundary regular fixed points of $f$ in the Gromov boundary. To do so, we introduce the notion of stable dilation at a boundary regular fixed point of the Gromov boundary, whose value is related to the dynamical behaviour of the fixed point. This theory applies in particular to holomorphic self-maps of bounded domains $\Omega \subset\subset \mathbb{C}^q$, where $\Omega$ is either strongly pseudoconvex, convex finite type, or pseudoconvex finite type with $q=2$, and solves several open problems from the literature. We extend results of holomorphic self-maps of the disc $\mathbb{D}\subset \mathbb{C}$ obtained by Bracci and Poggi-Corradini. In particular, with our geometric approach we are able to answer a question, open even for the unit ball $\mathbb{B}^q\subset \mathbb{C}^q$, namely that for holomorphic parabolic self-maps any escaping backward orbit with bounded step always converges to a point in the boundary.
Keywords: holomorphic dynamics, non-expanding maps, Gromov hyperbolicity, horofunctions, boundary fixed points
Published in DiRROS: 08.01.2025; Views: 27; Downloads: 8
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36. Varicella-zoster virus recapitulates its immune evasive behaviour in matured hiPSC-derived neurospheroidsJonas Govaerts, Elise Van Breedam, Helena Motaln, Boris Rogelj, Peter Ponsaerts, 2024, original scientific article Abstract: Varicella-zoster virus (VZV) encephalitis and meningitis are potential central nervous system (CNS) complications following primary VZV infection or reactivation. With Type-I interferon (IFN) signalling being an important first line cellular defence mechanism against VZV infection by the peripheral tissues, we here investigated the triggering of innate immune responses in a human neural-like environment. For this, we established and characterised 5-month matured hiPSC-derived neurospheroids (NSPHs) containing neurons and astrocytes. Subsequently, NSPHs were infected with reporter strains of VZV (VZVeGFP-ORF23) or Sendai virus (SeVeGFP), with the latter serving as an immune-activating positive control. Live cell and immunocytochemical analyses demonstrated VZVeGFP-ORF23 infection throughout the NSPHs, while SeVeGFP infection was limited to the outer NSPH border. Next, NanoString digital transcriptomics revealed that SeVeGFP-infected NSPHs activated a clear Type-I IFN response, while this was not the case in VZVeGFP-ORF23-infected NSPHs. Moreover, the latter displayed a strong suppression of genes related to IFN signalling and antigen presentation, as further demonstrated by suppression of IL-6 and CXCL10 production, failure to upregulate Type-I IFN activated anti-viral proteins (Mx1, IFIT2 and ISG15), as well as reduced expression of CD74, a key-protein in the MHC class II antigen presentation pathway. Finally, even though VZVeGFP-ORF23-infection seems to be immunologically ignored in NSPHs, its presence does result in the formation of stress granules upon long-term infection, as well as disruption of cellular integrity within the infected NSPHs. Concluding, in this study we demonstrate that 5-month matured hiPSC-derived NSPHs display functional innate immune reactivity towards SeV infection, and have the capacity to recapitulate the strong immune evasive behaviour towards VZV.
Keywords: interferon signalling, neurospheroids, antigen presentation, stress granules, structural integrity
Published in DiRROS: 08.01.2025; Views: 26; Downloads: 10
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37. Ciklični preizkusi armiranobetonskih stebrovMiha Kramar, Matija Gams, Andrej Anžlin, Matjaž Dolšek, 2016, original scientific article Abstract: V članku so prikazane eksperimentalne preiskave potresne odpornosti armiranobetonskih stebrov, ki so bile opravljene na Zavodu za gradbeništvo Slovenije (ZAG) z namenom, da se določi vpliv negotovosti oz. razpršenosti materialnih karakteristik in izvedbe na potresni odziv stebrov. Da bi zajeli kar se da veliko virov negotovosti, smo naročili izdelavo 12 stebrov, ki so jih izdelala tri različna podjetja na podlagi istega načrta in nominalno enakih materialov. Poleg materialnih karakteristik smo analizirali vpliv preklopa vzdolžne armature na potresni odziv stebrov. Stebre smo preizkusili s ciklično horizontalno obtežbo, ki smo jo povečevali do porušitve. Raziskave so pokazale, da so med proizvajalci v resnici velike razlike, ki pa so opazne predvsem pri odzivu stebrov po doseženi največji nosilnosti. Največji vpliv na duktilnost ima izvedba armaturnega detajla stremen. Z raziskavo smo potrdili tudi nekatere že znane odvisnosti in opazili, da bi lahko bil pomemben parameter potresne odpornosti tudi lokacija prvega stremena nad temeljem.
Keywords: armiranobetonski steber, potresni odziv, negotovosti, nosilnost, duktilnost, stremenska armatura
Published in DiRROS: 08.01.2025; Views: 25; Downloads: 8
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38. Provenance analysis of Roman stone artefacts from sedimentary rocks from the archaeological site near Mošnje, NW SloveniaSnježana Miletić, Sabina Dolenec, Judita Lux, Andrej Šmuc, Nina Zupančič, 2016, original scientific article Abstract: This study deals with the macroscopic and microfacies characterisation of Roman stone artefacts excavated in
2006 from a Roman villa rustica near Mošnje (NW Slovenia) with the aim of defining their provenance. A total of 28 representative finds (querns, mortars, whetstones, tooled and rounded stones, a fragment of stone slab, mosaic tesserae and two architectural elements - one with a relief) made of clastic and carbonate sedimentary rocks were examined. Comparison was made with rock samples taken from quarries and gravel bars close to the archaeological site, as well as from larger distance to the site. The majority of artefact sampled is composed of Upper Palaeozoic quartz sandstones, which are found as pebbles in gravel bars close to the archaeological site; while 2 samples were from Quaternary coarse grained clastic rocks which can be found in local glacio-fluvial Sediments. Other finds were made of four different Mesozoic shallow-water limestones which outcrop in different areas of Central and SW Slovenia. The nearest Lower Jurassic biopelmicritic limestones are found at the western periphery of Ljubljana in Podutik. Cretaceous miliolid limestones and biocalcarenitic limestones with rudists are common in the successions of the Dinaric Carbonate Platform in SW Slovenia (for example, on the Trieste-Komen Plateau), NE Italy and SW Croatia. This indicates that the limestones for architectural elements, stone mortars and tesserae were brought to Mošnje from distant locations. Smaller stone tools are likely to have been made at the location of the archaeological site from material gathered locally, mostly pebbles from clastic rocks, which were accessible and suitable for tooling.
Keywords: arheološke izkopanine, rimljani, sedimentne kamnine, apnenec, mikrofacies, izvor kamnine, arheološko najdišče, Slovenija
Published in DiRROS: 08.01.2025; Views: 26; Downloads: 10
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40. Hybrid nanostructures of ▫$Fe_3O_4$▫ and Au prepared via coprecipitation and ultrasonic spray pyrolysisLan Kresnik, Peter Majerič, Darja Feizpour, Klementina Pušnik Črešnar, Rebeka Rudolf, 2024, original scientific article Abstract: The coupled processes of coprecipitation and ultrasonic spray pyrolysis (USP) were used to synthesize Fe3O4-Au hybrid nanostructures. The first coprecipitation method enabled the synthesis of Fe3O4 nanoparticles by mixing iron salts’ ions (Fe2+ and Fe3+) and ammonia as the base, and USP was used as the coating process of the Fe3O4 nanoparticles with Au. The formatted hybrid nanostructures consist of Fe3O4 nanoparticles that have Au on their surface in the form of gold nanoparticles (AuNPs). AuNPs have a crystalline structure and range in size from 10 to 200 nm. Additional characterization techniques, including ICP-OES, TEM, SEM, EDS, DLS, zeta potential, and room temperature magnetic hysteresis loops, were used to determine the chemical, physical, and magnetic properties of the Fe3O4 nanoparticles and hybrid nanostructures. It was found that USP produces separate AuNPs too (not just on the Fe3O4 surface), suggesting a bimodal formation of AuNPs. The zeta potential of the Fe3O4 nanoparticles showed poor stability (−15 mV), indicating a high tendency to aggregate, and the zeta potential of the hybrid nanostructures was also very low (≅0), which, comparatively means even worse stability. The saturation magnetization of the Fe3O4 nanoparticles was 35 emu/g, which is relatively lower than that of bulk Fe3O4, while the saturation magnetization of the hybrid nanostructures was significantly lower (0.1 emu/g) compared to the Fe3O4 nanoparticles.
Keywords: magnetic NPs, gold NPs, transmission electron microscopy, magnetic properties
Published in DiRROS: 08.01.2025; Views: 26; Downloads: 12
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