81. In vitro and in vivo correlation of skin and cellular responses to nucleic acid deliveryMaša Omerzel, Katarina Žnidar, A. Sales Conniff, Tanja Jesenko, Boštjan Markelc, Jared Tur, Nina Semenova, Kristopher Kohena, Simona Kranjc Brezar, Loree C. Heller, Maja Čemažar, 2022, original scientific article Abstract: Skin, the largest organ in the body, provides a passive physical barrier against infection and contains elements of the innate and adaptive immune systems. Skin consists of various cells, including keratinocytes, fibroblasts, endothelial cells and immune cells. This diversity of cell types could be important to gene therapies because DNA transfection could elicit different responses in different cell types. Previously, we observed the upregulation and activation of cytosolic DNA sensing pathways in several non-tumor and tumor cell types as well in tumors after the electroporation (electrotransfer) of plasmid DNA (pDNA). Based on this research and the innate immuno- genicity of skin, we correlated the effects of pDNA electrotransfer to fibroblasts and keratinocytes to mouse skin using reverse transcription real-time PCR (RT-qPCR) and several types of protein quantification. After pDNA electrotransfer, the mRNAs of the putative DNA sensors DEAD (AspGlu-Ala-Asp) box polypeptide 60 (Ddx60), absent in melanoma 2 (Aim2), Z-DNA binding protein 1 (Zbp1), interferon activated gene 202 (Ifi202), and interferon-inducible protein 204 (Ifi204) were upregulated in keratinocytes, while Ddx60, Zbp1 and Ifi204 were upregulated in fibroblasts. Increased levels of the mRNAs and proteins of several cytokines and chemokines were detected and varied based on cell type. Mouse skin experiments in vivo confirmed our in vitro results with increased expression of putative DNA sensor mRNAs and of the mRNAs and proteins of several cytokines and chemokines.
Keywords: DNA sensors, cytokines, electrotransfer, skin
Published in DiRROS: 06.09.2022; Views: 582; Downloads: 301
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82. Diversity, migration routes, and worldwide population genetic structure of Lecanosticta acicola, the causal agent of brown spot needle blightMarili Laas, Kalev Adamson, Irene Barnes, Josef Janoušek, Martin S. Mullett, Katarína Adamčíková, Mitsuteru Akiba, Ludwig Beenken, Helena Bragança, Timur S. Bulgakov, Barbara Piškur, 2022, original scientific article Abstract: Lecanosticta acicola is a pine needle pathogen causing brown spot needle blight that results in premature needle shedding with considerable damage described in North America, Europe, and Asia. Microsatellite and mating type markers were used to study the population genetics, migration history, and reproduction mode of the pathogen, based on a collection of 650 isolates from 27 countries and 26 hosts across the range of L. acicola. The presence of L. acicola in Georgia was confirmed in this study. Migration analyses indicate there have been several introduction events from North America into Europe. However, some of the source populations still appear to remain unknown. The populations in Croatia and western Asia appear to originate from genetically similar populations in North America. Intercontinental movement of the pathogen was reflected in an identical haplotype occurring on two continents, in North America (Canada) and Europe (Germany). Several shared haplotypes between European populations further suggests more local pathogen movement between countries. Moreover, migration analyses indicate that the populations in northern Europe originate from more established populations in central Europe. Overall, the highest genetic diversity was observed in south-eastern USA. In Europe, the highest diversity was observed in France, where the presence of both known pathogen lineages was recorded. Less than half of the observed populations contained mating types in equal proportions. Although there is evidence of some sexual reproduction taking place, the pathogen spreads predominantly asexually and through anthropogenic activity.
Keywords: brown spot needle blight, Lecanosticta acicola, pones, Pinus spp., popularion structure
Published in DiRROS: 12.08.2022; Views: 576; Downloads: 284
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85. Early-warning signals of individual tree mortality based on annual radial growthMaxime Cailleret, Vasilis Dakos, Steven Jansen, Elisabeth M.R. Robert, Tuomas Aakala, Mariano M. Amoroso, Joe A. Antos, Christof Bigler, Harald Bugmann, Marco Caccianaga, Katarina Čufar, Tom Levanič, 2019, original scientific article Abstract: Tree mortality is a key driver of forest dynamics and its occurrence is projected to increase in the future due to climate change. Despite recent advances in our understanding of the physiological mechanisms leading to death, we still lack robust indicators of mortality risk that could be applied at the individual tree scale. Here, we build on a previous contribution exploring the differences in growth level between trees that died and survived a given mortality event to assess whether changes in temporal autocorrelation, variance, and synchrony in time-series of annual radial growth data can be used as early warning signals of mortality risk. Taking advantage of a unique global ring-width database of 3065 dead trees and 4389 living trees growing together at 198 sites (belonging to 36 gymnosperm and angiosperm species), we analyzed temporal changes in autocorrelation, variance, and synchrony before tree death (diachronic analysis), and also compared these metrics between trees that died and trees that survived a given mortality event (synchronic analysis). Changes in autocorrelation were a poor indicator of mortality risk. However, we found a gradual increase in interannual growth variability and a decrease in growth synchrony in the last %20 years before mortality of gymnosperms, irrespective of the cause of mortality. These changes could be associated with drought-induced alterations in carbon economy and allocation patterns. In angiosperms, we did not find any consistent changes in any metric. Such lack of any signal might be explained by the relatively high capacity of angiosperms to recover after a stress-induced growth decline. Our analysis provides a robust method for estimating early-warning signals of tree mortality based on annual growth data. In addition to the frequently reported decrease in growth rates, an increase in inter-annual growth variability and a decrease in growth synchrony may be powerful predictors of gymnosperm mortality risk, but not necessarily so for angiosperms.
Keywords: tree mortality, ring-width, forest, growth, resilience indicators, drought, biotic agents, variance
Published in DiRROS: 20.07.2022; Views: 618; Downloads: 401
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86. Genska terapija v onkologiji, prvi razvojni koraki v SlovenijiMaja Čemažar, Tanja Jesenko, Maša Omerzel, Boštjan Markelc, Urška Kamenšek, Simona Kranjc Brezar, Špela Kos, Urša Lampreht Tratar, Katarina Žnidar, Andrej Renčelj, Urška Matkovič, Teja Valant, Kristina Levpušček, Živa Modic, Tilen Komel, Tim Božič, Urša Kešar, Barbara Starešinič, Katja Uršič Valentinuzzi, Monika Savarin, Primož Strojan, Gorana Gašljević, Maja Ota, Aleš Grošelj, Črt Jamšek, Rosana Hudej, Matjaž Peterka, Franc Smrekar, Barbara Hubad, Marjan Hosta, Jaka Kužnik, Alojz Hosta, Damijan Miklavčič, Matej Reberšek, Aleksandra Cvetkoska, Anja Zajc, Janja Dermol-Černe, Nataša Tozon, Nina Milevoj, Alenka Nemec Svete, Gregor Serša, 2022, professional article Abstract: Genska terapija postaja čedalje bolj zanimiva tudi v onkologiji. Med aplikacijami je morda najzanimivejša imunostimulacija. Pripravimo lahko plazmidno DNA, ki nosi zapis za različne imunostimulatorne molekule, ki jih vnesemo v celice tumorjev ali normalnih tkiv. Ta tkiva postanejo proizvajalci teh molekul, ki lahko delujejo lokalno ali pa se izločajo tudi sistemsko v krvni obtok. Ker plazmidna DNA ne prehaja celične membrane, so potrebni dostavni sistemi, virusni ali nevirusni. V naših študijah uporabljamo predvsem nevirusni dostavni sistem – elektroporacijo. Interlevkin 12 (IL-12) je eden od zanimivih citokinov, za katerega je znano protitumorsko delovanje s spodbujanjem imunskega odziva in antiangiogenim delovanjem. Namen projekta SmartGene.si je bil pripraviti plazmid z zapisom za interlevkin 12 (plazmid phIL12) in pripraviti vse potrebno za njegovo klinično testiranje za zdravljenje kožnih tumorjev. V konzorciju smo združili moči s partnerji z akademskega in industrijskega področja. Treba je bilo pripraviti plazmid za uporabo v humani onkologiji po zahtevah Evropske agencije za zdravila (EMA). Za prijavo klinične študije na Javno agencijo za zdravila in medicinske pripomočke (JAZMP) smo morali izvesti tudi vse neklinične raziskave o varnosti in učinkovitosti zdravila. Nato je bilo treba razviti postopek priprave zdravila, zagotoviti primerne prostore za pripravo in izvedbo postopka priprave zdravila. V treh letih smo dosegli vse te zastavljene cilje in dobili dovoljenje za izvajanje klinične študije na kožnih tumorjih, ki ga je izdala JAZMP na osnovi pozitivnega mnenja Komisije Republike Slovenije za medicinsko etiko. Zdaj poteka klinična študija faze I preizkušanja plazmida phIL12 na kožnih tumorjih glave in vratu z namenom preveriti varnost in sprejemljivost genskega elektroprenosa plazmida v tumorje. Cilj študije je prav tako določiti primeren odmerek zdravila, ki bi ga v nadaljnji klinični študiji uporabili kot adjuvantno zdravljenje k ablativnim terapijam, kot sta radioterapija ali elektrokemoterapija.
Keywords: genska terapija, interlevkin-12, plazmidna DNA, elektroprenos genov, rak kože
Published in DiRROS: 01.07.2022; Views: 1190; Downloads: 240
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87. Posodobljena priporočila diagnostike in zdravljenja diferenciranega raka ščitniceNikola Bešić, Alenka Grošel, Andraž Perhavec, Andrej Doma, Andreja A. Schwarzbartl-Pevec, Andreja Klevišar Ivančič, Andrej Vogrin, Barbara Vidergar-Kralj, Barbara Gazić, Barbara Perić, Blaž Krhin, Cvetka Grašič-Kuhar, Edvard Pirnat, Ivana Žagar, Jerca Blazina, Katarina Zevnik, Katica Bajuk-Studen, Katja Zaletel, Margareta Strojan Fležar, Marko Hočevar, Marko Kokalj, Marta Dremelj, Nebojša Glumac, Olga Blatnik, Simona Gaberšček, Snežana Pavlović Djokić, Ulrika Klopčič, Veronika Kloboves-Prevodnik, Zorica Čekić, 2022, professional article Keywords: rak ščitnice, diferencirani rak, zdravljenje, smernice
Published in DiRROS: 01.07.2022; Views: 812; Downloads: 244
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88. Building of the Al-Containing secondary raw materials registry for the production of low CO2 mineral binders in South-Eastern European regionGorazd Žibret, Klemen Teran, Lea Žibret, Katarina Šter, Sabina Dolenec, 2021, original scientific article Abstract: The bottleneck in the process for increasing production of low CO2 mineral binders, based on BCSA (belite sulfoaluminate) clinkers, is the availability of Al-rich raw materials. For that purpose, a new registry of Al-containing secondary mineral residues (industrial and mine waste) has been developed and is presented in this paper. The methodology of creating the registry consists of three main steps: Gathering ideas, consolidation of ideas, and implementation. In order to achieve this, the following methodology was adopted: Analysis of similar registries by potential end-users and seeking potential solutions and tools to be used, and conducting 3 rounds of stakeholder consultations via workshops in order to determine crucial parameters and features the registry needs to contain. The key discussion points were about which data the registry needs to contain, who shall be the potential users, and what are the stakeholderʼs expectations from the registryʼs portal. Potential individual registry variables were identified as being relevant/irrelevant or available/unavailable, and potential solutions for the registrys sustainability were explored. Each Al-rich waste/residue data entry is divided into 10 slots, describing legal status, location, quantities, chemical (REE included), mineralogical, physical and radiological properties, life-cycle assessment, additional data, and data relevancy. The registry will act as a matchmaking tool between producers/holders of Al-rich secondary raw materials and potential producers of cement clinkers.
Keywords: cement, belite sulfoaluminate clinker, waste, by--product, deležniki
Published in DiRROS: 10.06.2022; Views: 528; Downloads: 319
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89. The valorisation of selected quarry and mine waste for sustainable cement production within the concept of circular economyEmilija Fidanchevski, Katarina Šter, Maruša Mrak, Ljiljana Kljajević, Gorazd Žibret, Klemen Teran, Bojan Poletanovic, Monika Fidanchevska, Sabina Dolenec, Ildikó Merta, 2022, original scientific article Abstract: The cement industry could potentially consume large amounts of solid industrial waste in order to improve its sustainability. The suitability of selected quarry and mine waste as secondary raw materials (SRM) was examined for the sustainable production of cement following the concept of a circular economy. The chemical, mineralogical, and radiological characterization of SRM was conducted in this study. Its potential use in low-carbon and low-energy belite-sulfoaluminate cement was investigated by incorporating the examined SRM into cement clinker. Various characterization methods were used to characterize the cement, including X-ray powder diffraction (XRD), thermal analysis (DTA/TG), and isothermal calorimetry. Depending on the chemical composition of the waste, lower or higher amounts were allowed to be incorporated into the raw clinker mixture for a targeted clinker phase composition. Among the samples, differences were observed in the phase composition of synthesized clinkers, which slightly influenced the reactivity of the cement but did not significantly change the compressive strength of the final product.
Keywords: mineralne surovine, rudarski odpadki, rudniki, kamnolomi, cement, klinker, krožno gospodarstvo, BCSA clinker, cement, circular economy, clinker, mine wastes, quarry waste
Published in DiRROS: 10.06.2022; Views: 605; Downloads: 318
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90. Pan-European phylogeography of the European roe deer (Capreolus capreolus)Kamila Plis, Magdalena Niedziałkowska, Tomasz Borowik, Johannes Lang, Mike Heddergott, Juha Tiainen, Aleksey Bunevich, Nikica Šprem, Ladislav Paule, Aleksey A. Danilkin, Marina Kholodova, Elena Zvychaynaya, Nadezhda Kashinina, Boštjan Pokorny, Katarina Flajšman, 2022, original scientific article Abstract: To provide the most comprehensive picture of species phylogeny and phylogeography of European roe deer (Capreolus capreolus), we analyzed mtDNA control region (610 bp) of 1469 samples of roe deer from Central and Eastern Europe and included into the analyses additional 1541 mtDNA sequences from GenBank from other regions of the continent. We detected two mtDNA lineages of the species: European and Siberian (an introgression of C. pygargus mtDNA into C. capreolus). The Siberian lineage was most frequent in the eastern part of the continent and declined toward Central Europe. The European lineage contained three clades (Central, Eastern, and Western) composed of several haplogroups, many of which were separated in space. The Western clade appeared to have a discontinuous range from Portugal to Russia. Most of the haplogroups in the Central and the Eastern clades were under expansion during the Weichselian glacial period before the Last Glacial Maximum (LGM), while the expansion time of the Western clade overlapped with the Eemian interglacial. The high genetic diversity of extant roe deer is the result of their survival during the LGM probably in a large, contiguous range spanning from the Iberian Peninsula to the Caucasus Mts and in two northern refugia.
Keywords: Capreolus capreolus, expansion, mitochondrial DNA, the Last Glacial Maximum refugia, the Quaternary history, Phylogenetics
Published in DiRROS: 26.05.2022; Views: 604; Downloads: 417
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