1. CRISPR/Cas-mediated plant genome editing : outstanding challenges a decade after implementationTeodoro Cardi, Jana Murovec, Allah Bakhsh, Vladislava Galović, Tjaša Lukan, Kubilay Yıldırım, Milica Zlatković, Katrijn Van Laere, 2023, review article Abstract: The discovery of the CRISPR/Cas genome-editing system has revolutionized our understanding of the plant genome. CRISPR/Cas has been used for over a decade to modify plant genomes for the study of specific genes and biosynthetic pathways as well as to speed up breeding in many plant species, including both model and non-model crops. Although the CRISPR/Cas system is very efficient for genome editing, many bottlenecks and challenges slow down further improvement and applications. In this review we discuss the challenges that can occur during tissue culture, transformation, regeneration, and mutant detection. We also review the opportunities provided by new CRISPR platforms and specific applications related to gene regulation, abiotic and biotic stress response improvement, and de novo domestication of plants.
Keywords: CRISPR applications, CRISPR platforms, gene regulations, mutant detection, plant regeneration
Published in DiRROS: 05.08.2024; Views: 85; Downloads: 129
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2. Maristem - stem cells of marine/aquatic invertebrates : from basic research to innovative applicationsLoriano Ballarin, Baruch Rinkevich, Kestin Bartscherer, Artur Burzynski, Sebastien Cambier, Matteo Cammarata, Isabelle Domart-Coulon, Damjana Drobne, Juanma Encinas, Uri Frank, Anne-Marie Geneviere, Bert Hobmayer, Helike Löhelaid, Daniel Lyons, Pedro Martinez, Paola Oliveri, Lorena Perić, Stefano Piraino, Andreja Ramšak, Sebastian Rakers, Fabian Rentzsch, Amalia Rosner, Tiago Henriques da Silva, Ildiko Somorjai, Sherif Suleiman, Ana Varela Coelho, 2018, original scientific article Abstract: The “stem cells” discipline represents one of the most dynamic areas in biomedicine. While adult marine/aquatic invertebrate stem cell (MISC) biology is of prime research and medical interest, studies on stem cells from organisms outside the classical vertebrate (e.g., human, mouse, and zebrafish) and invertebrate (e.g., Drosophila, Caenorhabditis) models have not been pursued vigorously. Marine/aquatic invertebrates constitute the largest biodiversity and the widest phylogenetic radiation on Earth, from morphologically simple organisms (e.g., sponges, cnidarians), to the more complex mollusks, crustaceans, echinoderms, and protochordates. These organisms contain a kaleidoscope of MISC-types that allow the production of a large number of novel bioactive-molecules, many of which are of significant potential interest for human health. MISCs further participate in aging and regeneration phenomena, including whole-body regeneration. For years, the European MISC-community has been highly fragmented and has established scarce ties with biomedical industries in an attempt to harness MISCs for human welfare. Thus, it is important to (i) consolidate the European community of researchers working on MISCs; (ii) promote and coordinate European research on MISC biology; (iii) stimulate young researchers to embark on research in MISC-biology; (iv) develop, validate, and share novel MISC tools and methodologies; (v) establish the MISC discipline as a forefront interest of biomedical disciplines, including nanobiomedicine; and (vi) establish collaborations with industries to exploit MISCs as sources of bioactive molecules. In order to fill the recognized gaps, the EC-COST Action 16203 “MARISTEM” has recently been launched. At its initial stage, the consortium unites 26 scientists from EC countries, Cooperating countries, and Near Neighbor Countries.
Keywords: aging, bioactive molecules, blue biotechnology, cancer, cell culture, COST Action, Europe, marine/aquatic invertebrates, regeneration, stem cells
Published in DiRROS: 24.07.2024; Views: 323; Downloads: 99
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3. Effect of ionizing radiation on human skeletal muscle precursor cellsMihaela Jurdana, Maja Čemažar, Katarina Pegan, Tomaž Marš, 2013, published scientific conference contribution Abstract: Background. Long term effects of different doses of ionizing radiation on human skeletal muscle myoblast proliferation, cytokine signalling and stress response capacity were studied in primary cell cultures.Materials and methods. Human skeletal muscle myoblasts obtained from muscle biopsies were cultured and irradiated with a Darpac 2000 X-ray unit at doses of 4, 6 and 8 Gy. Acute effects of radiation were studied by interleukin - 6 (IL-6) release and stress response detected by the heat shock protein (HSP) level, while long term effects were followed by proliferation capacity and cell death.Results. Compared with non-irradiated control and cells treated with inhibitor of cell proliferation Ara C, myoblast proliferation decreased 72 h post-irradiation, this effect was more pronounced with increasing doses. Post-irradiation myoblast survival determined by measurement of released LDH enzyme activity revealed increased activity after exposure to irradiation. The acute response of myoblasts to lower doses of irradiation (4 and 6 Gy) was decreased secretion of constitutive IL-6. Higher doses of irradiation triggered a stress response in myoblasts, determined by increased levels of stress markers (HSPs 27 and 70).Conclusions. Our results show that myoblasts are sensitive to irradiation in terms of their proliferation capacity and capacity to secret IL-6. Since myoblast proliferation and differentiation are a key stage in muscle regeneration, this effect of irradiation needs to be taken in account, particularly in certain clinical conditions.
Keywords: myoblasts, irradiation, proliferation, interleukin 6, muscle regeneration, apoptosis
Published in DiRROS: 03.04.2024; Views: 269; Downloads: 147
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4. Autologous platelet and extracellular vesicle-rich plasma as therapeutic fluid : a reviewKaja Troha, Domen Vozel, Matevž Arko, Apolonija Bedina Zavec, Drago Dolinar, Matej Hočevar, Zala Jan, Matic Kisovec, Boštjan Kocjančič, Ljubiša Pađen, Manca Pajnič, Samo Penič, Anna Romolo, Neža Repar, Vesna Spasovski, Nejc Steiner, Vid Šuštar, Aleš Iglič, Damjana Drobne, Ksenija Kogej, Saba Battelino, Veronika Kralj-Iglič, 2023, review article Keywords: platelet-rich plasma, platelets, centrifugation, extracellular vesicles, small cellular particles, wound healing, regeneration, fresh-frozen plasma, intercellular signaling peptides and proteins
Published in DiRROS: 26.01.2024; Views: 517; Downloads: 247
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6. Causes and consequences of large-scale windthrow on the development of fir-beech forests in the Dinaric mountainsMatjaž Čater, Ajša Alagić, Mitja Ferlan, Jernej Jevšenak, Aleksander Marinšek, 2023, original scientific article Abstract: We investigated several aspects of windthrow that are relevant to our understanding and management of forest ecosystems. As an example, we used an extreme event in December 2017, when the strongest storm in recent history occurred in the Slovenian Dinaric High Karst. We examined influential factors such as soil properties, wind speed, precipitation and ecological consequences for the affected forests. Soil properties were measured around standing and fallen silver fir trees at all three research sites. Tree species composition in the regeneration was observed on plots with chemical and acoustic ungulate deterrents and on control plots without deterrents. Economic estimates of yield loss due to damage were calculated at the national level. A model of the potential threat from windthrow was also developed based on data collected from windthrow events and meteorological data over the past 20 years. Our results indicate that soil depth and mineral fraction depth were similar at sites with and without damaged trees and were not the determining factors for tree toppling. Plots with acoustic deterrents showed the most effective regeneration development, the least decline in silver fir and the greatest increase in noble hardwood seedlings, while plots with chemical deterrents showed the least browsing damage. The estimated economic loss of €16.1 million is 6.6% less than the harvest under normal conditions. The economic loss was relatively low due to the nature of the storm, with the predominant type of damage being uprooted trees with no damaged trunks. The windthrow hazard model revealed that a large number of consecutive events with strong winds in each section weakened the stand, which was subsequently knocked down during the next extreme wind and rainfall event.
Keywords: natural forest regeneration, windthrow, economic loss, browsing, ungulate deterrents, forest soil properties, potential threat model
Published in DiRROS: 27.06.2023; Views: 1496; Downloads: 721
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7. Engineered combinatorial cell device for wound healing and bone regenerationLucija Kadunc, Duško Lainšček, Rok Gašperšič, Petra Sušjan, Uroš Kovačič, Miha Butinar, Boris Turk, Roman Jerala, Iva Hafner Bratkovič, 2023, original scientific article Abstract: Growth factors are the key regulators that promote tissue regeneration and healing processes. While the effects of individual growth factors are well documented, a combination of multiple secreted growth factors underlies stem cell–mediated regeneration. To avoid the potential dangers and laborintensive individual approach of stem cell therapy while maintaining their regeneration-promoting effects based on multiple secreted growth factors, we engineered a “mix-and-match” combinatorial platform based on a library of cell lines producing growth factors. Treatment with a combination of growth factors secreted by engineered mammalian cells was more efficient than with individual growth factors or even stem cell–conditioned medium in a gap closure assay. Furthermore, we implemented in a mouse model a device for allogenic cell therapy for an in situ production of growth factors, where it improved cutaneous wound healing. Augmented bone regeneration was achieved on calvarial bone defects in rats treated with a cell device secreting IGF, FGF, PDGF, TGF-β, and VEGF. In both in vivo models, the systemic concentration of secreted factors was negligible, demonstrating the local effect of the regeneration device. Finally, we introduced a genetic switch that enables temporal control over combinations of trophic factors released at different stages of regeneration mimicking the maturation of natural wound healing to improve therapy and prevent scar formation.
Keywords: wound healing, bone regeneration, growth factors
Published in DiRROS: 01.06.2023; Views: 718; Downloads: 290
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8. Structure, development and growth of selection forests at the Granata research siteMatej Reščič, Andrej Bončina, 2007, original scientific article Abstract: The Granata research site was established for the study of structure, growth and regeneration of silver fir-European beech single stem selection forest. Three phytocoenological relevés were carried out, all trees ?5 cm diameter at breast height (d.b.h.) were measured by full callipering, tree growth was analysed for sample trees, regeneration of tree species was registered on 63 sampling plots. In addition, historical data from past forest inventories werestudied. In the period from 1952 to 2003, the share of silver fir has decreased from 86% to 26%, the share of Norway spruce increased significantly,whereas the share of beech and sycamore has slightly increased. The current growing stock amounts to 350 m3 ha-1, with large-size diameter trees (d.b.h. ?50 cm) representing 51% of total growing stock. The site is divided into 16 stand patches with significant differences in tree species composition and diameter distribution. Regeneration is sufficient with silver fir prevailing in total number of seedlings (42%). 23% of all seedlings are damaged as a result of game browsing.
Keywords: selection forest, plenterwald, selection system management, stand structure, tree species composition, regeneration, diameter distribution
Published in DiRROS: 12.07.2017; Views: 5048; Downloads: 2122
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9. Biodiversity of types of ectomycorrhizae in a norway spruce stands on PokljukaUrša Vilhar, Igor Smolej, Tadeja Trošt Sedej, Lado Kutnar, Hojka Kraigher, 2004, original scientific article Abstract: Types of ectomycorrhizae were studied in soil cores from a young regeneration center in an autochthonous Norway spruce stand on Pokljuka (Triglav National Park, 1200 m.a.s.l.). Soil cores of equal volume (274 ml, 0 - 18 cm deep) weretaken from 33 sampling plots. In the samples all the roots were counted and types of ectomycorrhizae briefly characterized. From these data diversity indices (species diversity (d) and Shannon- Weaver index of diversity (H)) were calculated. Interactions among mycorrhizae, light regime and survival of spruce seedlings were studied. Out of about 50,000 root tips approximately 1 %were non-mycorrhizal, 63 % were old unviable mycorrhizae and 36 % were identifiable ectomycorrhizal root tips, forming 27 different types of ectomycorrhizae. Sixteen types of ectomycorrhizae were briefly characterized. The Shannon diversity index for types of ectomycorrhizae was high (3.13) with respect to the above-ground diversity of vegetation (1.7). The direct site factor was shown to be negatively correlated to ~Piceirhiza cornuta. The diffuse site factor was negatively correlated to Cortinarius sp. (obtusus type) and positively correlated to Inocybe sp. The ground vegetation cover waspositively correlated to Piceirhiza gelatinosa and the total vegetation cover to Elaphomyces sp.
Keywords: ectomycorrhizae, types of ectomycorrhizae, Norway spruce, natural regeneration, Pokljuka
Published in DiRROS: 12.07.2017; Views: 4554; Downloads: 1925
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10. Analysis of the influece of ungulates on the regeneration of Dinaric fir-beech forests in the research site Trnovec in the Kočevje forest managementregionKristjan Jarni, Dušan Robič, Andrej Bončina, 2004, original scientific article Abstract: In the period 1970-2000, 152 fenced-in areas were built in the Kočevje Forest Management Region with an aim to protect tree seedlings and saplings from ungulatesć activity and to monitor the influence of roe and red deer on natural regeneration. The average surface area of fenced areas is 0.71 ha. Using the pair comparison technique (fenced vs. unfenced areas), the structureand the composition of the natural regeneration of tree species as well as complete shrub and herb vegetation were analysed in the research site Trnovec. Furthermore, the vegetation was investigated using the Braun-Blanquetmethod. The research results show significant differences between fenced and unfenced areas, both in tree species composition and in theheight structure of the sapling community. In fenced areas the total numberof saplings taller than 50 cm is higher and an increase is also evident in the number of saplings of silver fir Abies alba, sycamore Acer pseudoplatanus, elm Ulmus glabra and other minor tree species. There are also significant differences in species composition and in the abundance of plant species in the herb layer. The results show that natural regeneration of Dinaric fir-beech forests is successful, provided the influence of ungulates is excluded.
Keywords: natural forest regeneration, Fagus sylvatica, roe deer, fir-beech forest, Abies alba, fenced area, seedling browsing, Kočevje forest region, research site Trnovec
Published in DiRROS: 12.07.2017; Views: 4795; Downloads: 2104
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