1281. Production of volatile moth sex pheromones in transgenic Nicotiana benthamiana plantsRubèn Mateos Fernández, Elena Moreno Gimenéz, Silvia Gianoglio, Alfredo Quijano-Rubio, Jose Gavaldá-García, Lucía Estellés, Alba Rubert, José Luis Rambla, Marta Vazquez-Vilar, Estefanía Huet, Asunción Fernández-del-Carmen, Ana Espinosa-Ruiz, Mojca Juteršek, Sandra Vacas, Ismael Navarro Fuertes, Vicente Navarro-Llopis, Jaime Primo, Diego Orzaez, 2021, original scientific article Abstract: Plant-based bioproduction of insect sex pheromones has been proposed as an innovative strategy to increase the sustainability of pest control in agriculture. Here, we describe the engineering of transgenic plants producing (Z)-11-hexadecenol (Z11-16OH) and (Z)-11-hexadecenyl acetate (Z11-16OAc), two main volatile components in many Lepidoptera sex pheromone blends. We assembled multigene DNA constructs encoding the pheromone biosynthetic pathway and stably transformed them into Nicotiana benthamiana plants. The constructs contained the Amyelois transitella AtrΔ11 desaturase gene, the Helicoverpa armigera fatty acyl reductase HarFAR gene, and the Euonymus alatus diacylglycerol acetyltransferase EaDAct gene in different configurations. All the pheromone-producing plants showed dwarf phenotypes, the severity of which correlated with pheromone levels. All but one of the recovered lines produced high levels of Z11-16OH, but very low levels of Z11-16OAc, probably as a result of recurrent truncations at the level of the EaDAct gene. Only one plant line (SxPv1.2) was recovered that harboured an intact pheromone pathway and which produced moderate levels of Z11-16OAc (11.8 μg g-1 FW) and high levels of Z11-16OH (111.4 μg g-1). Z11-16OAc production was accompanied in SxPv1.2 by a partial recovery of the dwarf phenotype. SxPv1.2 was used to estimate the rates of volatile pheromone release, which resulted in 8.48 ng g-1 FW per day for Z11-16OH and 9.44 ng g-1 FW per day for Z11-16OAc. Our results suggest that pheromone release acts as a limiting factor in pheromone biodispenser strategies and establish a roadmap for biotechnological improvements. Published in DiRROS: 19.07.2024; Views: 404; Downloads: 202 Full text (1,99 MB) This document has many files! More... |
1282. A mini-TGA protein modulates gene expression through heterogeneous association with transcription factorsŠpela Tomaž, Marko Petek, Tjaša Lukan, Karmen Pogačar, Katja Stare, Erica Teixeira Prates, Daniel A. Jacobson, Jan Zrimec, Gregor Bajc, Matej Butala, Maruša Pompe Novak, Ajda Taler-Verčič, Aleksandra Usenik, Dušan Turk, Anna Coll Rius, Kristina Gruden, 2022, original scientific article Abstract: TGA (TGACG-binding) transcription factors, which bind their target DNA through a conserved basic region leucine zipper (bZIP) domain, are vital regulators of gene expression in salicylic acid (SA)-mediated plant immunity. Here, we investigated the role of StTGA2.1, a potato (Solanum tuberosum) TGA lacking the full bZIP, which we named a mini-TGA. Such truncated proteins have been widely assigned as loss-of-function mutants. We, however, confirmed that StTGA2.1 overexpression compensates for SA-deficiency, indicating a distinct mechanism of action compared with model plant species. To understand the underlying mechanisms, we showed that StTGA2.1 can physically interact with StTGA2.2 and StTGA2.3, while its interaction with DNA was not detected. We investigated the changes in transcriptional regulation due to StTGA2.1 overexpression, identifying direct and indirect target genes. Using in planta transactivation assays, we confirmed that StTGA2.1 interacts with StTGA2.3 to activate StPRX07, a member of class III peroxidases (StPRX), which are known to play role in immune response. Finally, via structural modeling and molecular dynamics simulations, we hypothesized that the compact molecular architecture of StTGA2.1 distorts DNA conformation upon heterodimer binding to enable transcriptional activation. This study demonstrates how protein truncation can lead to distinct functions and that such events should be studied carefully in other protein families. Published in DiRROS: 18.07.2024; Views: 432; Downloads: 166 Full text (1,37 MB) This document has many files! More... |
1283. Hyperthermia as a potential cornerstone of effective multimodality treatment with radiotherapy, cisplatin and PARP inhibitor in IDH1-mutated cancer cellsMohammed Khurshed, Elia Prades-Sagarra, Sarah Al Saleh, Peter Sminia, Johanna W Wilmink, Remco J. Molenaar, Hans Crezee, Cornelis J. F. van Noorden, 2022, original scientific article Abstract: Mutations in the isocitrate dehydrogenase 1 (IDH1MUT) gene occur in various types of malignancies, including ~60% of chondrosarcomas, ~30% of intrahepatic cholangiocarcinomas and >80% of low-grade gliomas. IDH1MUT are causal in the development and progression of these types of cancer due to neomorphic production of the oncometabolite D-2-hydroxyglutarate (D-2HG). Intracellular accumulation of D-2HG has been implicated in suppressing homologous recombination and renders IDH1MUT cancer cells sensitive to DNA-repair-inhibiting agents, such as poly-(adenosine 5′-diphosphate–ribose) polymerase inhibitors (PARPi). Hyperthermia increases the efficacy of DNA-damaging therapies such as radiotherapy and platinum-based chemotherapy, mainly by inhibition of DNA repair. In the current study, we investigated the additional effects of hyperthermia (42 °C for 1 h) in the treatment of IDH1MUT HCT116 colon cancer cells and hyperthermia1080 chondrosarcoma cancer cells in combination with radiation, cisplatin and/or a PARPi on clonogenic cell survival, cell cycle distribution and the induction and repair of DNA double-strand breaks. We found that hyperthermia in combination with radiation or cisplatin induces an increase in double-strand breaks and cell death, up to 10-fold in IDH1MUT cancer cells compared to IDH1 wild-type cells. This vulnerability was abolished by the IDH1MUT inhibitor AGI-5198 and was further increased by the PARPi. In conclusion, our study shows that IDH1MUT cancer cells are sensitized to hyperthermia in combination with irradiation or cisplatin and a PARPi. Therefore, hyperthermia may be an efficacious sensitizer to cytotoxic therapies in tumors where the clinical application of hyperthermia is feasible, such as IDH1MUT chondrosarcoma of the extremities. Keywords: isocitrate dehydrogenase, PARP, hyperthermia, D‐2‐hydroxyglutarate, radiotherapy, cisplatin Published in DiRROS: 18.07.2024; Views: 327; Downloads: 219 Full text (2,26 MB) This document has many files! More... |
1284. Towards better-informed dispersal probabilities in historical biogeography : arachnids as a model lineageMatjaž Kuntner, Eva Turk, 2022, original scientific article Abstract: Historical biogeography is an integrative scientific field critical for testing evolutionary hypotheses pertinent to organismal distributions, but despite recent theoretical and analytical advances, biogeographic reconstructions continue to struggle with accuracy and rigor. Most modern studies include the three elements needed for historical biogeographic inference, namely a time-calibrated phylogeny, contemporary taxonomic distributions, and estimations of organismal dispersal probabilities. The latter, we argue, are particularly vague, and historical biogeography would greatly benefit from dispersal probability estimations that are better informed and biologically meaningful. To achieve that goal, next-generation biogeography should ideally consider: a) dispersal-related traits; b) ecology; c) geological histories; and d) geographical factors. We briefly recap the three case studies on spiders that have pioneered this approach. Due to their old age and mega-diversity—considering both phylogenetic and life style diversity—arachnids are an ideal animal lineage for modern biogeographic research. There is no reason, however, that the concept should not be applied to all life. Further modifications of the proposed concept and, particularly, methodological implementation are needed to render this biogeographic framework widely useful. Keywords: next-generation biogeography, ancestral areas, dispersal biology, geological history, vicariance, extinction, arachnids Published in DiRROS: 18.07.2024; Views: 274; Downloads: 133 Full text (1,10 MB) This document has many files! More... |
1285. Multi-platform study of the extreme bloom of the barrel jellyfish Rhizostoma pulmo (Cnidaria: Scyphozoa) in the northernmost gulf of the Mediterranean Sea (Gulf of Trieste) in April 2021Nydia Catalina Reyes Suárez, Valentina Tirelli, Laura Ursella, Matjaž Ličer, Massimo Celio, Vanessa Cardin, 2022, original scientific article Abstract: On 7 April 2021, an exceptional bloom of the scyphomedusa Rhizostoma pulmo was observed in the Gulf of Trieste (Italy). Blooms of this species in the northern Adriatic Sea have been reported since the late 1800s: the density of jellyfish observed in 2021 reached more than 10 specimens per square metre. We analyse the bloom from a multi-platform approach using observations and model data at different timescales. We attempt to explain the intensity of the bloom as a consequence of thermohaline and hydrodynamical conditions in the gulf. Meteo-oceanographic conditions that may have contributed to the exceptional aggregation of jellyfish observed along the northernmost coast of the Adriatic Sea are discussed in detail. Specifically, our results indicate that this bloom was enabled by (1) the presence of a high number of jellyfish in the gulf, probably linked to the anomalously warm sea conditions in spring 2020 and winter 2021, which may have favoured a longer reproductive period and enhanced survival of adult R. pulmo, respectively; and (2) strong wind events, such as the bora wind for the Gulf of Trieste, which enhanced upwelling and mixing processes in the gulf, bringing the jellyfish from the deeper waters to the surface and clustering them along the coast. Keywords: jellyfishes, Gulf of Trieste, Mediterranean Sea, hydrobiology, marine biology Published in DiRROS: 18.07.2024; Views: 327; Downloads: 188 Full text (10,49 MB) This document has many files! More... |
1286. First application of the AMBI index to the macrobenthic soft-bottom community of Terra Nova Bay (Ross Sea, Southern Ocean)Cristina Munari, Ángel Borja, Cinzia Corinaldesi, Eugenio Rastelli, Marco Lo Martire, Valentina Pitacco, Michele Mistri, 2022, original scientific article Abstract: The assemblages of marine benthic organisms and sediment characteristics were investigated in the coastal area between the Mario Zucchelli Antarctic Research Station and Adelie Cove in Terra Nova Bay (Ross Sea, Southern Ocean) during the 2015 summer season. Sediment samples were taken from 11 stations at depths between 25 and 140 m. The dominance of sand characterised sites, and the biochemical composition of the sedimentary organic matter resulted in very variable between the different sites. A total of 142 taxa were identified, with Annelida (68 taxa) and Arthropoda (35 taxa) constituting the main macrobenthic groups. The benthic community at deeper stations showed higher species richness and lower dominance compared to the shallower stations. For the first time in Antarctica, we also investigated the response of the AZTI’s Marine Biotic Index (AMBI) to the organic gradient. Of the 142 taxa found, 97 were not listed in the AMBI library, and we were able to assign as many as 88 taxa to an ecological group. All of these new species were added to the new AMBI species list. AMBI showed a good response to the organic gradient. Keywords: antarctic benthos, ecological quality assessment, indexes, Ross Sea Published in DiRROS: 18.07.2024; Views: 308; Downloads: 229 Full text (1,56 MB) This document has many files! More... |
1287. Expression of inducible factors reprograms CAR-T cells for enhanced function and safetyAnže Smole, Alexander Benton, Mathilde A. Poussin, Monika A. Eiva, Claudia Mezzanotte, 2022, original scientific article Abstract: Despite the success of CAR-T cell cancer immunotherapy, challenges in efficacy and safety remain. Investigators have begun to enhance CAR-T cells with the expression of accessory molecules to address these challenges. Current systems rely on constitutive transgene expression or multiple viral vectors, resulting in unregulated response and product heterogeneity. Here, we develop a genetic platform that combines autonomous antigen-induced production of an accessory molecule with constitutive CAR expression in a single lentiviral vector called Uni-Vect. The broad therapeutic application of Uni-Vect is demonstrated in vivo by activation-dependent expression of (1) an immunostimulatory cytokine that improves efficacy, (2) an antibody that ameliorates cytokine-release syndrome, and (3) transcription factors that modulate T cell biology. Uni-Vect is also implemented as a platform to characterize immune receptors. Overall, we demonstrate that Uni-Vect provides a foundation for a more clinically actionable next-generation cellular immunotherapy. Keywords: CAR-T cells, TCR, inducible, transcription factor, NFAT, single lentiviral expression system, IL-6, IL-12, CRS, armored Published in DiRROS: 18.07.2024; Views: 340; Downloads: 65 Link to file |
1288. The seven grand challenges in arachnid scienceMatjaž Kuntner, 2022, original scientific article Abstract: This perspective identifies the grand challenges in arachnid science: 1. Grasp the arachnid species diversity. There is a need to accelerate taxonomic research to obtain a sense of arachnid species diversity, however, at the same time, taxonomy needs to increase its quality, rigor, and repeatability. 2. Standardize arachnid systematics research. A solid phylogenetic definition and morphological diagnosis of Arachnida and its composing subgroups, usually treated at the rank of order, are needed. Studies should aim to stabilize and standardize phylogenetic efforts at all levels of hierarchy, and systematists should adopt criteria for higher level ranks in arachnid classification. 3. Interpret arachnid trait evolution through omics approaches. Among the field’s grand challenges is to define the genetic diversity encoding for the diverse arachnid traits, including developmental, morphological and ecological characteristics, biomaterials such as silks, venoms, digestive fluids, or allergens and bioproducts that cause diseases. Comparative genomics, transcriptomics, and proteomics will provide the empirical basis for biotechnology to modify arachnid genomes to fit numerous applications. 4. Facilitate biotechnological applications of arachnid molecules and biomaterials. Among the grand field challenges is to define potential applications of arachnid bioproducts from therapeutics to industry. New natural and biodegradable products, e.g. from spider silks, should ease our burden on ecosystems. 5. Utilize arachnids as models in ecological and biogeographic research. Biodiversity inventory sampling and analytical techniques should be extended from spiders to other arachnid groups. Spiders and their webs could be used as environmental DNA samplers, measuring or monitoring ecosystems’ overall biodiversity. Arachnids are excellent models to address biogeographical questions at the global to local scales. 6. Disentangle evolutionary drivers of arachnid diversity. Among the field grand challenges is a more precise evaluation to what extent the emergence of arachnid phenotypes is shaped by classical selection processes, and under what conditions, if any, sexual conflict needs to be invoked. 7. Define effective conservation measures for arachnids in the light of global changes. Effective conservation measures in arachnology should integrate the data from phylogenetic diversity, physiology, ecology, biogeography, and global change biology. Keywords: diversity, systematics, arachnids, evolution, biomaterials Published in DiRROS: 18.07.2024; Views: 276; Downloads: 137 Full text (709,51 KB) This document has many files! More... |
1289. Status and trends in the rate of introduction of marine non-indigenous species in European seasArgyro Zenetos, Konstantinos Tsiamis, Marika Galanidi, Natacha Carvalho, Cátia Bartilotti, Borut Mavrič, Martina Orlando-Bonaca, 2022, original scientific article Abstract: nvasive alien species are a major worldwide driver of biodiversity change. The current study lists verified records of non-indigenous species (NIS) in European marine waters until 2020, with the purpose of establishing a baseline, assessing trends, and discussing appropriate threshold values for good environmental status (GES) according to the relevant European legislation. All NIS records were verified by national experts and trends are presented in six-year assessment periods from 1970 to 2020 according to the European Union Marine Strategy Framework Directive. Altogether, 874 NIS have been introduced to European marine waters until 2020 with the Mediterranean Sea and North-East Atlantic Ocean hosting most of the introductions. Overall, the number of new introductions has steadily increased since 2000. The annual rate of new introductions reached 21 new NIS in European seas within the last six-year assessment period (2012–2017). This increase is likely due to increased human activities and research efforts that have intensified during the early 21st century within European Seas. As Europe seas are not environmentally, nor geographically homogenous, the setting of threshold values for assessing GES requires regional expertise. Further, once management measures are operational, pathway-specific threshold values would enable assessing the effectiveness of such measures. Keywords: non-indigenous species, European seas, regional seas, MSFD, good environmental status, validation, uncertainties Published in DiRROS: 18.07.2024; Views: 307; Downloads: 313 Full text (2,13 MB) This document has many files! More... |
1290. Small but voracious : invasive generalist consumes more zooplankton in winter than native planktivoreŽiga Ogorelec, Alexander Brinker, Dietmar Straile, 2022, original scientific article Abstract: In recent years, Lake Constance has experienced an invasion and domination of three-spined stickleback (Gasterosteus aculeatus) in the pelagic zone, which has coincided with a decline in the native whitefish (Coregonus wartmanni) population. Similar massive invasions of sticklebacks into pelagic zones have been recognized also in marine areas or small lakes worldwide. However, their diet overlaps with native species is rarely evaluated, especially in the winter season, which often presents a bottleneck for fish survival. In this study, we compared the diet of pelagic sticklebacks with the diet of the substantially larger native whitefish in different seasons, to evaluate the threat of the recent stickleback invasion on whitefish populations. By monthly sampling of zooplankton and both fish species diets, we could demonstrate that sticklebacks select similar prey throughout most of the year and consume more prey than whitefish during the winter. With relations between prey availability and prey selection, interspecific and intraspecific seasonal diet variability and indices like a prey-specific index of relative importance, we discuss the importance of zooplankton species traits and abundance for whitefish and stickleback predation. This study shows that sticklebacks, despite their small size, represent a serious potential diet competitor to native planktivorous fish. Sticklebacks quickly adapt to new environments, and thus we advocate precautions regarding their introduction into similar lakes as Lake Constance, as this could cause irreversible ecological changes. Keywords: Bythotrephes longimanus, Lake Constance, overwintering strategy, pelagic whitefish diet, planktivory, predator size, seasonal prey selection, stickleback invasion Published in DiRROS: 18.07.2024; Views: 352; Downloads: 220 Full text (4,43 MB) This document has many files! More... |