1. 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, izvirni znanstveni članek Povzetek: 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.
Ključne besede: CAR-T cells, TCR, inducible, transcription factor, NFAT, single lentiviral expression system, IL-6, IL-12, CRS, armored
Objavljeno v DiRROS: 18.07.2024; Ogledov: 6; Prenosov: 3
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2. The seven grand challenges in arachnid scienceMatjaž Kuntner, 2022, izvirni znanstveni članek Povzetek: 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.
Ključne besede: diversity, systematics, arachnids, evolution, biomaterials
Objavljeno v DiRROS: 18.07.2024; Ogledov: 15; Prenosov: 6
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3. 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, izvirni znanstveni članek Povzetek: 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.
Ključne besede: non-indigenous species, European seas, regional seas, MSFD, good environmental status, validation, uncertainties
Objavljeno v DiRROS: 18.07.2024; Ogledov: 2; Prenosov: 5
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4. Small but voracious : invasive generalist consumes more zooplankton in winter than native planktivoreŽiga Ogorelec, Alexander Brinker, Dietmar Straile, 2022, izvirni znanstveni članek Povzetek: 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.
Ključne besede: Bythotrephes longimanus, Lake Constance, overwintering strategy, pelagic whitefish diet, planktivory, predator size, seasonal prey selection, stickleback invasion
Objavljeno v DiRROS: 18.07.2024; Ogledov: 10; Prenosov: 8
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5. The freshwater balance of the Adriatic Seastudy : a sensitivity studyMartin Vodopivec, Klodian Zaimi, Álvaro Judice Peliz, 2022, izvirni znanstveni članek Povzetek: The Adriatic Sea is a narrow semi-enclosed basin with considerable freshwater inflow, connected to the saltier Mediterranean Sea through a narrow strait. The northern and central parts of the basin are characterized by a shallow shelf, while the southern part features a pit exceeding 1,200 m in depth. We conducted a series of modeling experiments over a 16-year period (2000–2015) using different runoff configurations and different sources of atmospheric forcing to investigate the sensitivity of the Adriatic Sea circulation and hydrology to freshwater balance and heat loss. Our results show that the shelf salinity and the salinity of the surface layers in the southern Adriatic are part of a self-amplifying loop involving dense water formation, water exchange with the Mediterranean Sea, and salty water intrusions to the shelf. Therefore, the characteristics of the basin and the water circulation are highly sensitive to the freshwater budget and heat losses. River discharge is subject to large interannual variations and is poorly known for many of the Adriatic freshwater sources. To improve the accuracy of the freshwater budget, we created a new climatology for three Albanian rivers and modulated the monthly climatological discharge in accordance with the rivers Po and Isonzo (Soča). Evaporation, precipitation, and heat losses vary strongly among the available atmospheric reanalyses and we show that the choice of atmospheric forcing has a substantial impact on the hydrology and circulation of the Adriatic Sea.
Objavljeno v DiRROS: 18.07.2024; Ogledov: 5; Prenosov: 6
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6. The cytotoxic effects of cannabidiol and cannabigerol on glioblastoma stem cells may mostly involve GPR55 and TRPV1 signallingTamara Lah Turnšek, Bernarda Majc, Metka Novak, Ajda Sušnik, Barbara Breznik, Andrej Porčnik, Roman Bošnjak, Aleksander Sadikov, Marta Malavolta, Selma Halilčević, Jernej Mlakar, Roby Zomer, 2022, izvirni znanstveni članek Povzetek: Glioblastoma (GBM) is one of the most aggressive cancers, comprising 60–70% of all gliomas. The large G-protein-coupled receptor family includes cannabinoid receptors CB1, CB2, GPR55, and non-specific ion receptor protein transporters TRPs. First, we found up-regulated CNR1, GPR55, and TRPV1 expression in glioma patient-derived tissue samples and cell lines compared with non-malignant brain samples. CNR1 and GPR55 did not correlate with glioma grade, whereas TRPV1 negatively correlated with grade and positively correlated with longer overall survival. This suggests a tumour-suppressor role of TRPV1. With respect to markers of GBM stem cells, preferred targets of therapy, TRPV1 and GPR55, but not CNR1, strongly correlated with different sets of stemness gene markers: NOTCH, OLIG2, CD9, TRIM28, and TUFM and CD15, SOX2, OCT4, and ID1, respectively. This is in line with the higher expression of TRPV1 and GPR55 genes in GSCs compared with differentiated GBM cells. Second, in a panel of patient-derived GSCs, we found that CBG and CBD exhibited the highest cytotoxicity at a molar ratio of 3:1. We suggest that this mixture should be tested in experimental animals and clinical studies, in which currently used Δ9-tetrahydrocannabinol (THC) is replaced with efficient and non-psychoactive CBG in adjuvant standard-of-care therapy.
Ključne besede: glioblastoma, glioma, cannabigerol, cannabidiol, cannabinoid receptors, stem cells
Objavljeno v DiRROS: 18.07.2024; Ogledov: 3; Prenosov: 5
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7. A small bacteriophage protein determines the hierarchy over co-residential jumbo phage in Bacillus thuringiensis serovar israelensisAnja Pavlin, Anže Lovše, Gregor Bajc, Jan Otoničar, Amela Kujović, Živa Lengar, Ion Gutiérrez-Aguirre, Rok Kostanjšek, Janez Konc, Nadine Fornelos, Matej Butala, 2022, izvirni znanstveni članek Povzetek: Bacillus thuringiensis serovar israelensis is the most widely used biopesticide against insects, including vectors of animal and human diseases. Among several extrachromosomal elements, this endospore-forming entomopathogen harbors two bacteriophages: a linear DNA replicon named GIL01 that does not integrate into the chromosome during lysogeny and a circular-jumbo prophage known as pBtic235. Here, we show that GIL01 hinders the induction of cohabiting prophage pBtic235. The GIL01-encoded small protein, gp7, which interacts with the host LexA repressor, is a global transcription regulator and represses the induction of pBtic235 after DNA damage to presumably allow GIL01 to multiply first. In a complex with host LexA in stressed cells, gp7 down-regulates the expression of more than 250 host and pBtic235 genes, many of which are involved in the cellular functions of genome maintenance, cell-wall transport, and membrane and protein stability. We show that gp7 homologs that are found exclusively in bacteriophages act in a similar fashion to enhance LexA’s binding to DNA, while likely also affecting host gene expression. Our results provide evidence that GIL01 influences both its host and its co-resident bacteriophage.
Objavljeno v DiRROS: 18.07.2024; Ogledov: 7; Prenosov: 4
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8. Genome-informed design of a LAMP assay for the specific detection of the strain of ‘Candidatus Phytoplasma asteris’ phytoplasma occurring in grapevines in South AfricaŠpela Alič, Marina Dermastia, Johan Burger, Matthew Dickinson, Gerhard Pietersen, Gert Pietersen, Tanja Dreo, 2022, izvirni znanstveni članek Povzetek: Grapevine yellows is one of the most damaging phytoplasma-associated diseases worldwide. It is linked to several phytoplasma species, which can vary regionally due to phytoplasma and insect-vector diversity. Specific, rapid, and reliable detection of the grapevine yellows pathogen has an important role in phytoplasma control. The purpose of this study was to develop and validate a specific loop-mediated isothermal amplification (LAMP) assay for detection of a distinct strain of grapevine ‘Candidatus Phytoplasma asteris’ that is present in South Africa, through implementation of a genome-informed test design approach. Several freely available, user-friendly, web-based tools were coupled to design the specific LAMP assays. The criteria for selection of the assays were set for each step of the process, which resulted in four experimentally operative LAMP assays that targeted the ftsH/hflB gene region, specific to the aster yellows phytoplasma strain from South Africa. A real-time PCR was developed, targeting the same genetic region, to provide extensive validation of the LAMP assay. The validated molecular assays are highly specific to the targeted aster yellows phytoplasma strain from South Africa, with good sensitivity and reproducibility. We show a genome-informed molecular test design and an efficient validation approach for molecular tests if reference and sample materials are sparse and hard to obtain.
Ključne besede: aster yellows, fruit, LAMP, molecular detection, pathogendetection, phytoplasma, Prokaryotes
Objavljeno v DiRROS: 17.07.2024; Ogledov: 22; Prenosov: 4
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9. Where and how : new insight for brown algal forest restoration in the AdriaticMartina Orlando-Bonaca, Gilda Savonitto, Valentina Asnaghi, Domen Trkov, Valentina Pitacco, Milijan Šiško, Tihomir Makovec, Petra Slavinec, Ana Lokovšek, Saul Ciriaco, Mariachiara Chiantore, Sara Kaleb, Emmanuelle Patricia Descourvières, Marina Srijemsi, Annalisa Falace, 2022, izvirni znanstveni članek Povzetek: Several anthropogenic factors are responsible for the decline of Cystoseira sensu lato (hereafter Cystoseira) forests along Mediterranean coasts. Some Cystoseira species are already regionally extinct, and their decline has been widely recorded. Sustainable and efficient techniques for the restoration of Cystoseira are needed. In this context, the objectives of this study were i) to analyse the reproductive traits of three populations of Gongolaria barbata from three nearby donor sites in the northern Adriatic Sea, assessing the differences in their reproductive potential and reproductive success; and ii) to evaluate the outplanting success in terms of the effectiveness of G. barbata restoration, in relation to the different donor and receiving sites (Miramare MPA and in front of the Marine Biology Station Piran - MBSP) and implemented methods (ex situ and hybrid method combining a mesocosm cultivation and a suspended culture in the field). After 2 weeks of cultivation in mesocosms, half of the tiles with germlings were transported to the receiving sites and placed on suspended lantern nets (hybrid method), which were later (after 3 months) transferred to the seabed on concrete plates with protective cages. The remaining tiles were placed on the seabed on concrete plates with protective cages after a 4-week culture (ex situ method). At both sites, lantern nets and plates were randomly placed at 3 m depth. Thallus length was measured monthly in each treatment. Seedlings in suspended culture showed lower performance at the Miramare MPA, most likely due to the unfavourable environmental conditions. The satisfactory results obtained at MBSP demonstrate the efficiency of the hybrid method and confirm its potential to reduce the cost and time required for cultivation. Since unpredictable climatic events pose the greatest threat to restoration performance, these challenges must be considered when establishing new restoration practices. Moreover, herbivore regulation is extremely urgent and should be planned and implemented on a larger regional scale.
Ključne besede: canopy-forming algae, restoration, ex situ, cultivation, Mediterranean Sea
Objavljeno v DiRROS: 17.07.2024; Ogledov: 36; Prenosov: 2
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10. Assessment of different experimental setups to determineviral filtration efficiency of face masksArijana Filipić, Katja Fric, Maja Ravnikar, Polona Kogovšek, 2022, izvirni znanstveni članek Povzetek: Abstract
As a result of the COVID-19 pandemic, many new materials and masks came onto the market. To determine their suitability, several standards specify which properties to test, including bacterial filtration efficiency (BFE), while none describe how to determine viral filtration efficiency (VFE), a property that is particularly important in times of pandemic. Therefore, we focused our research on evaluating the suitability and efficiency of different systems for determining VFE. Here, we evaluated the VFE of 6 mask types (e.g., a surgical mask, a respirator, material for mask production, and cloth masks) with different filtration efficiencies in four experimental setups and compared the results with BFE results. The study included 17 BFE and 22 VFE experiments with 73 and 81 mask samples tested, respectively. We have shown that the masks tested had high VFE (>99% for surgical masks and respirators, ≥98% for material, and 87–97% for cloth masks) and that all experimental setups provided highly reproducible and reliable VFE results (coefficient of variation < 6%). Therefore, the VFE tests described in this study can be integrated into existing standards for mask testing.
Ključne besede: face masks, virus filtration efficiency, bacterial filtration efficiency, EN 14683:2019+AC:2019, air sampler
Objavljeno v DiRROS: 17.07.2024; Ogledov: 16; Prenosov: 3
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