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Mechanism of action, potency and efficacy : considerations for cell therapies
Carl G. Simon Jr., Erich H. Bozenhardt, Christina M. Celluzzi, David Dobnik, Melanie L. Grant, Uma Lakshmipathy, Thiana Nebel, 2024, izvirni znanstveni članek

Povzetek: One of the most challenging aspects of developing advanced cell therapy products (CTPs) is defining the mechanism of action (MOA), potency and efficacy of the product. This perspective examines these concepts and presents helpful ways to think about them through the lens of metrology. A logical framework for thinking about MOA, potency and efficacy is presented that is consistent with the existing regulatory guidelines, but also accommodates what has been learned from the 27 US FDA-approved CTPs. Available information regarding MOA, potency and efficacy for the 27 FDA-approved CTPs is reviewed to provide background and perspective. Potency process and efficacy process charts are introduced to clarify and illustrate the relationships between six key concepts: MOA, potency, potency test, efficacy, efficacy endpoint and efficacy endpoint test. Careful consideration of the meaning of these terms makes it easier to discuss the challenges of correlating potency test results with clinical outcomes and to understand how the relationships between the concepts can be misunderstood during development and clinical trials. Examples of how a product can be “potent but not efficacious” or “not potent but efficacious” are presented. Two example applications of the framework compare how MOA is assessed in cell cultures, animal models and human clinical trials and reveals the challenge of establishing MOA in humans. Lastly, important considerations for the development of potency tests for a CTP are discussed. These perspectives can help product developers set appropriate expectations for understanding a product’s MOA and potency, avoid unrealistic assumptions and improve communication among team members during the development of CTPs.
Ključne besede: cell therapy product, efficacy endpoint test, mechanism of action, potency test, metrology
Objavljeno v DiRROS: 27.05.2024; Ogledov: 41; Prenosov: 23
.pdf Celotno besedilo (4,26 MB)
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Genotoxicity and heating performance of VxFe3-xO4 nanoparticles in health applications
Beatriz Sanz-Sagué, Amaia Sáenz-Hernández, Bojana Žegura, Alja Štern, Katja Kološa, Iza Rozman, 2024, izvirni znanstveni članek

Povzetek: The applications of magnetic nanoparticles (MNPs) as biocatalysts in different biomedical areas have been evolved very recently. One of the main challenges in this field is to design affective MNPs surfaces with catalytically active atomic centres, while producing minimal toxicological side effects on the hosting cell or tissues. MNPs of vanadium spinel ferrite (VFe2O4) are a promising material for mimicking the action of natural enzymes in degrading harmful substrates due to the presence of active V5+ centres. However, the toxicity of this material has not been yet studied in detail enough to grant biomedical safety. In this work, we have extensively measured the structural, compositional, and magnetic properties of a series of VxFe3-xO4 spinel ferrite MNPs to assess the surface composition and oxidation state of V atoms, and also performed systematic and extensive in vitro cytotoxicity and genotoxicity testing required to assess their safety in potential clinical applications. We could establish the presence of V5+ at the particle surface even in water-based colloidal samples at pH 7, as well as different amounts of V2+ and V3+ substitution at the A and B sites of the spinel structure. All samples showed large heating efficiency with Specific Loss Power values up to 400 W/g (H0 = 30 kA/m; f = 700 kHz). Samples analysed for safety in human hepatocellular carcinoma (HepG2) cell line with up to 24h of exposure showed that these MNPs did not induce major genomic abnormalities such as micronuclei, nuclear buds, or nucleoplasmic bridges (MNIs, NBUDs, and NPBs), nor did they cause DNA double-strand breaks (DSBs) or aneugenic effects—types of damage considered most harmful to cellular genetic material. The present study is an essential step towards the use of these type of nanomaterials in any biomedical or clinical application.
Ključne besede: magnetic nanoparticles, vanadium ferrite, cytotoxicity, genotoxicity, specific power absorption, cell viability
Objavljeno v DiRROS: 23.05.2024; Ogledov: 59; Prenosov: 40
.pdf Celotno besedilo (8,91 MB)
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Too hot to handle : effects of water temperature on the early life stages of Gongolaria barbata (Fucales)
Ana Lokovšek, Valentina Pitacco, Annalisa Falace, Domen Trkov, Martina Orlando-Bonaca, 2024, izvirni znanstveni članek

Povzetek: Gongolaria barbata plays a crucial role as a habitat-forming Fucales species in the Mediterranean Sea, thriving in shallow, sheltered coastal regions, where it exhibits optimal growth in a temperature range of 10 to 25 °C. In the northern Adriatic Sea, a semi-enclosed part of the Mediterranean, there has been a remarkable increase in seawater temperatures in recent decades, often exceeding 28 °C in summer. These high temperatures pose a significant threat to the vulnerable early life stages of G. barbata. This study delves into the effects of four temperatures (15, 18, 24, and 28 °C) on the growth of G. barbata over its first 16 days, closely monitoring mortality, deformities, and overall survival. Our experiments reveal that higher temperatures can result in deformities and increased mortality of germlings. Notably, a temperature of 28 °C resulted in the death of all germlings within the first week, whereas those exposed to 24 °C survived until the second week, albeit with significant deformities prior to death. In contrast, germlings cultivated at 15 and 18 °C exhibited normal development with minimal deformities. These results highlight the susceptibility of the early life stages of G. barbata to temperature-induced stress and provide valuable insights into the potential consequences of rising seawater temperatures in the Mediterranean.
Ključne besede: sea ​​temperature, early stages of growth, deformations, macroalgae, climate change, Mediterranean Sea
Objavljeno v DiRROS: 17.05.2024; Ogledov: 86; Prenosov: 176
.pdf Celotno besedilo (2,43 MB)
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Two new species of Elaphoidella (Copepoda, Harpacticoida) from subterranean waters in Northeast Thailand, with a record of a gynandromorphic specimen and an up-to-date key to Elaphoidella species from Southeast Asia
Chaichat Boonyanusith, Anton Brancelj, La-orsri Sanoamuang, 2024, izvirni znanstveni članek

Povzetek: Two new species of copepods of the genus Elaphoidella Chappuis, 1929 were discovered in a cave and a spring in northeastern Thailand. The first species, E. phuphamanensis sp. nov., belongs to species-group VII sensu Lang. It is most similar to E. turgisetosa Petkovski, 1980 in the armament of the male third exopod of the fourth swimming leg and the shape and armament of the fifth swimming leg in both sexes. However, it is easily distinguished from other congeners by the segmentation of the first swimming leg, the endopod of the fourth swimming leg, and the armature of the third exopod of swimming legs 2–4 in both sexes. The second species, E. propecabezasi sp. nov., is located in species-group I sensu Lang, where the male does not have a transformed seta on the third exopod of the fourth swimming leg and the female fifth swimming leg has four baseoendopodal robust setae, unequal in length. It is most similar to E. cabezasi Petkovski, 1982 and E. paraaffinis Watiroyram, Sanoamuang and Brancelj, 2017 in having the same armature formula as endopods 1–2 of female swimming legs 1–4. However, the ornamentation of the anal operculum, the shape of the caudal ramus, and the armature of the fifth swimming leg in both sexes distinguish them from each other. A rare gynandromorphic specimen of E. propecabezasi sp. nov. was recorded, and a revised key to Elaphoidella species in Southeast Asia is provided.
Ključne besede: cave-dwelling copepods, endemic distribution, groundwater habitats, intersex, morphology, zoology, Southeast Asia
Objavljeno v DiRROS: 17.05.2024; Ogledov: 82; Prenosov: 236
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Environmental factors influence cross-talk between a heat shock protein and an oxidative stress protein modification in the lizard Gallotia galloti
Edward Gilbert, Anamarija Žagar, Marta López-Darias, Rodrigo Megía-Palma, Karen A. Lister, Max Dolton Jones, Miguel A. Carretero, Nina Guerra Serén, Pedro Beltran-Alvarez, Katharina C. Wollenberg-Valero, 2024, izvirni znanstveni članek

Povzetek: Better understanding how organisms respond to their abiotic environment, especially at the biochemical level, is critical in predicting population trajectories under climate change. In this study, we measured constitutive stress biomarkers and protein post-translational modifications associated with oxidative stress in Gallotia galloti, an insular lizard species inhabiting highly heterogeneous environments on Tenerife. Tenerife is a small volcanic island in a relatively isolated archipelago off the West coast of Africa. We found that expression of GRP94, a molecular chaperone protein, and levels of protein carbonylation, a marker of cellular stress, change across different environments, depending on solar radiation-related variables and topology. Here, we report in a wild animal population, cross-talk between the baseline levels of the heat shock protein-like GRP94 and oxidative damage (protein carbonylation), which are influenced by a range of available temperatures, quantified through modelled operative temperature. This suggests a dynamic trade-off between cellular homeostasis and oxidative damage in lizards adapted to this thermally and topologically heterogeneous environment.
Objavljeno v DiRROS: 17.05.2024; Ogledov: 85; Prenosov: 109
.pdf Celotno besedilo (1,83 MB)
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Identification of epigenetically regulated genes involved in plant-virus interaction and their role in virus-triggered induced resistance
Régis L. Corrêa, Denis Kutnjak, Silvia Ambrós, Mónica Bustos, Santiago F. Elena, 2024, izvirni znanstveni članek

Povzetek: Background: Plant responses to a wide range of stresses are known to be regulated by epigenetic mechanisms. Path-ogen-related investigations, particularly against RNA viruses, are however scarce. It has been demonstrated that Arabi-dopsis thaliana plants defective in some members of the RNA-directed DNA methylation (RdDM) or histone modi-fication pathways presented differential susceptibility to the turnip mosaic virus. In order to identify genes directly targeted by the RdDM-related RNA Polymerase V (POLV ) complex and the histone demethylase protein JUMONJI14 (JMJ14) during infection, the transcriptomes of infected mutant and control plants were obtained and integrated with available chromatin occupancy data for various epigenetic proteins and marks. Results: A comprehensive list of virus-responsive gene candidates to be regulated by the two proteins was obtained. Twelve genes were selected for further characterization, confirming their dynamic regulation during the course of infection. Several epigenetic marks on their promoter sequences were found using in silico data, raising confidence that the identified genes are actually regulated by epigenetic mechanisms. The altered expression of six of these genes in mutants of the methyltransferase gene CURLY LEAF and the histone deacetylase gene HISTONE DEACETYLASE 19 suggests that some virus-responsive genes may be regulated by multiple coordinated epigenetic complexes. A temporally separated multiple plant virus infection experiment in which plants were transiently infected with one virus and then infected by a second one was designed to investigate the possible roles of the identified POLV- and JMJ14-regulated genes in wild-type (WT ) plants. Plants that had previously been stimulated with viruses were found to be more resistant to subsequent virus challenge than control plants. Several POLV- and JMJ14-regulated genes were found to be regulated in virus induced resistance in WT plants, with some of them poisoned to be expressed in early infection stages. Conclusions: A set of confident candidate genes directly regulated by the POLV and JMJ14 proteins during virus infection was identified, with indications that some of them may be regulated by multiple epigenetic modules. A sub-set of these genes may also play a role in the tolerance of WT plants to repeated, intermittent virus infections.Keywords Biotic stress, Defense priming, Epigenetics, Histone modifications, Induced resistance, Potyvirus, RNA-directed DNA methylation.
Ključne besede: biotic stress, defense priming, epigenetics, histone modifications, induced resistance, Potyvirus, RNA-directed DNA methylation
Objavljeno v DiRROS: 17.05.2024; Ogledov: 78; Prenosov: 769
.pdf Celotno besedilo (6,66 MB)
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First report of collar and root rot of lettuce caused by Plectosphaerella cucumerina in Serbia
Milica Mihajlović, Jovana Hrustić, Ana Vučurović, Mila Grahovac, Dragana Budakov, Tatjana Dudaš, Brankica Pešić, 2024, drugi znanstveni članki

Povzetek: In March 2021, unusual plant stuning, collar, and wet root rot of lettuce (Lactuca sativa L.) during the rosette stage was observed in two commercial fields in Serbia (44°58'N, 20°32'E; 44°45'N, 20°43'E). Disease incidence in the fields (≈ 0.9 ha each) was approximately 15 and 20%, respectively. Initial above-ground symptoms were yellowing and wilting of leaves, while below-ground symptoms were collar, wet root rot, and lesions becoming necrotic. Eventually, whole plants wilted, collapsed, and died. A total of 35 symptomatic plants were collected from the fields, and diseased tissues were cut into small pieces, surface sterilized, and plated on potato dextrose agar (PDA). Isolation resulted in 20 morphologically uniform monoconidial isolates. The isolates formed white to creamy colonies, gradually becoming salmon pink, slimy, or moist in appearance, with sparse aerial mycelia. Numerous hyphal coils with conidiophores and hyaline, smooth-surfaced, ellipsoid to ovoid, septate or aseptate conidia were formed (4.5 to 10.1×1.2 to 3.7 μm (n = 100)). To confirm the species identity, the internal transcribed spacer (ITS) region and the D1/D2 region of a selected representative isolate 13-3-c were amplified and sequenced by using primer pairs ITS1/ITS4 (White et al. 1990) and N1/N2 (O’Donnell and Gray 1995), respectively. The sequences were deposited in GenBank (ITS: OR880564 and D1/D2: OR880567). Sequence analysis revealed 100% nucleotide identity with P. cucumerina isolates from different countries deposited in the NCBI GenBank, including isolate MH860704 (Vu et al. 2019) (ITS region) and isolate KY662256 (Su et al. 2017) (D1/D2 region). Neighbor-joining analysis was conducted based on the combined ITS and D1/D2 regions, and the tree was constructed with the substitution models (1,000 bootstrap). The combined phylogeny confirmed that the sequences shared a common clade with P. cucumerina. Hence, morphological, microscopic, and molecular characterization confirmed the pathogen as P. cucumerina (Palm et al., 1995; Carlucci et al., 2012). In a pathogenicity assay, 10 isolates were tested. Five 30-day-old lettuce plants (cv. Majska Kraljica) per isolate were root-dipped in the conidial suspensions (1×105 conidia/ml). The 10 inoculated plants were transplanted into 1 L pots containing sterile substrate (Floragard, Germany). Plants treated with sterile distilled water were used as controls. Plants were maintained in a greenhouse at 25 to 28°C under a 12-hour photoperiod (Cai et al., 2021). Four weeks after inoculation, stunting, chlorosis, and wilting of plants were observed, while collars and roots exhibited typical decaying symptoms. No symptoms were observed on the control plants. The pathogen was reisolated from symptomatic tissue as previously described. Koch's postulates were completed by confirming the identity of reisolates based on morphological features. To our knowledge, this is the first report of P. cucumerina on lettuce or any other crop in Serbia. P. cucumerina is already known as a pathogen of lettuce and other hosts grown in many countries worldwide, as well as in some European countries (Belgium, England, Italy, the Netherlands, and Switzerland) (Zhang et al. 2019). This emerging pathogen may cause significant economic losses in lettuce production in Serbia and in the entire Balkan region. Our results may help to develop effective management strategies based on accurate and timely identification and regular pathogen monitoring.
Ključne besede: detection, identification, salat, plant diseases
Objavljeno v DiRROS: 17.05.2024; Ogledov: 67; Prenosov: 29
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A global database of dissolved organic matter (DOM) concentration measurements in coastal waters (CoastDOM v1)
Christian Lønborg, Cátia Carreira, Gwenaël Abril, Katja Klun, Tinkara Tinta, 2024, izvirni znanstveni članek

Povzetek: Measurements of dissolved organic carbon (DOC), nitrogen (DON), and phosphorus (DOP) concentrations are used to characterize the dissolved organic matter (DOM) pool and are important components of biogeochemical cycling in the coastal ocean. Here, we present the first edition of a global database (CoastDOM v1; available at, Lønborg et al., 2023) compiling previously published and unpublished measurements of DOC, DON, and DOP in coastal waters. These data are complemented by hydrographic data such as temperature and salinity and, to the extent possible, other biogeochemical variables (e.g. chlorophyll a, inorganic nutrients) and the inorganic carbon system (e.g. dissolved inorganic carbon and total alkalinity). Overall, CoastDOM v1 includes observations of concentrations from all continents. However, most data were collected in the Northern Hemisphere, with a clear gap in DOM measurements from the Southern Hemisphere. The data included were collected from 1978 to 2022 and consist of 62 338 data points for DOC, 20 356 for DON, and 13 533 for DOP. The number of measurements decreases progressively in the sequence DOC > DON > DOP, reflecting both differences in the maturity of the analytical methods and the greater focus on carbon cycling by the aquatic science community. The global database shows that the average DOC concentration in coastal waters (average ± standard deviation (SD): 182±314 µmolC L−1; median: 103 µmolC L−1) is 13-fold higher than the average coastal DON concentration (13.6 ± 30.4 µmol N L−1; median: 8.0 µmol N L−1), which is itself 39-fold higher than the average coastal DOP concentration (0.34 ± 1.11 µmol P L−1; median: 0.18 µmol P L−1). This dataset will be useful for identifying global spatial and temporal patterns in DOM and will help facilitate the reuse of DOC, DON, and DOP data in studies aimed at better characterizing local biogeochemical processes; closing nutrient budgets; estimating carbon, nitrogen, and phosphorous pools; and establishing a baseline for modelling future changes in coastal waters.
Ključne besede: global database, dissolved organic matter, coastal waters, marine biology
Objavljeno v DiRROS: 17.05.2024; Ogledov: 77; Prenosov: 209
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Annual recurrence of prokaryotic climax communities in shallow waters of the North Mediterranean
Mauro Celussi, Vincenzo Manna, Elisa Banchi, Viviana Fonti, Matteo Bazzaro, Vesna Flander-Putrle, Katja Klun, Martina Kralj, Neža Orel, Tinkara Tinta, 2024, izvirni znanstveni članek

Povzetek: In temperate coastal environments, wide fluctuations of biotic and abiotic factors drive microbiome dynamics. To link recurrent ecological patterns with planktonic microbial communities, we analysed a monthly-sampled 3-year time series of 16S rRNA amplicon sequencing data, alongside environmental variables, collected at two stations in the northern Adriatic Sea. Time series multivariate analyses allowed us to identify three stable, mature communities (climaxes), whose recurrence was mainly driven by changes in photoperiod and temperature. Mixotrophs (e.g., Ca. Nitrosopumilus, SUP05 clade, and Marine Group II) thrived under oligotrophic, low-light conditions, whereas copiotrophs (e.g., NS4 and NS5 clades) bloomed at higher temperatures and substrate availability. The early spring climax was characterised by a more diverse set of amplicon sequence variants, including copiotrophs associated with phytoplankton-derived organic matter degradation, and photo-auto/heterotrophic organisms (e.g., Synechococcus sp., Roseobacter clade), whose rhythmicity was linked to photoperiod lengthening. Through the identification of recurrent climax assemblages, we begin to delineate a typology of ecosystem based on microbiome composition and functionality, allowing for the intercomparison of microbial assemblages among different biomes, a still underachieved goal in the omics era.
Ključne besede: marine microbial ecology, coastal seas, microbial community analyses, marine biology, marine ecology
Objavljeno v DiRROS: 17.05.2024; Ogledov: 94; Prenosov: 111
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Discovering unknown Madagascar biodiversity : integrative taxonomy of raft spiders (Pisauridae: Dolomedes)
Kuang-Ping Yu, Matjaž Kuntner, 2024, izvirni znanstveni članek

Povzetek: Madagascar is a global biodiversity hotspot, but its biodiversity continues to be underestimated and understudied. Of raft spiders, genus Dolomedes Latreille, 1804, literature only reports two species on Madagascar. Our single expedition to humid forests of eastern and northern Madagascar, however, yielded a series of Dolomedes exemplars representing both sexes of five morphospecies. To avoid only using morphological diagnostics, we devised and tested an integrative taxonomic model for Dolomedes based on the unified species concept. The model first determines morphospecies within a morphometrics framework, then tests their validity via species delimitation using COI. It then incorporates habitat preferences, geological barriers, and dispersal related traits to form hypotheses about gene flow limitations. Our results reveal four new Dolomedes species that we describe from both sexes as Dolomedes gregoric sp. nov., D. bedjanic sp. nov., D. hydatostella sp. nov., and D. rotundus sp. nov. The range of D. kalanoro Silva & Griswold, 2013, now also known from both sexes, is expanded to eastern Madagascar. By increasing the known raft spider diversity from one valid species to five, our results merely scratch the surface of the true Dolomedes species diversity on Madagascar. Our integrative taxonomic model provides the framework for future revisions of raft spiders anywhere.
Ključne besede: raft spiders, integrative taxonomic model, Madagascar, biodiversity, integrative taxonomy, zoology
Objavljeno v DiRROS: 16.05.2024; Ogledov: 100; Prenosov: 99
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