1. Early stress detection in forest trees using a nanobody-functionalized electrochemical biosensor for ascorbate peroxidaseClaudia D'Ercole, Rossella Svigelj, Tanja Mrak, Ario De Marco, 2025, izvirni znanstveni članek Opis: Forest environments are exposed to multiple stressful factors of both abiotic and biotic nature such as heavy metal contamination, drought, or pest infestations which may lead to their massive decline. We designed a comprehensive approach for isolating, producing and functionalizing reagents suitable for the affordable detection of forest plant stress biomarkers with the aim to provide quantitative data to assess plant stress fluctuation and, possibly, to design mitigation strategies. We first optimized a panning protocol to recover nanobodies targeting shared sequences that could cross-react with both Pisum sativum and Populus nigra ascorbate peroxidase (APX). After their production as recombinant constructs and their extensive biophysical and biochemical characterization, such reagents were exploited as the immunocapture element of an electrochemical biosensor conceived as a potential point-of-care device. Such biosensor could detect both pea and poplar APX in leaf extracts and could be used to clearly discriminate between control and heavy metal-stressed poplar plants based on their APX activity, even before the appearance of any phenotypic symptom. The combination of fast and inexpensive reagent production with the development of portable diagnostics opens the opportunity for large-scale, on-site surveys of forest trees. Ključne besede: plant stress, scavengers, diagnostics, nanobodies, biosensors Objavljeno v RUNG: 11.04.2025; Ogledov: 217; Prenosov: 2
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3. Development of molecular methods for the comprehensive evaluation of stress conditions affecting forest trees : dissertationClaudia D'Ercole, 2025, doktorska disertacija Ključne besede: protein complexes, adhirons, nanobodies, plant stress, immunoreagents, protoplasts, ascorbate peroxidase, dissertations Objavljeno v RUNG: 04.02.2025; Ogledov: 656; Prenosov: 9
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4. Stress and morphology in the Italian verbsElena Guerzoni, 2024, druge monografije in druga zaključena dela Opis: In this paper I present an analysis of stress in Italian verbs. A preliminary observation reveals that
inflection affects stress position on verbs in an apparently unpredictable fashion. Although, at first
sight, the variability of stress placement in Italian verbs seems to preclude a systematic account, this
paper shows that once the morphological complexity of verbs is factored in, it is in fact the one and
the same algorithm that assigns stress in this entire word class. Ključne besede: Italian verb, tense, stress, phonology, morphology Objavljeno v RUNG: 31.07.2024; Ogledov: 1306; Prenosov: 0 Gradivo ima več datotek! Več... |
5. Placement and ordering of the (en)cliticsFranc Marušič, Petra Mišmaš, Rok Žaucer, 2024, samostojni znanstveni sestavek ali poglavje v monografski publikaciji Ključne besede: clitics, enclitics, Slavic, syntax, Slovenian, Slovak, Czech, Bulgarian, Macedonian, BCMS, Polish, clitic cluster, stress Objavljeno v RUNG: 31.05.2024; Ogledov: 1623; Prenosov: 8
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6. Instruct-ERIC network : biophysical characterization of antigen-nanobody complexesClaudia D'Ercole, 2024, objavljeni povzetek znanstvenega prispevka na konferenci Opis: Forest environments are exposed to multiple stressful factors of both abiotic and biotic nature which may lead to their massive decline [1]. Understanding the molecular mechanisms of specific stress conditions and monitoring the fluctuations of reliable forest plant biomarkers with affordable methods would be instrumental for assessing stress levels over the time. Ascorbate peroxidase (APX) represents a suitable plant biomarker. APX is a hydrogen peroxide-scavenging enzyme the critical role of which has been described in several plants, both herbaceous and woody. Its activity generally increases under oxidative stress during which its peroxide detoxifying function is part of the wider ascorbate-glutathione cycle [2]. The development of reagents to detect such fluctuations would help the evaluation of plant physiological conditions. In this study, nanobodies (Nbs) targeting APX have been identified. Nbs correspond to the variable domain of heavy chain-only antibodies derived from camelids. They are small (15 kDa), stable, and can be easily produced in bacteria fused to different protein tags according to the downstream applications [3]. After their isolation by biopanning against soluble APX, they have been produced and underwent a biophysical characterization in combination with their antigen (APX-Nb complex) to identify the best binders in terms of stability and affinity. The protein complex characterization was supported by Instruct-ERIC and mainly performed at the BIOCEV institute of Prague. Data from Mass Photometry and Dynamic Light scattering evidenced the formation of the protein complexes, whereas the preliminary data of Hydrogen-Deuterium Exchange Mass Spectrometry, performed with the aim of identifying the residues involved in the paratope/epitope interface, were insufficient to clarify the issue and rather suggested that the interaction has low affinity. This indication was then confirmed by ELISA assay. The combination of multiple methods allowed a comprehensive sample characterization which will require further structural analyses to provide a complete picture of the APX-Nb complex.
[1] G. Marie. B. C. M. B. C. Walters, “Forest decline and tree mortality in a southeastern Ohio oak-hickory forest,” Ohio Journal of Science , vol. 97, 1997.
[2] O. Chew, J. Whelan, and A. H. Millar, “Molecular Definition of the Ascorbate-Glutathione Cycle in Arabidopsis Mitochondria Reveals Dual Targeting of Antioxidant Defenses in Plants,” Journal of Biological Chemistry, vol. 278, no. 47, 2003, doi: 10.1074/jbc.M307525200.
[3] S. Muyldermans, “A guide to: generation and design of nanobodies,” FEBS J, vol. 288, no. 7, pp. 2084–2102, Apr. 2021, doi: 10.1111/febs.15515. Ključne besede: nanobody, ascorbate peroxidase, plant stress, protein complex, biophysical methodologies Objavljeno v RUNG: 31.05.2024; Ogledov: 1687; Prenosov: 0 Gradivo ima več datotek! Več... |
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8. Stress and morphology in the Italian verb systemElena Guerzoni, 2000, druge monografije in druga zaključena dela Opis: In this paper I present an analysis of stress in Italian verbs. A preliminary comparison between nouns and adjectives on the one hand and verbs on the other reveals that these the two classes of words differ in an interesting way when it comes to stress assignment: while on a noun or adjective stress can be predicted to a large extent on the basis of a stress rule that fully coincides with the main stress rule of Classical Latin, inflection affects stress position on verbs in an apparently unpredictable fashion. Although, at first sight, this difference seems to preclude a unified account of stress in the language, this paper shows that once the additional morphological complexity of verbs is factored in, it is in fact the same algorithm that assigns stress in both word classes. Ključne besede: Italian verb, tense, stress Objavljeno v RUNG: 27.02.2024; Ogledov: 2703; Prenosov: 0 Gradivo ima več datotek! Več... |
9. Database of the Western South Slavic verb HyperVerb 1.0Franc Marušič, Rok Žaucer, Petra Mišmaš, Boban Arsenijević, Marko Simonović, Stefan Milosavljević, Katarina Gomboc Čeh, Jelena Simić, 2022, zaključena znanstvena zbirka raziskovalnih podatkov Opis: Slavic, verb, Ključne besede: Slavic, verb, inflection, theme vowel, Slovenian, BCMS, Bosnian, Croatian, Serbian, verbal morphology, verbal stress pattern, verbal inflection Objavljeno v RUNG: 12.09.2022; Ogledov: 3081; Prenosov: 54
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10. Unraveling the Role of Major Facilitator Superfamily (MFS) Transporters in Plant Abiotic Stress ToleranceChristina Paparokidou, 2014, končno poročilo o rezultatih raziskav Opis: The yields of worldwide crop production are negatively affected by adverse environmental conditions, such as drought, salinity, heavy metal contamination and nutrient depletion in cultivated soils (Yan et al., 2013; Golldack et al., 2011; Yadav, 2010; Kobayashi and Nishizawa, 2012). This reduced crop production constitutes a major problem for food sustainability world-wide (Spiertz, 2013). Indeed, one of the major challenges for plant biotechnology will be to satisfy the increased demand for food on one hand, and to compensate for the loss of crop production on the other. Thus, the discovery of new plant genes that are able to cope with these conditions is critical and expected not only to elucidate the molecular mechanisms underlying crop abiotic stress, but also to pioneer genetic engineering strategies for improved crop productivity.
The goal of this project was to identify novel genes belonging to the Major Facilitator Superfamily (MFS) of plant membrane transporters with potential major roles in conferring abiotic stress tolerance, namely to heavy metal, osmotic and iron deficiency stress. We selected three Arabidopsis thaliana MFS transporter genes, provisionally called MFS10, MFS11 and MFS12, which based on publicly available microarray data display specific expression patterns suggesting roles in plant tolerance to different abiotic stresses.
RT-PCR analyses showed that in fact the MFS10 gene is highly induced by cadmium (Cd), selenium (Se), salt and mannitol, while the MFS11 gene is upregulated by abscisic acid (ABA) and glucose, and the MFS12 gene by iron (Fe) deficiency, ABA and glucose. Furthermore, subcellular localization of fluorescent reporter fusions indicated that the MFS10 and MFS11 gene products are both plasma membrane localized transporters, while the encoded product of the MFS12 gene appears to be an endoplasmic reticulum localized transporter. Finally, reverse genetics using a null mutant allele for the MFS10 gene demonstrated that it functions as a regulator of plant responses to hyperosmotic (drought and salt) stress. Ključne besede: Major Facilitator Superfamily (MFS), abiotic stress tolerance, heavy metals, osmotic stress, ABA stress, iron deficiency, T-DNA insertion lines, RT-PCR, confocal microscopy. Objavljeno v RUNG: 03.05.2022; Ogledov: 2726; Prenosov: 0 Gradivo ima več datotek! Več... |