Publications

Abstract Currently, salinity and heat are two critical threats to crop production and food security which are being aggravated by the global climatic instability. In this scenario, it is imperative to understand plant responses to simultaneous exposure to different stressors and the cross-talk between underlying functional mechanisms. Thus, in this study, the physiological and biochemical responses of tomato plants (Solanum lycopersicum L.) to the combination of salinity (100 mM NaCl) and heat (42 degrees C; 4 h/day) stress were evaluated. After 21 days of co-exposure, the accumulation of Na+ in plant tissues was superior when salt-treated plants were also exposed to high temperatures compared to the individual saline treatment, leading to the depletion of other nutrients and a harsher negative effect on plant growth. Despite that, neither oxidative damage nor a major accumulation of reactive oxygen species took place under stress conditions, mostly due to the accumulation of antioxidant (AOX) metabolites alongside the activation of several AOX enzymes. Nonetheless, the plausible allocation of resources towards the defense pathways related to oxidative and osmotic stress, along with severe Na toxicity, heavily compromised the ability of plants to grow properly when the combination of salinity and heat was imposed.

Abstract Aluminum (Al) toxicity limits crops growth and production in acidic soils. Compared to roots, less is known about the toxic effects of Al in leaves. Al subcellular compartmentalization is also largely unknown. Using rye (Secale cereale L.) Beira (more tolerant) and RioDeva (more sensitive to Al) genotypes, we evaluated the patterns of Al accumulation in leaf cell organelles and the photosynthetic and metabolic changes to cope with Al toxicity. The tolerant genotype accumulated less Al in all organelles, except the vacuoles. This suggests that Al compartmentalization plays a role in Al tolerance of Beira genotype. PSII efficiency, stomatal conductance, pigment biosynthesis, and photosynthesis metabolism were less affected in the tolerant genotype. In the Calvin cycle, carboxylation was compromised by Al exposure in the tolerant genotype. Other Calvin cycle-related enzymes, phoshoglycerate kinase (PGK), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), triose-phosphate isomerase (TPI), and fructose 1,6-bisphosphatase (FBPase) activities decreased in the sensitive line after 48 h of Al exposure. Consequentially, carbohydrate and organic acid metabolism were affected in a genotype-specific manner, where sugar levels increased only in the tolerant genotype. In conclusion, Al transport to the leaf and compartmentalization in the vacuoles tolerant genotype's leaf cells provide complementary mechanisms of Al tolerance, protecting the photosynthetic apparatus and thereby sustaining growth.

Abstract Purpose Soil is a complex open system covering various physical and chemical attributes. In soil testing, multivariate analysis (MVA) has an important application because it allows the interpretation of a large amount of data for the design of relevant environmental scenarios. The purpose of this research is to summarize recent applications of MVA for identifying soil types or characteristics and for predicting soil attributes with a critical evaluation. Methods Based on a comprehensive search of the available database, in this review, we have provided updated information on the most representative classification and regression MVA applied in the past decade in soil surveys. Regression MVA were compared in terms of applicability, efficiency, and predictive power of different soil attributes. Results Principal component analysis (PCA) allows the grouping of soils into independent clusters according to their differences in texture or physicochemical composition, which may mirror local or regional environmental signatures. PCA is also used to reduce the dimensionality of spectral data before their application in regression MVA. Partial least square regression (PLSR) is the most commonly applied regression MVA for predicting soil attributes after the correlation of spectra (e.g., Vis-NIR) vs. conventional analysis results. The resulting PLSR models, evaluated by correlation coefficient (R-2) and root mean squared error (RMSE), can be valid for the estimation of several soil attributes (e.g., organic carbon, clay). Conclusions Application of regression MVA may have limitations in predicting some soil attributes. Objective interpretation of the dynamic nature of soils requires the selection of representative samples as well as appropriate MVA, which can have significant potential in an effective soil survey.

Abstract Since metal contamination compromises crop growth and food safety, eco-friendly responses to prevent this are needed. This study provided an integrative evaluation of the potential of wrack (macroalgae debris) to increase the tolerance of Hordeum vulgare L. (barley plant) to copper (Cu).Plants were grown in a soil mixed with 219 mg Cu kg(-1) with/without 2% (m/m) wrack for 14 days. Copper impaired all growth-related parameters. Wrack application counteracted most of these negative impacts and lowered metal accumulation in roots. Metal exposure increased reactive oxygen species content [superoxide anion (O-2(center dot-)) and hydrogen peroxide [H2O2]) and lipid peroxidation degree, which was evaluated through the quantification of malondialdehyde, while co-treatment with wrack partially reverted some of these effects. The non-enzymatic antioxidant (AOX) system was mainly activated by Cu, with accumulated glutathione (GSH), ascorbate (AsA) and phenols, mostly in roots, while proline content was reduced. Wrack protective action was through a modulation of GSH and AsA redox state and enhanced ascorbate peroxidase. The results suggest wrack's potential to alleviate Cu-induced phytotoxicity, which probably relies on the reduction of Cu-induced oxidative stress through a more efficient activity of AOX metabolites and by limiting Cu absorption and bioaccumulation, especially in roots of barley plants.

Abstract Plant biostimulants such as seaweed extracts, present a sustainable alternative to agrochemicals. Moreover, the accumulation of beach-cast seaweed (wrack), due to climate change and anthropic pressures, is expected to increase in the coming decades. Thus, from a perspective of circular economy, based on the valorisation of an organic residue, this study aimed at achieving an elemental and biochemical characterisation of beach wrack and understanding the effects of an aqueous extract prepared from this residue on the mitigation of metal-induced stress, using barley (Hordeum vulgare L.) as a model. The quantification of wrack's macro- and micronutrients showed that K, Ca and Na were the most abundant elements, being this composition similar to that of other organic fertilisers. Furthermore, despite the studied wrack having lower values of photosynthetic pigments, amino acids and sugars, higher amounts of phenols and flavonoids (8.35 +/- 0.24 and 3.95 +/- 1.22 mg g(-1) dry matter respectively) were detected when compared to freshly collected seaweeds. This work highlighted that wrack showed potential as a fertiliser-through increasing root biomass (63%) and leaf biometry (up to 45%) in plants treated with 5.0 and 7.5 g L-1 wrack extract alone-also being a possible cost-effective, eco-friendly and sustainable biostimulant to mitigate the phytotoxic effects of Cu, since plants treated with 7.5 g wrack L-1 showed an increase of 34% in leaf length, when compared to seedlings exposed only to Cu.

Abstract Throughout their life cycle, plants are subjected to a variety of environmental constraints, including abiotic stresses. The present study aimed at characterizing the responses of the two tomato cultivars Gold Nugget (GN) and Purple Calabash (PC) exposed to a combination of nickel (Ni) and drought. The following hypotheses were pursued: (i) the activation of responses to one stressor eases further adjustments to a second stressor; and (ii) the two tomato cultivars are differentially susceptible to drought and heavy metal-stress. Besides biometrical evaluations, the distribution of Ni in tissues and the redox homeostasis in both cultivars were compared in response to Ni-stress, polyethylene glycol (PEG)-induced drought, and to their combination. Regarding single stresses, Ni caused more harmful effects to plants than PEG-induced drought, in terms of growth inhibition and production of reactive oxygen species. Ni was mostly accumulated in the roots. The GN cultivar promptly activated antioxidant defenses under Ni-stress, while, in PC, such antioxidants were more strongly induced under combined stress. Stress co-exposure led to a drastic proline accumulation, resembling a signal of stress sensitivity. Overall, the GN cultivar seemed to be less susceptible to the combined stress than PC, as it could activate stronger antioxidant defenses under single Ni toxicity, possibly easing further adjustments demanded by the later co-exposure to drought. This study showed that the two cultivars of the same species had different levels of perception and responsiveness to Ni-induced stress, which translated into different susceptibilities to the combined exposure to PEG-induced drought. [GRAPHICS] .

Abstract One of the major assets of anticancer nanomedicine is the ability to co-deliver drug combinations, as it enables targeting of different cellular populations and/or signaling pathways implicated in tumorigenesis and thus tackling tumor heterogeneity. Moreover, drug resistance can be circumvented, for example, upon co-encapsulation and delivery of doxorubicin and sphingolipids, as ceramides. Herein, the impact of short (C6) and long (C18) alkyl chain length ceramides on the nature of drug interaction, within the scope of combination with doxorubicin, was performed in bulk triple-negative breast cancer (TNBC) cells, as well as on the density of putative cancer stem cells and phenotype, including live single-cell tracking.& nbsp;C6-or C18-ceramide enabled a synergistic drug interaction in all conditions and (bulk) cell lines tested. However, differentiation among these two ceramides was reflected on the migratory potential of cancer cells, particularly significant against the highly motile MDA-MB-231 cells. This effect was supported by the down-regulation of the PI3K/Akt pathway enabled by C6-ceramide and in contrast with C18-ceramide. The decrease of the migratory potential enabled by the targeted liposomal combinations is of high relevance in the context of TNBC, due to the underlying metastatic potential.& nbsp;Surprisingly, the nature of the drug interaction assessed at the level of bulk cancer cells revealed to be insufficient to predict whether a drug combination enables a decrease in the percentage of the master regulators of tumor relapse as ALDH(+)/(high) putative TNBC cancer stem cells, suggesting, for the first time, that it should be extended further down to this level.

Abstract Artificial receptors that mimic their natural biological counterparts have several advantages, such as lower production costs and increased shelf-life stability/versatility, while overcoming the ethical issues related to raising antibodies in animals. In this work, the proposed tailor-made molecularly imprinted polymer (MIP)-allergen receptors aimed at substituting or even transcending the performance of biological antibodies. For this purpose, a MIP was proposed as an artificial antibody for the recognition of hazelnut Cor a 14-allergen. The target protein was grafted onto the conducting polypyrrole receptor film using gold screen-printed electrodes (Au-SPE). The electrochemical assessment presented a linear response for the dynamic range of 100 fg mL(-1)-1 mu g mL(-1) and a LOD of 24.5 fg mL(-1), as determined by square wave voltammetry from the calibration curves prepared with standards diluted in phosphate buffer. Surface plasmon resonance (SPR) was used as a secondary transducer to evaluate the performance of the Cor a 14-MIP sensor, enabling a linear dynamic range of 100 fg mL(-1) - 0.1 mu g mL(-1) and a LOD of 18.1 fg mL(-1). The selectivity of the tailored-made Cor a 14-MIP was tested against potentially cross-reactive plant/animal species based on the rebinding affinity (Freundlich isotherm-K-F) of homologues/similar proteins, being further compared with custom-made polyclonal anti-Cor a 14 IgG immunosensor. Results evidenced that the MIP mimics the biorecognition of biological antibodies, presenting higher selectivity (only minor cross-reactivity towards walnut and Brazil nut 2S albumins) than the Cor a 14/anti-Cor a 14 IgG immunosensor. The application of electrochemical Cor a 14-MIP sensor to model mixtures of hazelnut in pasta enabled quantifying hazelnut down to 1 mg kg(-1) (corresponding to 0.16 mg kg(-1) of hazelnut protein in the matrix). To the best of our knowledge, Cor a 14-MIP is the first sensor based on an artificial/synthetic biorecognition platform for the specific detection of hazelnut allergens, while presenting high-performance parameters with demonstrated application in food safety management.

Abstract Graphene and its derivatives are generally portrayed as electron transfer enhancers that effectively boost the electrochemical response of classic electrodes for applications in renewable energy, electronics, or analysis (amongst others). However, a number of fundamental studies have challenged this view. In certain reports, not only could no beneficial effect be demonstrated, but the opposite was concluded. If we want to advance towards a more rational design of high-performance electrode devices, these discrepancies need to be cleared and the fundamental aspects of electron transfer reactions through graphene-electrodes further understood. The present study contributes to this cause by exploring the relationships between the structure and morphological appearance of graphene films and their electrochemical performance in fundamental proof-of-concept experiments. The results unveil that important differences in the structure and morphology of the films (which are tightly related to the composition and load of graphene materials) govern the electrochemical response of the modified electrodes. Thereby, a possible explanation for the apparently contradictory conclusions reported in the literature is provided.

Abstract Optosensing chitosan-based membranes have been applied for the detection of heavy metals, especially in drinking water. The novelty of this study is based on the use of sulphated polysaccharides, in such optosensing membranes, aiming at an improved analytical performance. The sulphated polysaccharides, such as ulvan, fucoidan and chondroitin sulfate, were extracted from by-products and wastes of marine-related activities. The membranes were developed for the analysis of aluminum. The variation in the visible absorbance of the sensor membranes after the contact between the chromophore and the aluminum cation was studied. The membranes containing sulphated polysaccharides showed improved signals when compared to the chitosan-only membrane. As for the detection limits for the membranes containing ulvan, fucoidan and chondroitin sulfate, 0.17 mg L-1, 0.21 mg L-1 and 0.36 mg L-1 were obtained, respectively. The values were much lower than that obtained for the chitosan-only membrane, 0.52 mg L-1, which shows the improvement obtained from the sulphated polysaccharides. The results were obtained with the presence of CTAB in analysis solution, which forms a ternary complex with the aluminum cation and the chromophore. This resulted in an hyperchromic and batochromic shift in the absorption band. When in the presence of this surfactant, the membranes showed lower detection limits and higher selectivity.