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Bioactive potential of hazelnut husks was determined as a function of their cultivar source and extraction solvent. Hazelnut husks from four hazelnut cultivars (Butler, Grada de Viseu, Lansing and Morell) were picked in a hazelnut orchard at harvest and extracted with five solvents with different polarity: water, methanol, acetone, ethyl acetate and hexane. Phenolics were identified by HPLC–DAD and antioxidant activity was determined by three complementary methods: DPPH, FRAP and inhibition of lipid peroxidation. A total of 11 phenolics were identified in studied cultivars and grouped in five main classes namely, ellagitannin (ellagic acid), benzoic acids (gallic acid, protocatechuic acid and vanillic acid), flavonols (kaempferol-3,7-O-diglucoside, kaempferol-3-O-[6-acetylglucoside]-7-O-glucoside, kaempferol-3-O-[6acetylglucoside]-7-O-rhamnoside and quercetin-3-O-rutinoside), flavone (luteolin-7-O-rutinoside) and flavan-3-ol (epicatechin). Cultivar and extraction solvent influenced significantly (p

Background Various strategies are needed to mitigate the negative impact on or to increase fruit quality. The effect of spraying kaolin (K), Ascophyllum nodosum (An) and salicylic acid (SA), in trees with and without irrigation, on quality and sensorial attributes of hazelnut (Grada de Viseu cultivar) was investigated during two consecutive years (2016 and 2017) in a commercial orchard located in Moimenta da Beira, Portugal. Results The treatments affected positively the biometric parameters nut and kernel weight, length, width, thickness and volume as well as the vitamin E level, antioxidant activity and content of some individual phenolics, such as protocatechuic acid, gallocatechin, catechin and epicatechin. The levels of amino acids in hazelnut kernels decreased in all the assayed treatments, while the kernel colour and sensorial attributes were not affected by the treatments. Hazelnut physical properties (nut and kernels), chemical and phytochemical composition and antioxidant activities were positively related. Conclusions The application of K, An and SA improved the hazelnut tree response to climate change, without compromising the hazelnut chemical and sensorial quality. Furthermore, due to the similar observations for the same treatments with and without irrigation, it can be stated that K, An and SA can be efficient and cost-effective tools to mitigate summer stress in rain-fed orchards. © 2020 Society of Chemical Industry.

Chestnut fruits, from Castanea sativa Miller species, have been previously characterized mainly to study the nutritional and biochemical parameters, with mineral contents receiving less attention from the researchers, although these minerals are essential for several mechanisms in human body. In this study, a detailed elemental compositional analysis was performed using Proton Induced X-ray Emission (PIXE) to characterize two varieties of Castanea sativa Miller chestnuts (Longal and Judia). This study comprises the composition of the outer shells (pericarp), inner shells (episperm) and the edible fruits. From our knowledge, this is the first time that the presence of up to 20 elements on the shells and the fruits are reported. Furthermore, by PIXE it was possible to quantify them, allowing significant improvements in the assessment of chestnuts composition. Essential and nonessential elements were quantified on a food item that is used mainly for human consumption but it is also incorporated in animal feeding. Authors acknowledge R. Pinheiro for sample preparation. This work was developed within the Coordinated Research Project D61024 ‘‘Development of New Applications of Machine Generated Food Irradiation Technologies’’ financed by the International Atomic Energy Agency (IAEA). IST authors gratefully acknowledge the FCT (Fundação para a Ciência e a Tecnologia) support through the UID/Multi/04349/ 2013 and UID/FIS/50010/2013 projects. info:eu-repo/semantics/publishedVersion

The aim of this work was to study the impact of ionizing radiation (gamma radiation and electron beam, 1–3 kGy) on the antioxidants of the strawberry, through evaluation of the total phenolic and L-ascorbic acid contents, antioxidant activity, and cytotoxicity (293 T and A549 cell lines) after storage at 4 °C for 15 days. Results showed that both gamma and electron beam radiation could preserve the phenolic content of strawberries during the storage. Both irradiation treatments induced the degradation on L-ascorbic acid content and indicated to increase the antioxidant activity mainly on strawberries treated at 2 kGy. Regarding cytotoxicity assay, non-tumor 293 T cells were more sensitive to strawberry extracts than A549 cells and ionizing radiation had no impact on the viability of both cell lines. These results pointed out that gamma and electron-beam radiation at 2 kGy can be used as safe and environmental friendly post-harvest treatments of strawberries to preserve their bioactive properties.

Portobello variety of Agaricus bisporus mushrooms, appreciated for its taste, makes it desirable to be eaten fresh and also as flour in soups and gravies. Gamma and electron-beam radiation at four doses (1, 2, 5, and 10 kGy) were used to analyze its preservation effect on Portobello mushroom flour. A proximate analysis, as well as the impact on fatty acids, tocopherols, soluble sugars, organic acids, and ergosterol profiles, were performed every 3 months, during a storage period of 1 year. Gamma rays preserved mannitol (most abundant soluble sugar) over the 12 months, while electron beam radiation preserved organic acids. No significant changes were sought for any radiation type, and the slight changes extracted from the estimated marginal means reveal a tendency for irradiation as having preserving effects of nutrients and other important molecules. Thus, both irradiation types, up to 10 kGy are suitable for preservation of A. bisporus Portobello flour. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/AGR/00690/2020), C2TN (UID/Multi/04349/2013) for R.V.C. Cardoso’s grant (SFRH/BD/137436/2018), M. Carocho’s research contract. L. Barros and A. Fernandes also thank FCT’s funding through the institutional scientific employment program-contracts. This work is funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural R . Acknowledgments are also due European Agricultural Fund for Rural Development (EAFRD), through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). info:eu-repo/semantics/publishedVersion

Zearalenone (ZEA) is produced in cereals by different species of Fusarium, being a non-steroidal estrogenic mycotoxin. Despite having a low acute toxicity, ZEA strongly interferes with estrogen receptors. Gamma-radiation has been investigated to eliminate mycotoxins from food and feed, showing promising results. The present study aims to investigate the gamma-radiation effect on ZEA at different moisture conditions and to evaluate the cytotoxicity and estrogenicity of the irradiated ZEA. Different concentrations of dehydrated ZEA and aqueous solutions of ZEA were exposed to gamma-radiation doses ranging from 0.4 to 8.6 kGy and the mycotoxin concentration determined after exposure by high performance liquid chromatography (HPLC) with fluorescence detection. Following this, the cytotoxicity of irradiated samples was assessed in HepG2 cells, by measuring alterations of metabolic activity, plasma membrane integrity and lysosomal function, and their estrogenicity by measuring luciferase activity in HeLa 9903 cells. Gamma-radiation was found to be effective in reducing ZEA, with significant increases in degradation with increased moisture content. Furthermore, a reduction of cytotoxicity with irradiation was observed. ZEA estrogenicity was also increasingly reduced with increasing radiation doses, but mainly in aqueous solutions. These results suggest reduction of ZEA levels and of its toxicity in food and feed commodities may be achieved by irradiation., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, C2TN (UIDB/04349/2020) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte and by the INIA project RTA2012-00053-00-00., info:eu-repo/semantics/publishedVersion

The preserving capacity of gamma radiation (10 kGy) on the chemical, nutritional and antioxidant components of Aloysia citrodora Paláu, Melissa officinalis L., Melittis melissophyllum L. and Mentha piperita L., stored for 12 and 18 months, was evaluated. Despite the maintenance of the main characteristics during the first 12 months of storage, the additional 6 months induced several significant changes in individual compounds. In general, the analyzed species reacted dissimilarly throughout time, but it was possible to verify that the fatty acids, tocopherols and antioxidant capacity presented the most significant changes after 18 months of storage, inclusively in samples submitted to gamma radiation. In fact, the applied treatment (10 kGy) did not seem to be effective to prevent the decrease of free sugars, organic acids and tocopherols, especially considering the 18 months period. On the other hand, the evolution in color parameters indicated a greener color (yet slightly more yellow) among irradiated samples. Likewise, gamma radiation had a positive effect on oleic acid, β-carotene bleaching inhibition (in infusions), DPPH scavenging activity and reducing power (in methanolic extracts). Nevertheless, it might be generally concluded that gamma radiation is less suitable than electron-beam to maintain the characteristics of dried herbs during extended storage periods. The authors are grateful to the Ministry of Agriculture, Portugal (Project PRODER/FEADER/EU, AROMAP), for financial support of the work and E. Pereira grant. Authors also thank Foundation for Science and Technology (FCT, Portugal) for financial support to CIMO (UID/ AGR/00690/2013), REQUIMTE (UID/QUI/50006/2013 - POCI/01/ 0145/FEDER/007265), C2TN (RECI/AAG-TEC/0400/2012 and UID/ Multi/04349/2013 projects), J.C.M. Barreira (BPD/72802/2010) and L. Barros (SFRH/BPD/107855/2015) grants. The authors are also grateful to “MaisErvas - Aromáticas e Medicinais” for providing the plant material. info:eu-repo/semantics/publishedVersion

Ochratoxin A (OTA) is one of the main mycotoxins that can be found in food. The use of gamma radiation is a technique for preserving food that may exert some effects on mycotoxins. OTA was irradiated in its dry form, in aqueous and in methanolic solutions, and in wheat flour, grape juice and wine. Additionally, the toxicity of OTA irradiated in water was tested. In aqueous solutions, more than 90% of the OTA was degraded by -radiation doses 2.5 kGy, and a 2-fold reduction in OTA cytotoxicity was observed. In food matrices, the elimination of OTA by -radiation was found more difficult, as radiation doses of 30 kGy eliminate at most 24% of the OTA. Higher moisture content of food matrices did not substantially increase OTA elimination. It is concluded that OTA is very sensitive to irradiation in water solutions but resistant in its dry form and in food matrices., Thalita Calado received support through a grant SFRH/BD/79364/2011 from the Portuguese Foundation for Science and Technology (FCT –Portugal).LuísAbrunhosawassupportedbygrantUMINHO/BPD/51/ 2015 from project UID/BIO/04469/2013 financed by FCT/MEC (OE). This study was supported by the FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-010145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte; and under the scope of the projects RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and RTA2012-00053-00-00 INIA. The C2TN/IST authors gratefully acknowledge their FCT support through projects RECI/AAG-TEC/0400/2012 and UID/Multi/04349/ 2013. The authors are grateful to Luis Alté for his excellent technical assistance., info:eu-repo/semantics/publishedVersion

The aim of this work was to perform a comprehensive study on the effect of gamma radiation on strawberries by evaluating its effect on physicochemical, microbiological, sensory acceptance, phenolic and flavonoid contents, and antioxidant activity over a storage period of 14 days at 4 °C. The results showed that gamma irradiation significantly decreased the proliferation of molds and yeasts at 1, 2 and 3 kGy up to seven days of storage. However, a significant decrease of firmness, redness and chroma was noticed with gamma radiation doses. Moreover, sensory scores revealed that gamma radiation had no significant effect on strawberries compared to the non-treated, the dose of 2 kGy seemed to be the best dose to maintain the acceptance sensory quality of strawberries for the panel. A positive relation between different physicochemical parameters and sensory descriptors was illustrated with Principal Compounds Analysis. The gamma radiation treatment and refrigerated storage indicated to have an increasing trend on the bioactive content of strawberries. Based on the obtained results, the dose of 2 kGy point out to be the optimal treatment to maintain the safety and global quality of strawberries with extended shelf-life.

The adsorption of four phenolic compounds (gallic acid, protocatechuic acid, vanillic acid and syringic acid) is investigated using a synthesized mesoporous carbon on both single and multi-component synthetic solutions. Some correlation of the adsorption capacity of the carbon and the nature of adsorbate could be made, except for gallic acid whose concentration decrease seems to be not exclusively due to adsorption but also to polymerization reaction. In the multi-component mixture, negative effects in the adsorption capacity are observed probably due to competition for the active centers of the adsorbent surface. In desorption studies, ethanol presents better performance than water and acetonitrile. Vanillic acid is the compound with the higher adsorption and interestingly it is then possible to desorb a relatively high amount of it from the adsorbent, which may represent a possibility for a selective recovery of vanillic acid. These results present a potential way to treat the wastewater from the cork industry.

Gamma-irradiated sheep cream (1 kGy) was used to produce butter that was analysed during refrigerated storage up to 90 days in shelf-life extension studies including sensory, microbiological and water content assessments. The water content of the sheep butter made with irradiated fermented cream was 10% higher than that of control butter. A slower increase in bacterial load during storage was detected for the butter manufactured with irradiated fermented sheep cream compared with control butter; however, fungi increased at similar rates in both cream-irradiated and control butters during storage. The sensory evaluation suggested, in general, the acceptance of the irradiated samples by the panellists. The microbiocidal potential of gamma radiation of fermented sheep cream was highlighted, but was not reflected in extension of shelf-life of the butter produced. The traditional manufacturing process itself was found to be a major vehicle for microbial contamination of this dairy product, preventing its shelf-life extension.

Olive pomace is an environmentally detrimental waste from olive oil industry, containing large amounts of bioactive compounds that might be used by the food industry. In this work, the effects of gamma radiation on phenolic compounds and bioactive properties (antioxidant, antimicrobial activities and hepatotoxicity) of Crude Olive Pomace (COP) and Extracted Olive Pomace (EOP) extracts were evaluated. Hydroxytyrosol was the main phenolic compound identified in both olive pomace extracts (24–25 mg/g). The gamma radiation treatment of olive pomace improved at least 2-fold the extractability of phenolic compounds. Moreover, results suggested that gamma radiation at 5 kGy increased the antioxidant activity in EOP, while keeping the ability to protect erythrocytes against oxidation-induced haemolysis. Gamma radiation at 5 kGy could be a suitable technology for olive oil pomaceswaste valorization, contributing to enhance extraction of phenolic compounds and bioactive properties, especially when applied on extracted material. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support to C2TN UID/Multi/04349/2019, CIMO UID/AGR/00690/2019, J. Madureira [SFRH/BD/136506/2018]; L. Barros, J. Pinela and M.I. Dias thank the national funding by FCT, P.I., through the institutional scientific employment programcontract; FEDER-Interreg España-Portugal program for financial support through the project 0377_Iberphenol_6_E; “UCASUL - União de Cooperativas Agrícolas do Sul” agro industrial cooperative for providing the samples; to Technological Unit of Radiosterilization (University of Lisbon) for the samples irradiation; and to Molecular Materials Synthesis Laboratory (responsible Researcher Dr. Dulce Belo, C2TN) for the solvent evaporations. info:eu-repo/semantics/publishedVersion

Human epidermal growth factor 2 (HER2) is a ligand-free tyrosine kinase receptor of the HER family that is overexpressed in some of the most aggressive tumours. Although it is known that HER2 dimerization involves a specific region of its extracellular domain, the so-called &ldquo, dimerization arm&rdquo, the mechanism of dimerization inhibition remains uncertain. However, uncovering how antibody interactions lead to inhibition of HER2 dimerization is of key importance in understanding its role in tumour progression and therapy. Herein, we employed several computational modelling techniques for a molecular-level understanding of the interactions between HER and specific anti-HER2 antibodies, namely an antigen-binding (Fab) fragment (F0178) and a single-chain variable fragment from Trastuzumab (scFv). Specifically, we investigated the effects of antibody-HER2 interactions on the key residues of &ldquo, from molecular dynamics (MD) simulations of unbound HER (in a total of 1 &micro, s), as well as ScFv:HER2 and F0178:HER2 complexes (for a total of 2.5 &micro, s). A deep surface analysis of HER receptor revealed that the binding of specific anti-HER2 antibodies induced conformational changes both in the interfacial residues, which was expected, and in the ECDII (extracellular domain), in particular at the &ldquo, which is critical in establishing protein&ndash, protein interface (PPI) interactions. Our results support and advance the knowledge on the already described trastuzumab effect on blocking HER2 dimerization through synergistic inhibition and/or steric hindrance. Furthermore, our approach offers a new strategy for fine-tuning target activity through allosteric ligands.

Olive pomace, an environmentally detrimental residue generated during olive oil extraction, contains bioactive compounds in demand by the food industry. To valorize this waste product a suitable yield for the extraction process is required. Heat-assisted extraction of bioactive compounds from olive pomace was optimized by a circumscribed central composite design and response surface methodology. Our previous studies indicated that irradiation could improve 2.4-fold the extractability of the main phenolic compounds from olive pomace. The effect of extraction time, temperature and solvent concentration on the yield of polyphenols from irradiated olive pomace at 5 kGy was tested. Hydroxytyrosol-1- -glucoside, hydroxytyrosol, tyrosol and caffeic acid were quantified by High Performance Liquid Chromatography to calculate the total polyphenol content. The optimal general conditions by RSM modeling were extraction time of 120 min, temperature of 85 C, and 76% of ethanol in water. Using these selected conditions, 19.04 1.50 mg/g dry weight, 148.88 8.73 mg/g extract of total polyphenols were obtained, representing a yield of 13.7%, which was consistent with the value predicted by the model. This work demonstrated the potential of residues from the olive oil industry as a suitable alternative to obtain compounds that could be used as ingredients for the food industry. This work was funded by the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to C2TN (UIDB/04349/2020), CIMO (UIDB/00690/2020), J.M. (SFRH/BD/136506/2018); L.B. thank the national funding by FCT, P.I., through the institutional scientific employment program-contract; and the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project OliveBIOextract (NORTE-01-0247-FEDER-049865). The GIP-USAL is funded by the Strategic Research Programs for Units of Excellence (ref CLU-2018-04) and Consejería de Educación de la Junta de Castilla y León (Project SA093P20). info:eu-repo/semantics/publishedVersion

Irradiation is a feasible and safe decontamination technique, being applied to several types of foods including edible and medicinal plants. The aim of this study was to evaluate the effects of different gamma radiation doses (1, 5 and 10 kGy) on the individual profile of phenolic compounds determined by HPLC-DAD-ESI/MS, and the bioactive potential (cytotoxic, virucidal, and antimicrobial activities) of Aloysia citrodora Paláu (lemon verbena), Mentha × piperita L. (peppermint) and Thymus vulgaris L. (thyme). The observed cytotoxic activity varied with the plant and with the applied dose, being higher in Thymus vulgaris irradiated with 10 kGy. The virucidal activity was also dependent on the radiation dose, but was preserved with irradiation treatment. Gamma rays had no effect on the antimicrobial activity of the studied plants. Otherwise, the effects of gamma radiation on the phenolic profile were heterogeneous, with an increase in some compounds and decrease in others, depending on the species and on the radiation dose. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and the FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), C2TN (UID/Multi/04349/2013), L. Barros and R. C. Calhelha contracts. The authors are grateful to the FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. info:eu-repo/semantics/publishedVersion

The main objectives of the present work were as follows: (a) to develop an analytical procedure to quantify free sugars on chestnuts based on green-chemistry principles and (b) to evaluate the effect of hot air convective drying (50 °C along 10 hr) on the carbohydrate profile of sliced chestnuts (Longal and Judia varieties). The analytical method allowed the quantification of several sugars in chestnut extracts, with low detection limits and good precision. In fresh, Judia variety had higher sucrose, fructose, and glucose contents than Longal, together with amylose. Nevertheless, when applying hot air convective drying to chestnut slices, a significant increase in fructose contents was observed with time, particularly after 10 hr drying in Longal variety. Nonetheless, a significant increase in glucose content was observed on both varieties, probably related to starch thermal hydrolysis. Practical applications: An environment-friendly methodology for free sugars quantification, based on ethanol/water extraction (green solvents), ion-exchange high-performance liquid chromatography separation with water as mobile phase, and evaporative light scattering detection, was developed. The method allowed the quantification of raffinose, sucrose, glucose, and fructose in chestnut extracts, with detection limits of 6 μg for sucrose and 0.3 μg for other sugars. Good precision was achieved with intra-day and inter-day coefficients of variation below 5%. Regarding chestnuts' drying, hot air convective drying might be applied to chestnut slices but it may induce modifications on the carbohydrate composition of the samples, particularly when long drying periods are applied, requiring some attention because it might induce both visual and sensory changes. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), C2TN (UID/Multi/ 04349/2013), L. Barros and J. Barreira contracts, and A. Fernandes post-doc grant (SFRH/BPD/114753/2016). This work is funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural®; and to FEEI through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). This work was also developed within the Coordinated Research Project D61024 “Development of New Applications of Machine Generated Food Irradiation Technologies” financed by the International Atomic Energy Agency (IAEA). info:eu-repo/semantics/publishedVersion

The wastes generated during the olive oil extraction process, even if presenting a negative impact for the environment, contain several bioactive compounds that have considerable health benefits. After suitable extraction and purification, these compounds can be used as food antioxidants or as active ingredients in nutraceutical and cosmetic products due to their interesting technological and pharmaceutical properties. The aim of this review, after presenting general applications of the different types of wastes generated from this industry, is to focus on the olive pomace produced by the two-phase system and to explore the challenging applications of the main individual compounds present in this waste. Hydroxytyrosol, tyrosol, oleuropein, oleuropein aglycone, and verbascoside are the most abundant bioactive compounds present in olive pomace. Besides their antioxidant activity, these compounds also demonstrated other biological properties such as antimicrobial, anticancer, or anti-inflammatory, thus being used in formulations to produce pharmaceutical and cosmetic products or in the fortification of food. Nevertheless, it is mandatory to involve both industries and researchers to create strategies to valorize these byproducts while maintaining environmental sustainability. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to C2TN (UIDB/04349/2020), CIMO (UIDB/00690/2020) and Joana Madureira (SFRH/BD/136506/2018). Lillian Barros thank the national funding by FCT, P.I., through the institutional scientific employment program-contract. The authors thank the European Regional Development Fund (ERDF), through the Incentive System to Research and Technological development within the Portugal2020 Competitiveness and Internationalization Operational Program for the mobilizing project “BIOMA” (POCI- 01-0247-FEDER-046112). The Agroenvironment Unit is supported by Junta de Castilla y León (Escalera de Excelencia CLU-2018-04) co-funded by the P.O. FEDER of Castilla y León 2014–2020. info:eu-repo/semantics/publishedVersion

The suitability of post-packaging gamma radiation treatment for preserving total folates or vitamin B9 in watercress (Nasturtium officinale R. Br.) and buckler sorrel (Rumex induratus Boiss. & Reut.) during storage at 4 °C was evaluated. Comparable amounts of total folates were found in fresh, non-stored samples of both species. In watercress, the irradiation treatment of up to 5 kGy reduced the loss of total folates caused by 7 days of storage. In turn, the 12-day storage period did not affect the total folate content of buckler sorrel (while the 2 kGy dose decreased the initial levels), evidencing that packaging and refrigeration are enough for preservation. These results suggest that the suitability of post-packaging irradiation for preserving total folates may depend not only on the applied dose but also on the plant matrix under analysis. In addition, new data useful to complete food composition tables or databases is provided. The authors are grateful to the Foundation for Science and Technology ( FCT , Portugal) and FEDER under Programme PT2020 for financial support to CIMO ( UID/AGR/00690/2013 ), REQUIMTE/LAQV ( UID/QUI/50006/2013 – POCI/01/0145/FERDER/007265) and C2TN ( UID/Multi/04349/2013 ) and the Grant of J. Pinela (UID/AGR/00690/2013_DNAABN); and to the ALIMNOVA Research Group (Project UCM-252/2017). info:eu-repo/semantics/publishedVersion

In this work, we study degradation of clofibric acid (CFA) in aqueous solution using either ionizing radiation from a $^{60}$Co source or a non-thermal plasma produced by discharges in the air above the solution. The results obtained with the two technologies are compared in terms of effectiveness of CFA degradation and its by-products. In both cases the CFA degradation follows a quasi-exponential decay in time well modelled by a kinetic scheme which considers the competition between CFA and all reaction intermediates for the reactive species generated in solution as well as the amount of the end product formed. A new degradation law is deduced to explain the results. Although the end-product CO$_2$ was detected and the CFA conversion found to be very high under the studied conditions, HPLC analysis reveals several degradation intermediates still bearing the aromatic ring with the chlorine substituent. The extent of mineralization is rather limited. The energy yield is found to be higher in the gamma radiation experiments., 25 pages, 7 figures

Slaughterhouse industry produces large volumes of polluted wastewater, which cause negative impacts on the environment. The objective of this study was to assess the effect of electron-beam irradiation on the ecotoxicity of slaughterhouse effluents with absorbed doses up to 35 kGy. Two acute toxicity assays were applied to evaluate the efficiency of irradiation onto toxicity of wastewater. The exposed living-organisms were a luminescent bacteria Vibrio fischeri, and a freshwater microcrustacean Daphnia similis. Also, the total organic carbon was analysed in order to determine any possible organic carbon removal after irradiation. The ecotoxicological results evidenced that both living-organisms were suitable for the measurements. Therefore, the results demonstrated the toxicity of the effluent and its similarity for both organisms as well as the potential of radiation to reduce these effects. The 35 kGy dose was very effective for reducing toxic effects of slaughterhouse wastewater for daphnids suggesting that ionizing radiation could be used as a tool for removing toxic charge of such effluents. The type of contamination presented by the effluent justify the needs for alternatives of treatment.

Hexaploid tritordeum (HchHchAABB; 2n = 42) results from the cross between Hordeum chilense (HchHch; 2n = 14) and cultivated durum wheat (Triticum turgidum ssp. durum (AABB; 2n = 28). Morphologically, tritordeum resembles the wheat parent, showing promise for agriculture and wheat breeding. Start Codon Targeted (SCoT) polymorphism is a recently developed technique that generates gene-targeted markers. Thus, we considered it interesting to evaluate its potential for the DNA fingerprinting of newly synthesized hexaploid tritordeums and their respective parents. In this study, 60 SCoT primers were tested, and 18 and 19 of them revealed SCoT polymorphisms in the newly synthesized tritordeum lines HT27 and HT22, respectively, and their parents. An analysis of the presence/absence of bands among tritordeums and their parents revealed three types of polymorphic markers: (i) shared by tritordeums and one of their parents, (ii) exclusively amplified in tritordeums, and (iii) exclusively amplified in the parents. No polymorphism was detected among individuals of each parental species. Three SCoT markers were exclusively amplified in tritordeums of lines HT22 and HT27, being considered as polyploidization-induced rearrangements. About 70 % of the SCoT markers of H. chilense origin were not transmitted to the allopolyploids of both lines, and most of the SCoTs scored in the newly synthesized allopolyploids originated from wheat, reinforcing the potential use of tritordeum as an alternative crop., This study was supported by the International Consortium HY-WHEAT (project P-KBBE/AGR-GPL/0002/2010) funded by: COMPETE program (Programa Operacional Factores de Competitividade); Quadro de Referência Estratégico Nacional (QREN); European Union (Fundo Europeu de Desenvolvimento Regional); and the Portuguese Foundation for Science and Technology (FCT).

Parchment documents are part of our cultural heritage and, as historical artifacts that they are, should be preserved. The aim of this study was to validate an appropriate methodology to characterize the bioburden of parchment documents, and to assess the growth and gamma radiation inactivation patterns of the microbiota present in that material. Another goal was to estimate the minimum gamma radiation dose ( D min ) to be applied for the decontamination of parchment as an alternative treatment to the current toxic chemical and non-chemical decontamination methods. Two bioburden assessment methodologies were evaluated: the Swab Method (SM) and the Destructive Method (DM). The recovery efficiency of each method was estimated by artificial contamination, using a Cladosporium cladosporioides spore suspension. The parchment samples' microbiota was typified using morphological methods and the fungal isolates were identified by ITS-DNA sequencing. The inactivation pattern was assessed using the DM after exposure to different gamma radiation doses, and using C. cladosporioides as reference. Based on the applied methodology, parchment samples presented bioburden values lower than 5×10 3 CFU/cm 2 for total microbiota, and lower than 10 CFU/cm 2 for fungal propagules. The results suggest no evident inactivation trend for the natural parchment microbiota, especially regarding the fungal community. A minimum gamma radiation dose ( D min ) of 5 kGy is proposed for the decontamination treatment of parchment. Determining the minimal decontamination dose in parchment is essential for a correct application of gamma radiation as an alternative decontamination treatment for this type of documents avoiding the toxicity and the degradation promoted by the traditional chemical and non-chemical treatments.

The incorporation of plants in industry requires high standards of hygiene, being the raw materials obligatorily decontaminated. Irradiation is increasingly applied as a decontamination technique in several countries. This work aimed to evaluate the effects of gamma radiation on the phenolic composition, cytotoxicity, virucidal and antimicrobial properties of Aloysia citrodora L. (lemon verbena) and Mentha x piperita L. (peppermint) infusions. Phenolic compounds profile was obtained by HPLC-DAD-ESI/MS and cytotoxicity was ev aluated in human tumour and non-tumour cell lines; in virucidal efficacy, MNV-1 and HAdV-5 viral titers were determined by plaque assay in Raw264.7 and A549 cell lines, respectively; and the antibacterial potential was evaluated against Gram-negative and Gram-positive bacteria. Radiation treatment caused a significant increase in total phenolic compounds of both plants, while the virucidal efficacy was depend on the plant, the virus, and absorbed dose. S. aureus, with MIC and MBC values of 5 mg/mL, was the only bacteria sensible to the infusions of non-irradiated and 1 kGy irradiated lemon verbena and peppermint samples; gamma radiation appears to cause no alteration in the antibacterial potential of the studied plants. Peppermint infusion (irradiated at 10 kGy) showed the highest cytotoxic potential in all tumour cell lines. The authors are grateful to the Foundation for Science andTechnology (FCT, Portugal) and FEDER under Programe PT2020for financial support to CIMO (UID/AGR/00690/2013), LSRE(Project UID/EQU/50020/2013), C2TN (RECI/AAG-TEC/0400/2012;UID/Multi/04349/2013), L. Barros (SFRH/BPD/107855/2015) andR.C. Calhelha (SFRH/BPD/BPD/68344/2010) grants. The authors arealso grateful to Ministry of Agriculture, Portugal (Project AROMAP– PRODER/FEADER/EU), for financial support of the work and E.Pereira and Andreia I. Pimenta grants, and to “MaisErvas – Aromáti-cas e Medicinais” company for providing the samples. info:eu-repo/semantics/publishedVersion

Human Epidermal Growth Factor Receptor 2 (HER2) is, among EGFR family, one of the most relevant members as it remains overexpressed on tumor cells and provides resistance to well-studied anti-HER2 monoclonal antibody, Trastuzumab (Herceptin®), or tyrosine kinase inhibitor. Furthermore, HER2 plays a key role in the HER family due the interaction with other HER receptors via a complex signaling network to regulate cell growth, differentiation and survival. In this work, we have employed computational modelling and Molecular Dynamic (MD) simulations to attain a deeper understanding of the interaction of specific anti-HER2 antibodies and HER2. The dynamic behavior of HER2 receptor in complex with F0178 and scFv from Trastuzumab was investigated by two replicas of 0.5 µs MD simulations for each system as well as for the individual ones. A variety of structural, energetic and dynamic characteristics ranging from pairwise interactions formation to covariance analyses were performed to the 2 bundle complexes. Our aim was to understand the all-atom details of these intermolecular couplings, fundamental for the development of new therapies.

This study aimed to determine the effect of gamma radiation on the preservation of phenolic compounds and on decontamination of dry herbs in terms of ochratoxin A (OTA) and aflatoxin B1 (AFB1), using Aloysia citrodora Paláu as a case study. For this purpose, artificially contaminated dry leaves were submitted to gamma radiation at different doses (1, 5, and 10 kGy; at dose rate of 1.7 kGy/h). Phenolic compounds were analysed by HPLC-DAD-ESI/MS and mycotoxin levels were determined by HPLC-fluorescence. Eleven phenolic compounds were identified in the samples and despite the apparent degradation of some compounds (namely verbasoside), 1 and 10 kGy doses point to a preservation of the majority of the compounds. The mean mycotoxin reduction varied between 5.3% and 9.6% for OTA and from 4.9% to 5.2% for AFB1. It was not observed a significant effect of the irradiation treatments on mycotoxin levels, and a slight degradation of the phenolic compounds in the irradiated samples was observed. The authors are grateful to Ministry of Agriculture, Portugal (Project AROMAP— PRODER/FEADER/EU), for financial support of the work and E. Pereira grant. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support to CIMO (Pest-OE/AGR/UI0690/2015) and L. Barros grant (SFRH/BPD/107855/2015). Also, to POCI-01-0145-FEDER- 006984 (LA LSRE-LCM), funded by FEDER, through POCI-COMPETE2020 and FCT. C2TN/IST are also gratefully to FCT for financial support through the RECI/AAG-TEC/0400/2012 and UID/Multi/04349/2013 projects. The authors are also grateful to “MaisErvas—Aromáticas e Medicinais” for samples providing. The GIP-USAL is financially supported by the Spanish Government through the project AGL2015-64522-C2-2-R. info:eu-repo/semantics/publishedVersion

A comprehensive assessment of the toxicity and antioxidant activity of cork boiling wastewater and the effects of gamma radiation on these parameters was performed. Antioxidant activity was evaluated using different methodologies as DPPH radical scavenging activity, reducing power and inhibition of ß-carotene bleaching. The results have shown that gamma radiation can induce an increase on the antioxidant activity of cork boiling wastewater. Toxicity tests were performed to access the potential added value of the irradiated wastewaters and/or minimization of the impact for discharge in the environment. Two different methods for toxicity evaluation were followed, bacterial growth inhibition test and cytotoxicity assay, in order to predict the behavior of different cells (prokaryotic and eukaryotic) in the presence of cork wastewater. Non-treated cork boiling wastewater seemed to be non-toxic for prokaryotic cells (Pseudomonas fluorescens and Bacillus subtilis) but toxic for eukaryotic cells (A549 human cells and RAW264.7 mouse cells). The gamma radiation treatment at doses of 100 kGy appeared to increase the toxicity of cork compounds for all tested cells, which could be related to a toxic effect of radiolytic products of cork compounds in the wastewaters. This work was supported by Fundação para a Ciência e a Tecnologia (FCT, Portugal) through financial support of RECI/AAG-TEC/0400/2012 “Application of Ionizing Radiation for a Sustainable Environment” project. C2TN/IST authors gratefully acknowledge the FCT support through the UID/Multi/04349/2013 project. The authors are also grateful to International Atomic Energy Agency (Contract No. 16513) and to the cork industry (Amorim S.A) for allowing us to collect all the samples necessary for the accomplishment of this work. info:eu-repo/semantics/publishedVersion

For decades, the suitability and safety of various kinds of packaging materials in food irradiation treatment have been under question. In this chapter, food packaging materials authorized to be in contact with food during irradiation are described. The impact of ionizing radiation due to cross-linking and chain scission on the mechanical and barrier properties of authorized polymers is also discussed. One of the major safety concerns related to food irradiation is the formation of radiolysis products (RPs) in the polymer and their subsequent potential migration into the irradiated food. A list of numerous RPs is provided and their safety assessments are discussed. Besides its bactericidal effect, irradiation can be used to develop biodegradable polymers and ameliorate the release of active agents from active packaging. Finally, the radiosensitization effect of irradiation combined with antimicrobial edible coatings and films on pathogenic bacteria in various types of foods is demonstrated.

Adenovirus is the most prevalent enteric virus in waters worldwide due to its environmental stability, which leads to public health concerns. Mitigation strategies are therefore required. The aim of this study was to assess the inactivation of human adenovirus type 5 (HAdV-5) by gamma radiation in aqueous environments. Various substrates with different organic loads, including domestic wastewater, were inoculated with HAdV-5 either individually or in a viral pool (with murine norovirus type 1 [MNV-1]) and were irradiated in a Cobalt-60 irradiator at several gamma radiation doses (0.9 to 10.8 kGy). The infectivity of viral particles, before and after irradiation, was tested by plaque assay using A549 cells. D 10 values (dose required to inactivate 90% of a population or the dose of irradiation needed to produce a 1 log 10 reduction in the population) were estimated for each substrate based on virus infectivity inactivation exponential kinetics. The capability of two detection methods, nested PCR and enzyme-linked immunosorbent assay (ELISA), to track inactivated viral particles was also assessed. After irradiation at 3.5 kGy, a reduction of the HAdV-5 titer of 4 log PFU/ml on substrates with lower organic loads was obtained, but in highly organic matrixes, the virus titer reduction was only 1 log PFU/ml. The D 10 values of HAdV-5 in high organic substrates were significantly higher than in water suspensions. The obtained results point out some discrepancies between nested PCR, ELISA, and plaque assay on the assessments of HAdV-5 inactivation. These results suggest that the inactivation of HAdV-5 by gamma radiation, in aqueous environments, is significantly affected by substrate composition. This study highlights the virucidal potential of gamma radiation that may be used as a disinfection treatment for sustainable water supplies. IMPORTANCE Human adenovirus (HAdV) is the most prevalent of the enteric viruses in environmental waters worldwide. The purposes of this study are to provide new insights on the inactivation of enteric virus by gamma irradiation and to introduce new concepts and reinforce the benefits and utility of radiation technologies as disinfection processes. This may be an effective tool to guarantee the reduction of viral pathogens and to contribute to public health and sustainable water supplies.

The use of ionizing radiation dates back to many years ago, and is accredited for application in different foods with several purposes. It has been increasingly used in many countries for the treatment of aromatic plants. Thymus vulgaris L. (thyme) is a plant commonly used by food, pharmaceutical and cosmetic industries representing a natural source of several bioactives such as phenolic compounds. The aim of this work was to evaluate the effects of gamma radiation on the antioxidant activity (measured through the free radical scavenging activity, reducing power and lipid peroxidation inhibition) and phenolic compounds profile (obtained by HPLC-DAD-ESI/MS) of infusions prepared from irradiated thyme. The results showed that gamma irradiation at the dose of 10 kGy improved the free radical scavenging activity, reducing power and lipid peroxidation inhibition capacity of the studied infusions, while increasing significantly the concentrations of methyleriodictyol-O-pentosylhexoside, luteolin-7-O-glucoside, eriodictyol and total flavonoids content. Thus, gamma radiation could be considered as a suitable treatment to be used in Thymus vulgaris L., herein validated for its bioactive parameters. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programe PT2020 for financial support to CIMO (UID/AGR/00690/2013), LSRE (Project UID/EQU/50020/2013), C2TN (RECI/AAG-TEC/0400/2012), L. Barros (SFRH/BPD/107855/2015) and R.C. Calhelha (SFRH/BPD/ BPD/68344/2010) grants. The authors are also grateful to Ministry of Agriculture, Portugal (Project AROMAP - PRODER/FEADER/EU), for financial support of the work and E. Pereira grant, and to Pragmático Aroma Lda company for providing the “MaisErvas - Aromáticas e Medicinais” samples.

Wastewater from cork processing industry present high levels of organic compounds such as phenolics that must be degraded before discharge into the municipal sewer or into public water courses. The aim of this work was to find out if gamma radiation treatment could increase the biodegradability of recalcitrant compounds using a microbial consortium and a mixed solution of four phenolic acids as a model. Chemical and microbiological analyses were performed in non-irradiated (0 kGy) and irradiated (100 kGy) mixed phenolic acids cultures during incubation time. A preliminary HPLC and GC-MS analysis were performed to detect the major phenolic compounds in cork wastewater samples. Results indicated the presence of gallic, protocatechuic, vanillic and syringic acids in cork boiling water and gallic, protocatechuic and vanillic acids in sediment tank samples. The Total Phenolic content (TP) of mixed phenolic acids cultures during incubation time indicated a decrease of 38% for 100 kGy samples. The HPLC analysis suggested that the radiolytic products of syringic and vanillic acids are protocatechuic and gallic acids. The CFU counts pointed out to a decreasing tendency along the incubation time for phenolic acids cultures (0 kGy and 100 kGy) suggesting a non-degradation trend. The selected microbial consortium was not able to metabolize the phenolic compounds solutions at the used conditions. This could be due to the detected radiolytic degradation dynamics of the phenolic acids considering the antimicrobial activity of these compounds.

The effects of conventional and inert-gas enriched atmospheres, as well as those caused by different γ-irradiation doses, on buckler sorrel (Rumex induratus) leaves quality were evaluated and compared after 12 days of storage at 4 °C. The green colour of the abaxial surface of the leaves was relatively stable, as well as the carbohydrates content and the calorific value. The storage time decreased the amounts of fructose, glucose, trehalose, α-tocopherol, and β-tocopherol and increased the levels of total organic acids and δ-tocopherol. The total tocopherols content was higher in air-packaged non-irradiated leaves, antioxidant compounds that may have contributed to the preservation of polyunsaturated fatty acids (PUFA). Some antioxidant properties were also favoured during storage. It was found that the overall postharvest quality of buckler sorrel leaves is better maintained with the argon-enriched atmospheres, while the 6 kGy dose was a suitable option to preserve PUF+A during cold storage. This study highlights the suitability of the applied postharvest treatments and the interest to include buckler sorrel leaves in contemporary diets. info:eu-repo/semantics/publishedVersion

A preliminary study using gamma radiation on slaughterhouse wastewater samples was carried out. Chemical oxygen demand (COD), biochemical oxygen demand (BOD) and total suspended solids (TSS) results were obtained at a dose rate of 0.9 kGy h−1. A decrease of COD, BOD and colour was observed after irradiation at high absorbed doses. The microbiological results, following irradiation in the same conditions, correlated with the BOD results. The results obtained highlight the potential of this technology for wastewater treatment.