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The antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are considered new classes of water contaminants due to their potential adverse effects on aquatic ecosystems and human health. This paper describes the susceptibility evaluation of Escherichia coli, isolated from the influent and effluent of the biological reactors in two wastewater treatment plants (WWTP1 and WWTP2), and the evaluation of the disinfection efficiency of the UV radiation system in WWTP2. The exposure to different families of antibiotics was evaluated, namely, macrolides (erythromycin, azithromycin, clarithromycin), quinolones (ofloxacin, ciprofloxacin), nitroimidazoles (metronidazole), sulfanilamides (sulfamethoxazole) and trimethoprim, the latter is used in combination with sulfamethoxazole due to its synergistic effect. The results of the antimicrobial susceptibility tests, using disc diffusion (Kirby–Bauer) method, showed the occurrence of E. coli strains resistance. The analysis performed indicated an overall resistance (considering both influents and effluents) in WWTP1 and WWTP2, of respectively: 33.3% and 37.5% to erythromycin; 0.0% and 4.0% to azithromycin; 25.0% and 29.2% to clarithromycin; 12.5% and 4.2% to ofloxacin; 16.7% and 4.2% to ciprofloxacin; 29.2% and 12.5% to trimethoprim; 41.7% and 12.5% to sulfamethoxazole. A variability of resistance was observed along the studied period, from WWTP1 and WWTP2, and from influent to effluent. Disinfection by UV demonstrated good performance, achieving in some samples 100% removal of E. coli that has growth in TBX agar. However, a proper supervision is needed in order to achieve the allowed limits concerning the number of bacteria. The results from this work contribute to a better awareness of ARB dissemination from wastewater treatment plants to the aquatic environment.

Background Colony plate counting still remains the “gold standard” procedure for the identification of viable Listeria monocytogenes cells and to address microbiological quality assurance of foodstuff. However, this classical method requires an analysis time superior to the current demands in food industry. Hence, new time-enhanced methods based on improved cultural techniques, nucleic acid detection and immunological assays have raised as new validated alternatives. Nevertheless, those rapid methods still require specific and expensive equipment and imply a considerable workload. Scope and approach In the last years, electrochemical biosensors and bioassays have been intensively investigated, emerging as excellent alternatives to surpass the disadvantages of the conventional and standard rapid methods. They provide the desired fast, sensitive and selective response, towards a portable, cost-effective and user-friendly performance. Therefore, in this work, a comprehensive review about the foundations, current achievements and limitations described over the past fifteen years for electrochemical biosensing of Listeria monocytogenes in food products is presented. Key findings and conclusions The latest innovations rely on the use of low-cost electrochemical transducers, integration of novel (nano)materials and incorporation of new bioreceptors in the sensing strategy. Single-cell detection and intelligent packaging are also growing trends. Regardless of the remaining challenges, that still need to be overcome, electrochemical biosensing seems to have a role as one of the promising strategies to lead the future of foodborne pathogen analysis.

The aim of this study was to evaluate the influence of lyophilization and drying in stove on phenolic compounds content and the biological activity of Apis mellifera and Trigona spinipes pollens produced in Brazil. In general, the bee pollen produced by T. spinipes presented highest antioxidant activity in dried and fresh samples assessed either by test of plasma ferric reduction capacity (FRAP) and free radical scavenging assay (DPPH) methods. For A. mellifera bee pollen the antioxidant activity was higher on the fresh samples. Nevertheless, b-carotene bleaching assay (BCB) and linoleic acid content were higher in T. spinipes samples, mainly in the fresh ones. Higher antioxidant activity was owing to higher content in phenolic compounds. Lyophilization method was the best for phenolic compounds’ conservation for both species. The bee pollen of both species has a high amount of flavonoids: kaempferol-3-O-glucoside was the most abundant in A. mellifera while for T. spinipes the most prevalent was resorcylic acidþepicatechin. All extracts presented antibacterial activity against Saphylococcus aureus (ATCC 43300)TM, (ESA 83138150), (ESA 32), Pseudomonas aeruginosa (ATCC 15442)TM, Pseudomonas aeruginosa (MRC.4) and (MRC.10). The methods used for storage influenced the biological properties of bee pollen from both species. Regarding the content of phenolic compounds, differences were observed amongst the pollen types: for A. mellifera these were best preserved with lyophilization, while for T. spinipes the three storage methods were equivalent. The authors are grateful to the Coordenac¸~ao de Aperfeic¸oamento de Pessoal de N ıvel Superior – CAPES (88881.062167/2014-01 for M.L.M.E.). A. Pascoal would like to thank Foundation for Science and Technology (FCT, Portugal), Programa Operacional Potencial Humano (POPH), and European Union (EU) for his Postdoctoral grant SFRH/BPD/ 91380/2012. info:eu-repo/semantics/publishedVersion

Efficiently detecting peanut traces in food products can prevent severe allergic reactions and serious health implications. This work presents the development of an electrochemical dual immunosensor for the simultaneous analysis of two major peanut allergens, Ara h 1 and Ara h 6, in food matrices. A sandwich immunoassay was performed on a dual working screen-printed carbon electrode using monoclonal antibodies. The antibody–antigen interaction was detected by linear sweep voltammetry through the oxidation of enzymatically deposited silver, which was formed by using detection antibodies labeled with alkaline phosphatase and a 3-indoxyl phosphate/silver nitrate mixture as the enzymatic substrate. The assay time was 2 h 20 min, with a hands-on time of 30 min, and precise results and low limits of detection were obtained (Ara h 1: 5.2 ng·mL−1, Ara h 6: 0.017 ng·mL−1). The selectivity of the method was confirmed through the analysis of other food allergens and ingredients (e.g., hazelnut, soybean and lupin). The dual sensor was successfully applied to the analysis of several food products and was able to quantify the presence of peanuts down to 0.05% (w/w). The accuracy of the results was confirmed through recovery studies and by comparison with an enzyme-linked immunosorbent assay. Tracking food allergens is of utmost importance and can be performed using the present biosensor in a suitable and practical way.

Groundnuts, like peanuts, are commonly integrated into the Mediterranean dietary pattern, and their consumption has been recommended worldwide. However, reported cases of peanut allergy have increased and, therefore, commercial food tracking is essential, since in extreme cases peanut intake causes anaphylaxis. Efficient detection of peanut traces in food samples can be achieved using electrochemical immunosensors that benefit from their advantageous features such as high selectivity and sensitivity, low cost, and rapid detection. Because disposable screen-printed electrodes can be connected to portable devices, they can be used for in situ allergen analysis. In the present work, a voltammetric immunosensor was developed to quantify a major peanut allergen, Ara h 1, using screen-printed carbon electrodes (SPCE) as transducers. Distinct carbon-based nanomaterials were tested, and nanodiamonds were selected for the nanostructuration of the transducer’s surface. A sandwich-type immunoassay was performed on the nanodiamond-coated SPCEs using an alkaline phosphatase-labelled detection antibody and a mixture containing an enzymatic substrate (3-indoxyl phosphate) and silver nitrate. The immunological interaction was detected through the (linear sweep) voltammetric stripping of the enzymatically deposited silver. A linear concentration range was established between 25 and 500 ng·mL−1 (ip = (0.027 ± 0.001) [Ara h 1] + (1.41 ± 0.31), r = 0.994, n = 5), with a sensitivity of 0.342 µA·mL·ng-1·cm-2. The limits of detection (LOD) and quantification (LOQ) were 0.78 and 2.6 ng·mL−1, respectively. Other allergens, non-target proteins and food product ingredients were tested to assess the selectivity of the immunosensor. Its applicability was evaluated by analysing a set of breakfast cereals, cookies, and energetic and cereal bars.

This study describes the possibilities of valorising a waste stream that originates from apple wood by mapping the reducing capacity and phenolic profile from extracts derived from apple tree (Malus domestica). This study evaluated the efficiency of warm solvent extraction (WSE) and ultrasound-assisted extraction (UAE) techniques for extracting antioxidant phenolic compounds from the bark and core wood of an apple tree cultivated in the north-eastern part of Belgium. Furthermore, the influence of the pre-treatment technique, namely, fresh, oven-dried, and freeze-dried samples, respectively, on the yield of polyphenols was studied. Fresh bark extract obtained by UAE—the most efficient extraction technique—employing acetone 60% v/v contains the highest levels of phenolic compounds as well as the highest antioxidant activity. High-performance liquid chromatographic analysis shows that phloridzin is the major compound of the identified polyphenol markers present in bark and core wood extracts. Based on the obtained results, it may be possible to produce a polyphenolic extract from apple wood at an industrial scale without extensive costs or altering the antioxidant properties. This study reveals the potential of apple tree wood residues valorisation through the recovery of phenolic compounds for food, pharmaceutical, and cosmetic applications., Annick Boeykens is a beneficiary of a PWO (‘Projectmatig Wetenschappelijk Onderzoek’) Grant, provided to Odisee by the Flem-ish Government, for the investigation project ‘Phenolic compounds in by-products’. Manuela M. Moreira (SFRH/BPD/97049/2013) wishes to acknowledge Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior for funding her postdoctoral fellowship by means of a POPH-QREN—Tipologia 4.1—Formação Avançada. The financial support from FCT/MEC through national funds and cofi-nanced by FEDER, under the Partnership Agreement PT2020 through the project UID/QUI/50006/2013—POCI/01/0145/FEDER/007265 and the project 6818—Transnational Cooperation, Agreement between Por-tugal (FCT) and Serbia (MSTD) are also acknowledged.

Specific monitoring of cystatin C (CysC) levels in biological fluids is critical for diagnosis, treatment and mechanistic understanding of a spectrum of diseases, particularly chronic kidney disease (CKD). Despite evidences that CysC correlates with the high risk and/or progression of CKD, its use in clinical practice is still scarce. In this context, we report the development of a simple and sensitive immunosensor for the detection of CysC. The biosensor combines the technology of cost-effective screen-printed electrodes with the high specificity of a sandwich immunoassay. Optimized conditions showed that the sensor operates in a linear range between 10 and 100 ng mL-1, with a detection limit and a sensitivity of 6.0 ng mL-1 and 6.4 ± 0.3 μA ng mL-1 cm-2, respectively. Moreover, the sensor provided precise results (RSD ≤ 6.2%) and the quantification of CysC in CKD serum samples revealed to be in agreement with the values obtained by a particle-enhanced nephelometric immunoassay. In this light, the proposed immunosensor qualifies for clinical application, constituting a step forward in the development of fast, sensitive and cost-effective diagnostic tools that can improve the current medical care settings of CKD patients., This work was supported by European Union through projects Norte-01-0145-FEDER-000024 and Norte-01-0145-FEDER-000011, co-funded by FEDER in the scope of CCDR-N and NORTE2020 Partnership Agreement. This work also received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e a Tecnologia) through project UID/QUI/50006/2013.

Children, an important vulnerable group, spend most of their time at schools (up to 10 h per day, mostly indoors) and the respective air quality may significantly impact on children health. Thus, this work reviews the published studies on children biomonitoring and environmental exposure to particulate matter (PM) and polycyclic aromatic hydrocarbons (PAHs) at school microenvironments (indoors and outdoors), major sources and potential health risks. A total of 28, 35, and 31% of the studies reported levels that exceeded the international outdoor ambient air guidelines for PM10, PM2.5, and benzo(a)pyrene, respectively. Indoor and outdoor concentrations of PM10 at European schools, the most characterized continent, ranged between 7.5 and 229 μg/m3 and 21–166 μg/m3, respectively; levels of PM2.5 varied between 4 and 100 μg/m3 indoors and 6.1–115 μg/m3 outdoors. Despite scarce information in some geographical regions (America, Oceania and Africa), the collected data clearly show that Asian children are exposed to the highest concentrations of PM and PAHs at school environments, which were associated with increased carcinogenic risks and with the highest values of urinary total monohydroxyl PAH metabolites (PAH biomarkers of exposure). Additionally, children attending schools in polluted urban and industrial areas are exposed to higher levels of PM and PAHs with increased concentrations of urinary PAH metabolites in comparison with children from rural areas. Strong evidences demonstrated associations between environmental exposure to PM and PAHs with several health outcomes, including increased risk of asthma, pulmonary infections, skin diseases, and allergies. Nevertheless, there is a scientific gap on studies that include the characterization of PM fine fraction and the levels of PAHs in the total air (particulate and gas phases) of indoor and outdoor air of school environments and the associated risks for the health of children. There is a clear need to improve indoor air quality in schools and to establish international guidelines for exposure limits in these environments. Keywords: Biomarkers, Children exposure, Health effects, Particulate matter (PM), Polycyclic aromatic hydrocarbons (PAHs)

Olea europaea cultivar, native in the Mediterranean basin, has expanded worldwide, mainly due to the olive oil industry. This expansion is attributed to the benefits of olive oil consumption, since this product is rich in nutritional and bioactive compounds. However, the olive industry generates high amounts of wastes, which could be related to polluting effects on soil and water. To minimize the environmental impact, different strategies of revalorization have been proposed. In this sense, the aim of this work was to develop high cosmetic value added oleuropein-enriched extracts (O20 and O30), a bioactive compound from olive byproducts, performing a comprehensive characterization using high performance liquid chromatography coupled to mass spectrometry and evaluate their bioactivity by in vitro assays. A total of 49 compounds were detected, with oleuropein and its derivatives widely found in O30 extract, whereas iridoids were mainly detected in O20 extract. Moreover, 10 compounds were detected for the first time in olive leaves. Both extracts demonstrated strong antioxidant and antiradical activities, although O30 showed higher values. In addition, radical oxygen and nitrogen species scavenging and enzyme inhibition values were higher in O30, with the exception of HOCl and hyaluronidase inhibition assays. Regarding cell viability, olive byproduct extracts did not lead to a decrease in keratinocytes viability until 100 µg/mL. All data reported by the present study reflect the potential of industrial byproducts as cosmetic ingredients.

The biggest obstacle to the treatment of diseases that affect the central nervous system (CNS) is the passage of drugs across the blood-brain barrier (BBB), a physical barrier that regulates the entry of substances into the brain and ensures the homeostasis of the CNS. This review summarizes current research on lipid-based nanoparticles for the nanoencapsulation of neuroprotective compounds. A survey of studies on nanoemulsions (NEs), nanoliposomes/nanophytosomes and solid lipid nanoparticles (SLNs)/nanostructured lipid carriers (NLCs) was carried out and is discussed herein, with particular emphasis upon their unique characteristics, the most important parameters influencing the formulation of each one, and examples of neuroprotective compounds/extracts nanoencapsulated using these nanoparticles. Gastrointestinal absorption is also discussed, as it may pose some obstacles for the absorption of free and nanoencapsulated neuroprotective compounds into the bloodstream, consequently hampering drug concentration in the brain. The transport mechanisms through which compounds or nanoparticles may cross BBB into the brain parenchyma, and the potential to increase drug bioavailability, are also discussed. Additionally, factors contributing to BBB disruption and neurodegeneration are described. Finally, the advantages of, and obstacles to, conventional and unconventional routes of administration to deliver nanoencapsulated neuroprotective drugs to the brain are also discussed, taking into account the avoidance of first-pass metabolism, onset of action, ability to bypass the BBB and concentration of the drug in the brain.

The role of milk and dairy products in supplying iodine to pregnant women is unknown in Portugal. The aim of this study was to evaluate the association between milk and dairy product consumption and the iodine status of pregnant women in the IoMum cohort of the Oporto region. Pregnant women were recruited between 10 and 13 weeks of gestation, when they provided a spot urine sample and information on lifestyle and intake of iodine-rich foods. Urinary iodine concentration (UIC) was determined by inductively coupled plasma MS. A total of 468 pregnant women (269 iodine supplement users and 199 non-supplement users) were considered eligible for analysis. Milk (but not yogurt or cheese) intake was positively associated with UIC, in the whole population (P = 0·02) and in the non-supplement users (P = 0·002), but not in the supplement users (P = 0·29). In non-supplement users, adjusted multinomial logistic regression analysis showed that milk consumption

Lack of knowledge about iodine has been suggested as a risk factor for iodine deficiency in pregnant women, but no studies have addressed this issue in Portugal. So, the aim of this study was to investigate iodine knowledge among Portuguese pregnant women and its association with iodine status. IoMum, a prospective observational study, included 485 pregnant women recruited at Centro Hospitalar e Universitário de S. João, Porto, between the 10th and 13th gestational weeks. Partial scores for knowledge on iodine importance, on iodine food sources or on iodised salt were obtained through the application of a structured questionnaire. Then, a total iodine knowledge score was calculated and grouped into low, medium and high knowledge categories. Urinary iodine concentration (UIC) was measured in spot urine samples by inductively coupled plasma MS. Of the pregnant women, 54 % correctly recognised iodine as important to neurocognitive development, 32 % were unable to identify any iodine-rich food and 71 % presented lack of knowledge regarding iodised salt. Of the women, 61 % had a medium total score of iodine knowledge. Knowledge on iodine importance during pregnancy was positively associated with iodine supplementation and also with UIC. Nevertheless, median UIC in women who correctly recognised the importance of iodine was below the cut-off for adequacy in pregnancy (150 µg/l). In conclusion, knowledge on iodine importance is positively associated with iodine status. Despite this, recognising iodine importance during pregnancy may not be sufficient to ensure iodine adequacy. Literacy-promoting actions are urgently needed to improve iodine status in pregnancy. This work was supported by National Funds through FCT – Fundação para a Ciência e a Tecnologia/MCTES within CINTESIS, R&D Unit (reference UID/IC/4255/2019) and REQUIMTE (reference UIDB/info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB/50006/2020/PT) and through FCT postdoctoral fellowship (V. C. F., grant number SFRH/BPD/109153/2015).

Sustainability arises as a challenge in the last decades and the cosmetic field is not an exception. The search for new cosmetic active principles extracted from food byproducts through eco-innovative technologies has emerged as a novel concept. However, the European Regulation is very restrictive and different assays should be performed to guarantee the products' efficacy and safety. Considering a multidisciplinary approach, this paper aims to compare chestnut shell extracts prepared by three different green techniques – Subcritical Water Extraction (SWE), Ultrasound-Assisted Extraction (UAE) and Supercritical Fluids Extraction (SFE) – regarding antimicrobial activity, hyaluronidase and elastase inhibitory activities, in-vitro cytotoxicity on dermal cells (HaCaT and HFF-1) and skin permeation (using Franz cells coupled to human skin). The best extract was tested on skin and ocular 3D models and through an in-vivo patch test to evaluate its irritancy and safety. The results demonstrated the antimicrobial properties of the extracts (particularly against Staphylococcus aureus) as well as the capacity to inhibit the hyaluronidase (IC50 = 0.76–54.36 mg/mL) and elastase (33.56–82.70% at 0.4 mg/mL) activities. Regarding in-vitro cell assays, UAE and SWE extracts revealed the best outcomes. Between 0.1 and 100 μg/mL, none of the extracts conducted to a decrease of cell viability in both cell lines (HaCaT and HFF-1). The ex-vivo assay proved that SWE extract led to a higher polyphenol's permeation (1061.6 μg/g dw), being selected for further assays. Ellagic acid was the major polyphenol that permeated the human skin (732.1–847.0 μg/g dw). The 3D skin and ocular models evidenced absence of irritation by SWE extract, without decreasing cell viability and releasing IL-1α. Finally, an in-vivo patch test confirmed the absence of irritation in humans based on the acute irritation index of 0.50 achieved 24 h after the patch removal. To the best of our knowledge, this is the first study that validated by in-vitro and in-vivo assays, and in accordance with the European Regulation 1223/2009, a new cosmetic ingredient extracted from chestnut shells., The authors' kindly thanks to Sortegel (Sortes, Portugal) for the samples. This research was funded by FCT/MCTES through national funds (UIDB/50006/2020) and project PTDC/ASP-AGR/29277/2017 - Castanea sativa shells as a new source of active ingredients for Functional Food and Cosmetic applications: a sustainable approach, financially supported by national funds by FCT/MCTES and co-supported by Fundo Europeu de Desenvolvimento Regional (FEDER) throughout COMPETE 2020 - Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-029277). This work was also supported by Project UIDB/04033/2020 (CITAB-Center for the Research and Technology of Agro-Environmental and Biological Sciences). Diana Pinto (SFRH/BD/144534/2019) is grateful for the PhD grant financed by FCT/MCTES and POPH-QREN and supported by funds from UE and Fundo Social Europeu (FSE) through Programa Operacional Regional Norte. Francisca Rodrigues is thankful for her contract (CEECIND/01886/2020) financed by FCT/MCTES—CEEC Individual 2020 Program Contract.

Hardy kiwifruit (Actinidia arguta) is a highly appreciated exotic fruit endowed with outstanding bioactive compounds. The present work proposes to characterize the pulp from A. arguta organic fruits, emphasizing its radicals scavenging capacity and effects on intestinal cells (Caco-2 and HT29-MTX). The physicochemical properties and phenolic profile were also screened. The total phenolic and flavonoid contents (TPC and TFC, respectively) of pulp were 12.21 mg GAE/g on dry weight (DW) and 5.92 mg CE/g DW, respectively. A high antioxidant activity was observed (FRAP: 151.41 μmol FSE/g DW; DPPH: 12.17 mg TE/g DW). Furthermore, the pulp did not induce a toxic effect on Caco-2 and HT29-MTX cells viability up to 1000 μg/mL. Regarding in vitro scavenging capacity, the pulp revealed the highest scavenging power against NO. (IC50 =3.45 μg/mL) and HOCl (IC50 =12.77 μg/mL). These results emphasize the richness of A. arguta fruit pulp to be used in different food products.

This work focused on evaluating the possibility of using vineyard pruning wastes from two Portuguese Vitis vinifera varieties, Touriga Nacional (TN) and Tinta Roriz (TR), as new potential ingredients for the nutraceutical industry. An environmentally friendly extraction technique, namely subcritical-water extraction (SWE), was employed. The overall results indicate that phenolic acids were the major class of compounds quantified, being gallic acid the principal one. The highest value for total phenolic content (TPC) was obtained for the TR extract at 250 °C (181 ± 12 mg GAE/g dw). In terms of antioxidant activity, the DPPH values for the extracts obtained at 250 °C were approximately 4-fold higher than the ones obtained at 125 °C, with TR extract presenting the highest value (203 ± 22 mg TE/g dw). Thus, the TR extract obtained through SWE at 250 °C was selected to evaluate the scavenging activity and the in vitro effects on cells due to the best results achieved in the previous assays. This extract presented the ability to scavenge reactive oxygen species (O2●-, HOCl and ROO●). No adverse effects were observed in HFF-1 viability after exposure to extract concentrations below 100 μg/mL. This work demonstrated that vine-canes extracts could be a potential ingredient to nutraceutical industry

The majority of works about vine-canes are focused on the evaluation of their chemical composition and antioxidant potential. To the best of our knowledge, the possible applications of produced extracts in cosmetic formulations have never been explored. The aim of the present study was to evaluate the antioxidant properties of vine-canes subcritical water extracts for use as active ingredients in the cosmetic industry. For that, the phenolic content and antioxidant activity of six vine-cane varieties, namely Alvarinho and Loureiro from the Minho region and Touriga Nacional and Tinta Roriz (TR) from both the Douro and D&atilde, o regions, were evaluated through spectrophotometric and chromatographic methods. All extracts presented similar antioxidant activity and the highest phenolic content was reported for TR variety from the Douro region (33.7 &plusmn, 1.9 mg GAE/g dw). The capacity of vine-cane extracts to capture reactive oxygen species superoxide (O2&minus, ) was also studied, with the highest IC50 value being obtained for Loureiro variety (56.68 &plusmn, 2.60 &micro, g/mL). Furthermore, no adverse effects on HaCaT and HFF-1 dermal cell lines in concentrations below 100 and 1000 g/mL, respectively, were determined. Finally, Loureiro vine-cane extract was incorporated into a topical formulation, and physical and microbiological properties were within expected values, demonstrating that vine-canes extracts can be successfully incorporated in cosmetic products.

In this work, the effectiveness of green zero-valent iron nanoparticles (gnZVIs) for the removal of the antibiotic sulfadiazine (SDZ) from water via adsorption and reduction was tested. Additionally, the effectiveness of this material as a catalyst for the Fenton and photo-Fenton processes was also investigated. This represents the first study concerning the use of gnZVIs for the degradation of a sulfonamide antibiotic. The results obtained indicate that gnZVIs were able to remove up to 58% of SDZ via adsorption and up to 69% via adsorption plus reduction using a SDZ/Fe3+ molar ratio of 1:61.6. Furthermore, gnZVIs showed strong effectiveness as a catalyst for the Fenton and photo-Fenton reactions, with complete SDZ removal in 8 h and 5 min, respectively, using a SDZ/Fe3+/H2O2 molar ratio of 1:38.4:38.4. These results demonstrate that the use of gnZVIs constitutes an attractive and potential alternative technology for water remediation, reducing environmental impact and operational costs.

This study gives new insights to understand the type of interactions between Ginkgo biloba L. and Scutellaria baicalensis Georgi, two Chinese medicinal plants with well documented neuroprotective effects, on three targets in Alzheimer's disease (AD): acetylcholinesterase (AChE) and butyrylcholnesterase (BuChE) inhibition and hydrogen peroxide scavenging. Individual samples, binary mixtures with different proportions of both plant species, and also a commercial multicomponent combination containing both plants together with unroasted Coffea arabica L. and quercetin-3-O-rutinoside were used to perform this in vitro evaluation. Sample phenolic profiles were also determined by HPLC-DAD, showing the presence of several flavonoid glycosides, phenolic acids and a methylxanthine. In order to investigate the possible synergism/antagonism interaction, data obtained were analyzed by CompuSyn software. The results showed that G. biloba and S. baicalensis alone display better activities than in mixtures, most of the interactions exhibiting different degrees of antagonism. A slight synergism interaction was only observed for the commercial multicomponent mixture tested against H2O2. Further analysis was carried out to understand which compounds could be responsible for the antagonistic interaction. Seventeen single pure compounds present in all extracts were tested against AChE inhibition, most of them displaying weak or no activity. Only caffeine had a remarkable activity. Five different binary and quaternary mixture compositions were design to deepen the interaction between these compounds, revealing mainly phenolic acid-flavonoid, flavonoid-flavonoid and methylxanthine-flavonoid-phenolic acid antagonistic interactions. These results clearly show that, for the targets evaluated, there is no potentiation of the neuroprotective effect by combining S. baicalensis and G. biloba extracts.

Chestnut processing industry generates large amounts of by-products, including leaves, burs and shells that are a source of bioactive compounds. The purpose of this study was to establish an ultrasound-assisted extraction (UAE) of phenolic and antioxidant compounds from industrial chestnut shells. A central composite design (CCD) was conducted to analyze the effects of time (4&ndash, 46 min) and temperature (34&ndash, 76 &deg, C) in the antioxidant activity (2,2&prime, azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP)) and total phenolic compounds (TPC) of chestnut shells extracts. The optimal extraction conditions were obtained at 70 &deg, C for 40 min. The optimal extract was characterized regarding phenolic profile, radical scavenging capacity, and effects on intestinal and dermal cell lines. The optimal extract revealed high amounts of ellagic acid (40.4 &micro, g/mg dw), followed by caffeic acid derivative (15.4 &micro, g/mg dw) and epigallocatechin (15.3 &micro, g/mg dw). Indeed, the extract exhibited the highest scavenging efficiencies against NO● (IC50 = 0.1 &micro, g/mL) and HOCl (IC50 = 0.7 &micro, g/mL) and did not conducted to a decrease on HaCaT and HFF-1 viability up to 100 &mu, g/mL. Oppositely, a decrease on Caco-2 and HT29-MTX viability was observed. This study suggests that UAE could be a sustainable option to valorize chestnut shells as raw material for different industries.

This study aimed to develop new canned chub mackerel products incorporating edible seaweeds (Ascophyllum nodosum, Fucus spiralis, Saccorhiza polyschides, Chondrus crispus, Porphyra sp. and Ulva sp.) harvested in the Portuguese North-Central coast, with simultaneous sensory improvement and minerals enrichment. Two processes were compared, namely the addition of seaweeds in i) the canning step and ii) in the brining step (as the replacement for salt). The concentrations of four macrominerals (Na, K, Ca and Mg), chloride, and twelve trace elements (Co, Cu, Fe, I, Li, Mn, Mo, Rb, Se, Sr, V and Zn) were determined by high-resolution continuum source flame atomic absorption spectrometry (HR-CS-FAAS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Results showed that canned chub mackerel incorporating C. crispus and F. spiralis was found to be the preferred sensory option, also exhibiting contents enriched with Cl, Co, Cu, Fe, I, Li, Mg, Mn, Mo, Na, Rb, Se, and Sr. This effect was more pronounced when both seaweed species were added to replace the salt added in the brining step., This work received financial support from project iCanSea—Conservas com macroalgas para diferenciação nutricional e sensorial, sponsored by Portugal 2020 under the Programa Operacional Competitividade e Internacionalização (Ref.3171/Portugal 2020). The Fundação para a Ciência e a Tecnologia (FCT) and the FEDER, under Programme PT2020 (Project UID/QUI/50006/2019) and the project Qualidade e Segurança Alimentar—uma abordagem (nano)tecnológica (NORTE-01-0145-FEDER-000011) are also acknowledged for the financial funding.

In vitro methods have gained rising importance in ecotoxicology due to ethical concerns. The aim of this study was to assess the single and combined in vitro effects of gold, as nanoparticle (AuNPs) and ionic (Au+) form, and the pharmaceutical gemfibrozil (GEM). Sparus aurata liver organ culture was exposed to gold (4 to 7200 μg·L−1), GEM (1.5 to 15,000 μg·L−1) and combination 80 μg·L−1 gold +150 μg·L−1 GEM for 24 h. Endpoints related with antioxidant status, peroxidative/genetic damage were assessed. AuNPs caused more effects than Au+, increasing catalase and glutathione reductase activities and damaging DNA and cellular membranes. Effects were dependent on AuNPs size, coating and concentration. GEM damaged DNA at an environmentally relevant concentration, 1.5 μg·L−1. Overall, the effects of the combined exposures were higher than the predicted, based on single exposures. This study showed that liver culture can be a useful model to study contaminants effects.

An electrochemical magnetic immunosensing strategy was developed for the determination of HER2-ECD, a breast cancer biomarker, and breast cancer cells in human serum. A sandwich assay was performed on carboxylic acid-functionalized magnetic beads (MBs) using a screen-printed carbon electrode (SPCE) as transducer surface. The affinity process was detected using electroactive labels; core/shell streptavidin-modified CdSe@ZnS Quantum Dots (QDs). Cd2+ ions, released from the QDs, were determined by differential pulse anodic stripping voltammetry (DPASV). An assay time of 90 min, with an actual hands-on time of about 20 min, a linear range between 0.50–50 ng·mL−1 of HER2-ECD and a limit of detection of 0.29 ng·mL−1 were achieved. Analysis of live breast cancer cells was also performed using the optimized assay. Breast cancer cell lines SK-BR-3 (a HER2-positive cell line), MDA-MB-231 (a HER2-negative cell line) and MCF-7 (a cell line with low HER2 expression) were tested. The selectivity of the assay towards SK-BR-3 cells was confirmed. A concentration-dependent signal that was 12.5× higher than the signal obtained for the HER2-negative cells (MDA-MB-231) and a limit of detection of 2 cells·mL−1 was obtained.

Cooks and kitchen workers at restaurants are continuously exposed to cooking fumes released during food processing. There is limited information about kitchen and barbecue grill workers’ occupational exposure. This work determined the levels of 1-hydroxypyrene and 3-hydroxybenzo(a)pyrene in the post-shift urines of barbecue grill workers during a regular working period. Levels of both polycyclic aromatic hydrocarbon (PAH) metabolites were extracted by solid-phase extraction and quantified by high-performance liquid chromatography with fluorescence detection. Overall, median concentrations of 1-hydroxypyrene were significantly higher during the period when workers were exposed than in the days out of duty (0.068 versus 0.050 µmol/mol creatinine; p = 0.003). Total exposure to 1-hydroxypyrene was well below the benchmark level of 0.5 µmol/mol creatinine proposed by the American Conference of Governmental Industrial Hygienists. The PAH biomarker of exposure to carcinogens, 3-hydroxybenzo(a)pyrene, was only detected during the working period (0.123–2.07 nmol/mol creatinine). Findings achieved in this study suggest that grill workers are occupationally exposed to PAHs and thus are more vulnerable to the potential health risks associated with long-term exposures. However, more studies are needed with other biomarkers of exposure to PAHs to better estimate the occupational exposure of barbecue grill workers.

This work characterizes the dimension and the exceptionality of 2017 large- and mega-fires that occurred in the center region of Portugal through the assessment of their impact on the ambient levels of particulate matter (PM10 and PM2.5), retrieved from local monitoring stations, and the associated public health risks. PM10 and PM2.5 concentrations were increased during the occurrence of large fires and megafires, with daily concentrations exceeding the European/national guidelines in 7&ndash, 14 and 1&ndash, 12 days of 2017 (up to 704 &micro, g/m3 for PM10 and 46 &micro, g/m3 for PM2.5), respectively. PM10 concentrations were correlated with total burned area (0.500 &lt, r &lt, 0.949, p &gt, 0.05) and with monthly total burned area/distance2 (0.500 &lt, 0.667, 0.05). The forest fires of 2017 took the life of 112 citizens. A total of 474 cases of hospital admissions due to cardiovascular diseases and 3524 cases of asthma incidence symptoms per 100,000 individuals at risk were assessed due to exposure to 2017 forest fires. Real-time and in situ PM methodologies should be combined with protection action plans to reduce public health risks. Portuguese rural stations should monitor other health-relevant pollutants (e.g., carbon monoxide and volatile organic compounds) released from wildfires to allow performing more robust and comprehensive measurements that will allow a better assessment of the potential health risks for the exposed populations.

Firefighters represent one of the riskiest occupations, yet due to the logistic reasons, the respective exposure assessment is one of the most challenging. Thus, this work assessed the impact of firefighting activities on levels of urinary monohydroxyl-polycyclic aromatic hydrocarbons (OHPAHs; 1-hydroxynaphthalene, 1-hydroxyacenaphthene, 2-hydroxyfluorene, 1-hydroxyphenanthrene, 1-hydroxypyrene, 3-hydroxybenzo(a)pyrene) and genotoxic/oxidative-effect biomarkers (basal DNA and oxidative DNA damage) of firefighters from eight firehouses. Cardiac frequency, blood pressure and arterial oxygen saturation were also monitored. OHPAHs were determined by liquid-chromatography with fluorescence detection, while genotoxic/oxidative-effect biomarkers were assessed by the comet assay. Concentrations of total OHPAHs were up to 340% higher (p ≤ 0.05) in (non-smoking and smoking) exposed workers than in control subjects (non-smoking and non-exposed to combat activities); the highest increments were observed for 1-hydroxynaphthalene and 1-hydroxyacenaphthene (82-88% of ∑OHPAHs), and for 2-hydroxyfluorene (5-15%). Levels of biomarker for oxidative stress were increased in non-smoking exposed workers than in control group (316%; p ≤ 0.001); inconclusive results were found for DNA damage. Positive correlations were found between the cardiac frequency, ∑OHPAHs and the oxidative DNA damage of non-smoking (non-exposed and exposed) firefighters. Evidences were raised regarding the simultaneous use of these biomarkers for the surveillance of firefighters' health and to better estimate the potential short-term health risks. Highlights: Urinary PAH metabolites and genotoxic/oxidative effects were assessed in firefighters; Levels of total PAH metabolites were increased up to 340% in exposed subjects; Urinary 1OHNaph+1OHAce, and 2OHFlu were the compounds with the highest increments; Firefighting and tobacco smoke raised the oxidative DNA damage in exposed subjects; Cardiac frequency, PAH metabolites and DNA damage can be used for health surveillance. This work received financial support from European (FEDER funds through COMPETE) and National funds through Fundação para a Ciência e Tecnologia for projects UID/QUI/50006/2019 and PCIF/SSO/ 0017/2018, and in the scope of the P2020 Partnership Agreement Project POCI-01-0145-FEDER-006939 (LEPABE). K. Slezakova and S. Costa were supported by the grants SFRH/BPD/105100/2014 and SFRH/BPD/100948/2014, respectively. info:eu-repo/semantics/publishedVersion

In addition to environmental pollution issues, social concerns about the sustainability, safety, and quality of conventionally grown fruits and vegetables have been increasing. In order to evaluate if there were any microbiological differences between samples of organic and conventional lettuce, a wide range of parameters were tested, including pathogens and indicator organisms: the enumeration of Escherichia coli; the detection of Salmonella spp.; the detection/enumeration of Listeria monocytogenes; the enumeration of lactic acid bacteria, Pseudomonas spp. yeasts and molds, and Enterobacteriaceae. This study also evaluated the chemical safety of the lettuce samples, quantifying the nitrate concentration and 20 pesticides (14 organochlorine and 6 organophosphorus pesticides). Significant differences (p < 0.05) between the conventional and organic samples were only detected for the counts of total microorganisms at 30 °C. Pathogens were absent in all the samples. The analytical method, using the quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach for pesticide extraction, was suitable for detecting the targeted analytes; the limit of quantification (LOQ) was between 0.6 and 1.8 µg/kg (lower than the Maximum Residue Levels (MRLs) established by EU legislation). In three organic lettuce samples, one organochlorine pesticide (α-HCH) was observed below the MRLs. For the samples analyzed and for the parameters investigated, except for the total mesophilic counts, the organic and conventional lettuces were not different., This research was supported by the Horizon 2020 project SafeConsume (Grant Agreement No. 727580) and by National Funds from FCT—Fundação para a Ciência e a Tecnologia through projects UID/Multi/50016/2019 and UIDB/50006/2020. Virgínia Cruz Fernandes thanks FCT for the financial support through a Post-doctoral fellowship (SFRH/BPD/109153/2015).

Humans are exposed every day to assorted environmental pollutants namely, polychlorinated biphenyls (PCB), organochlorine pesticides (OCP), brominated flame-retardants (BFR), polycyclic aromatic hydrocarbons (PAH), synthetic musks, heavy metals (e.g. cadmium) and plastic additives (e.g. bisphenol A, BPA). Besides environmental persistence, biomagnification and bioaccumulative properties, these pollutants are classified as endocrine disruptors (EDs), metabolic disruptors, neurologic disruptors. These compounds affect the normal function of several mechanisms in the human body being linked to human health issues as cancer development (e.g. breast, uterine and endometrial), miscarriage, birth defects, premature delivery and infertility. In order to prevent future health issue of women and possible progeny, the assessment of EDs accumulated is essential, particularly in adipose tissue. These samples have been referred as the ideal matrix to establish over time accumulation and long-term exposure of persistent and non-persistent lipophilic EDs. However, the invasive sample collection procedure and methodology processing discourages the usage of this matrix for biomonitoring studies. In this review, a Web of Science search without any publishing year restriction on the analysis in adipose tissue of PCB, OCP, BPA, cadmium, BFR and synthetic musks was performed. A total of 313 studies were found, 158 were European studies from which the studies with data on women EDs accumulation were selected for detailed analysis (n = 90). The results were structured and presented in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The present paper is an overview on the existent EDs analytical methods and levels accumulated in women adipose tissue, with the correspondent health implications across Europe. The limits of detection and quantification were compared and a discussion with results obtained about the presence of ED was performed.

Screening and early diagnosis are crucial to increase the success of cancer patients’ treatments and improve the survival rate. To contribute to this success, distinct electrochemical immunosensing platforms were developed for the analysis of the ExtraCellular Domain of the Human Epidermal growth factor Receptor 2 (HER2‐ECD) through sandwich assays on nanomaterial‐modified screen‐printed carbon electrodes (SPCEs). The most promising platforms showed to be SPCEs modified with (i) gold nanoparticles (AuNPs) and (ii) multiwalled carbon nanotubes combined with AuNPs. The antibody‐antigen interaction was detected using a secondary antibody labelled with alkaline phosphatase and 3‐indoxyl phosphate and silver ions as the enzymatic substrate. The electrochemical signal of the enzymatically generated metallic silver was recorded by linear sweep voltammetry. Under the optimized conditions, linear calibration plots were obtained between 7.5 and 50 ng/mL and the total assay time was 2 h 20 min, achieving LODs of 0.16 ng/mL (SPCE‐MWCNT/AuNP) and 8.5 ng/mL (SPCE‐AuNP), which are well below the established cut‐off value of 15 ng/mL for this cancer biomarker., Maria Freitas is grateful to FCT‐Fundação para a Ciência e a Tecnologia for her PhD grant (SFRH/BD/111942/2015), financed by POPH‐QREN‐Tipologia 4.1‐Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior. This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT) through project UID/QUI/50006/2013.

Aims Considering some fatty acids (FA) have anti-inflammatory effects and by other hand obesity and menopause are associated with pro-inflammation, we aimed to investigate the impact of different FA on metabolic parameters among postmenopausal women obese/overweight. Additionally, since after menopause it takes some years for hormonal changes came to an end, the years past menopause (YPM) and the use of hormonal replacement therapy (HRT) were considered. Methods Blood from 50 Portuguese Caucasian postmenopausal women was collected. FA plasma profile was evaluated by Gas Chromatography–FID (GC-FID). Metabolic parameters analyses were performed in a certified Clinical Analysis Laboratory. Regressions analyses were used to find correlations between FA and metabolic parameters. Results FA were significantly correlated with several metabolic parameters. For women in menopause less than 6 years more correlations were obtained. Unsaturated cis FA (UFAcis), ω3, ω6, Polyunsaturated cis FA (PUFA) and Short-Chain FA (SCFA) were linked with anti-inflammatory activity whereas PUFAtrans, Monounsaturated cis FA (MUFAcis), Saturated FA(SFA) and ω9 were associated with pro-inflammatory pathways. Conclusion FA have a role in both processes: protective and inflammatory, in postmenopausal obese/overweight women. Thus, the understanding of the inflammatory pathways can begin a new perspective in the follow-up for this high metabolic risk population.

In this contribution an electrochemical study is described for the first time of lipid peroxidation and the role of antioxidant on lipid protection using large unilamellar vesicles (LUVs). In order to simulate the cell membrane, LUVs composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were used. A vesicle-modified electrode was constructed by immobilizing DOPC LUVs onto carbon paste electrodes (CPEs). Lipid peroxidation was studied electrochemically by incubating the vesicle-modified electrodes with hydroxyl (HO) radicals generated via the Fenton reaction. Oxidative damage induced by HO was verified by using square wave voltammetry (SWV) and was indirectly measured by the increase of electrochemical peak current to [Fe(CN)6]4- which was used as the electrochemical label. Ascorbic acid (AA) was used as the antioxidant model in order to study its efficacy on free radical scavenging. The decrease of the electrochemical signal confirms the protective key role promoted by AA in the prevention of lipid peroxidation in vesicles. Through microscopy, it was possible to observe morphologic modification on vesicle structures after lipid peroxidation in the presence or absence of AA., This work was financial supported by Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Río Cuarto, and Agencia Nacional de Promoción Científica y Técnica, Marie Curie Actions, International Research Staff Exchange Scheme FP7-PEOPLE-2013-IRSES (612545), and by the European Union (FEDER funds through COMPETE) and National Funds (FCT-Fundação para a Ciência e a Tecnologia) through UID/QUI/50006/2013 and through grants no. PEst-C/EQB/LA0006/2013 and FCOMP-01-0124-FEDER-037285. Fátima Barroso is grateful to FCT by the grant SFRH/BPD/78845/2011, respectively financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior.is gratefully acknowledged. M. A. L. thanks from CONICET for a research post-doctoral fellowship.

The incidence of breast cancer has been increasing over the years. To control and monitor this disease several tumor biomarkers have been proposed for early diagnosis, patient follow-up and/or treatment guidance. The only serum breast cancer biomarker in current use is the cancer antigen 15-3 (CA 15-3). In this work a molecularly imprinted polymer (MIP)-based electrochemical (voltammetric) sensor to monitor breast cancer was developed, based on direct surface imprinting of CA 15-3 on a screen-printed gold electrode (Au-SPE). The imprinting was performed in two steps: (1) adsorption of CA 15-3 on the surface of the Au-SPE and (2) electropolymerization of 2-aminophenol around the adsorbed protein. After extraction of the imprinted protein voltammetric analysis was conducted using hexacyanoferrate(II/III) as redox probe, measuring the signals before and after protein binding. The sensor was characterized by voltammetric techniques and electrochemical impedance spectroscopy, and the analytical responses of imprinted and non-imprinted polymer sensors were studied. A linear relationship between the peak current intensity of the redox probe and the logarithm of CA 15-3 concentration was established between 5 and 50 U mL−1, achieving a limit of detection of 1.5 U mL−1. The prepared MIP-sensor provides fast (15 min) analysis and is cheap, easy to prepare, disposable and could easily be integrated in small portable point-of care devices., João Paulo Grosso Pacheco is grateful to FCT-Fundação para a Ciência e a Tecnologia for a Pos-Doc. grant (SFRH/BPD/101419/2014), financed by POPH-QREN-Tipologia 4.1-Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior. Maria Freitas is grateful to FCT (Fundação para a Ciência e a Tecnologia) for her PhD grant (SFRH/BD/111942/2015). This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT) through project UID/QUI/50006/2013. The authors are greatly indebted to all financing sources.

The purpose of this study was to optimize the extraction of antioxidants from Salicornia ramosissima by-product through Response Surface Methodology (RSM). The impacts of time (10–60 min) and temperature (40–80 °C) in total phenolic content (TPC) and antioxidant/antiradical activity were assessed. The optimal extract was attained at the binomial 80 °C/10 min and characterized regarding phenolic composition, pigments quantification, radicals quenching capacity, anti-acetylcholinesterase (AChE) activity, and cell viability effects. The optimal extract was characterized by low carotenoids content (8.15 μg/g on dry weight (dw)) and high amounts of hydroxycinnamic acid derivatives (23.12 mg/g dw). A remarkable scavenging efficiency was observed, particularly for HOCl (IC50 = 27.61 μg/mL) and ONOO− (IC50 = 47.32 μg/mL). No adverse effects were detected on HaCaT and HFF-1 viability up to 1000 μg/mL. Additionally, the neuroprotective properties of the optimal extract were demonstrated by AChE inhibition (32.34% at 1000 μg/mL). This study encourages the sustainable recovery of antioxidants from the undervalued biowaste of S. ramosissima as a new raw material for different industries.

Tracking unreported allergens in commercial foods can avoid acute allergic reactions. A 2-step electrochemical immunosensor was developed for the analysis of the peanut allergen Ara h 1 in a 1-h assay (&lt, 15 min hands-on time). Bare screen-printed carbon electrodes (SPCE) were used as transducers and monoclonal capture and detection antibodies were applied in a sandwich-type immunoassay. The short assay time was achieved by previously combining the target analyte and the detection antibody. Core/shell CdSe@ZnS Quantum Dots were used as electroactive label for the detection of the immunological interaction by differential pulse anodic stripping voltammetry. A linear range between 25 and 1000 ng·mL−1 (LOD = 3.5 ng·mL−1), an adequate precision of the method (Vx0 ≈ 6%), and a sensitivity of 23.0 nA·mL·ng−1·cm−2 were achieved. The immunosensor was able to detect Ara h 1 in a spiked allergen-free product down to 0.05% (m/m) of peanut. Commercial organic farming cookies and cereal and protein bars were tested to track and quantify Ara h 1. The results were validated by comparison with an ELISA kit.

The scientific community is increasingly concerned about the presence of pharmaceuticals in the aquatic environment, which is a consequence of their high consumption and inefficient removal by wastewater-treatment plants. The search for an effective and sustainable tertiary treatment is therefore needed to enhance their removal. For this purpose, the combination of electrochemical and adsorption processes into three-dimensional (3D) electrochemical systems has been proposed. In this study, a 3D system was studied to remove carbamazepine, an antiepileptic, consumed in high doses and very persistent in the environment. The influences of the following parameters on its removal were evaluated: anode and cathode materials and distance between them, electrolyte (NaCl) concentration and pH, and the (carbon-based) adsorbent material used as the particulate electrode. The obtained results demonstrated that the introduction of the particulate electrode improved the removal efficiency. This can be attributed to the simultaneous occurrence of different phenomena, such as adsorption/electrosorption, electrocoagulation, oxidation, and catalytic degradation.

BACKGROUND For obtaining bioactive fractions from plant material subcritical water is notably advantageous in comparison to any other technique. Excellent solvating properties and selectivity combined with finely tuned reactivity of subcritical water enable exploitation of plants potentials. Herein subcritical water extraction was used for recovery of bioactive compounds from leaves, roots and fruits of Sambucus ebulus L. Obtained extracts were characterized in terms of biological and chemical fingerprints. RESULTS AND DISSCUSION Results obtained by using several antioxidant assays that focused on different mechanisms showed that subcritical water extracts of Sambucus ebulus were proved to be powerful antioxidants. Enzyme inhibitory effects were tested against α-amylase, α-glucosidase and tyrosinase, and the results showed anti-diabetic potential of the extracts as well as its possible use in case of skin disorders. Antiproliferative properties were detected on three cell lines (A-549; LS-174T; HeLa) and showed prominent cytotoxicity against all tumor cell lines. Obtained inhibitory concentrations were in the range 0.58-8.10 µg/mL. Generally, the leaves SCW extracts exhibited stronger biological activities with higher levels of phenolic compounds compared to roots and fruits of Sambucus ebulus. Gallic acid, catechin and caffeic acid were identified as major components in these extracts and these components seem to relate with observed biological activity. CONCLUSION The obtained results suggest that Sambucus ebulus has a high potential for preparing new phyto-pharmaceuticals and functional food ingredients.

Widespread use of pharmaceuticals and suboptimal wastewater treatment have led to increased levels of these substances in aquatic ecosystems. Lipid-lowering drugs such as gemfibrozil, which are among the most abundant human pharmaceuticals in the environment, may have deleterious effects on aquatic organisms. We examined the genotoxicity of gemfibrozil in a fish species, the gilthead seabream (Sparus aurata), which is commercially important in southern Europe. Following 96-h waterborne exposure, molecular (erythrocyte DNA strand breaks) and cytogenetic (micronuclei and other nuclear abnormalities in cells) endpoints were measured. Gemfibrozil was positive in both endpoints, at environmentally relevant concentrations, a result that raises concerns about the potential genotoxic effects of the drug in recipient waters.

Pilocarpus microphyllus Stapf ex Wardlew (Rutaceae), popularly known as jaborandi, is a plant native to the northern and northeastern macroregions of Brazil. Several alkaloids from this species have been isolated. There are few reports of antibacterial and anthelmintic activities for these compounds. In this work, we report the antibacterial and anthelmintic activity of five alkaloids found in P. microphyllus leaves, namely, pilosine, epiisopilosine, isopilosine, epiisopiloturine and macaubine. Of these, only anthelmintic activity of one of the compounds has been previously reported. Nuclear magnetic resonance, HPLC and mass spectrometry were combined and used to identify and confirm the structure of the five compounds. As regards the anthelmintic activity, the alkaloids were studied using in vitro assays to evaluate survival time and damaged teguments for Schistosoma mansoni adult worms. We found epiisopilosine to have anthelmintic activity at very low concentrations (3.125 μg mL-1 ); at this concentration, it prevented mating, oviposition, reducing motor activity and altered the tegument of these worms. In contrast, none of the alkaloids showed antibacterial activity. Additionally, alkaloids displayed no cytotoxic effect on vero cells. The potent anthelmintic activity of epiisopilosine indicates the potential of this natural compound as an antiparasitic agent. Copyright © 2017 John Wiley & Sons, Ltd.

The cultivation of genetically modified organisms (GMO) continues to expand worldwide. Still, many consumers express concerns about the use of GMO in food or feed, and many countries have legislated on labelling systems to indicate the presence of GMO in commercial products. To deal with the increased number of GMO events and to address related regulations, alternative detection methods for GMO inspection are required. In this work, a genosensor based on Surface Plasmon Resonance under continuous flow was developed for the detection and quantification of a genetically modified soybean (event GTS 40-3-2). In a single chip, the simultaneous detection of the event-specific and the taxon-specific samples were achieved, whose detection limits were 20 pM and 16 pM, respectively. The reproducibility was 1.4%, which supports the use of the chip as a reliable and cost-effective alternative to other DNA-based techniques. The results indicate that the proposed method is a versatile tool for GMO quantification in food and feed samples. This work received financial support from the European Union (FEDER funds through COMPETE), National Funds (FCT, Fundação para a Ciência e a Tecnologia) through project UID/QUI/ 50006/2013 and cofinanced by FEDER funds. Alexandra Plácido is grateful for the FCT grant (SFRH/BD/97995/2013) financed by POPH-QREN (subsidized by FSE and MCTES).

A green synthetic route was developed to prepare silver nanoparticles (AgNPs) in aqueous solution for biological applications. Eschweilenol C, a compound derivative ellagic acid was identified as the main constituent of the aqueous fraction of the ethanolic extract of Terminalia fagifolia Mart. by NMR analysis. In the green synthesis, the ethanolic extract of T. fagifolia and its aqueous fraction were used to promote silver reduction and nanoparticle stabilization. The synthesized AgNPs presented a spherical or polygonal morphology shape by TEM analysis and AgNPs showed high levels of antioxidant and considerable antibacterial and antifungal activities. Synthesized nanoparticles presented significant antioxidant activity by sequestration of DPPH and ABTS radicals, in addition to iron reduction (FRAP assay) and measurement of antioxidant capacity in ORAC units, in addition, AgNP synthesized with the aqueous fraction also demonstrated antioxidant potential in microglial cells. Gram-positive and Gram-negative bacteria were susceptible to growth inhibition by the nanoparticles, among which the AgNPs formed by the ethanolic extract was the most effective. The data obtained by AFM images suggested that AgNPs could lead to the lysis of bacteria and subsequent death. The antifungal assays showed high efficiency against yeasts and dermatophytes. This work represents the first description of antifungal activity by AgNPs against Fonsecaea pedrosoi, the etiologic agent of chromoblastomycosis. In relation to biocompatibility, the AgNPs induced lower haemolysis than AgNO3., We thank Herbert Kogler and Reinhard Wimmer for the identification of Eschweilenol C. The NMR laboratory at Aalborg University is supported by the Obel Family, SparNord and Carlsberg foundations.The authors are grateful to Carla Eiras (LIMAV/CT/UFPI) and to FCT and EU for financial support through project UID/QUI/50006/2013– POCI-01-0145-FEDER-007265 from COMPETE and projectNORTE-01-0145-FEDER-000011 from COMPETE. Thanks to Andreia Pinto for help with the TEM measurements at Instituto de Medicina Molecular (IMM). This work was supported by the Histology and Comparative Pathology Laboratory of the IMM

To our knowledge, this is the first study in Portugal for the analysis of both antibiotics and psychiatric drugs in river waters from Douro and Leça rivers and its sediments. Samples were extracted using solid phase extraction and QuEChERS procedures and the analysis was performed using liquid chromatography with tandem mass spectrometry. Higher number of pharmaceuticals and higher concentrations were found in the Leça river for the two matrices under study. The highest detection frequency for the twenty-seven pharmaceuticals was observed for fluoxetine with 83.3% in both matrices. None of the studied antibiotics were detected in water collected in the Douro river however six antibiotics (azithromycin, ciprofloxacin, clarithromycin, moxifloxacin, ofloxacin, and trimethoprim) were found in the Leça river. Further, the analysis of sediments exhibits the occurrence of sulfamethoxypyridazine in the Douro river and azithromycin in the Leça river. The highest concentration was observed in Leça river for azithromycin in the river water (2,819 ng/L) and sediments (43.2 ng/g). Carbamazepine, citalopram, fluoxetine, sertraline, trazodone, and venlafaxine were the psychiatric drugs detected in river waters and sediments with more diazepam as water river contamination. Concentrations ranged frommethod detection limit (MDL)) to 2.0 ng/L (river waters) andMDL to 0.251 ng/g (sediments) in Douro river and between MDL to 354 ng/L (river waters) andMDL to 6.35 ng/g (sediments) in Leça river. To identify the possible sources of pollution more monitoring studies should be performed along the studied rivers.

Three-level Box-Behnken experimental design with three factors (pH, temperature and antibiotic initial concentration) combined with response surface methodology (RSM) was applied to study the removal of Metronidazole and Sulfamethoxazole by walnut shell based activated carbon. This methodology enabled to identify the effects of the different factors studied and their interactions in the response of each antibiotic. The relationship between the independent variable (sorption capacity) and the dependent variables (pH, temperature and antibiotic concentration) was adequately modelled by second-order polynomial equation. The pH factor exerted a significant but distinct influence on the removal efficiency of both antibiotics. The removal of Metronidazole is favoured by increasing pH values, with the maximum value obtained for pH 8 - upper limit of the study domain; while Sulfamethoxazole displays a maximum value around 5.5, with a decrease in the extent of adsorption as the pH increases. The best conditions, predicted by the model, for the removal of the antibiotic Sulfamethoxazole (106.9 mg/g) are obtained at a temperature of 30 °C, initial concentration of 40 mg/L and a pH value of 5.5. For the antibiotic Metronidazole, the highest removal value (127 mg/g) is expected to occur at the maximum levels attributed to each of the factors (pH = 8, Cin = 40 mg/L, T = 30 °C). The results of isotherm experiments (at 20 °C and pH 6) displayed a good agreement with the models predictions. The maximum sorption capacity, estimated by the Langmuir model, was 107.4 mg/g for Metronidazole and 93.5 mg/g for Sulfamethoxazole., This work was financially supported by the projects POCI-01-0145-FEDER-006939 (LEPABE), POCI-010145-FEDER-007265 (REQUIMTE/LAQV) funded by the European Regional Development Fund (ERDF), through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) and by national funds, through FCT - Fundação para a Ciência e a Tecnologia (LEPABE - UID/EQU/00511/2013; REQUIMTE/LAQV - UID/QUI/50006/2013) and NORTE-01-0145-FEDER-000005 – LEPABE-2-ECO-INNOVATION, supported by North Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).

Chronic diseases are one of the main causes of death worldwide mainly due to lifestyle factors, such as the high consumption of processed foods, low consumption of plants, and lack of exercise. In the last decade, different studies suggest the use of dietary supplements to attenuate many of the pathophysiological processes involved in the development of some of these diseases. Cross-sectional and prospective cohorts and other studies have shown in some situations an association between nutritional supplements diet and a lower prevalence and incidence of chronic diseases, such as cardiovascular disease, cancer, metabolic syndrome, diabetes, and neurodegenerative diseases as well as a reduced overall mortality. However, the number of clinical studies is very limited and in some cases controversial. The present chapter aims to review the role of a variety of dietary supplements in treating some of these chronic diseases through evaluation of the evidence-based effects of nutritional supplements on human health, from cohort studies to clinical trials and meta-analysis.

Submitted by Manuel Brandão (recipp@sc.ipp.pt) on 2020-07-08T11:56:07Z No. of bitstreams: 1 ART_DELERUE_2019_18.pdf: 2997327 bytes, checksum: af474a9e2419c63cafe457462ffda344 (MD5) Approved for entry into archive by Ana Maria de Sousa Rebelo (amsr@isep.ipp.pt) on 2020-07-10T14:08:56Z (GMT) No. of bitstreams: 1 ART_DELERUE_2019_18.pdf: 2997327 bytes, checksum: af474a9e2419c63cafe457462ffda344 (MD5) Made available in DSpace on 2020-07-10T14:08:56Z (GMT). No. of bitstreams: 1 ART_DELERUE_2019_18.pdf: 2997327 bytes, checksum: af474a9e2419c63cafe457462ffda344 (MD5) Previous issue date: 2019 This research was supported through the COMPETE – Operational Competitiveness Program and national funds through FCT – Foundation for Science and Technology, under the project “NANOAu – Effects of Gold Nanoparticles to Aquatic Organisms” (FCT PTDC/MAR-EST/3399/2012) (FCOMP-01-0124-FEDER-029435), through FCT/MCTES through national funds (PIDDAC), the cofounding by FEDER, within the PT2020 Partnership Agreement and Compete 2020 to CESAM (UID/AMB/50017 – POCI-01-0145-FEDER-007638) and UID/QUI/50006/2013. A. Barreto has a doctoral fellowship from FCT (SFRH/BD/97624/2013); L. G. Luis had a fellowship from FCT (BI/UI88/6881/2014). MO has financial support of the program Investigator FCT, co-funded by the Human Potential Operational Programme and European Social Fund (IF/00335(2015)). info:eu-repo/semantics/publishedVersion

Given their therapeutic activity, natural products have been used in traditional medicines throughout the centuries. The growing interest of the scientific community in phytopharmaceuticals, and more recently in marine products, has resulted in a significant number of research efforts towards understanding their effect in the treatment of neurodegenerative diseases, such as Alzheimer's (AD), Parkinson (PD) and Huntington (HD). Several studies have shown that many of the primary and secondary metabolites of plants, marine organisms and others, have high affinities for various brain receptors and may play a crucial role in the treatment of diseases affecting the central nervous system (CNS) in mammalians. Actually, such compounds may act on the brain receptors either by agonism, antagonism, allosteric modulation or other type of activity aimed at enhancing a certain effect. The current manuscript comprehensively reviews the state of the art on the interactions between natural compounds and brain receptors. This information is of foremost importance when it is intended to investigate and develop cutting-edge drugs, more effective and with alternative mechanisms of action to the conventional drugs presently used for the treatment of neurodegenerative diseases. Thus, we reviewed the effect of 173 natural products on neurotransmitter receptors, diabetes related receptors, neurotrophic factor related receptors, immune system related receptors, oxidative stress related receptors, transcription factors regulating gene expression related receptors and blood-brain barrier receptors., The author A.R.S. is grateful to the authors C.G. and J.M.R for the scientific assistance and suggestions shared throughout the supervision of her B.Sc. final project. The author A.R.S. also acknowledges the Department of Biology (DB) and the Centro de Biologia Molecular e Ambiental [Centre of Molecular and Environmental Biology] (CBMA) from School of Sciences (EC), University of Minho (UM), Braga, Portugal, by providing all the conditions leading to the B.Sc. in Biochemistry. The corresponding author C.G. is grateful to Fundação para a Ciência e a Tecnologia [Foundation for Science and Technology, FCT I.P.] (FCT I.P.) for the FCT Investigator (IF/01332/2014/CP1255/CT0001). The author J.M.R. acknowledges the CBMA and the Instituto de Ciência e Inovação para a Biosustentabilidade [Institute of Science and Innovation for Biosustainability] (IBS) from University of Minho (UM), Braga, Portugal, where he recently carried out his activities as invited assistant researcher and professor. The author J.M.R. is also grateful to the Laboratório Associado para a Química Verde (LAQV) [Green Chemistry Laboratory] from REQUIMTE – Rede de Química e Tecnologia [REQUIMTE – Chemistry and Technology Network], as well as to the Department of Chemistry and Biochemistry (DCB) from the Faculty of Sciences from University of Porto (FCUP), Porto, Portugal, where he is currently researcher. Regarding to the author J.M.R., this work was supported by the strategic programmes UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) (Research project entitled “EcoAgriFood: Innovative green products and processes to promote Agri-Food BioEconomy”) and PTDC/SAUNUT/30448/2017 (POCI-01-0145-FEDER-030448) (Research project entitled “Poly4CD: Action of Dietary Polyphenols in Preventing Celiac Disease”) funded by Portuguese national funds through Fundação para a Ciência e Tecnologia [Foundation for Science and Technology] (FCT-I.P.)/Ministério da Ciência, Tecnologia e Ensino Superior [Ministry of Science, Technology and Higher Education] (MCTES), and Fundo Europeu de Desenvolvimento Regional [European Regional Development Fund] (FEDER), under the scope of the COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI) [COMPETE2020 – Competitiveness and Internationalization Operational Program] and by COST Action 18101 “SOURDOMICS”, supported by COST (European Cooperation in Science and Technology). C. G. and C. D.-M. are grateful for the financial support from FCT/MCTES through national funds (UID/QUI/50006/2019). C. G. and C. D.-M. would also like to thank the EU and FCT for funding through the projects: DESignBIOtecHealth - New Technologies for three Health Challenges of Modern Societies: Diabetes, Drug Abuse and Kidney Diseases (Portugal2020, Norte-01-0145-FEDER-000024) and project PTDC/OCE-ETA/30240/2017- SilverBrain - From sea to brain: Green neuroprotective extracts for nanoencapsulation and functional food production (POCI-01-0145-FEDER-030240); and to the REQUIMTE for the project “Sea_4_Brain_Food”. All the authors acknowledge the CBMA and IB-S by the financial support provided specifically for this open access publication.

During kiwiberry production, different by-products are generated, including leaves that are removed to increase the fruit’s solar exposure. The aim of this work was to extract bioactive compounds from kiwiberry leaf by employing microwave-assisted extraction (MAE). Compatible food solvents (water and ethanol) were employed. The alcoholic extract contained the highest phenolic and flavonoid contents (629.48 mg of gallic acid equivalents (GAE) per gram of plant material on dry weight (dw) (GAE/g dw) and 136.81 mg of catechin equivalents per gram of plant material on dw (CAE/g dw), respectively). Oppositely, the hydroalcoholic extract achieved the highest antioxidant activity and scavenging activity against reactive oxygen and nitrogen species (IC50 = 29.10 μg/mL for O2•−, IC50 = 1.87 μg/mL for HOCl and IC50 = 1.18 μg/mL for •NO). The phenolic profile showed the presence of caffeoylquinic acids, proanthocyanidin, and quercetin in all samples. However, caffeoylquinic acids and quercetin were detected in higher amounts in the alcoholic extract, while proanthocyanidins were prevalent in the hydroalcoholic extract. No adverse effects were observed on Caco-2 viability, while the highest concentration (1000 µg/mL) of hydroalcoholic and alcoholic extracts conducted to a decrease of HT29-MTX viability. These results highlight the MAE potentialities to extract bioactive compounds from kiwiberry leaf.

The peptide PcL342-354C was obtained from the Cry1Ab16 toxin present in Bacillus thuringiensis ("Computational Modeling Deduced Three Dimensional Structure of Cry1Ab16 Toxin from B. thuringiensis AC11" (Kashyap, 2012) [1]). In this data article, we report the synthesis and characterization of the PcL342-354C peptide by MALDI-TOF/TOF mass spectrometry. In addition, the preparation of layer-by-layer films is shown based on interspersion of this peptide with both polyethylenimine (PEI) and poly(sodium 4-styrenesulfonate) (PSS), self-assembled on ITO (indium tin oxide) electrodes. The morphology of the ITO/PEI/PSS/PcL342-354C film was analyzed using atomic force microscopy (AFM). We also evaluated the effect of the number of bilayers in ITO/PEI/(PSS/PcL342-354C)n on the morphology of the film using AFM amplitude images. Further details about this study were published elsewhere, "Layer-by-layer films containing peptides of the Cry1Ab16 toxin from B. thuringiensis for potential biotechnological applications," (Plácido et al., 2016) [2]. Fil: Plácido, Alexandra. Instituto Superior de Engenharia Do Porto; Portugal Fil: de Oliveira Farias, Emanuel Airton. Universidade Federal do Piauí; Brasil Fil: Marani, Mariela Mirta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina Fil: Gomes Vasconcelos, Andreanne. Universidade Federal do Piauí; Brasil Fil: Leite, José R. S. A.. Universidade Federal do Piauí; Brasil Fil: Delerue Matos, Cristina. Instituto Superior de Engenharia Do Porto; Portugal