Enhancing curcumin bioaccessibility through different nanoformulations

5th International Conference on Food Digestion[Excerpt] Introduction: Curcumin, a natural polyphenolic phytochemical, is known for its wide range of biological activities, however it has an extremely low water solubility as well as a low bioavailability, which limit its application as a bioactive ingredient in food. The use of delivery systems at nanoscale such as nanoemulsions (NE) and solid lipid nanoparticles (SLN) has been reported as a promising mean of improving the lipophilic bioactive compounds’ bioavailability and their physical and chemical stability. However, the knowledge of the behaviour of different nanoformulations as well as the fate of bioactive compounds encapsulated within them in the gastrointestinal (GI) tract is of utmost importance to either assess their safety for human consumption and to produce tailored delivery systems (i.e. with optimized bioactivity). The aim of this work was the evaluation of the behaviour of two different bio-based nanoformulations (NE and SLN) incorporating curcumin when submitted to an in vitro digestion and the assessment of their cytotoxicity. [...]Ana C. Pinheiro and Joana T. Martins acknowledge the Foundation for Science and Technology (FCT) for their fellowships (SFRH/BPD/101181/2014 and SFRH/BPD/89992/2012). This work was supported by Portuguese Foundation for Science and Technology (FCT) under the scope of the Project PTDC/AGR-TEC/5215/2014, of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) 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.info:eu-repo/semantics/publishedVersio

Bio-based nanocarriers incorporating curcumin bioaccessibility and cell viability evaluation

Book of Abstracts of CEB Annual Meeting 2017[Excerpt] For decades, curcumin (Cur), a natural polyphenol product derived from turmeric (Curcuma longa) has been considered one of the most promising bioactive compounds due to its health benefits such as anti-inflammatory, antioxidant and anticarcinogenic properties. However, Cur application as functional compound in food products has been limited due to light, heat, and oxidation sensitive and mainly, to poor aqueous solubility which limit its bioavailability [1]. To increase Cur bioaccessibility and consequently, increase bioavailability, several carriers have been investigated, particularly nanocarriers. Among the various nanocarriers described in the literature, lipid-based nanocarriers may offer a promising tool to increase the stability, efficacy and safety of lipophilic compounds, namely Cur [2]. Moreover, the understanding of Cur-loaded nanocarriers’ behaviour under gastrointestinal (GI) conditions is fundamental to produce safe and customized nanocarriers with optimized bioactivity for oral consumption. The aim of this study was to comparatively analyze the impact of two different lipid nanocarriers incorporating Cur - solid lipid nanoparticles (SLN) and nanoemulsions (NE) – on bioaccessibility and Caco-2 cells viability. [...]info:eu-repo/semantics/publishedVersio

Mechanisms underlying dioxygen reduction in laccases. Structural and modelling studies focusing on proton transfer

<p>Abstract</p> <p>Background</p> <p>Laccases are enzymes that couple the oxidation of substrates with the reduction of dioxygen to water. They are the simplest members of the multi-copper oxidases and contain at least two types of copper centres; a mononuclear T1 and a trinuclear that includes two T3 and one T2 copper ions. Substrate oxidation takes place at the mononuclear centre whereas reduction of oxygen to water occurs at the trinuclear centre.</p> <p>Results</p> <p>In this study, the CotA laccase from <it>Bacillus subtilis </it>was used as a model to understand the mechanisms taking place at the molecular level, with a focus in the trinuclear centre. The structures of the holo-protein and of the oxidised form of the apo-protein, which has previously been reconstituted <it>in vitro </it>with Cu(I), have been determined. The former has a dioxygen moiety between the T3 coppers, while the latter has a monoatomic oxygen, here interpreted as a hydroxyl ion. The UV/visible spectra of these two forms have been analysed in the crystals and compared with the data obtained in solution. Theoretical calculations on these and other structures of CotA were used to identify groups that may be responsible for channelling the protons that are needed for reduction of dioxygen to water.</p> <p>Conclusions</p> <p>These results present evidence that Glu 498 is the only proton-active group in the vicinity of the trinuclear centre. This strongly suggests that this residue may be responsible for channelling the protons needed for the reduction. These results are compared with other data available for these enzymes, highlighting similarities and differences within laccases and multicopper oxidases.</p

Classificação Geotécnica do maciço rochoso da Pedreira do Monte das Flores – Évora (Portugal)

No âmbito da atualização do processo de homologação da Pedreira do Monte das Flores - Évora, como fornecedora de balastro ferroviário, procedeu-se à classificação geotécnica do maciço rochoso existente no local. Esta exploração possui 96,90 ha de área arrendada estando 70,70 ha concessionados à exploração. Geologicamente, a área estudada pertence à Zona de Ossa-Morena (Maciço de Évora), enquanto parte integrante do Maciço Ibérico que constitui o setor mais ocidental e contínuo da Cadeia Orogénica Varisca na Europa. Também os eventos tectonotérmicos alpinos se fizeram sentir neste maciço de forma atenuada, permitindo a preservação da história geológica mesozóica [Moita, 2008]. A unidade geológica onde se insere a exploração é constituída por litótipos ígneos e metamórficos de idade precâmbrica e paleozóica [Andrade et al., 1976], denotando-se um domínio das formações de rochas eruptivas. Na área estudada ocorre um afloramento de quartzodiorito e granodiorito de grão médio, não porfiróide, onde os minerais mais representativos da rocha são o feldspato potássico, a plagioclase e o quartzo, sem evidenciarem qualquer orientação à vista desarmada [Moita, 2008]. Neste estudo fez-se a descrição geotécnica da qualidade do maciço rochoso que ocorre na Pedreira do Monte das Flores. Por se tratar de um sistema de classificação generalista e correntemente utilizado na avaliação do comportamento geomecânico dos maciços rochosos, utilizou-se a Descrição Geotécnica Básica (“Basic Geotechnical Description”- BGD), proposta pela Sociedade Internacional de Mecânica das Rochas [ISRM, 1981]. O principal objetivo da aplicação desta classificação foi o de efetuar um zonamento geotécnico do maciço rochoso, quer do local atualmente em exploração, quer da área contígua para onde se prevê o alargamento da corta da exploração, com base no reconhecimento geológico e, na amostragem efetuada em locais selecionados para posterior realização de ensaios laboratoriais de caracterização mecânica. Este reconhecimento geológico de superfície permitiu identificar o tipo de rocha presente, determinar as características estruturais e caracterizar a alteração do maciço rochoso, assim como, definir as diferentes famílias de fraturas presentes e quantificar o espaçamento entre as descontinuidades nestas famílias. Com o objetivo de determinar as características mecânicas, tais como, a resistência à compressão uniaxial e o ângulo de atrito das fraturas, foi realizada uma campanha de ensaios laboratoriais e de campo, nomeadamente, o ensaio de resistência à compressão uniaxial, complementado pelo ensaio de carga pontual, de modo a determinar o valor daquela resistência. Realizou-se também o ensaio com o martelo de Schmidt dado ser um ensaio simples e rápido na caracterização de materiais [Pinho, 2003], que permite estimar o valor da resistência à compressão uniaxial dos planos das descontinuidades do maciço rochoso (JCS), nos diferentes locais de amostragem. O parâmetro JCS, o coeficiente de rugosidade da descontinuidade (JRC) e a tensão efectiva normal (’n ), foram necessários para obter o ângulo de atrito das diaclases (Øpico), de acordo com o método proposto pela Sociedade Internacional de Mecânica das Rochas [ISRM, 1978]. O estudo realizado permitiu concluir que o maciço apresenta boa qualidade, em regra, homogénea relativamente às suas características geológicas e geotécnicas. No entanto, distinguem-se duas zonas, ZG1 e ZG2, com base em pequenas diferenças nos valores da resistência à compressão uniaxial da rocha e do ângulo de atrito das fraturas

Microfluidic-assisted electrospinning, an alternative to coaxial, as a controlled dual drug release system to treat inflammatory arthritic diseases

Inflammatory arthritic diseases are characterized by a persistent inflammation of the synovial tissues where tumor necrosis factor alpha (TNFα) and interleukin-6 (IL-6) pro-inflammatory cytokines are over-expressed, leading to progressive musculoskeletal disability. Methotrexate (MTX), a disease-modifying-anti-rheumatic drug (DMARD) commonly applied in their treatment, can be used in combination with biological-DMARDs as anti-TNFα antibody to improve the treatments efficacy. However, their systemic administration comes with severe side-effects and limited therapeutic efficacy due to their off-target distribution and short half-life. To overcome such limitations, encapsulation of clinically relevant concentrations of MTX and anti-TNFα antibody into polycaprolactone (PCL) or poly(vinyl-alcohol) (PVA) microfluidic-assisted or coaxial electrospun fibrous meshes is proposed as local controlled dual drug release systems. Release studies show that microfluidic-assisted electrospinning meshes encapsulating both drugs achieved higher concentrations than coaxials. Biological assays using human articular chondrocytes (hACs) and monocytic cells (THP-1 cell line) demonstrate that fibrous meshes encapsulating the drugs are non-toxic. The systems' efficacy is proved by a significant decrease of TNFα and IL-6 concentrations in conditioned medium of lipopolysaccharide (LPS)-stimulated THP-1 cells, especially in the presence of microfluidic-assisted electrospun meshes, when compared with THP-1 conditioned medium (59.5% and 83.9% less, respectively). Therefore, microfluidic-assisted electrospinning fibrous meshes with encapsulating drugs represent an alternative to coaxial, as a local therapy for inflammatory arthritis diseases.This work was supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, for the Ph.D grant of Catarina Silva (UMINHO/BD/33/2016; NORTE-08-5369-FSE-000012), and by the Portuguese Science and Technology Foundation (FCT) for the cells project Cells4_ID (PTDC/BTM-SAL/28882/2017)

Behavioural stress responses predict environmental perception in European sea bass (Dicentrarchus labrax)

Individual variation in the response to environmental challenges depends partly on innate reaction norms, partly on experience-based cognitive/emotional evaluations that individuals make of the situation. The goal of this study was to investigate whether pre-existing differences in behaviour predict the outcome of such assessment of environmental cues, using a conditioned place preference/avoidance (CPP/CPA) paradigm. A comparative vertebrate model (European sea bass, Dicentrarchus labrax) was used, and ninety juvenile individuals were initially screened for behavioural reactivity using a net restraining test. Thereafter each individual was tested in a choice tank using net chasing as aversive stimulus or exposure to familiar conspecifics as appetitive stimulus in the preferred or non preferred side respectively (called hereafter stimulation side). Locomotor behaviour (i.e. time spent, distance travelled and swimming speed in each tank side) of each individual was recorded and analysed with video software. The results showed that fish which were previously exposed to appetitive stimulus increased significantly the time spent on the stimulation side, while aversive stimulus led to a strong decrease in time spent on the stimulation side. Moreover, this study showed clearly that proactive fish were characterised by a stronger preference for the social stimulus and when placed in a putative aversive environment showed a lower physiological stress responses than reactive fish. In conclusion, this study showed for the first time in sea bass, that the CPP/CPA paradigm can be used to assess the valence (positive vs. negative) that fish attribute to different stimuli and that individual behavioural traits is predictive of how stimuli are perceived and thus of the magnitude of preference or avoidance behaviour.European Commission [265957]; Portuguese Fundacao para a Ciencia e Tecnologia (FCT) [FRH/BPD/72952/2010]; FCT [SFRH/BD/80029/2011

Differential effects of food restriction and warming in the two-spotted goby: impaired reproductive performance and stressed offspring

Climate change is a growing threat to marine organisms and ecosystems, and it is already modifying ocean properties by, for example, increasing temperature and decreasing pH. Increasing water temperature may also lead to an impairment of primary productivity and an overall depletion of available zooplankton. Understanding how the crossover between warming and zooplankton availability impacts fish populations has paramount implications for conservation and mitigation strategies. Through a cross factorial design to test the effects of ocean temperature and food availability in a temperate marine teleost, Pomatochistus flavescens, we showed that hindered feeding impacted sheltering and avoidance behaviour. Also, low food availability impaired fish reproduction, particularly male reproduction, as the expression of cyp11b1, a gene with a pivotal role in the synthesis of the most important fish androgen, 11-ketotestosterone, was significantly reduced under a low food regime. In contrast, temperature alone did not affect reproductive success, but offspring showed increased saturated fatty acid content (embryos) and increased lipid peroxidation (larvae). Altogether, food availability had a stronger effect on fitness, showing that coping with elevated temperatures, an ability that may be expected in shallow-water fish, can be indirectly impacted, or even overwhelmed, by the effects of ocean warming on primary productivity and downstream ecological processes.info:eu-repo/semantics/publishedVersio

Stromal MED12 exon 2 mutations in complex fibroadenomas of the breast

© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. http://creativecommons.org/licenses/by-nc/4.0/This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.Aims: Here we explore the presence of mediator complex subunit 12 (MED12) exon 2 and telomerase reverse transcriptase (TERT) promoter hotspot mutations in complex fibroadenomas (CFAs) of the breast. Methods: The stromal components from 18 CFAs were subjected to Sanger sequencing of MED12 exon 2 and the TERT promoter hotspot loci. The epithelial and stromal components of two MED12 mutated CFAs were subjected to laser capture microdissection, and Sanger sequencing of MED12 exon 2, TERT promoter and PIK3CA exons 9 and 20, separately. Results: MED12 exon 2 mutations were identified in the stroma of 17% of CFAs. The analyses of epithelial and stromal components, microdissected separately, revealed that MED12 mutations were restricted to the stroma. No TERT promoter or PIK3CA mutations in exons 9 and 20 were detected in analysed CFAs. Conclusions: Like conventional fibroadenomas, MED12 exon 2 mutations appear to be restricted to the stromal component of CFAs, supporting the notion that CFAs are stromal neoplasms.This study was funded by the Breast Cancer Research Foundation. BW is funded by a Cycle for Survival grant, CS by a Fundação para a Ciência e Tecnologia grant (SFRH/BDE/110544/2015). FP is partially funded by a K12 CA184746 grant. The research reported in this paper was supported in part by a Cancer Centre Support Grant of the National Institutes of Health/National Cancer Institute (grant No P30CA008748).info:eu-repo/semantics/publishedVersio

RiskBenefit4EU project – a strategy for risk-benefit assessment of foods in Portugal

O balanço entre os riscos e benefícios para a saúde resultante do consumo de alimentos é um importante contributo para apoiar a definição de políticas de saúde e a promoção da literacia dos consumidores. No âmbito do projeto RiskBenefit4EU, financiado pela European Food Safety Authority e coordenado pelo Departamento de Alimentação e Nutrição do Instituto Nacional de Saúde Doutor Ricardo Jorge, o presente trabalho pretende descrever a estratégia implementada para a capacitação das equipas portuguesas em avaliação de risco-benefício de alimentos (RBA). Concretizada pelos parceiros do Institut National de la Recherche Agronomique e da Technical University of Denmark, a capacitação consiste em três atividades principais: 1) Formação teórica, focando os conceitos-chave para a avaliação de RBA; 2) Formação prática, aplicando os conceitos adquiridos e as metodologias transmitidas a um estudo de caso; e 3) Missões científicas, de curta duração, para formação avançada em domínios específicos da avaliação de RBA. No que diz respeito à formação prática, e com o objetivo de consolidar os conhecimentos adquiridos em avaliação de RBA, está previsto o desenvolvimento de um estudo de caso português sobre alimentos à base de cereais habitualmente consumidos por crianças. A estratégia de capacitação seguida neste projeto servirá de modelo para outras equipas e países, contribuindo para a disseminação de uma cultura de avaliação de RBA nas vertentes toxicológica, microbiológica e nutricional a nível internacional.The balance of risks and health benefits from food consumption constitutes a crucial topic to consumer literacy and health policy-makers. Through the RiskBenefit4EU project, funded by the European Food Safety Authority, and coordinated by the Food and Nutrition Department of Portuguese National Institute of Health Dr. Ricardo Jorge, the present work intends to identify the applied strategy to capacitate the Portuguese teams for the development and implementation of risk-benefit assessment (RBA) in food. The training of the Portuguese team is being accomplished by the French National Institute for Agricultural Research and the Technical University of Denmark members, through three main capacity building activities: 1) Theoretical training, focusing on the key concepts for RBA; 2) Practical training, applying the concepts acquired and the methodologies transmitted to a concrete case study; and 3) Short-term scientific missions for advanced training in specific areas of RBA. In order to complete the training of the Portuguese teams and consolidate the knowledge acquired in RBA, a Portuguese case study on cereal-based foods usually consumed by children is planned. The training strategy followed in this project will contribute as a model of capacity building for disseminating a culture of risk-benefit assessment in the toxicological, microbiological and nutritional aspects at the international level.Trabalho desenvolvido no âmbito do projeto “RiskBenefit4EU – Partnering to strengthen the risk-benefit assessment within EU using a holistic approach” financiado pela EFSA Partnering Grants (Grant Agreement Number A/EFSA/AFSCO/2017/01 – GA02).info:eu-repo/semantics/publishedVersio

Can we predict personality in fish? searching for consistency over time and across contexts

The interest in animal personality, broadly defined as consistency of individual behavioural traits over time and across contexts, has increased dramatically over the last years. Individual differences in behaviour are no longer recognised as noise around a mean but rather as adaptive variation and thus, essentially, raw material for evolution. Animal personality has been considered evolutionary conserved and has been shown to be present in all vertebrates including fish. Despite the importance of evolutionary and comparative aspects in this field, few studies have actually documented consistency across situations in fish. In addition, most studies are done with individually housed fish which may pose additional challenges when interpreting data from social species. Here, we investigate, for the first time in fish, whether individual differences in behavioural responses to a variety of challenges are consistent over time and across contexts using both individual and grouped-based tests. Twenty-four juveniles of Gilthead seabream Sparus aurata were subjected to three individual-based tests: feed intake recovery in a novel environment, novel object and restraining and to two group-based tests: risk-taking and hypoxia. Each test was repeated twice to assess consistency of behavioural responses over time. Risk taking and escape behaviours during restraining were shown to be significantly consistent over time. In addition, consistency across contexts was also observed: individuals that took longer to recover feed intake after transfer into a novel environment exhibited higher escape attempts during a restraining test and escaped faster from hypoxia conditions. These results highlight the possibility to predict behaviour in groups from individual personality traits.European Commission [265957 COPEWELL]; European Social Fund of Andalusia; Foundation for Science and Technology, Portugal [SFRH/BPD/77210/2011]info:eu-repo/semantics/publishedVersio