Anabela Marisa Azul

Development, optimization and application of an analytical methodology by ultra performance liquid chromatography-tandem mass spectrometry for determination of amanitins in urine and liver samples

Amanitins, highly toxic cyclopeptides isolated from various Amanita species, are the most potent poisons accounting for the hazardous effects on intestinal epithelium cells and hepatocytes, and probably the sole cause of fatal human poisoning. The present study was focused on the development, optimization and application of an analytical methodology by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), following urine and liver sample preparation by protein precipitation with organic solvents, and solid phase extraction (SPE) procedure, for the determination of the amatoxins, α- and β-amanitin. Linearity, detection and quantification limits, selectivity, sensitivity, intra and inter-assay precision and recovery were studied, in order to guarantee reliability in the analytical results. The developed method proved to be specific and selective, with LOD (Limit of Detection) values for α- and β-amanitin of 0.22 and 0.20 ng mL(-1) in urine and 10.9 and 9.7 ng g(-1) in liver, respectively. LOQ (Limit of Quantification) values ranged from 0.46 to 0.57 ng mL(-1) in urine and 12.3-14.7 ng g(-1) in tissue, for both amanitins. Linearity, in the range of 10.0-200.0 ng mL(-1) or ng g(-1), shows that coefficients of correlation were greater than 0.997 for α-amanitin and 0.993 for β-amanitin. Precision was checked at three levels during three consecutive days with intra-day and inter-day coefficients of variation not greater than 15.2%. The extraction recovery presents good results for the concentrations analyzed, with values ranging from 90.2 to 112.9% for both matrices. Thus, the proposed analytical method is innovative, presents a high potential in the identification, detection and determination of α- and β-amanitins in urine and tissue samples, as well as in other biological samples, such as kidney and mushrooms.

Fungal fruitbodies and soil macrofauna as indicators of land use practices on soil biodiversity in Montado

The impacts of land use on soil biodiversity are still poorly understood, although soil fungi and macrofauna are recognized to provide benefits to ecosystems. Here, we tested whether land use practices used to control shrub density influences the fruiting macromycetes (ectomycorrhizal-forming fungi—ECMF—and saprobes) and soil macrofauna diversity and abundance in Montado ecosystems. To address this influence, we conducted a 2-years’ period monitoring of fungi fruitbodies and macrofauna in sixteen experimental plots in Montado landscape in southern Portugal. A total of 4,881 frutibodies (57 taxa of ECMF and 64 taxa of saprobic fungi) and 3,667 soil invertebrates (73 species and morphospecies) were monitored in the experimental plots. There was greater losses in sporocarps production and taxa composition, particularly the ECMF, in plots where shrub density was controlled by permanent grazing (Ca) or involving cutting practices followed by soil tillage (M), in comparison with cutting practices with no soil tillage (Cu) and the control (C). The ECMF Laccaria laccata and Xerocomus subtomentosus exhibited a close relation with C and Cu plots while the saprobes, e.g., Entoloma conferendum, were associated to Ca and M plots. Most species associated to Cu plots were present in C plots during the 2 years, but not in Cu after the cutting practices (in the second year of sampling). Regarding soil macrofauna, higher values of taxa and species richness were observed in C and Cu plots in the first year of sampling. The ant species Aphaenogaster senilis and several Staphylinid morphospecies exhibited a close relation with M plots, whilst most spider families were directly associated to C and Cu plots. After the shrub cutting practices, higher values of taxa and species richness of soil macrofauna were observed in M and Ca plots; the presence of species with a high competitive ability to colonize disturbed areas faster might explain the results. Contrary to the frutibodies production and diversity, species richness and abundance within soil macrofauna were identical between Cu and C in 2004. Thus, fruiting macromycetes and soil macrofauna diversity and abundance in Montado’s, appear highly sensitive to land use and somewhat reflected a trend of severity to the current shrub management practices.

Mycorrhizal types in the Mediterranean Basin: safety teaching and training

More than 90% of vascular plants in terrestrial ecosystems form symbiotic root-fungal associations known as mycorrhiza; in Mediterranean biomes all known mycorrhizal types co-exist. However, the understanding of mycorrhizal diversity and structure and their ecophysiological impact in soil productivity and conservation has been shown to be very poor among biology graduates. Two major reasons were detected: lack of (1) reliable information available and (2) teacher training. Our experience reveals that teachers are discouraged from practicals that involve long protocols and/or toxic chemicals. Despite this, we observed their willingness to be involved in studies of fungal diversity and function in natural communities, based on training with simple, safe methods. We confirmed that our training courses were important not only for acquisition of new information and development of skills, but also that they stimulated the planning of a number of different practicals linked to themes taught during the year.

Morphotyping and Molecular Methods to Characterize Ectomycorrhizal Roots and Hyphae in Soil

At the interface between plants and soils, ectomycorrhizal (ECM) fungi explore soils, acquire resources, transfer resources to plants, and acquire carbon from plants. Mycorrhizas enhance plant survival, nutrition and growth and play key roles in ecosystems processes such as decomposition, nutrient cycling, soil carbon storage, productivity and sustainability. Mycorrhizas are critical for plant colonization of new soils (e.g. mine spoils, volcanic deposits, glacial moraines). ECM diversity ensures plant reestablishment after disturbance and can enhance survival and growth of trees in reforestation. ECM fungi can promote fine root development as well as produce antibiotics, hormones and vitamins. Mycorrhizal associations may help protect roots from pathogens and moderate effects of heavy metals and toxins. Many environmental problems may be alleviated by mycorrhizas – problems such as pollution, erosion, soil degradation, climate change, degradation of natural resources, and poor land use management. Mycorrhizal abilities to carry out important functions are linked to diversity. ECM diversity is large and documented in many ecosystems, particularly coniferous ecosystems (Gehring et al. 1998; Goodman and Trofymow 1998; Kranabetter and Wylie 1998; van der Heijden et al. 1998; Stendell et al. 1999; Bidartondo et al. 2000). This ECM diversity has been based on surveys of fruiting bodies, but is now based on more recent methods – morphotyping (microscopic observations) and phylotyping (molecular characterization). An advantage of fruiting body surveys is ease of collection and identification based on morphology; a disadvantage is the assumption that fungi fruiting in an area also form ectomycorrhizas on nearby roots. Clearly identification of ectomycorrhizas on roots is preferable. However there are difficulties in ECM identification – complex sampling design and

Quercirhiza flavocystidiata + Quercus suber L.

Resumen del trabajo presentado a la 62nd Annual Conference on Magnetism and Magnetic Materials (MMM), celebrada en Pittsburgh (USA) del 6 al 10 de noviembre de 2017.Financial support 224 from Fundacao para a Ciencia e a Tecnologia (Lisboa, Portugal) grants 225 (PTDC/CVT/2006/66114 and UID/CVT/00276/2013)Tese de doutoramento em Ciencias da Saude, apresentada a Faculdade de Medicina da Universidade de CoimbraTrabajo presentado en el 8o Congreso de la Asociacion Iberica de Endocrinologia Comparada (AIEC), celebrado en Madrid del 5 al 7 de septiembre de 2011.Trabajo presentado en la International Conference and Exposition Aquaculture Europe 2017, celebrada en Dubrovnik del 17 al 20 de octubre de 2017.O presente estudo centra-se na analise das paisagens de poder que materializam as dinâmicas sociais e territoriais ocorridas nos finais da Idade do Ferro e inicios de epoca Romana no Noroeste Peninsular. Enquadra-se numa necessidade de se desenvolverem novas e inovadoras metodologias de trabalho, aliadas a uma robustez teorica e a um rigor empirico, cujas interpretacoes permitam a criacao de um novo conhecimento e que tenha aplicabilidade a outros contextos. Esta investigacao foi desenvolvida numa optica da Arqueologia da Paisagem. Assim, adoptou-se uma perspectiva diacronica e uma metodologia interdisciplinar, procurando-se ampliar as metodologias tradicionalmente utilizadas neste tipo de estudos, sobretudo com o recurso as tecnologias geoespaciais, na tentativa de se caracterizar a estruturacao do povoamento e territorio neste periodo e area geografica. O objectivo principal deste estudo reside na tentativa de caracterizacao sociopolitica das comunidades do Noroeste Peninsular, particularmente na sua area Meridional, nos finais da Idade do Ferro e inicios de epoca Romana. Tomaremos como caso de estudo a regiao do Alto Tâmega e Cavado. Trata-se de uma zona de fronteira e de hibridizacao cultural pelo menos desde a Idade do Ferro, localizando-se justamente na transicao entre a area dos oppida ocidentais e a dos pequenos castros orientais. Nesta zona registam-se processos de intensificacao economica, embora mais centrados na exploracao de recursos minerais, ao inves da exploracao e intensificacao agricola, dando lugar a uma estruturacao territorial bastante especifica e a formas particulares de expressao da desigualdade. Procuraremos analisar as dinâmicas territoriais e os processos de continuidade e de mudanca social, bem como as diferentes economias politicas e estrategias de poder, numa epoca onde tem lugar complexas politicas de interaccao cultural, particularmente com o mundo mediterrânico, bem como uma crescente negociacao social e identitaria. Procuraremos definir e clarificar os diferentes modelos de povoamento e as diferentes formas de estruturacao territorial em epoca pre-Romana e Romana. A base fundamental deste trabalho sera a objectivacao dos criterios locacionais e territoriais de cada povoado, com vista a se discriminarem as decisoes sociais que motivaram a eleicao de um determinado sitio, ja que distintos modelos locacionais e territoriais poderao associar-se a diferentes contextos culturais e a distintas formas de organizacao territorial e sociopolitica. Todavia, sera de ressaltar que este trabalho tera sempre um sentido comparativo, procurando-se analisar a variabilidade, mais do que uma reconstrucao positiva das condicoes que possam ter influenciado a localizacao dos sitios. Alem disso, partimos da diversidade regional e da heterogeneidade cultural, ja que mais que um todo homogeneo com uma evolucao unilinear, devemos antes, segundo uma perspectiva pos-colonial, contemplar uma multiplicidade de experiencias historicas particulares a diferentes escalas antes e depois da chegada de Roma. Apos os primeiros contactos com os Romanos, documenta-se uma estruturacao complexa do povoamento do Noroeste, que se manifesta no pleno desenvolvimento de grandes povoados tipo oppidum, que actuariam como verdadeiros lugares centrais do ponto de vista politico. Este fenomeno, que se desenvolve anteriormente a criacao generalizada de novos assentamentos romanos, e arqueologicamente mais visivel na parte Meridional da Galiza e no Norte de Portugal, onde os processos de ?oppidizacao? sao mais massivos e prematuros. Contudo, este processo de sinecismo e de hierarquizacao do povoamento podera ser consequencia de dinâmicas anteriores, uma vez que nao podemos negligenciar os complexos particularismos historicos das comunidades indigenas, tal como parece suceder em outros territorios europeus. Evidentemente que o contacto e a pressao Romana pode ter acelerado e precipitado todo este processo, embora talvez nao se tenha tratado de um processo unidireccional e monocausal. E necessario percepcionar os oppida em relacao estrutural a toda uma serie de fenomenos sociais, politicos e culturais que ocorrem no sul da Callaecia entre os seculos II a.C. e I d.C..This research has been developed on the behalf of a Doctoral (M3.1.2/F/025/2007) and a Post-Doctoral (M3.1.7/F/005/2011) Research Project, both supported by the Regional Fund for Science (FRC) of the Azorean Regional Government. This work was partially funded by Portuguese National Funds through FCT – Fundacao para a Ciencia e Tecnologia (Foundation for Science and Technology), under the project “PTDC/AAC-AMB/098786/2008”.Tese de Doutoramento em Ciencias da Saude, Universidade do Minho Escola de Ciencias da Saude.Trabajo presentado en la European Geosciences Union General Assembly, celebrada en Viena (Austria), del 8 al 13 de abril de 2018Este trabajo ha sido parcialmente financiado por la Fundacao para a Ciencia e Tecnologia, Ministerio da Educacao e Ciencia, Republica Portiguesa (SFRH/BD/44089/2008) y los proyectos: “Origins and Spread of Agriculture in the western Mediterranean region (ERC-2008-AdG 230561)” y “Stable isotopes in Mediterranean natural and agricultural ecosystems: from a mechanistic understanding of isotope fractionation processes in plants to the application in paleoenvironmental research (DGI CGL2009- 13079-C02-01)”.Trabajo presentado en el 18th International Symposium on Pollutant Responses in Marine Organisms (PRIMO 18), celebrado en Trondheim (Noruega) del 24 al 27 de mayo de 2015.VI Simposio Internacional de Ciencias del Mar - VI International Symposium of Marine Sciences (ISMS 2018), 20- 22 June 2018, Vigo.-- 1 pageResumen del poster presentado a la XXXV Reunion Bienal de la Real Sociedad Espanola de Fisica y al 25o Encuentro Iberico de Ensenanza de la Fisica, celebrados en Gijon (Espana) del 13 al 17 de julio de 2015.-- et al.Trabajo presentado en el I Congresso Luso-Extremadurense de Ciencias e Tecnologia, celebrado en Evora (Portugal), del 20 al 21 de octubre de 2017The 2nd International Symposium on Submarine Canyons (INCISE2014), 29th September-1st October 2014, Edinburgh.-- 1 page, 1 figureTrabajo presentado en la 10th International Hydrogenase Conference celebrada en Szeged (Hungria) del 8 al 12 de julio de 2013.An overview of 13C nuclear magnetic resonance (NMR) spectroscopy methods and their applications in the study of the metabolism of brain cells in vitro and in the in vivo brain is presented as well as their implications for modern molecular imaging techniques. Various topics will be discussed, such as general properties of the 13C NMR spectrum, 13C NMR spectroscopy acquisition protocols, determination of fractional 13C enrichment, 13C(2H) NMR methodologies, and the use of 13C hyperpolarized substrates for NMR spectroscopy and imaging. Some illustrative applications are described, both in vitro and in vivo.This study was financially supported by Fundacao Para a Ciencia e a Tecnologia (SFRH7 BPD/5560/2001) and HP BIODIBERIA program at the Real Jardin Botanico (Madrid).Conferencia invitada. -- 7th Conference on Elementary Processes in Atomic Systems,Průhonice, Prague, September 3-6, 2017.-- https://www.jh-inst.cas.cz/tchem/cepas2017/about-cepas-2017/The current international socioeconomic context has been emphasising the need of a new set of individual competencies in order to succeed in the professional world. A new concept of employability has also emerged, with a person-centred approach, which reflects the need to deal effectively with career-related changes (Fugate, Kinicki, & Ashforth, 2004). In a psychosocial perspective, it is assumed an individual ability to adapt to social expectations and to behave according to the changing conditions (Savickas & Porfeli, 2012). On the other hand, the last decade represented a period of profound transformations in the European space of higher education, driven by Bologna guidelines. Higher education institutions are expected to adapt new ways of work organization, promoting new core competencies in future professionals. In this context, the paper will describe a research project which aims to identify and analyse the impact factors on the employability of Portuguese graduates, namely, individual characteristics, curricular experiences during graduation courses and also employer’s characteristics. This project represents an attempt to a closer and necessary articulation between higher education practices and labour market requirements.Trabajo presentado en el MedCLIVAR Conference: Bridging the Mediterranean Climates, celebrado en Belgrado (Serbia) del 17 al 21 de septiembre de 2018Trabajo presentado por Mariana Reis Sobreiro para optar al grado de Doctor por la Universidad de Salamanca y realizado en el Instituto de Biologia Molecular y Celular del Cancer.13 paginas, 7 tablas, 1 figuras, 1 apendice. Special thanks go to Ana Mouro, Joao Luis Mouro, Miguel Porto, Nuno Pires and Sara Dias for assisting in the fieldwork, and Nuno Gomes and associated workers for the EDM loan and field aid.Trabajo presentado en en 16th International Symposium on Fish Nutrition and Feeding (ISFNF 2014), celebrado en Cairns (Australia) del 25 al 30 de mayo de 2014.Tese desenvolvida no quadro do Programa Doutoral em Ciencias e Engenharia do Ambiente, com o financiamento de uma bolsa de doutoramento (SFRH/BD/42168/2007) da Fundacao para a Ciencia e a Tecnologia (FCT) e o apoio dos projectos EROSFIRE II (PTDC/AGR-CFL/70968/2006), e ”FIREREG”(PTDC/AGR-CFL/099420/2008), ambos financiados pela Fundacao para a Ciencia e a Tecnologia (FCT) e o FSE no âmbito do III Quadro Comunitario de Apoio.Editors: A. Campar Almeida, Ana M. S. Bettencourt, D. Moura, Sergio Monteiro-Rodrigues and Maria Isabel Caetano Alves. 29 pp.Trabajo presentado en el 10o Congreso de la Asociacion Iberica de Endocrinologia Comparada (AIEC), celebrado en Castellon (Espana), del 23 al 25 de septiembre de 2015Trabajo presentado en Aquaculture Europe 2019, celebrado en Berlin (Alemania), del 7 al 10 de octubre de 2019Tese de Candidatura ao grau de Doutor em Ciencias Biomedicas submetida ao Instituto de Ciencias Biomedicas de Abel Salazar da Universidade do Porto por Ana Maria Bastos Barros Neto e realizado em el Centro Andaluz de Biologia del Desarrollo, centro mixto CSIC-UPO.Trabajo presentado en el EMS Annual Meeting: European Conference for Applied Meteorology and Climatology, celebrado en Dublin (Irlanda), de 4 al 8 de septiembre de 2017

Early effects of fire on herbaceous vegetation and mycorrhizal symbiosis in high altitude grasslands of Natural Park of Estrela Mountain (PNSE)

Severe fires over the last decade have transformed the high altitude grassland biogenetic reserve in the Natural Park of Estrela Mountain (PNSE) of Portugal. Yet, there is little information about the impacts of the fires above and below ground. We assessed the species composition, richness and frequency of the herbaceous vegetation. We characterized the mycorrhizal symbiosis and analysed the soil properties in three cervunal areas of which one was destroyed by a severe fire in 2003. The results indicate that plant community in high altitude grassland of PNSE are rather poor in species but the number may increase after fire. The most remarkable change in the herbaceous vegetation after fire was the abrupt increase of Festuca trichophylla in the burnt area, to the detriment and abrupt decrease of Nardus stricta. Arbuscular mycorrhizas (AM) were the most widespread symbioses in all three grasslands. AM colonization of F. trichophylla was significantly lower in burnt grassland but the AM colonization in N. stricta increased. There was also a higher phosphorous content in soil of the burnt grassland. We put forward a hypothesis about the impacts of recurrent fires above and below ground and discuss its implications to functioning of high altitude ecosystems.

Protecting Soil Biodiversity and Soil Functions: Current Status and Future Challenges

Living soils are fundamental for human life as we know it. The top layer of Earth’s crust, essentially composed of minerals, water and air, harbours an immense variety of organisms, from plants to microorganisms, which qualifies it as a living system. Soil biodiversity is the main actor underlying the provision of services that are essential for regulating, providing and supporting human life. The increasing level of human activity has been subjecting soil to multiple pressures, resulting in soil degradation and biodiversity decline, hence deterioration in the system’s capability to render those ecosystem services. Growing concern on this resource’s misuse has led to a series of conventions and strategies targeting its conservation (such as the Thematic Strategy for Soil Protection in the European Union (EC) 2006) and advocating for specific protection measures that can ensure a sustainable use of soil. These measures mainly focus on extending our knowledge on how soil functions but also on developing monitoring programmes that can detect trends and changes in soil biodiversity. This demand boosted research on soil ecology over the last decades, with significant increases of scientific knowledge on its structural and functioning complexity. However, there are still some gaps and needs to be addressed in order to design adequate measures for soil protection. In this chapter we review the main advances in soil ecological research and monitoring and further discuss the status of current strategies towards soil protection and sustainability. Moreover, we present here a strategy, consisting of three action lines, for effectively contributing to soil protection. It is based on monitoring and mapping, experimentation and raising awareness towards soil issues, which hopefully can change the way we perceive and use soil, this very dynamic but non-renewable resource at the human life time scale.

Use of lignocellulosic wastes of pecan (Carya illinoinensis) in the cultivation of Ganoderma lucidum

BACKGROUND The wastes of pecan nut (Carya illinoinensis (Wangenh.) K. Koch) production are increasing worldwide and have high concentrations of tannins and phenols. AIMS To study the biodegradation of lignocellulosic wastes of pecan used as solid substrate for the cultivation of the white-rot fungus Ganoderma lucidum (Curtis) P. Karst. METHODS Six formulations of pecan wastes were used as solid substrate: pecan shells (PS100), pecan pericarp (PP100), pecan wood-chips (PB100), and the combinations PS50+PP50, PB50+PS50 and PB50+PP50. The substrates were inoculated with a wild strain of G. lucidum collected in the Iberian Peninsula. The biodegradation capability of G. lucidum was estimated by using the mycelial growth rate, the biological efficiency, the production and the dry biological efficiency. RESULTS Notably, all solid substrates were suitable for G. lucidum growth and mushroom yield. The best performance in mushroom yield was obtained with PB100 (55.4% BE), followed by PB50+PP50 (31.7% BE) and PB50+PS50 (25.4% BE). The mushroom yield in the substrates containing pecan wood-chips (PB) was significantly higher. CONCLUSIONS Our study is leading the way in attempting the cultivation of G. lucidum on lignocellulosic pecan waste. These results show an environmentally friendly alternative that increases the benefits for the global pecan industry, especially in rural areas, and transforms biomass into mushrooms with nutraceutical properties and biotechnological applications.

Handbook of Climate Change Communication: Vol. 1

This comprehensive handbook provides a unique overview of the theory, methodologies and best practices in climate change communication from around the world. It fosters the exchange of information, ideas and experience gained in the execution of successful projects and initiatives, and discusses novel methodological approaches aimed at promoting a better understanding of climate change adaptation. Addressing a gap in the literature on climate change communication and pursuing an integrated approach, the handbook documents and disseminates the wealth of experience currently available in this field. Volume 1 of the handbook provides a unique description of the theoretical basis and of some of the key facts and phenomena which help in achieving a better understanding of the basis of climate change communication, providing an essential basis for successful initiatives in this complex field.

Mitochondrial Follies: A Short Journey in Life and Energy

Communicating through comics can be an effective multi-layered strategy. The narrative of this comic transports the reader to a realistic, if abbreviated, view of mitochondrial biology blended with the changing daily routines of Lara. Lara is a fictionalized patient with an undisclosed metabolic disorder, who takes her health into her own hands, becoming interested in metabolism and mitochondria to better understand the processes by which living organisms convert food into energy at the cellular level. Moreover, her goal is also to communicate this fascinating world to friends and colleagues, in a way that may also be useful for scientists and the general public. The comic combines the discoveries of real individuals who have greatly contributed to the knowledge of mitochondria, namely Peter Mitchell, Hans Adolf Krebs and Lynn Margulis, with fictional characters, such as Lara and George, directly associated to the narrative. The interdisciplinary nature of graphic narration reflects the blending of text and scientific facts alongside visual information (both realistic and caricature-like) and critical-thinking-based dialogues and actions, that was enriched by the collaborative work between the researchers who developed the concept and the illustrator who brought it to life.