The Study of Tissue-Specific DNA Methylation as a Method for the Epigenetic Discrimination of Forensic Samples

In forensic sciences, the serological methods used to determine which body fluid was collected from the crime scene are merely presumptive or labor intensive since they rely on protein detection or on microscopic identification of cells. Given that certain forensic cases may need the precise identification of a body fluid to determine criminal contact, such is the example of a suspected sexual assault of a minor; certainty in the body fluid of origin may depict a precise picture of the events. The identification of loci that show differences in methylation according to the tissue of origin can aid forensic analysts in determining the origin of a DNA sample. The process of DNA methylation occurs naturally in the genome of living organisms and consists in the presence of a methyl group on the carbon 5 of a cytosine, which is typically followed by a guanine (CpG). Analyzing patterns of DNA methylation in body fluids collected from a crime scene is preferential to the analysis of proteins or mRNA since the same extracted DNA used for STR typing can be used for DNA methylation analysis. We have validated and identified loci able to discriminate blood, saliva, semen and vaginal epithelia. In the current study, we have also established the minimum amount of DNA able to provide reliable results using methodologies such as pyrosequencing and high-resolution melt (HRM) analysis for the different markers identified. Lastly, we performed an alternative bioinformatic analysis of data collected using an array that studied methylation in over 450,000 individual cytosines on the human genome. We were able to sort the locations that showed potentially higher methylation differences between body fluids and investigated over 100 of them using HRM analysis. The results of that study, allowed the identification of three new loci able to distinguish blood and two new loci able to distinguish saliva and vaginal epithelia, respectively. The use of DNA methylation patterns to aid forensic investigations started with a publication in 2010, therefore each small contribution such as this work may, similarly to what occured in the biochemistry field, result in the discovery of a method able to put the technology in the hands of forensic analysts

M-commerce: motivations and dynamics from Generation Z e-buyers

Generation Z is recognized as the generation that does not know a world without the internet, grows side to side with innovation and demands more from the market. Brands need to work for new digital updates, constant innovative systems, and interactive processes to captivate these consumers. Understanding the behaviors towards different digital platforms by Generation Z exposes the way this generation looks to the m-commerce platforms. This study aims to explore the shopping motivations and dynamics of Generation Z e-buyers from a cross-country panorama, contributing insights to e-retailers creating effective marketing strategies. The research intends to identify which factors impact the adoption of mobile shopping applications from Generation Z individuals. The study proposes a modified version of the UTAUT2 conceptual model, by adding the constructs Brand/Seller Reputation and Trust, using Gender and Nationality as moderators’ variables. The data for this study includes a sample of 600 responses from Generation Z individuals within Portugal and Italy. The outcomes of this study found that Habitat was the highest predictor of behavioral intentions, followed by Trust and Brand/Seller Reputation. In contrast, Social Influence, Facilitating Conditions, and Hedonic Motivations have low effects. Additionally, the variables Gender and Nationality do not demonstrate significant variations between the groups. The study’s results formulate solid implications for e-retailers.A Geração Z é reconhecida como a geração que não conhece um mundo sem a internet, nasceu lado a lado com a inovação e exige cada vez mais do mercado. As marcas devem trabalhar em prol de novos progressos digitais, de uma inovação constante nos seus sistemas e por processos interativos que captem esta geração de consumidores. Compreender o comportamento da Geração Z nas diferentes plataformas digitais, expõe como esta geração observa as plataformas de comércio móvel. O estudo tem como objetivo explorar as motivações e as dinâmicas de compra dos indivíduos da Geração Z através de uma perspetiva de dois países, proporcionando "insights" aos retalhistas "online" para a criação de estratégias de marketing eficientes. A pesquisa pretende identificar quais os fatores que impactam a adoção de aplicações de compra móveis pelos indíviduos da Geração Z. O estudo propõe uma versão modificada do modelo conceptual UTAUT2, que incluem os novos fatores Confiança e Reputação da Marca/Retalhista, utilizando o Género e a Nacionalidade como variáveis moderadoras. Os dados para este estudo incluem uma amostra de 600 respostas de indivíduos da Geração Z de Portugal e Itália. Através dos resultados obtidos descobriu-se que o Hábito foi o fator que mais contribuía para a intenção de compra, seguindo-se a Reputação da Marca/Retalhista e a Confiança. Contrariamente, as Influências Sociais, Condições Facilitadoras e Motivações Hedónicas tiveram efeitos reduzidos. Adicionalmente, as variáveis Género e Nacionalidade não demonstraram variações significativas entre os grupos. Os resultados do estudo geraram implicações significativas para os retalhistas presentes "online"

Aquaculture rainbow trouts are contaminated with multidrug-resistant Enterobacteriaceae species carrying clinically relevant antibiotic resistance genes

[Resumo][Abstract

Health and wellness tourism in pandemic times: paradigms shifts and sustainability challenges

The immediate consequences of coronavirus pandemic on tourism sector, triggered significant paradigm shifts on demand and also in the value of healthand healthcare as synonymous of security in tourist destinations, conditioning its image, promotion and the economic development of the territories. This special issue is focused on the challenges and opportunities prompted by this new reality in the specific field of Health and Wellness Tourism,concerning its impacts on related businesses and in worldwide destinations that has a solid tradition related with Thermal Baths, Spas and even withMedical services and facilities regarding tourist purposes. The importance of this approach is due to the fact this kind of Tourism figures as an important factor for the sustainable management of local communitiesand for the promotion of tourist destinations, contributing to the preservation and conservation of natural and even cultural heritage at the regional level.The aim of this special issue is to present the management and marketing strategies, development opportunities, innovative solutions and good practicesexamples that may outcome from these challenging circumstances, presenting full papers, case studies and research reports, based on a sustainabilityperspective. These approaches are developed in literature, regarding different contexts and countries, constituting a solid body of research on the topic.Sheldon and Park (2009) and Voigt and Pforr (2014) made an approach to wellness in a destination perspective, considering different implications fordestination development, management and competitiveness (Prideaux et al., 2014; Smith & Puczkó, 2014), the sustainable regional development(Pechlaner et al., 2014; Wray & Weiler, 2014) and introducing the concept of health region (Pforr et el., 2014). Also Peris-Ortiz & Álvarez-García (2015)analysed Health and Wellness Tourism as an important market segment, analysing it in different dimensions through several European case studies aboutregional development and innovative strategies on Thermalism (Araújo et al., 2015; Costa, Quintela & Mendes, 2015; Joukes & Costa, 2015). In whatconcerns to the actual context of coronavirus pandemic, Berini (2020) organized a sum of different studies regarding the impacts on Tourism and travelsector (Keller, 2020; Richards, 2020), focusing on marketing strategies during Covid-19 sanitary crisis (Andreini & Mangiò, 2020), on the sustainabilitydimension (Holden, 2020; Wells, 2020; Bougleux, 2020) and also forecasting behaviours and social impacts of this phenomenon (Peretta, 2020; Alberti &Perrotta, 2020; Heuwinkel, 2020). Health and Wellness Tourism has been an important factor for the economic development of territories, for the sustainable management of localcommunities and for the promotion of tourist destinations, contributing to the preservation and conservation of natural and even cultural heritage.Considering the importance and ancient tradition of Health and Wellness Tourism in many European (Belgium, France, Germany, Hungary, Portugal,Turkey, Spain) and worldwide countries (Australia, Canada, Chile, India, Peru, Thailand, United States) mainly due to their richness and abundance of hotsprings, it figures demanding to analyse the economic impacts of Coronavirus pandemic in this activity. Bearing in mind the shift on Health and Healthcareparadigms, figuring as synonymous of security in tourist destinations, conditioning its image, promotion and development, this special issue is focused onthe sustainability challenges and opportunities this new reality is designing in the specific field of Health and Wellness Tourism.info:eu-repo/semantics/publishedVersio

Overcoming barriers: the local and the innovative dimensions of inclusive socio-educational practices

This paper discusses some results of a broader research, focusing on a set of eleven socio-educational practices aiming to overcome school failure and dropout, developed in Portugal, giving particular attention to the local and innovative dimensions. This research aims to understand the point of view of the several actors involved, about which factors, processes and relationships contribute the most to building such practices. Data was gathered through documental analysis and semi-structured interviews with those (institutionally) responsible for each practice under study, and was analysed using two instruments. From the point of view of the people responsible, the practices that contribute the most to overcoming school failure and dropout fall into one of four categories: Study Support (4 Practices), Student Grouping (3), Mediation (3) and Pedagogical Differentiation (1). Some practices mobilise resources; others interfere with learning and life contexts, in order to confront institutional, situational and dispositional barriers to participation and learning. Those practices seem to have an impact on school-family communication. Formal schooling, as well as the socio-cultural inclusion of youth from disadvantaged backgrounds, are seen as relevant; yet, we can observe a somewhat fragile involvement of families and communities in practices aimed at promoting their youth’s educational success.Este texto analiza algunos resultados de una investigación más amplia, centrada en un conjunto de once prácticas socioeducativas destinadas a superar el fracaso y el abandono escolar, desarrolladas en Portugal, prestando especial atención a las dimensiones locales e innovadoras. Su objetivo es comprender el punto de vista de los diversos actores involucrados, acerca de qué factores, procesos y relaciones contribuyen más a la construcción de tales prácticas. Los datos se recopilaron mediante análisis documental y entrevistas semiestructuradas con los responsables (institucionalmente) de cada práctica, y se analizaron utilizando dos instrumentos. Para las personas responsables, las prácticas que más contribuyen a superar el fracaso escolar y el abandono escolar se dividen en una de cuatro categorías: Apoyo al estudio (4 prácticas), Agrupación de estudiantes (3), Mediación (3) y Diferenciación pedagógica (1). Algunas prácticas movilizan recursos; otros interfieren con el aprendizaje y los contextos de la vida, para enfrentar las barreras institucionales, situacionales y disposicionales a la participación y el aprendizaje. Esas prácticas parecen tener un impacto en la comunicación entre la escuela y la familia. La escolarización formal, así como la inclusión sociocultural de jóvenes de entornos desfavorecidos, se consideran relevantes; sin embargo, se puede observar una participación algo frágil de las familias y las comunidades en las prácticas destinadas a promover el éxito educativo de sus jóvenes.This research is funded by National Funds through FCT - Foundation for Science and Technology (Portugal), in the context of project PTDC/MHCCED/3775/2014. This research is funded by CIEd – Research Centre on Education, project UID/CED/01661/2019, Institute of Education, University of Minho, through national funds of FCT/MCTES-PT

Multi-actor perspectives on successful and inclusive socio-educational practices: overcoming school failure and dropout in Portugal

Project "EDUPLACES: Practices, voices and pathways of inclusive education" seeks to identify, characterise and discuss socio-educational practices aimed at overcoming school failure and dropout. Teachers/professionals, parents, children/young people and institutional partners engaged in ten practices, located in four Portuguese municipalities, participated in interviews (10) and focus groups (37). Cross-analysis produced relevant results pertaining to the practices’ location, basis (school or community organisation) and philosophy (student grouping, study support, mediation or pedagogical differentiation). Additionally, it revealed some differences between groups of participants: individual change is an outcome more predominantly identified by parents and children/young people; teachers/professionals frequently address successful approaches to school-family-community interactions; partners express a generally positive outlook on the practices, instead of focusing on barriers.This research is funded by National Funds through FCT - Foundation for Science and Technology (Portugal), in the context of project PTDC/MHC-CED/3775/2014. This research is funded by CIEd – Research Centre on Education, project UID/CED/01661/2019, Institute of Education, University of Minho, through national funds of FCT/MCTES-PT

Mecanismos de inactivação do gene LRP1B em tumores da tiróide

Tese de mestrado, Biologia (Biologia Molecular Humana), 2009, Universidade de Lisboa, Faculdade de CiênciasOs tumores da tiróide são o tipo mais frequente de neoplasia endócrina e podem ocorrer na forma esporádica e familiar. O LRP1B é um potencial gene supressor de tumores cujo papel nunca foi investigado em tiróide. Os genes supressores de tumores podem ser inactivados por diversos mecanismos entre os quais mecanismos epigenéticos. A epigenética compreende um conjunto de mecanismos que regulam a expressão génica não envolvendo modificações na sequência nucleotídica. Os mecanismos epigenéticos envolvem metilação de DNA, modificações de histonas, RNAs não codificantes e posição do nucleossoma. O objectivo deste trabalho foi investigar, a nível molecular e funcional, os mecanismos epigenéticos de inactivação do LRP1B em linhas celulares e tumores da tiróide. Estudou-se o estado de metilação de um fragmento de 230pb da ilha CpG do LRP1B em 10 linhas celulares derivadas de carcinomas da tiróide e em tumores da tiróide (20 AFTs, 17 CFT, 24 CPT, 10 CAT e 12 CNMTF), por conversão de bissulfito seguido de sequenciação. Detectou-se metilação em 30% das linhas celulares e em 40% das lesões da tiróide. No entanto, nenhuma das tiróides normais estudadas revelou metilação da ilha CpG do LRP1B. Para avaliar a influência da metilação na expressão, realizaram-se tratamentos com agentes desmetilantes (5-Aza e siDNMT1). Observou-se re-expressão do LRP1B numa das linhas com metilação. Para investigar o papel da metilação de histonas, silenciou-se a HMT G9a (H3K9) através de siRNA, o que resultou na diminuição da expressão do LRP1B numa linha celular sem metilação de DNA. Em conclusão, encontrámos metilação do promotor do LRP1B especificamente em tumores da tiróide. Salienta-se a sua ausência total em amostras de tiróides normais. Elucidámos, em linhas celulares, alguns dos mecanismos de inactivação do LRP1B. Os nossos resultados sugerem a metilação do promotor do LRP1B como um biomarcador epigenético com elevada especificidade e baixa sensibilidade.Thyroid tumours are the most frequent type of endocrine neoplasia and can occur in both sporadic and familial forms. LRP1B is a potential tumour suppressor gene and its role has never been adressed in thyroid. Tumour suppressor genes can be inactivated by a number of different mechanisms, namely epigenetic events. Epigenetics referes to the mechanisms that regulate gene expression without modifying the underlying DNA sequence. Some epigenetic mechanisms should be highlighted such as DNA methylation, histone modifications, non coding RNAs and nucleosome position. The aim of this work was to investigate the role of epigenetic events underlying LRP1B inactivation in both thyroid cell lines and tumours, using a molecular and functional approach. A 230bp fragment of LRP1B CpG island was studied in 10 thyroid carcinoma-derived cell lines and 83 thyroid tumours (20 FTA, 17 FTC, 24 PTC, 10 ATC and 12 FNMTC), using bisulfite conversion followed by sequencing. It was observed methylation in 30% of the cell lines and in 40% of the thyroid lesions. None of the normal thyroid samples revealed methylation in LRP1B CpG island. In order to evaluate the role of methylation in gene expression we performed a treatment with demethylating agents (5-Aza and siDNMT1). The treatment led to the reexpression in a cell line with DNA methylation. The role of histone methylation was assessed with the silencing of HMT G9a (H3K9) using siRNA leading to a decrease in LRP1B expression in one cell line without DNA methylation. In summary, we found DNA methylation in LRP1B promoter region in the studied thyroid tumours. However we did not find any DNA methylation in normal thyroid samples. We clarified some of the LRP1B inactivating mechanisms in cell lines. Our results suggest LRP1B promotor DNA methylation as an epigenetic biomarker with high specificity and low sensibility

A comunicação nas perturbações do espectro do autismo : um estudo de caso

Trabalho de Projeto apresentado à Escola Superior de Educação do Instituto Politécnico de Castelo Branco para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Educação Especial – Domínio Cognitivo e Motor.As Perturbações do Espectro do Autismo (PEA) são uma perturbação global do desenvolvimento que afeta três áreas do desenvolvimento do indivíduo, nomeadamente da comunicação, da socialização e da imaginação, sendo esta característica comumente conhecida como Tríade de Dificuldades. O presente trabalho centrou-se, em particular, na área da comunicação, através do estudo de uma criança diagnosticada com PEA a frequentar uma escola do Ensino Básico pela primeira vez. O trabalho de projeto, de natureza qualitativa, seguiu uma metodologia de Estudo de Caso, através do qual foi realizada uma investigação e intervenção sobre a mesma criança, procurando desenvolver competências comunicativas e modificar comportamentos sociais e funcionais na mesma. Ao longo da investigação, e após uma avaliação inicial por meio de observações não-participante anotadas em Grelhas de Observação e Escala de Envolvimento da Criança, foi aplicado um projeto de intervenção na área da comunicação, sendo realizadas atividades várias que procuraram desenvolver competências comunicativas na criança em estudo, nomeadamente de leitura e facilitassem a motivação daquela. O estudo, que se prolongou ao longo de seis meses, foi apoiado por observações participante, as quais foram descritas em realizadas entrevistas às pessoas que diariamente conviviam e participavam no desenvolvimento da criança, nomeadamente a mãe, professoras e terapeutas, a partir das quais se procurou compreender as suas perspetivas acerca da criança e do meio escolar no qual aquela se encontrava inserida. Após a aplicação do projeto de intervenção foram realizadas novas observações não-participante, sendo os dados anotados em Grelhas de Observação e na Escala de Envolvimento da Criança. Por fim, foi realizada uma análise da intervenção e uma confrontação dos dados obtidos antes e após a aplicação do projeto. Os resultados demonstram que a criança apresentou um ligeiro desenvolvimento na área da comunicação e nas suas competências comunicativas, bem como na sua integração na nova escola, na qual se havia deparado com novos colegas, professores, auxiliares, espaços e rotinas.Autism Spectrum Disorder (ASD) is a development global disorder which affects three development areas of the human being, such as communication, socialization and imagination, characteristic also known as The Wing Triad. The present work is specifically about the area of communication, through the study of a child diagnosed with ASD, attending at a primary school for the first time. The project, which has a qualitative nature, has followed a Study Case methodology, through which an investigation and an intervention took place, looking for communication skills development and social and functional conduct changing at the child. Through the investigation, and after an evaluation made with observations noted in Observation Charts and in the The Leuven Involvement Scale for Young Children, an intervention project was applied in the communication area, through activities for the child communication skills development, appealing for the child motivation by the use of different strategies. The study, which lasted for six months, was supported by observations, described in detailed field notes. Interviews were also made to the people who daily interact and participate in the child development such as the mother, teachers and therapists, looking for their comprehension about the child and the scholar environment. After the project application, other observations took place, whose data was written down in Observation Charts and in the the Leuven Involvement Scale for Young Children. At last, an intervention analysis was made, as well as the confrontation of the written data of both observations. The results shows us that the child has revealed a development and an improvement in the communication area, in his communication skills, as well as in his integration at the new school, where there were news colleagues, teachers, assistants, rooms and routines