Design of a Novel Phage Display Vector to Improve Enrichment of Phages with Antigen-Specific Nanobodies and to Identify the Nanobody Clones of Highest Affinity

Although phage display-based enrichments became a standard procedure, they still suffer from some drawbacks. Nonspecific phage binding limits the enrichment that can be achieved per se-lection round and therefore, in most cases, at least three or four rounds are required to identify the antigen binding nanobodies (Nbs) from the library. Moreover, the release of the phages captured via their antigen-specific Nb on the immobilized antigen is accomplished most often, by a pH shock that will also release the a-specific absorbed phages, thereby generating an unwanted back-ground. However, the biggest shortcoming is the difficulty to identify the Nb of best binding affinity. We now have to ferment and purify all individual Nb clones to homogeneity and measure their affinity parameters one after the other. This is very tedious, certainly, as the identified Nbs often share a high degree of amino acid sequence identity, which makes it impossible to predict the one having the best affinity. The objective of this thesis aimed to modify the phage display vector so that one round of selection will be sufficient, while the affinity of all antigen-positive Nbs can be compared immediately after ELISA. Firstly, a vector containing the Calmodulin Binding Peptide (CBP) tag was created, by substitut-ing the hemagglutinin (HA) tag present in a pMECS-GG plasmid by the CBP tag. Moreover, 6 Nbs with well-known and variable kinetic binding rates were also inserted into the vector pMECS-CBP. Next, the vector was used in a phage display setting, where two mini libraries comprising the 6 Nbs were made. One library had the conventional pMECS vector while the other was made using the pMECS-CBP vector. Although an enrichment of 1000 times was achieved using pMECS, pMECS-CBP failed to show any enrichment, supporting the hypothesis that Nb-CBP encountered serious expression problems. To investigate the validity of this hypothesis, periplasmic expression of Nb-CBP was performed and compared with that of the Nb without the tag. While the Nb yielded almost 4 mg per liter of culture media, the Nb-CBP could only be obtained at 0.63 mg per liter culture. Moreover, the Nb-CBP couldn’t be detected on a western blot. Several methods were used, such as adding lysozyme to improve the periplasmic extraction step, expressing 5 different colonies to check if there was a homogeneity of expression between them. In all meth-ods, the amount produced always remained below 1 mg per liter and Nb-CBP protein couldn’t be detected in any step of expression by Coomassie stained SDS-PAGE or western blot. Additionally, a high yield expressing-protein (SIRPα) was cloned into the CBP vector to identify whether the expression problem was caused by the peptide tag itself or by the Nb-CBP combina-tion. Even though SIRPα was obtained at a yield of 8.7 mg/L of culture when expressed from a vector not containing the CBP, its expressed yield using the CBP vector, dropped to 0.7 mg/L of culture, indicating that the CBP tag is incompatible with good periplasmic expression. Lastly, a nucleotide alignment of our CBP tag with a previously reported one, revealed differences in two Arginine codons, two Alanine codons and one Lysine codon. Moreover, our sequence employed two codons, AGA and CGG, rarely used in E. coli, which might be linked to the re-duced expression levels that we observed during this thesis

Stratification and peer effects: an analysis of Lisbon public schools

Educational stratification has been a difficult subject to deal with having yet no study shown a quantitative measure of it. Using the idea of distribution comparison a measure based on parents’ education is built for the primary schools in Lisbon. Upon the confirmation that Lisbon is stratified, I use the measure of peer effects based on stratification and determine its impact on test scores, concluding that the existence of stratification improves scores of students in schools with more educated parents and decreases scores of students in schools with less educated parents. Moreover, using fixed effects I derive the conclusion that the measure of peers’ characteristics helps explain most of differences among schools.NSBE - UN

Liikunnan merkitys alakouluikäisen psykososiaaliseen hyvinvointiin : Opetustapahtuma 6.luokkalaisille

Lasten ja nuorten terveyttä uhkaavia tekijöitä ovat mielenterveysongelmat ja sosiaalisen kehityksen häiriintyminen. Näiden terveyttä uhkaavien tekijöiden lisääntyminen näkyy myös mielenterveyspalvelujen käytön lisääntymisenä Suomessa. Liikunnan lisäämiseen kannattaa panostaa, koska sillä on myönteisiä vaikutuksia sekä fyysiseen että psyykkiseen terveyteen. Opinnäytetyömme tarkoituksena oli pitää kuudennen luokan oppilaille opetustapahtuma, jonka avulla pyrimme lisäämään heidän tietoutta liikunnan positiivisista vaikutuksista ja sen merkityksestä hyvinvointiin, sekä pyrkiä motivoimaan koululaisia liikkumaan enemmän. Tavoitteenamme on edistää oppilaiden ja heidän perheidensä terveellisiä elintapoja, motivoida liikuntaan, vaikuttaa asenteisiin ja siten myös parantaa oppilaiden psyykkistä hyvää oloa, sekä kehittää sosiaalisia suhteita. Opinnäytetyössä tarkastelemme myös, miten terveydenhoitaja voi edistää oppilaiden liikunnallisuutta ja näin ehkäistä koululaisten psyykkisiä tai sosiaalisia ongelmia. Tavoitteena oli oman ammattitaidon kehittyminen ja tiedon saaminen tulevaisuuden työskentelyä varten sekä oppia edistämään ja tukemaan oppilaiden liikunnallisuutta sekä psyykkistä ja sosiaalista hyvinvointia. Yhteistyösopimus on allekirjoitettu Seinäjoella Törnävän alakoulun kanssa. Opinnäytetyö toteutettiin toiminnallisena opinnäytetyönä. Järjestimme kuudesluokkalaisille 45 minuutin liikuntatapahtuman ja opetustuokion. Kuudennen luokan oppilailta ja opettajilta saadun palautteen perusteella selvisi, että oppilaat ja opettajat pitivät järjestämästämme opetustapahtumasta. Opetustapahtuma oli ollut opettavainen ja hyödyllinen. Lisäksi yli puolet vastaajista kertoi saaneensa uusia vinkkejä liikuntamahdollisuuksista. Terveydenhoitajan rooli lasten terveyshaittojen ennaltaehkäisijänä ja normaalin kehityksen tukijana on merkittävä. Tärkeää on pyrkiä tunnistamaan varhain alkaneet psyykkiset häiriöt, koska ne ovat uhka yksilön tulevalle kehitykselle ja lapsuusiän normaali kehitys taas luo pohjan aikuisiän hyvälle mielenterveydelle.Children's and young people's health threats are mental health problems and disruption of social development. The health threat increase is also reflected in the growing usage of mental health services in Finland. More exercise is worth investing in, because it has a positive effect on both physical and mental health. The purpose of our study was to organize a teaching event for 6th grade students. We aim to increase their awareness on the positive effects of physical activity and its importance for well-being, as well as to try to motivate school children to exercise more. Our goal is to help promote knowledge for pupils and their families towards a healthy lifestyle and motivation for exercise, to influence attitudes, and thus also to improve students' mental well-being, and at the same time to develop social relationships. The aim was also to examine how public health nurses can promote students' physical activity and thus prevent school psychological or social problems. Another goal was personal professional development and gathering information on the condition of future work, as well as to learn to promote and support students' physical activity and mental and social well-being. The cooperation agreement was signed with Seinäjoki Törnävä primary school. This thesis was executed as a functional thesis. We arranged a 45 minute-exercise and teaching event for the sixth graders. Based on the feedback, it became clear that the students and teachers liked the organization of the teaching event. The teaching event was instructive and useful. In addition, more than half of the respondents said that they got new ideas for sports activities. The public health nurse’s role in children's health prevention and normal development is major. It is important to try to identify the early beginning to mental disorders, because they are a threat to an individual's future development and the normal development of childhood again lays the foundation for good adult mental health

Modelos de previsão de falência de empresas

Mestrado em ContabilidadeFalência é o termo que se dá quando uma empresa não consegue fazer face a todos os seus compromissos financeiros, como o pagamento dos salários aos colaboradores, o pagamento de impostos, o pagamento aos fornecedores, entre outros. O risco de falência depende de factores internos e externos, como a qualidade da gestão, o grau de concentração de clientes, a capacidade de financiamento ou até o próprio sector de actividade da empresa. A partir de informações contabilísticas expressas no balanço e na demonstração de resultados foram criados métodos para prever a falência, entre eles, o mais importante, o Z-Score de Altman que utilizou rácios financeiros, numa análise multivariável, os quais identificam empresas que iriam à falência num determinado período de tempo. Estes indicadores são cada vez mais utilizados pelos diversos interessados, como sejam, instituições financeiras, investidores, auditores, contabilistas e gestores, uma vez que servem de auxílio para tomar conhecimento se a empresa em questão é fiável para concessão de crédito ou para investimento nas suas acções

Os seniores na sociedade em rede: dinâmicas de promoção da inclusão e da literacia digitais em Portugal

A sociedade em rede surge associada a um conjunto de competências e predisposições tendencialmente menos presentes entre as gerações mais velhas, especialmente entre os menos qualificados. Não obstante, a adesão dos seniores às novas TIC tem sido crescente. Ao longo da última década, importantes dinâmicas de promoção da inclusão e da literacia digitais direcionadas para as gerações mais velhas foram surgindo em Portugal. Trata-se de respostas das políticas e das instituições à necessidade percebida de estimular a utilização das novas TIC pelos seniores e de melhorar as suas competências digitais, fatores cada vez mais preponderantes para viver e participar numa sociedade marcada pela informação e pelo conhecimento e para usufruir de todas as valências que esta oferece. Neste paper procura-se sistematizar as principais políticas e iniciativas de incentivo à literacia digital das gerações mais velhas desenvolvidas em Portugal nos últimos anos. Procura-se também refletir sobre os desafios e aspetos relevantes de uma promoção bem sucedida da inclusão e da literacia digitais para seniores.The network society is associated with a set of skills and predispositions tendentially less present among the older generations, especially the less qualified. However, new ICTs adoption by the seniors has been increasing. Over the last decade, important dynamics of digital inclusion and literacy promotion targeting the older people have been emerging in Portugal. These are responses from policies and institutions to the perceived need to stimulate and improve the relation between seniors and new networking technologies. ICT usage and skills are increasingly fundamental to functioning successfully in today’s information and knowledge based society and to take advantage of all the possibilities it offers. This paper aims to systematize the main policies and initiatives developed in Portugal in recent years to promote digital literacy of the older generations. It also seeks to reflect on the challenges and the key issues of a successful promotion of digital inclusion and literacy for seniors

Experiências de visita a um centro de ciência: o caso do público não-escolar do Pavilhão do Conhecimento - Ciência Viva

Os centros de ciência, criados no âmbito do Programa Ciência Viva, constituem-se como um dos novos contextos de divulgação de ciência e de promoção da cultura científica. Como contributo para um conhecimento mais aprofundado dos públicos destes museus interactivos de ciência e tecnologia e do seu relacionamento com estes espaços, foi desenvolvida uma pesquisa de carácter qualitativo, centrada no público não-escolar do pólo dinamizador da rede de centros Ciência Viva - o Pavilhão do Conhecimento. O objectivo principal desta pesquisa, que contemplou entrevistas a visitantes, foi identificar e interpretar diferentes experiências e estilos de visita entre o público não-escolar. Procurou-se perceber de que forma o contexto da visita, mas também outros factores relacionados com as trajectórias biográficas, influenciam essas experiências. O presente working paper apresenta os principais resultados e conclusões da investigação.Science centers, created within the framework of the Ciência Viva programme, constitute one of the new ways of expanding science to a wider public and of scientific culture promotion. In contribution to a more detailed knowledge of these interactive science and technology museums’ public and their relationship with these spaces, a qualitative research has been developed, focusing on the non-formal public of the main center of the Ciência Viva network – the Pavilion of Knowledge. The main focus of this research, which included several interviews to visitors, was to identify and interpret different experiences and visiting styles among the non-formal public. The goal was to understand in which way the context of the visit, as well as other factors related to their biographic trajectories, influences those experiences. The current working paper presents the main results and conclusions of the research

Experiências de visita a um centro de ciência: Um estudo qualitativo sobre o público não-escolar do Pavilhão do Conhecimento - Ciência Viva

A problemática central em que se baseia esta investigação relaciona-se com a denominada cultura científica. A ciência e a tecnologia ocupam um papel cada vez mais importante e decisivo nas sociedades contemporâneas. Todavia, têm-se registado défices assinaláveis de cultura científica na sociedade portuguesa, perante o que têm surgido novos contextos de divulgação de ciência. Os centros de ciência, criados no âmbito do programa Ciência Viva, são um deles. Estes são museus interactivos de ciência e tecnologia e representam um novo conceito de museologia. Importa pois perceber como está a ser recepcionada esta modalidade de divulgação de ciência. Como contributo para um conhecimento mais aprofundado dos públicos dos centros de ciência e do seu relacionamento com estes espaços, foi desenvolvida uma pesquisa de carácter qualitativo, centrada no público não-escolar do pólo dinamizador da rede de centros Ciência Viva - o Pavilhão do Conhecimento. O objectivo principal desta pesquisa, que passou por entrevistas a monitores e visitantes, foi identificar e interpretar diferentes experiências e estilos de visita entre o público nãoescolar. Procurou-se perceber de que forma o contexto da visita, mas também outros factores relacionados com as trajectórias biográficas, influenciam essas experiências. Captaram-se pontos em comum entre os visitantes, mas reteve-se também a sua heterogeneidade. Diferentes motivações, diferentes formas de actuar e de viver a visita, de apreciar e retirar proveitos daquele espaço, relacionadas com factores como a formação, a profissão, o interesse pela ciência, a composição do grupo de visita. Explicitou-se também a atractividade do centro de ciência para públicos menos típicos.The central problematics in which this investigation is based on is connected to the socalled scientific culture. Science and technology take on an increasingly important and decisive role in contemporary society. However, there are records of significant deficits in scientific culture in Portuguese society, which paved the road for new ways of expanding science to a wider public. Science centers, created within the framework of the Ciência Viva program, are one of them. They are interactive science and technology museums which represent a new concept of museology. Therefore, it is important to understand how this method of expanding science is being accepted. In contribution to a more detailed knowledge of the science centers’ public and their relationship with these spaces, a qualitative research has been developed, focusing on the non-formal public of the main center of the Ciência Viva network – the Pavilion of Knowledge. The main focus of this research, which included several interviews to explainers and visitors, was to identify and interpret different experiences and visiting styles among the non-formal public. The goal was to understand in which way the context of the visit, as well as other factors related to their biographic trajectories, influences those experiences. Common points between visitors have been observed, but also their heterogeneity. Different motivations, different ways of reacting and experiencing their visit, of appreciating and enjoying the space, which are connected to such factors as education, occupation, interest in science, visit group composition. The appeal of the science center to a less usual public was also focused in this research

Nuclear Medicine and Drug Delivery

Nuclear Medicine is a molecular‐imaging modality that diagnoses and treats diseases with very small amounts of radioactive materials, known as radiopharmaceuticals. Drug delivery refers to approaches, formulations, technologies, and systems for transporting a pharmaceutical compound in the body as needed to safely achieve its desired therapeutic effect. The use of nuclear medicine and radiopharmaceuticals allows studying the in vivo behavior of developed drug‐delivery systems, and pharmacoscintigraphy is, from the beginning, one of the most promising aspects of this medical specialty. In this chapter, we review the technologies, fundaments, rationales, and strategies more frequently used and present examples of their application in the development and evaluation of drug‐delivery systems

Processo de importação: enquadramento e análise

Mestrado em Línguas e Relações EmpresariaisO presente relatório de estágio visa, além de uma breve introdução à empresa em que se insere, analisar o enquadramento de um processo de importação realizado na empresa Mário Miranda de Almeida, S.A. Será apresentada uma breve descrição deste processo e posteriormente analisadas as barreiras e conclusões retiradas da sua realização. Todos os dados foram adquiridos na empresa Mário Miranda de Almeida, S.A., também conhecida por Orima. O referido estágio teve a duração de 5 meses, tendo decorrido entre os meses de Fevereiro e Junho do presente ano de 2010. Nomeado orientador do estágio é o Professor António Moreno, sendo a supervisora a Dra. Cecília Almeida. O referido relatório irá consistir na análise de um processo de importação que, como não foi possível concluir, será relacionado e concluído com um processo de importação fictício que realizei anteriormente. Tendo tido a possibilidade de trabalhar directamente no departamento de importação da empresa Orima, e sendo uma área directamente relacionada com o Mestrado em Línguas e Relações Empresariais, considerei que teria particular interesse abordar o assunto no relatório de estágio. Creio que a minha experiência poderá, de futuro, fazer perceber a correlação directa entre a formação académica adquirida ao longo da lincenciatura e posterior mestrado e a experiência prática de estágio. Além do propósito primário da exposição escrita – análise do processo de importação - esta será ainda constituída por uma breve contextualização da empresa Mário Miranda de Almeida, S.A, bem como das funções das quais fui incumbida ao longo dos 5 meses de experiência. Os principais objectivos do estágio, definidos em conjunto com a Dra. Cecília Almeida, consistiram numa inclusão no mundo empresarial, estabelecendo contacto a todos os níveis quer com o comércio internacional como nacional desenvolvido na empresa, e desenvolvendo, autonomamente, tarefas ligadas a estes processos. Por último, irei proceder a uma análise crítica dos resultados obtidos ao longo desta experiência, considerando a avaliação do estágio tanto para a empresa Orima como para mim enquanto estagiária em âmbito empresarial.This report on my training period, besides a brief introduction of the company in which it takes place, aims to analyse the framework of an import procedure which took place in the company Mário Miranda de Almeida S.A. A brief description of this procedure will be presented and subsequently the procedual barriers and conclusions learned from its implementation will be analysed. All the data was obtained at Mário Miranda de Almeida, S.A., also known as Orima. The abovementioned training period lasted 5 months, from February to June of the present year of 2010. The training leader is Professor António Moreno, and the training supervisor is Dr. Cecília Almeida. The report will include the analysis of an import procedure, which because it was not concluded, will be related to and concluded with a fictitious import procedure that had been done before. Having had the possibility of working directly in the import department of Orima and because it is directly related to my Master degree in Languages and Business Communication, I considered it would be of particular interest to address this issue in this report. I believe my experience may, in the future, make people realize the direct co-relation between academic knowledge acquired throughout the course and subsequent Masters degree, and the practical experience gained throughout the training period. Besides the primary purpose of this written report – analysis of an import procedure – a brief contextualization of the company Mário Miranda de Almeida, S.A. will also be included and the tasks I was responsible for during the 5 months of experience I went through. The main objectives of the training period, decided in conjunction with Dr. Cecília Almeida consisted on an inclusion in the business world, establishing contact at all levels with the national, as well as international, trade developed in the company, and accomplishing tasks related with these procedures autonomously. Finally, I will make a critical analysis to the results obtained through this experience, analysing the training period for the company Orima as well as for myself as a trainee in business context

Revisão Sistemática

Introdução: A doença pulmonar obstrutiva crónica é uma doença inflamatória dos pulmões e vias aéreas, cujo tratamento médico atualmente preconizado se baseia em broncodilatadores inalados. A presente dissertação tem como objetivo esclarecer o impacto da telerreabilitação no tratamento desta doença respiratória, nomeadamente no que respeita às seguintes variáveis: dispneia, fadiga, tolerância ao esforço, frequência de exacerbações e de hospitalizações, estado de saúde, qualidade de vida e grau de adesão/satisfação face ao programa. Métodos: Enquanto tipologia de estudo, foi realizada uma revisão sistemática. Foram pesquisados estudos clínicos randomizados publicados até ao dia 20 de dezembro de 2015, nas bases de dados MedLine, Embase, Lilacs, Cochrane e Scisearch, tendo sido utilizada como palavra-chave a expressão “telerehabilitation copd”. Dos 35 ensaios identificados, 14 foram selecionados. Destes, apenas 3 foram incluídos no estudo, tendo sido, posteriormente, submetidos a uma avaliação metodológica, segundo a escala de PEDro. Resultados: Registou-se um impacto positivo da telerreabilitação na maioria dos parâmetros avaliados. Não foi, contudo, possível aferir o impacto desta intervenção na frequência de exacerbações. Conclusão: A telerreabilitação é uma estratégia terapêutica válida, segura e adaptada às características individuais de cada paciente, que demonstrou apresentar um impacto claramente benéfico na maioria das manifestações clínicas da DPOC. Apresenta ainda uma importante componente de promoção de um estilo de vida saudável e fisicamente ativo, bem como de estimulação da participação ativa dos doentes no controlo da própria patologia.Introduction: The chronic obstructive pulmonary disease is an inflammatory disease of the lungs and airways, wherein the medical treatment currently envisaged is based on inhaled bronchodilators. The present study aims to clarify the impact of telerehabilitation on the treatment of this respiratory disease, in particular what concerns the following variables: dyspnoea, fatigue, tolerance to exertion, frequency of exacerbations and hospitalization, health condition, life quality and degree of compliance/satisfaction towards the programme. Methods: It was performed a systematic review. Random clinical studies published up to December 20th 2015 were researched in database from MedLine, Embase, Lilacs, Cochrane and Scisearch, using the keyword “telerehabilitation copd”. From the 35 identified assays, 14 were selected. From these, only 3 were included in the study and later on subject to a methodological assessment, according to the PEDro measure. Results: A positive impact of telerehabilitation was registered in most of the parameters assessed. However, it wasn’t possible to assess the impact of this intervention on the frequency of exacerbations. Conclusion: Telerehabilitation is a valid therapeutic strategy that is safe and adapted to the individual characteristics of each patient, which has shown a clearly beneficial impact on clinic manifestations of the disease. It also shows an important component of promoting a healthy and physically active lifestyle, as well as of fostering an active participation of the patients on the control of their own pathology