Imunorreatividade em material histológico arquivado durante um, quatro e sete anos

Quando aplicada no âmbito da Anatomia Patológica, a imuno-histoquímica tem constituído um poderoso meio de identificação/caracterização de várias estruturas histológicas, permitindo delinear prognóstico e terapêutica para várias patologias. Tendo em conta que as amostras histológicas analisadas podem ser conservadas ao longo de vários anos, interessa avaliar a manutenção da antigenicidade ao longo do tempo, de forma a garantir a qualidade final da técnica quando aplicada em material de arquivo. Assim, o principal objetivo deste trabalho foi comparar a imunorreatividade do material histológico arquivado durante um, quatro e sete anos. Foi utilizado material histológico de próstata, pulmão e mama, no qual se procedeu à imunomarcação de citoqueratinas (Clones AE1/AE3), CD34 e proteína p63, por método de multímero/HRP no sistema Ventana BenchMark Ultra®. Foi realizado um ensaio com recuperação antigênica por alta temperatura (RAAT) e outro sem esta etapa. As imunomarcações (n=162) foram classificadas por três avaliadores independentes num escore quantitativo final (escala 0-100). O par média/desvio-padrão do escore final para os casos com sete anos foi de 69,06/19,05, para os casos com quatro anos foi de 66,47/20,73 e para os casos com um ano foi de 69,08/19,35, não se tendo encontrado diferenças estatisticamente significativas. Os casos sem RAAT obtiveram um par média/desvio-padrão de 54,90/17,00, enquanto os casos com RAAT obtiveram 81,50/11,60, o que revelou diferenças estatisticamente significativas (p=0,000). Para os casos em estudo conclui-se que o fator “tempo de arquivo” não está associado a alterações da imunorreatividade. A importância da RAAT na obtenção de imunomarcação de qualidade sai fortemente realçada. ABSTRACT - When applied within the framework of Pathology, immunohistochemistry has been a powerful means of identification/characterization of various histological structures, allowing to outline prognosis and therapy for various diseases. Given that the analyzed histological samples can be preserved for several years, it is interesting to assess the retention of antigenicity over time in order to ensure the quality of the final technique, when applied to stored material. Thus, the main objective of this study was to compare the immunoreactivity of the histological material archived for one, four and seven years. It was used histological material from prostate, lung and breast, in which it was performed the immunostaining of cytokeratins (clones AE1/AE3), CD34 and p63 protein by the method of multimer/HRP system on a Ventana BenchMark Ultra®. It was conducted a test with heat induced epitope retrieval (HIER) and another one without this step. The stained slides (n=162) were classified by three independent assessors using a quantitative score (scale 0-100). The pair mean/standard deviation of the score for cases with seven years was 69,06/19,05, for cases with four years was 66,47/20,73 and for cases with one year was 69,08/19,35, which did not revealed any statistically significant differences. The cases without HIER had a couple mean/standard deviation of 54.90/17.00 while the cases with HIER obtained 81.50/11.60, which revealed statistically significant differences (p=0.000). For this case study it was concluded that the factor archive period is not associated with changes in immunoreactivity. The importance of HIER in obtaining high quality immunostaining comes out strongly highlighted.

Structural characterization and immunogenicity in wildtype and immune tolerant mice of degraded recombinant human interferon alpha2b

Purpose - To study the influence of protein structure on the immunogenicity in wildtype and immune tolerant mice of well-characterized degradation products of recombinant human interferon alpha2b (rhIFNα2b). Methods - RhIFNα2b was degraded by metal catalyzed oxidation (M), crosslinking with glutaraldehyde (G), oxidation with hydrogen peroxide (H) and incubation in a boiling water bath (B). The products were characterized with UV absorption, circular dichroism and fluorescence spectroscopy, gel permeation chromatography, reversed-phase HPLC, SDS-PAGE, Western blotting and mass spectrometry. The immunogenicity of the products was evaluated in wildtype mice and in transgenic mice immune tolerant for hIFNα2. Serum antibodies were detected by ELISA or surface plasmon resonance. Results - M-rhIFNα2b contained covalently aggregated rhIFNα2b with three methionines partly oxidized to methionine sulfoxides. G-rhIFNα2b contained covalent aggregates and did not show changes in secondary structure. H-rhIFNα2b was only chemically changed with four partly oxidized methionines. B-rhIFNα2b was largely unfolded and heavily aggregated. Native (N) rhIFNα2b was immunogenic in the wildtype mice but not in the transgenic mice, showing that the latter were immune tolerant for rhIFNα2b. The antirhIFNα2b antibody levels in the wildtype mice depended on the degradation product: M-rhIFNα2b > H-rhIFNα2b ~ N-rhIFNα2b >> B-rhIFNα2b; G-rhIFNα2b did not induce anti-rhIFNα2b antibodies. In the transgenic mice, only M-rhIFNα2b could break the immune tolerance. Conclusions - RhIFNα2b immunogenicity is related to its structural integrity. Moreover, the immunogenicity of aggregated rhIFNα2b depends on the structure and orientation of the constituent protein molecules and/or on the aggregate size.

Mould and yeast identification in archival settings: preliminary results on the use of traditional methods and molecular biology options in Portuguese archives

This project was developed to fully assess the indoor air quality in archives and libraries from a fungal flora point of view. It uses classical methodologies such as traditional culture media – for the viable fungi – and modern molecular biology protocols, especially relevant to assess the non-viable fraction of the biological contaminants. Denaturing high-performance liquid chromatography (DHPLC) has emerged as an alternative to denaturing gradient gel electrophoresis (DGGE) and has already been applied to the study of a few bacterial communities. We propose the application of DHPLC to the study of fungal colonization on paper-based archive materials. This technology allows for the identification of each component of a mixture of fungi based on their genetic variation. In a highly complex mixture of microbial DNA this method can be used simply to study the population dynamics, and it also allows for sample fraction collection, which can, in many cases, be immediately sequenced, circumventing the need for cloning. Some examples of the methodological application are shown. Also applied is fragment length analysis for the study of mixed Candida samples. Both of these methods can later be applied in various fields, such as clinical and sand sample analysis. So far, the environmental analyses have been extremely useful to determine potentially pathogenic/toxinogenic fungi such as Stachybotrys sp., Aspergillus niger, Aspergillus fumigatus, and Fusarium sp. This work will hopefully lead to more accurate evaluation of environmental conditions for both human health and the preservation of documents.

Sistema de localização de um veículo autónomo terrestre

Mestrado em Engenharia Electrotécnica e de Computadores.

Escoamento de superfície e vulnerabilidade às cheias no concelho da Ribeira Grande (São Miguel, Açores)

Dissertação de Mestrado, Geologia do Ambiente e Sociedade, 8 de Maio de 2015, Universidade dos Açores.

How new online communication tools and the availability of content influence the audiences and the creative process

We live in a changing world. At an impressive speed, every day new technological resources appear. We increasingly use the Internet to obtain and share information, and new online communication tools are emerging. Each of them encompasses new potential and creates new audiences. In recent years, we witnessed the emergence of Facebook, Twitter, YouTube and other media platforms. They have provided us with an even greater interactivity between sender and receiver, as well as generated a new sense of community. At the same time we also see the availability of content like it never happened before. We are increasingly sharing texts, videos, photos, etc. This poster intends to explore the potential of using these new online communication tools in the cultural sphere to create new audiences, to develop of a new kind of community, to provide information as well as different ways of building organizations’ memory. The transience of performing arts is accompanied by the need to counter that transience by means of documentation. This desire to ‘save’ events reaches its expression with the information archive of the different production moments as well as the opportunity to record the event and present it through, for instance, digital platforms. In this poster we intend to answer the following questions: which online communication tools are being used to engage audiences in the cultural sphere (specifically between theater companies in Lisbon)? Is there a new relationship with the public? Are online communication tools creating a new kind of community? What changes are these tools introducing in the creative process? In what way the availability of content and its archive contribute to the organization memory? Among several references, we will approach the two-way communication model that James E. Grunig & Todd T. Hunt (1984) already presented and the concept of mass self-communication of Manuel Castells (2010). Castells also tells us that we have moved from traditional media to a system of communication networks. For Scott Kirsner (2010), we have entered an era of digital creativity, where artists have the tools to do what they imagined and the public no longer wants to just consume cultural goods, but instead to have a voice and participate. The creativity process is now depending on the public choice as they wander through the screen. It is the receiver who owns an object which can be exchanged. Virtual reality has encouraged the receiver to abandon its position of passive observer and to become a participant agent, which implies a challenge to organizations: inventing new forms of interfaces. Therefore, we intend to find new and effective online tools that can be used by cultural organizations; the best way to manage them; to show how organizations can create a community with the public and how the availability of online content and its archive can contribute to the organizations’ memory.

Differences in Mismatch Negativity (MMN) response to Pure-tone and Speech sounds in normal subjects: an additional explanation

The relation of automatic auditory discrimination, measured with MMN, with the type of stimuli has not been well established in the literature, despite its importance as an electrophysiological measure of central sound representation. In this study, MMN response was elicited by pure-tone and speech binaurally passive auditory oddball paradigm in a group of 8 normal young adult subjects at the same intensity level (75 dB SPL). The frequency difference in pure-tone oddball was 100 Hz (standard = 1 000 Hz; deviant = 1 100 Hz; same duration = 100 ms), in speech oddball (standard /ba/; deviant /pa/; same duration = 175 ms) the Portuguese phonemes are both plosive bi-labial in order to maintain a narrow frequency band. Differences were found across electrode location between speech and pure-tone stimuli. Larger MMN amplitude, duration and higher latency to speech were verified compared to pure-tone in Cz and Fz as well as significance differences in latency and amplitude between mastoids. Results suggest that speech may be processed differently than non-speech; also it may occur in a later stage due to overlapping processes since more neural resources are required to speech processing.

Dossiê os livreiros e o seu património

Este dossiê representa a continuidade de um projecto que tem procurado debater publi­camente o património dos agentes ligados à produção e circulação do livro. Centrando-se nos espólios dos livreiros, a introdução problematizante e os depoimentos procuram dis­cutir o seu carácter invisível enquanto objecto de pesquisa, salvaguarda e divulgação. Há, então, uma reflexão que urge fazer, à semelhança do que ocorre com o património dos edi­tores (arquivos históricos e não só), no sentido de reconhecer o estatuto patrimonial e histó­rico dos acervos dos livreiros e das livrarias enquanto espaços fundamentais da construção da cultura impressa e da história do livro e da edição. Mas é também urgente conferir ao debate um cunho de acção e de cooperação inter-institucional que suscitem uma sensibili­dade política e o aparecimento de possibilidades concretas de salvamento, recolha e depó­sito sistemático de um dos mais fundamentais elementos da memória colectiva escrita. Os depoimentos referidos são de Fátima Ribeiro de Medeiros e dos Livreiros da Sá da Costa.

O Arquivo Audiovisual da SIC: a reutilização dos conteúdos da estação na produção diária de notícias

Relatório de estágio apresentado à Escola Superior de Comunicação Social como parte dos requisitos para obtenção de grau de mestre em Audiovisual e Multimédia.

Efémeros sinais

Este artigo apresenta três sítios em linha onde se pode detectar a presença do teatro na internet: a página da companhia teatral italiana Piccolo Teatro de Milano (www.piccoloteatro.org); a revista electrónica Didaskalia - Ancient Theatre Today (http://didaskalia.open.ac.uk); e o arquivo digital "Footlight Notes" (http://footlightnotes.tripod.com).

Sequencing educational resources with Seqins

Existing adaptive educational hypermedia systems have been using learning resources sequencing approaches in order to enrich the learning experience. In this context, educational resources, either expository or evaluative, play a central role. However, there is a lack of tools that support sequencing essentially due to the fact that existing specifications are complex. This paper presents Seqins as a sequencing tool of digital educational resources. Seqins includes a simple and flexible sequencing model that will foster heterogeneous students to learn at different rhythms. The tool communicates through the IMS Learning Tools Interoperability specification with a plethora of e-learning systems such as learning management systems, repositories, authoring and automatic evaluation systems. In order to validate Seqins we integrate it in an e-learning Ensemble framework instance for the computer programming learning domain.

TV Recommendation and Personalization Systems: Integrating Broadcast and Video On-demand Services

he expansion of Digital Television and the convergence between conventional broadcasting and television over IP contributed to the gradual increase of the number of available channels and on demand video content. Moreover, the dissemination of the use of mobile devices like laptops, smartphones and tablets on everyday activities resulted in a shift of the traditional television viewing paradigm from the couch to everywhere, anytime from any device. Although this new scenario enables a great improvement in viewing experiences, it also brings new challenges given the overload of information that the viewer faces. Recommendation systems stand out as a possible solution to help a watcher on the selection of the content that best fits his/her preferences. This paper describes a web based system that helps the user navigating on broadcasted and online television content by implementing recommendations based on collaborative and content based filtering. The algorithms developed estimate the similarity between items and users and predict the rating that a user would assign to a particular item (television program, movie, etc.). To enable interoperability between different systems, programs characteristics (title, genre, actors, etc.) are stored according to the TV-Anytime standard. The set of recommendations produced are presented through a Web Application that allows the user to interact with the system based on the obtained recommendations.

As determinantes da qualidade da informação financeira via imparidade de ativos

Dissertação de Mestrado apresentada ao Instituto de Contabilidade e Administração do Porto para a obtenção do grau de Mestre em Contabilidade e Finanças, sob orientação da Professora Doutora Cláudia Maria Ferreira Pereira

Políticas de Arquivo na Administração Pública

Comunicação apresentada no Ciclo de Debates "Pensar a Administração Pública", 8.ª Sessão - Políticas de arquivo na Administração Pública, organizada pelo INA, em Lisboa a 12 de novembro de 2014.

Gestão do conhecimento e cultura organizacional: barreiras e facilitadores

Este trabalho incide sobre a gestão do conhecimento e cultura organizacional, as suas barreiras os seus facilitadores na Parque Escolar E.P.E. Este estudo teve por base o método quadripolar. Várias foram as atividades ao longo deste trabalho, inicialmente foi recolhida a documentação interna, nomeadamente diplomas legais, regulamentos, manuais de procedimentos, manuais de formações internas, entre outros documentos, que serviram de base ao reconhecimento da instituição, a sua evolução estrutural e de funcionamento. Para identificar as barreiras e os facilitadores na recuperação da informação nos três principais meios para o efeito: arquivo físico, file system e aplicações informáticas foram aplicados inquéritos aos produtores/ utilizadores de informação da Parque Escolar, E.P.E. Com base neste estudo foi possível identificar qual o recurso de recuperação de informação que traz mais dificuldades na sua utilização, se existem documentos exclusivos em papel ou exclusivos em formato digital, se os mesmos são recuperáveis com facilidade. Foi possível averiguar se os colaboradores da Parque Escolar, E.P.E. consideram os documentos que constam no Arquivo Físico mais fidedignos do que os documentos em formato digital guardados no file system ou nas aplicações informáticas. Em relação às aplicações informáticas foi ainda possível averiguar se os colaboradores consideram uteis as suas atualizações, ou se demonstram alguma resistência à mudança, e se consideram que tiveram o acompanhamento necessário para compreender e aplicar as alterações. Com este estudo esperamos ter contribuído para dar uma maior visibilidade à temática da gestão do conhecimento e como a cultura organizacional pode influenciar, criando barreiras ou facilitadores.