Bilirubin directly disrupts membrane lipid polarity and fluidity, protein order, and redox status in rat mitochondria

Background/Aims: Unconjugated bilirubin (UCB) impairs crucial aspects of cell function and induces apoptosis in primary cultured neurones. While mechanisms of cytotoxicity begin to unfold, mitochondria appear as potential primary targets. Methods: We used electron paramagnetic resonance spectroscopy analysis of isolated rat mitochondria to test the hypothesis that UCB physically interacts with mitochondria to induce structural membrane perturbation, leading to increased permeability, and subsequent release of apoptotic factors. Results: Our data demonstrate profound changes on mitochondrial membrane properties during incubation with UCB, including modified membrane lipid polarity and fluidity (P , 0:01), as well as disrupted protein mobility(P , 0:001). Consistent with increased permeability, cytochrome c was released from the intermembrane space(P , 0:01), perhaps uncoupling the respiratory chain and further increasing oxidative stress (P , 0:01). Both ursodeoxycholate, a mitochondrial-membrane stabilising agent, and cyclosporine A, an inhibitor of the permeability transition, almost completely abrogated UCB-induced perturbation. Conclusions: UCB directly interacts with mitochondria influencing membrane lipid and protein properties, redox status, and cytochrome c content. Thus, apoptosis induced by UCB may be mediated, at least in part, by physical perturbation of the mitochondrial membrane. These novel findings should ultimately prove useful to our evolving understanding of UCB cytotoxicity.

The Gap Between Knowledge and Action: Obstacles, Restraints and Deficits During the Execution of Refurbishments

This article outlines the initial draft of a PhD project which investigates refurbishment or rehabilitation projects in two German cities. The study focuses on obstacles, restraints and deficits as well as factors of success, which can be identified during the execution of the refurbishments. Moreover the study examines the process of the refurbishment itself, the general conditions under which the refurbishments are being executed as well as the implementation of sustainability criteria. First the article gives a short summary of the theoretical considerations of the study. In this respect it shortly outlines the global conditions of urban development and conducting challenges for cities in the 21st century, guiding principles of a sustainable urban development as well as goals of sustainable refurbishments. Finally the article shortly describes the case studies and presents the initial results of the empirical work.

The sociological perspective on the knowledge-based society: assumptions, facts and visions

The paper will present the central discourse of the knowledge-based society. Already in the 1960s the debate of the industrial society already raised the question whether there can be considered a paradigm shift towards a knowledge-based society. Some prominent authors already foreseen ‘knowledge’ as the main indicator in order to displace ‘labour’ and ‘capital’ as the main driving forces of the capitalistic development. Today on the political level and also in many scientific disciplines the assumption that we are already living in a knowledge-based society seems obvious. Although we still do not have a theory of the knowledge-based society and there still exist a methodological gap about the empirical indicators, the vision of a knowledge-based society determines at least the perception of the Western societies. In a first step the author will pinpoint the assumptions about the knowledge-based society on three levels: on the societal, on the organisational and on the individual level. These assumptions are relied on the following topics: a) The role of the information and communication technologies; b) The dynamic development of globalisation as an ‘evolutionary’ process; c) The increasing importance of knowledge management within organisations; d) The changing role of the state within the economic processes. Not only the differentiation between the levels but also the revision of the assumptions of a knowledge-based society will show that the ‘topics raised in the debates’ cannot be considered as the results of a profound societal paradigm shift. However what seems very impressive is the normative and virtual shift towards a concept of modernity, which strongly focuses on the role of technology as a driving force as well as on the global economic markets, which has to be accepted. Therefore – according to the official debate - the successful adaptation of these processes seems the only way to meet the knowledge-based society. Analysing the societal changes on the three levels, the label ‘knowledge-based society’ can be seen critically. Therefore the main question of Theodor W. Adorno during the 16th Congress of Sociology in 1968 did not loose its actuality. Facing the societal changes he asked whether we are still living in the industrial society or already in a post-industrial state. Thinking about the knowledge-based society according to these two options, this exercise would enrich the whole debate in terms of social inequality, political, economic exclusion processes and at least the power relationship between social groups.

A coherent approach to non-integer order derivatives

Signal Processing, vol. 86, nº 10

Sub-notificação de reacções adversas medicamentosas : um problema com solução? : contribuição para o conhecimento da sub-notificação de RAMs em Portugal e implementação de medidas minimizadoras do impacte

RESUMO - Introdução: As Reacções Adversas Medicamentosas (RAMs) constituem um grave problema de Saúde Pública em termos da mortalidade e morbilidade provocadas, tendo também um impacto económico considerável nos Sistemas de Saúde. Os Sistemas de Notificacão Espontânea de RAMs são considerados como o método de vigilância de medicamentos mais eficaz, sendo a sub-notificação de RAMs uma das suas maiores limitações. Em termos globais, foi estimado que apenas 6% de todas as reacções adversas são notificadas. Portugal apresenta uma taxa de notificação de RAMs relativamente baixa quando comparada com os países mais notificadores da Europa. São objectivos deste estudo: 1) caracterizar as atitudes e os comportamentos dos médicos, dos farmacêuticos e dos enfermeiros em Portugal Continental relativamente à notificação de RAMs; e 2) caracterizar a efectividade de intervenções educacionais destinadas a reduzir a sub-notificação de RAMs. Métodos: Numa primeira fase será efectuado um estudo de caso-controlo em médicos, farmacêuticos e enfermeiros de Portugal Continental, a exercer actividade no Servico Nacional de Saúde (SNS), de modo a caracterizar as suas atitudes e comportamentos relativamente à notificação de RAMs. Como casos serão considerados os Profissionais de Saúde que notificaram pelo menos uma RAM num determinado período e os controlos os Profissionais de Saúde que não notificaram qualquer RAM nesse mesmo período, sendo estes útimos seleccionados aleatoriamente. O estudo será conduzido através de um questionário de auto-resposta, em que as questões relativas às atitudes e comportamentos são baseadas nos “sete pecados mortais” de Inman. Será utilizada uma Escala Visual Analógica para registar as respostas, podendo estas ir de zero (totalmente em desacordo) até 10 (totalmente de acordo). Será utilizada uma análise de regressão logística para determinar o odds ratio ajustado (ORadj) da notificação de RAMs para uma mudança de exposição correspondente ao range interquartil para cada atitude. Numa segunda fase, será efectuado ensaio aleatorizado controlado de cluster, para caracterizar a efectividade das intervenções educacionais realizadas sobre as causas identificadas na primeira parte do trabalho, com o intuito de reduzir a sub-notificação de RAMs. Com base em informacão de 2007 foram identificados 43 clusters dispersos pelas várias Regiões de Saúde. As intervenções educacionais são compostas por uma apresentação com uma hora de duração complementada por um folheto recordatório. Serão ainda realizados dois sub-estudos, em que o V1.0, Final 28Set09 viii Sub-notificação de RAMs em Portugal – Um problema com solução ? primeiro tentará caracterizar o efeito de contaminação entre Profissionais de Saúde e o segundo pretende caracterizar a duração do efeito das intervenções educacionais. Resultados a atingir: Pretende-se, com a implementação deste projecto, aumentar o número de notificações de RAMs pelos médicos, farmacêuticos e enfermeiros em cerca de 110%, de modo a atingir-se uma taxa de notificação de aproximadamente 300 notificações por milhão de habitantes por ano (i.e., multiplicar por 2,1 o número notificações existentes). -------------------ABSTRACT - Introduction: The Adverse Drug Reactions (ADRs) are a serious Public Health problem in terms of mortality and morbidity caused, being also an economic burden for the health systems. The Spontaneous Adverse Event Reporting Systems are considered as the most effective drug surveillance methods, in which the ADR under-reporting represents one of its biggest limitations. It was estimated that only 6% of all adverse reactions are notified globally. When comparing with high ADR reporting rate countries Portugal shows a low ADR reporting rate. This study aims to: 1) characterize the physicians, pharmacists and nurses attitudes and behaviours related to ADR under-reporting; 2) characterize the educational interventions effectiveness to decrease the ADRs under-reporting. Methods: During a first phase a case-control study will be conducted in physicians, pharmacists and nurses in Continental Portugal working in the National Health System (NHS) in order to characterize their attitudes and behaviours related to ADR reporting. The Healthcare Professionals that have reported at least one ADR during a determined period will be considered as the cases and those that have not reported any ADR during the same period will be considered as the controls. The controls will be randomly selected. The study will be conducted through a self-administered questionnaire in which the questions related to the attitudes and behaviours are based in the Inmans’s “seven mortal sins”. A Visual Analogue Scale will be used to record the responses. The responses can range from 0 (totally disagree) to 10 (totally agree). Logistic regression will be used to determine the ADR reporting adjusted odds ratio (ORadj) for a change in the exposure corresponding to the interquartile range for each attitude. In the second phase of the study a cluster-randomized controlled trial will be conducted to characterize the educational interventions effectiveness focused on the first phase identified causes with the aim to decrease the ADRs under-reporting. Based in 2007’s information 43 clusters have been identified throughout the several Health Regions. The educational interventions are composed of one hour presentation complemented by an informational leaflet. Two sub-studies will be also conducted in which the first one will try to characterize the contamination effect between the Healthcare Professionals and the second to characterize the educational interventions effect duration. V1.0, Final 28Set09 x Sub-notificação de RAMs em Portugal – Um problema com solução ? Outcome: With the project implementation an increase of the ADR notifications performed by the physicians, pharmacists and nurses by 110% is aimed in order to obtain approximately 300 notifications per million habitants per year (i.e., multiply by 2,1 the existent notifications).

Automatic cymbal classification

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia Informática

Congruences on monoids of order-preserving or order-reversing transformations on a finite chain

Glasgow Mathematical Journal, nº 47 (2005), pg. 413-424

Abelian kernels of monoids of order-preserving maps and of some of its extensions

Semigroup Forum vol. 68 (2004), p. 335–356

On the monoids of transformations that preserve the order and a uniform partition

Communications in Algebra

The interrogation of witnesses abroad in execution of a european investigation order: an examination from eyes of the defence

European Master Human Rights and Democratisation

Depositions of rulers in the later middle ages: on the theory of th "useless ruler" and its practical utilization

pp. 157-168

Question order and context effects on multi-items scales questions

Dissertação apresentada como requisito parcial para obtenção do grau de Mestre em Estatística e Gestão de Informação

Regulation of autoimmune neuroinflammation by stress-responsive genes

Dissertation presented to obtain the Ph.D degree in Biology

A study on reducing digital piracy: an analytical network process approach with benefits, opportunities, costs and risks analysis

Dissertação para obtenção do Grau de Mestre em Engenharia e Gestão Industrial

Comparison of different techniques for mouse transgenesis

RESUMO: O uso de ratinhos transgénicos em neurociências aumentou consideravelmente nos últimos anos devido ao crescente interesse em compreender o cérebro e a necessidade de solucionar situações clínicas do foro neurológico e psiquiátrico. Para esse efeito, diferentes métodos de produção de animais transgénicos têm sido testados. O objectivo desta tese foi comparar métodos de integração aleatória de um transgene no genoma de ratinhos em termos de eficiência, estabilidade da integração do transgene, número de animais e de horas de trabalho necessárias para cada método. Assim, foi comparado o método mais utilizado - microinjecção pronuclear (PNMI) - com duas outras técnicas cujo desempenho foi considerado promissor – a transferência génica através dos testículos por electroporação e transfecção por lentivírus in vivo. As três técnicas foram realizadas usando um gene repórter sob o controlo de um promotor constitutivo, e depois reproduzidas usando um gene de interesse de modo a permitir obtenção de um animal capaz de ser usado em experimentação laboratorial. O transgene de interesse utilizado codifica uma proteína de fusão correspondendo a uma variante da rodopsina (channelrhodopsin) fundida à proteína enhanced yellow fluorescente protein ((EYFP) resultando num produto designado ChR2-EYFP. Este animal transgénico apresentaria expressão deste canal iónico apenas em células dopaminergicas, o que, com manipulação optogenética, tornaria possivel a activação especifica deste grupo de neurónios e, simultaneamente, a observação do impacto desta manipulação no comportamento num animal em livre movimento. Estas ferramentas são importantes na investigação básica em neurociências pois ajudam a esclarecer o papel de grupos específicos de neurónios e compreender doenças como a doença de Parkinson ou a esquizofrenia onde a função de certos tipos de neurónios de encontra alterada. Quando comparados os três métodos realizados verifica-se que usando um gene repórter PMNI resulta em 31,3% de, a de animais transgénicos obtidos, a electroporação de testículos em 0% e a injecção de lentivírus em 0%. Quando usado o gene de interesse, os resultados obtidos são, respectivamente, 18,8%, 63,9% e 0%.--------------ABSTRACT: The use of transgenic mice is increasing in all fields of research, particularly in neuroscience, due to the widespread need of animal models to solve neurological and psychiatric medical conditions. Different methodologies have been tested in the last decades in order to produce such transgenic animals. The ultimate goal of this thesis is to compare different methods of random integration of a transgene in the genome of mice in terms of efficiency, stability of the transgene integration, number of animals required and the labour intensity of each technique. We compared the most used method – pronuclear microinjection (PNMI) – with two other promising techniques – Testis Mediated Gene Transfer (TMGT) by electroporation and in vivo lentiviral transfection. The three techniques were performed using a reporter gene – green fluorescent protein (GFP), whose transcription was driven by the constitutive cytomegalovirus (CMV) promoter. These three techniques were later reproduced using the tyrosine hydroxylase promoter (TH) and the neuronal manipulator, channelrhodopsin-2 fused to the enhanced yellow fluorescent reporter protein (ChR2-EYFP). The transgenic animal we sough to produce would express the light driven channel only in dopaminergic cells, making possible to specifically activate this group of neurons, while simultaneously observe the behaviour in a freely moving animal. This is a very important tool in basic neuroscience research since it helps to clarify the role of specific groups of neurons, map circuits in the brain, and consequently understand neurological diseases such as Parkinson’s disease or schizophrenia, where the function of certain types of neurons is affected. When comparing the three methods, it was verified that using a reporter gene PNMI resulted in 31.3% of transgenic mice obtained, testis electroporation in 0% and lentiviral injection in 0%. When using the gene of interest, the results obtained were, respectively, 18.8%, 63.9% and 0%.