Modulation of osteoclastogenesis by antihypertensive drugs

Despite its rigid structure, bone is a dynamic tissue that is in constant remodeling. This process requires the action of the bone-resorbing osteoclasts and the bone-synthesing osteoblasts. One of the adverse effects attributed to some antihypertensive agents is the ability to alter normal bone metabolism. However, their effective actions on human bone cells remain to be clarified. In this work, the effects of five calcium channel blockers, a class of antihypertensive drugs (AHDs), were investigated on osteoclastic differentiation. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood, and were maintained in the absence (control) or in the presence of 10-8-10-4 M of different AHDs (amlodipine, felodipine, diltiazem, lercanidipine and nifedipine). Cell cultures were characterized throughout a 21 day period for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors, and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. It was observed that the tested AHDs had the ability to differentially affect osteoclastogenesis. At low doses, amlodipine and felodipine caused an increase on osteoclastic differentiation, while the other drugs inhibited it. At higher doses, all the molecules caused a decrease on the process. The tested AHDs also showed different effects on the analysed signaling pathways. In conclusion, AHDs appeared to have a direct effect on human osteoclast precursor cells, affecting their differentiation. Interestingly, some of them increased while others inhibited the process. Unraveling the mechanisms beneath these observations might help to explain the adverse effects on bone tissue described for this drug class.

Habitat differentiation between groups of Risso's dolphins as evidence of coastal nursery areas

26th Annual Conference of the European Cetacean Society, Galway, Ireland 26-28 March 2012.

A criação de valor no mercado da saúde enquanto factor diferenciador para a negociação de preços e competitividade em contexto de crise mundial

A criação de valor no mercado da saúde enquanto factor diferenciador para a negociação de preços e competitividade em contexto de crise mundial Numa altura em que o sector da saúde é apontado como uma área crítica de custos, torna-se cada vez mais difícil orientar a contratação em saúde baseada em valor para os pacientes, ou seja, pelos resultados obtidos e não pelo volume de cuidados prestados. Pretendeu-se estudar a criação de valor no mercado da saúde enquanto factor diferenciador para a negociação de preços e competitividade em contexto de crise económica. Procedeu-se à comparação dos resultados operacionais de uma empresa enquanto prestadora de serviços de Oxigenoterapia ao domicílio, tendo por base duas estratégias diferentes: redução directa de preços ou manutenção de preços com criação de valor para o cliente. As propostas foram posteriormente apresentadas para avaliação e votação on-line por um grupo 8 gestores hospitalares. A proposta baseada em valor (Nº2) apresenta melhores resultados operacionais (41%) embora apresente maiores custos. No que se refere à votação das propostas e tendo em conta o cenário apresentado, metade dos gestores optaram pela proposta Nº1 (N=4) e outra metade pela proposta Nº2 (N=4). Contudo, a maioria dos gestores (N=7) consideraram a proposta Nº1 a mais competitiva em contexto de competição com mais fornecedores. Conclui-se que numa negociação de contratos de cuidados de saúde, uma proposta baseada em valor, pode garantir a manutenção dos preços. Todavia, mantendo-se uma situação económica de recessão e num cenário competitivo de vários fornecedores este tipo de propostas pode não eleita, pelo facto de aparentemente não representar ganhos imediatos para a instituição contratante.

Differential expression of the eukaryotic release factor 3 (eRF3/GSPT1) according to gastric cancer histological types

Background: There are now several lines of evidence to suggest that protein synthesis and translation factors are involved in the regulation of cell proliferation and cancer development. Aims: To investigate gene expression patterns of eukaryotic releasing factor 3 (eRF3) in gastric cancer. Methods: RNA was prepared from 25 gastric tumour biopsies and adjacent non-neoplastic mucosa. Real time TaqMan reverse transcription polymerase chain reaction (RT-PCR) was performed to measure the relative gene expression levels. DNA was isolated from tumour and normal tissues and gene dosage was determined by a quantitative real time PCR using SYBR Green dye. Results: Different histological types of gastric tumours were analysed and nine of the 25 tumours revealed eRF3/GSPT1 overexpression; moreover, eight of the 12 intestinal type carcinomas analysed overexpressed the gene, whereas eRF3/GSPT1 was overexpressed in only one of the 10 diffuse type carcinomas (Kruskal-Wallis Test; p , 0.05). No correlation was found between ploidy and transcript expression levels of eRF3/GSPT1. Overexpression of eRF3/GSPT1 was not associated with increased translation rates because the upregulation of eRF3/GSPT1 did not correlate with increased eRF1 levels. Conclusions: Overexpression of eRF3/GSPT1 in intestinal type gastric tumours may lead to an increase in the translation efficiency of specific oncogenic transcripts. Alternatively, eRF3/GSPT1 may be involved in tumorigenesis as a result of its non-translational roles, namely (dis)regulating the cell cycle, apoptosis, or transcription.

Q192R polymorphism of the paraoxonase-1 gene as a risk factor for obesity in Portuguese women

Introduction - Obesity became a major public health problem as a result of its increasing prevalence worldwide. Paraoxonase-1 (PON1) is an esterase able to protect membranes and lipoproteins from oxidative modifications. At the PON1 gene, several polymorphisms in the promoter and coding regions have been identified. The aims of this study were i) to assess PON1 L55M and Q192R polymorphisms as a risk factor for obesity in women; ii) to compare PON1 activity according to the expression of each allele in L55M and Q192R polymorphisms; iii) to compare PON1 activity between obese and normal-weight women. Materials and methods - We studied 75 healthy (35.9±8.2 years) and 81 obese women (34.3±8.2 years). Inclusion criteria for obese subjects were body mass index ≥30 kg/m2 and absence of inflammatory/neoplasic conditions or kidney/hepatic dysfunction. The two PON1 polymorphisms were assessed by real-time PCR with TaqMan probes. PON1 enzymatic activity was assessed by spectrophotometric methods, using paraoxon as a substrate. Results - No significant differences were found for PON1 activity between normal and obese women. Nevertheless, PON1 activity was greater (P<0.01) for the RR genotype (in Q192R polymorphism) and for the LL genotype (in L55M polymorphism). The frequency of allele R of Q192R polymorphism was significantly higher in obese women (P<0.05) and was associated with an increased risk of obesity (odds ratio=2.0 – 95% confidence interval (1.04; 3.87)). Conclusion - 55M and Q192R polymorphisms influence PON1 activity. The allele R of the Q192R polymorphism is associated with an increased risk for development of obesity among Portuguese Caucasian premenopausal women.

Modulation of translation factor's gene expression by histone deacetylase inhibitors in breast cancer cells

The histone deacetylase inhibitors sodium butyrate (NaBu) and trichostatin A (TSA) exhibit anti-proliferative activity by causing cell cycle arrest and apoptosis. The mechanisms by which NaBu and TSA cause apoptosis and cell cycle arrest are not yet completely clarified, although these agents are known to modulate the expression of several genes including cell-cycle- and apoptosis-related genes. The enzymes involved in the process of translation have important roles in controlling cell growth and apoptosis, and several of these translation factors have been described as having a causal role in the development of cancer. The expression patterns of the translation mechanism, namely of the elongation factors eEF1A1 and eEF1A2, and of the termination factors eRF1 and eRF3, were studied in the breast cancer cell line MCF-7 by real-time quantitative reverse transcription-polymerase chain reaction after a 24-h treatment with NaBu and TSA. NaBu induced inhibition of translation factors' transcription, whereas TSA caused an increase in mRNA levels. Thus, these two agents may modulate the expression of translation factors through different pathways. We propose that the inhibition caused by NaBu may, in part, be responsible for the cell cycle arrest and apoptosis induced by this agent in MCF-7 cells.

Effects of antihypertensive drugs in the differentiation and activation of human bone cells

O osso é um tecido metabolicamente ativo e a sua remodelação é importante para regular e manter a massa óssea. Esse processo envolve a reabsorção do material ósseo por ação dos osteoclastos e a síntese de novo material ósseo mediado pelos osteoblastos. Vários estudos têm sugerido que a pressão arterial elevada está associada a alterações no metabolismo do cálcio, o que leva ao aumento da perda de cálcio e da remoção de cálcio do osso. Embora as alterações no metabolismo ósseo sejam um efeito adverso associado a alguns fármacos antihipertensores, o conhecimento em relação a este efeito terapêutico ligado com os bloqueadores de canais de cálcio é ainda muito escasso. Uma vez que os possíveis efeitos no osso podem ser atribuídos à ação antihipertensiva dessas moléculas, ou através de um efeito direto nas atividades metabólicas ósseas, torna-se necessário esclarecer este assunto. Devido ao facto de que as alterações no metabolismo ósseo são um efeito adverso associado a alguns fármacos antihipertensores, o objetivo deste trabalho é avaliar o efeito que os bloqueadores dos canais de cálcio exercem sobre as células ósseas humanas, nomeadamente osteoclastos, osteoblastos e co-culturas de ambos os tipos celulares. Verificou-se que os efeitos dos fármacos antihipertensores variaram consoante o fármaco testado e o sistema de cultura usado. Alguns fármacos revelaram a capacidade de estimular a osteoclastogénese e a osteoblastogénese em concentrações baixas. Independentemente da identidade do fármaco, concentrações elevadas revelaram ser prejudiciais para a resposta das células ósseas. Os mecanismos intracelulares através dos quais os efeitos foram exercidos foram igualmente afetados de forma diferencial pelos diferentes fármacos. Em resumo, este trabalho demonstrou que os bloqueadores dos canais de cálcio utilizados possuem a capacidade de afetar direta- e indiretamente a resposta de células ósseas humanas, cultivadas isoladamente ou co-cultivadas. Este tipo de informação é crucial para compreender e prevenir os potenciais efeitos destes fármacos no tecido ósseo, e também para adequar e eventualmente melhorar a terapêutica antihipertensora de cada paciente.

Mapping non-suicidal self-injury in adolescence: Development and confirmatory factor analysis of the impulse, self-harm and sucidal ideation questionnaire for adolescents (ISSIQ-A)

Non-suicidal self-injury (NSSI) is the deliberate, self-inflicted destruction of body tissue without suicidal intent and an important clinical phenomenon. Rates of NSSI appear to be disproportionately high in adolescents and young adults, and is a risk factor for suicidal ideation and behavior. The present study reports the psychometric properties of the Impulse, Self-harm and Suicide Ideation Questionnaire for Adolescents (ISSIQ-A), a measure designed to comprehensively assess the impulsivity, NSSI behaviors and suicide ideation. An additional module of this questionnaire assesses the functions of NSSI. Results of Confirmatory Factor Analysis (CFA) of the scale on 1722 youths showed items' suitability and confirmed a model of four different dimensions (Impulse, Self-harm, Risk-behavior and Suicide ideation) with good fit and validity. Further analysis showed that youth׳s engagement in self-harm may exert two different functions: to create or alleviate emotional states, and to influence social relationships. Our findings contribute to research and assessment on non-suicidal self-injury, suggesting that the ISSIQ-A is a valid and reliable measure to assess impulse, self-harm and suicidal thoughts, in adolescence.

O endomarketing como factor de sucesso nas organizações: o impacto no empenhamento e na satisfação dos clientes internos da empresa PT PRO

A literatura descreve que as ações de endomarketing contribuem para a aproximação, interação e o bom relacionamento entre a empresa e os colaboradores. Sendo caracterizada como uma relação de troca, na qual a empresa fornece instrumentos capazes de promover o interesse dos colaboradores a participarem mais ativamente em toda a envolvente organizacional - através de ações que favoreçam o diálogo, sugestões, partilha de conhecimentos e informações, bem como relatos de situações corriqueiras ocorridas no dia-a-dia da empresa. Em contrapartida, à medida que o colaborador se sente valorizado pela empresa e pelos colegas, passa a perceber a importância do seu trabalho dentro da organização, permitindo que identifique o trabalho como algo prazeroso. Esta troca contribui para a produtividade e desenvolvimento da empresa, bem como a satisfação e empenhamento dos colaboradores no trabalho. Neste sentido, visando averiguar como esta situação é vivenciada na prática decidiu-se pela realização de um estágio, tendo como questão de estudo qual o impacto das ações de endomarketing no empenhamento e na satisfação dos clientes internos da PT PRO? Para responder a tal indagação foi realizado um estágio na empresa PT PRO, nomeadamente na Direção de Desenvolvimento de Mercado, Marketing e Formação. Apresentam-se ao longo do presente relatório a descrição das especificidades do estágio, tal como as principais ilações daí retiradas tendo através de pesquisas documentais (arquivo e documentos internos e Intranet), reuniões com a Direção de Marketing e com a tutora do estágio, tal como pela aplicação e análise de um inquérito por questionário aplicado aos colaboradores, chegando-se à conclusão que, de facto, as ações de endomarketing levadas a cabo pela empresa contribuem para o empenhamento e satisfação dos colaboradores.

Students' engagement in schools : differentiation and promotion

The objective of this work is to present elements of the project Student engagement in Schools (SES). The team consists of 10 researchers from six Universities. Student engagement in schools is a multidimensional construct that unites affective, behavioural, and cognitive dimensions of student adaptation in the school and has influence on students’outcomes. The team of researchers conceptualized two major studies, a differential study to analyze the relations between SES and contextual factors, personal factors, student’s outcomes, and a quasi-experimental study to analyze the effects on SES of a specific intervention programmes. In study 1, the sample size is around 600 students (150 6th graders, 150 7th graders, 150 9th graders, and 150 10th graders). We shall focus on years of school transition, with rural and urban populations, on different regions of the country, and on students with different family background. We shall conduct questionnaires with national and international scales. The study 2 will involve students in 7th and 9th grade, from four classes, two of the experimental group and two of the control group. Patterns of verbal communication between a teacher and students can influence the classroom environment and SES. This model of communication would result in more effective student management and more time on-task for learning.

A realidade virtual como factor de atracção para a engenharia

O objectivo da Realidade Virtual é simples de entender mas muito difícil de implementar: criar ambientes completamente indiferenciáveis do mundo real com os quais se possa interagir de um modo natural. Desde a criação do Sensorama por Morton Heiling em 1962, passando pela difusão do conceito pelo público geral na década de 90 até os dias de hoje, a evolução da Realidade Virtual tem sido constante. Este conjunto de tecnologias tem estado envolvido por uma certa descrença por parte da sociedade, motivada pelas grandes expectativas que lhe foram atribuídas e pelo estado de desenvolvimento do hardware aquando do seu auge. No entanto, actualmente assiste-se a um ressurgimento do seu interesse no público geral com a introdução de imagem estereoscópica no cinema ou o sucesso dos controladores da consola Nintendo Wii. Hoje em dia as suas aplicações são muito variadas: desde o treino de pilotos de avião ao tratamento de fobias, passando pela industria do entretenimento e a visita virtual de locais com interesse histórico ou turístico. O objectivo desta tese de mestrado é explorar uma área que ainda não tem sido muito abrangida pela Realidade Virtual e que cobre também aspectos educacionais e lúdicos de modo a ser um factor de atracção para os estudantes do ensino secundário: a simulação de instrumentos musicais. Para tal foi implementado um sistema capaz de simular instrumentos musicais de percussão (uma bateria) utilizando imagem estereoscópica, som posicional e interfaces com o utilizador realistas. Os resultados obtidos nas sessões de avaliação efectuadas por alunos recentemente ingressados no ensino superior demonstram que o sistema desenvolvido, bem como a inovação em interfaces do utilizador com os dispositivos electrónicos de uma forma geral, constituem um meio efectivo na sua motivação para a escolha de um curso na área da engenharia.

O contributo da implementação de um sistema de controlo interno robusto para as pequenas e médias empresas portuguesas: o caso das agências de meios e publicidade

Mestrado em Auditoria

Development and validation of a psychometric scale for assessing PA chest image quality: a pilot study

Purpose - To develop and validate a psychometric scale for assessing image quality perception for chest X-ray images. Methods - Bandura's theory was used to guide scale development. A review of the literature was undertaken to identify items/factors which could be used to evaluate image quality using a perceptual approach. A draft scale was then created (22 items) and presented to a focus group (student and qualified radiographers). Within the focus group the draft scale was discussed and modified. A series of seven postero-anterior chest images were generated using a phantom with a range of image qualities. Image quality perception was confirmed for the seven images using signal-to-noise ratio (SNR 17.2–36.5). Participants (student and qualified radiographers and radiology trainees) were then invited to independently score each of the seven images using the draft image quality perception scale. Cronbach alpha was used to test interval reliability. Results - Fifty three participants used the scale to grade image quality perception on each of the seven images. Aggregated mean scale score increased with increasing SNR from 42.1 to 87.7 (r = 0.98, P < 0.001). For each of the 22 individual scale items there was clear differentiation of low, mid and high quality images. A Cronbach alpha coefficient of >0.7 was obtained across each of the seven images. Conclusion - This study represents the first development of a chest image quality perception scale based on Bandura's theory. There was excellent correlation between the image quality perception scores derived using the scale and the SNR. Further research will involve a more detailed item and factor analysis.

Epigenetic and transcriptional signatures of stable versus plastic differentiation of proinflammatory gd T cell subsets

Two distinct subsets of γδ T cells that produce interleukin 17 (IL-17) (CD27(-) γδ T cells) or interferon-γ (IFN-γ) (CD27(+) γδ T cells) develop in the mouse thymus, but the molecular determinants of their functional potential in the periphery remain unknown. Here we conducted a genome-wide characterization of the methylation patterns of histone H3, along with analysis of mRNA encoding transcription factors, to identify the regulatory networks of peripheral IFN-γ-producing or IL-17-producing γδ T cell subsets in vivo. We found that CD27(+) γδ T cells were committed to the expression of Ifng but not Il17, whereas CD27(-) γδ T cells displayed permissive chromatin configurations at loci encoding both cytokines and their regulatory transcription factors and differentiated into cells that produced both IL-17 and IFN-γ in a tumor microenvironment.

Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells

Bone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.