Análise molecular e celular da resposta regenerativa da pele/escamas da dourada (Sparus aurata)

Dissertação mest., Engenharia Biológica, Universidade do Algarve, 2009Em peixes os mecanismos por detrás da regulação da pele e escamas face a alterações na homeostasia do cálcio estão pouco estudados. Esta tese pretende contribuir para a identificação dos transcriptos presentes na pele de peixe e a forma como eles alteram em resposta a uma modificação, através da remoção de uma grande percentagem de escamas; identificar as células características da escama utilizando de marcadores moleculares e avaliar as consequências morfológicas de remoção de escamas e a sua regeneração durante 3 e 7 dias em relação ao controlo. O organismo usado para este estudo foi a dourada (Sparus auratus), recentemente escolhido como um modelo para espécies aquícolas e para as quais existem numerosos recursos moleculares. A pele removida foi utilizada para extracção de RNA e análise histológica. O RNA total de pele/escama foi hibridado com um oligo-array de dourada, de forma a identificar diferenças de expressão entre genes dos grupo controlo e tratados. Da técnica de microarray obtiveram-se resultados reprodutíveis e os dados foram analisados estatisticamente, utilizando o programa SAM (Significance Analysis of Microarrays). No grupo de peixes 3 dias após a remoção de escamas um total de 247 sondas estavam diferencialmente expressas em relação ao controlo do dia 3, dessas 82 estão upreguladas e as restantes down-reguladas. Por outro lado, no grupo dos 7 dias após a remoção de escamas apenas 14 sondas tinham uma expressão diferencial. Para associar as alterações no transcriptoma com morfologia pele, foram realizados ensaios histoquímicos. Através da utilização de um marcador enzimático (TRAP) foi possível verificar a existência de osteoblastos nos tecidos controlo e tratado, mas não foram observadas mudanças significativas. Em relação à OSN foi realizada imunohistoquímica onde só foi detectado sinal no tecido controlo, com os marcadores pCNA e ASPIC foi detectado um aumento do sinal no tecido controlo para o tratado e para os tempos de amostragem

O DUPLO DA FICÇÃO

O presente artigo pretende realizar uma leitura do romance Sem nome, do escritor português Helder Macedo, considerando três pontos principais de análise: a elaboração de um narrador caprichoso que se filia a uma determinada tradição literária; a apropriação de referências intertextuais e o discurso metaficcional; a estrutura narrativa construída a partir de oposições ou antinomias, presentes em todos os níveis de seus romances

As erratas pensantes: uma leitura de Pedro e Paula, de Helder Macedo

O presente ensaio pretende realizar uma leitura do romance Pedro e Paula, do escritor português Helder Macedo, considerando a apropriação de referências intertextuais da obra de Machado de Assis. Para tanto, pretendemos interpretar Pedro e Paula a partir dos critérios adotados pelo próprio autor para interpretar as obras ficcionais de sua eleição, incluindo Machado de Assis.The present essay intends to undertake a reading of Pedro e Paula, the novel by the Portuguese author Helder Macedo, takinginto account the appropriation of intertextual references of Machado de Assis’ works. Thus, we intend to interpret Pedro e Paula from the criteria adopted by the very author to interpret the fictional works of his election, Machado de Assis included

application of lean methodologies

Poster presentationHealthcare acquired infections (HAI) are an important cause of morbidity and mortality. Hand hygiene (HH) is one of the most effective measures for preventing HAI. However, stirring healthcare workers to comply with HH remains a challenge. Two common barriers often found are forgetfulness and the lack of time. We hypothesize that the perception of lack of time and forgetfulness stems from the pressure of too much tasks in a short period of time. Therefore, it is rational to search for efficiency and minimize waste. This study aims at exploring the use of LEAN methodologies to improve nurses’ work processes in an ICU ward. A design science research approach was used. A questionnaire was applied to the nurses followed by an observational study of an 8-hour work shift to identify nurses’ activities, processes of care, and to describe HH compliance. Next, a value stream map (VSM) was designed and the analysis performed with the nurses, which helped to identify points of possible improvement in the process of care. After this, weekly LEAN workshops were held with the objective of improving workflow efficiency. From the initial observational study, we found that a nurse may take on average 16% of their work time using the information system, and the overall rate of HH was 63%. The full compliance to HH would amount to 13% of a nurse’s workload. Three processes were identified as the most relevant HH compliance drivers: the reorganization of essential supplies, the optimal provision of medicines and monitoring equipment automatic data collection. The participant nurses defined as objectives to reduce the supplies’ delivering time and establish a stock management system. To achieve it the reorganization of the storage with the ABC method, with the nurses deciding the layout of the storage space, was essential. The use of LEAN methods permitted to detect and address improvement opportunities with significant benefits. Moreover, nurses’ participation in the process enriched the experience, helping to design a customized intervention and heightened their sense of teamwork.publishersversionpublishe

Ultramorphology of digestive tract of Anticarsia gemmatalis (Hübner, 1818) (Lepidoptera: Noctuidae) at final larval development.

RESUMO: O trato digestivo dos insetos constitui uma importante barreira físico-química natural contra invasão de patógenos. Algumas larvas de lepidópteros são consideradas pragas agrícolas potenciais e sua biologia tem recebido muita atenção; no entanto, pouco se sabe sobre a morfologia do sistema digestivo. A análise morfológica do trato digestivo de Anticarsia gemmatalis em nível ultraestrutural é um método bastante eficaz para o estudo dos seus mecanismos de defesa. Os materiais foram fixados (solução de glutaraldeído 2,5%; 0.1M tampão fosfato, pH 7.3), pós-fixados (tetróxido de ósmio 1% no mesmo tampão), desidratados em ponto crítico, recobertos com ouro e analisados ao microscópio eletrônico de varredura 515-Philips. O trato digestivo de A. gemmatalis consiste de um tubo retilíneo de diâmetro e comprimento variável, subdividido em três regiões: intestino anterior formado pela cavidade bucal, faringe, esôfago e papo; o intestino médio que é a região mais longa do trato digestivo, sem aparente diferenciação morfológica ao longo do comprimento; e o intestino posterior que é diferenciado em piloro, íleo, cólon, e reto. Embora a morfologia geral do trato digestivo de A. gemmatalis seja bastante semelhante ao de outras espécies de Lepidoptera, o arranjo anatômico das camadas musculares do papo difere do descrito para larvas destes insetos. ABSTRACT: The digestive tract of insects is an important natural, physical, and chemical defense barrier against pathogen invasion. Certain lepidopteran caterpillars are serious pests of agricultural crops and their biology has received much attention, but little is known about the larval noctuid gut. The morphological analysis of the digestive tract in Anticarsia gemmatalis under scanning electron microscopy (SEM) is a good model for studies about its defense mechanism. The material was fixed (2,5% glutaraldehyde solution; 0.1M-phosphate buffer, pH 7.3), post-fixed (1% osmium tetroxide in the same buffer), dried at critical point, gold coated and analyzed in a SEM 515-Philips. A. gemmatalis digestive tract consists of a straight duct of varying length and diameter, subdivided in three main regions: the foregut formed by the oral cavity, pharynx, esophagus, and crop; the midgut that is the largest portion of the digestive tract

Numerical-experimental plastic-damage characterisation of additively manufactured 18ni300 maraging steel by means of multiaxial double-notched specimens

PTDC/EME/31307/2017Additive manufacturing (AM) has become a viable option for producing structural parts with a high degree of geometrical complexity. Despite such trend, accurate material properties, under diversified testing conditions, are scarce or practically non-existent for the most recent additively manufactured (AMed) materials. Such data gap may compromise component performance design, through numerical simulation, especially enhanced by topological optimisation of AMed components. This study aimed at a comprehensive characterisation of laser powder bed fusion as-built 18Ni300 maraging steel and its systematic comparison to the conventional counterpart. Multiaxial double-notched specimens demonstrated a successful depiction of both plastic and damage behaviour under different stress states. Tensile specimens with distinct notch configurations were also used for high stress triaxiality range characterisation. This study demonstrates that the multiaxial double-notched specimens constitute a viable option towards the inverse plastic behaviour calibration of high-strength additively manufactured steels in distinct state of stress conditions. AMed maraging steel exhibited higher strength and lower ductility than the conventional material.publishersversionpublishe

Regulation of Stanniocalcin Secretion by Calcium and PTHrP in Gilthead Seabream (Sparus aurata)

Calcium balance is of paramount importance for vertebrates. In fish, the endocrine modulators of calcium homeostasis include the stanniocalcin (STC), and some members of the parathyroid hormone (PTH) family, such as the PTH-related protein (PTHrP), acting as antagonists. STC is ubiquitously expressed in higher vertebrates. In turn, bony fish exhibit specific STC-producing glands named the corpuscles of Stannius (CS). Previous studies pointed to a calcium-sensing receptor (CaSR) involvement in the secretion of STC, but little is known of the involvement of other putative regulators. The CS provides a unique model to deepen the study of STC secretion. We developed an ex vivo assay to culture CS of fish and a competitive ELISA method to measure STC concentrations. As expected, STC released from the CS responds to CaSR stimulation by calcium, calcimimetics, and calcilytic drugs. Moreover, we uncover the presence (by PCR) of two PTHrP receptors in the CS, e.g., PTH1R and PTH3R. Thus, ex vivo incubations revealed a dose-response inhibition of STC secretion in response to PTHrP at basal Ca2+ concentrations. This inhibition is achieved through specific and reversible second messenger pathways (transmembrane adenylyl cyclases and phospholipase C), as the use of specific inhibitors highlights. Together, these results provide evidence for endocrine modulation between two antagonist hormones, STC and PTHrP