CCE Award Lecture - Cell Culture and Social Engineering

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Deepening Knowledge on CHO cells metabolism using multiple tracer substrates

Optimization of culture conditions through trial-and-error has led to much of the past progress on animal cell culture. Today, a renewed account of the underlying biologies through a “systems” view is providing a deeper knowledge of cellular physiology and laying the basis to reengineer desirable states of further increased culture performance. In this work, we took a comprehensive view of cellular metabolism by extensively profiling extracellular and intracellular metabolomes after 13C‑label supplementation in parallel labelling cultures of [1,2-13C]Glucose, [U-13C,15N]Asn, [U-13C,15N]Ser and [1-13C]Pyruvate. This integrative approach was used to trace metabolic rearrangements in different scenarios of asparagine and serine availability in GS-CHO cells. The absence of asparagine in the medium caused growth arrest, and was associated with a dramatic increase in pyruvate uptake, a higher ratio of pyruvate carboxylation to dehydrogenation and an inability for de novo asparagine synthesis. The release of ammonia and amino acids such as aspartate, glutamate and alanine were deeply impacted. This confirms asparagine to be essential for GS‑CHO cells as the main source of intracellular nitrogen as well as having an important anaplerotic role in TCA cycle activity. In turn, serine limitation also negatively affected culture growth while triggering its de novo synthesis, confirmed by label incorporation coming from pyruvate, and reduced glycine and formate secretion congruent with its role as a precursor in the metabolism of one‑carbon units. The results obtained suggest that feeding schemes of asparagine or serine should be tightly tuned to minimize by-product formation while assuring biosynthetic needs. Ongoing is the contextualization of the data from the parallel labelling cultures into a metabolic network model for detailed mapping of cellular fluxomes in each scenario. Overall, this work contributes to unfold important insights into GS-CHO cells metabolism, and can be used as a basis for exploring bioprocess optimization strategies

O contributo da utilização do QR Code no 1.º Ciclo do Ensino Básico

As Tecnologias da Informação e da Comunicação (TIC) têm-se tornado um elemento constituinte da aprendizagem. Neste sentido, os professores devem proporcionar, a todos os alunos, as mesmas oportunidades e condições a nível de literacia digital, nomeadamente através de novas e diferentes abordagens dos conteúdos. Para o efeito, foi realizada uma investigação que pretendeu estudar as potencialidades do «QR Code» no âmbito da Prática de Ensino Supervisionada tendo-se envolvido uma turma de alunos do 4º ano de escolaridade de uma Escola Básica de Castelo Branco. Os principais objetivos desta investigação assentaram na promoção da utilização das TIC, na reflexão das suas potencialidades do «QR Code», em contexto educativo e da sua integração nas planificações da Prática de Ensino Supervisionada. Foi realizada uma investigação de caráter qualitativo com um enfoque numa investigação-ação onde foram utilizadas técnicas de recolha de dados (observação participante, notas de campo, registos fotográficos, inquérito por questionário e a entrevista semiestruturada). Após a recolha, tratamento e análise dos dados aferiu-se que o uso da aplicação digital «QR Code» como recurso pedagógico, foi um elemento decisivo para a melhoria do desenvolvimento do vocabulário dos alunos, da produção textual, mais concretamente na redação das sinopses, na compreensão da leitura, na cooperação com os outros e no interesse sentido dos alunos perante o processo de ensino e aprendizagemInformation and Communication Technologies (ICT) have become a constituent element of learning. In this sense, teachers should provide all students with the same opportunities and conditions in terms of digital literacy, in particular through new and different approaches to content. For that purpose, an investigation was carried out that aimed to study the potential of the QR Code in the scope of Supervised Teaching Practice, involving a group of students from the 4th year of schooling at a Bran-co Castle Basic School. The main objectives of this research were to promote the use of ICT, to reflect on the potential of the QR Code in an educational context and to integrate it into the plans of the Supervised Teaching Practice. A qualitative research was carried out with a focus on an action research where data collection techniques were used (participant observation, field notes, photographic records, questionnaire survey and semi-structured interview). After the data collection, treatment and analysis, it was verified that the use of the digital application "QR Code" as a pedagogical resource was a decisive element for the improvement of the students' vocabulary development, of textual production, more concretely in the writing of Challenges 2019: Desafios da Inteligência Artificial, Artificial Intelligence Challenges 396 the synopses , in reading comprehension, in cooperation with others and in the students' sense of interest in the teaching and learning processinfo:eu-repo/semantics/publishedVersio

Part 1, X-ray structural characterisation of some complexes of second- and third-row transition metals ; Part 2, Crystallographic studies into the nature of two industrial bleaching agents

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Who wants a price discount? The differential effects of price discounts on luxury brands and fast­-fashion brands

The fashion industry is a significant contributor to the global economy, and marketing strategies, such as price promotions, are commonly used to attract customers and promote sales. However, the impact of these promotions on consumers' perceptions of product quality and value can vary depending on the type of product. This study aims to investigate the effect of price promotions on consumers' perceptions of quality and value for apparel products, as well as the impact on consumer’s self­esteem and social­-display­-self­-esteem, with a focus on the differences between luxury fashion and fast-fashion brands. The research employed a between­-subjects experimental design and collected and analyzed survey data to compare how price discounts impact fast­-fashion and luxury fashion brands in terms of their effects on consumers' perceptions and behavior. The findings revealed that price promotions have negative effects on the perceived value of luxury goods, but the inverse effect was observed on fast-­fashion brands. Regarding perceived quality, luxury-­fashion brands, which rely on their image and reputation to convey a sense of high quality, are more sensitive to the negative effects of price promotions, while fast­-fashion brands are less affected. Additionally, consumers in the luxury-­promotion category had higher self­-esteem than those in the fast­-fashion promotion category, and consumers had higher levels of social­-display-­self­esteem, when consuming luxury­-fashion brands. This dissertation provides theoretical and managerial implications for fashion retailers and marketing management in understanding the potential positive and negative effects of price promotions on apparel consumption for different brand categories.A indústria da moda contribui significativamente para a economia global, e neste setor, faz-se um uso constante de promoções para atrair clientes e aumentar as vendas. No entanto, o impacto destas promoções na perceção de qualidade e valor dos consumidores pode variar dependendo do tipo de produto. Este estudo visa investigar o efeito das promoções na perceção de qualidade e valor que os consumidores atribuem ao vestuário, bem como o impacto na sua autoestima, e na autoestima que resulta da exibição pública dos produtos, com foco nas diferenças entre marcas de luxo e fast-fashion. Foi utilizado um design experimental entre indivíduos, tendo sido recolhidos e analisados dados através de um questionário online, para comparar os efeitos de promoções nas perceções e comportamentos dos consumidores, nas diferentes categorias de marcas. As conclusões revelaram que as promoções têm efeitos negativos no valor percebido dos bens de luxo, mas o efeito inverso foi observado nas marcas de fast-fashion. Relativamente à qualidade percebida, as marcas de luxo, que dependem da sua imagem e reputação, são mais sensíveis aos efeitos negativos das promoções, do que as marcas de fast-fashion. Adicionalmente, os consumidores da categoria de promoção de luxo apresentaram níveis de autoestima superior aos da promoção de fast-fashion, e níveis mais elevados de autoestima resultante da exibição pública dos produtos ao consumirem marcas de luxo. Esta dissertação proporciona implicações teóricas e práticas de gestão na compreensão dos potenciais efeitos positivos e negativos das promoções no consumo de vestuário para diferentes categorias de marcas

Development of a battery management system interface for adapting second life batteries to energy storage

As technology progresses so does the need to adapt and repurpose components and practices that need to be improved for the sake of efficiency, environmental protection, financial sustainability and, overall, to obtain the best final product. With the advent of electric vehicles, surges the issue of the used batteries for vehicles, namely cars such as Tesla (Models X and S), VW (GTE, E-Golf and E-Up), Nissan (Leaf and NV200) and BMW i3. After its normal daily use the battery might not be suited anymore to be used again in a different vehicle since it requires Power, however, there is still an important and ever growing use case for said battery as an energy storage unit for domestic and industrial installations paired up with renewable energy injection. In order to safely and accurately provide energy there is a need of technical and theoretical expertise not only for installing the system but to regulate it too.As technology progresses so does the need to adapt and repurpose components and practices that need to be improved for the sake of efficiency, environmental protection, financial sustainability and, overall, to obtain the best final product. With the advent of electric vehicles, surges the issue of the used batteries for vehicles, namely cars such as Tesla (Models X and S), VW (GTE, E-Golf and E-Up), Nissan (Leaf and NV200) and BMW i3. After its normal daily use the battery might not be suited anymore to be used again in a different vehicle since it requires Power, however, there is still an important and ever growing use case for said battery as an energy storage unit for domestic and industrial installations paired up with renewable energy injection. In order to safely and accurately provide energy there is a need of technical and theoretical expertise not only for installing the system but to regulate it too

Enhanced process control of an integrated and scalable bioprocess for production and isolation of MSC-derived extracellular vesicles for cardiac repair

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Process intensification for production of a Peste des Petites Ruminants Virus (PPRV) vaccine

Peste des Petites Ruminants Virus (PPRV) is a highly contagious disease affecting small ruminants in Africa and Asian countries, with negative/significant economic impact. Aiming to eradicate the disease, a novel/scalable PPRV vaccine production process is clearly needed. Built upon work previously done at iBET, a new production process is herein proposed using Vero cells growing on microcarriers, serum-free medium (SFM) and stirred-tank bioreactors (STB). This includes a new method for cells detachment from microcarriers, and perfusion culture for reducing turnaround time. The PPRV vaccine production process was developed in 2L STB (BIOSTAT DCU-3, Sartorius) using Nigeria 75/1 strain. Engineering correlations (e.g. shear stress and Eddy size) were used to optimize culture conditions. Vero cells were adapted to grow in ProVeroTM-1 SFM (Sartorius). A new enzymatic and mechanical method for in situ cell detachment from microcarriers was designed. PBS washing, TrypLE Select and trypsin inhibitor concentrations, and stirring rates were the variables explored. Perfusion culture was evaluated in 2L STB (equipped with spin-filter) in order to reduce seed-train preparation time. PPRV were clarified using depth filtration (Sartopure PP2, Sartorius). Vero cells were adapted to ProVeroTM-1 SFM, reaching growth rates of 0.03 h-1 (similar to serum-containing cultures). The new in situ cell detachment method was successfully implemented, with yields above 80%; no impact on cell re-attachment or virus productivity was observed. A two-fold increase in maximum cell concentration was obtained using perfusion when compared to batch culture. Combining the new in situ cell detachment method with perfusion culture will enable the scale-up to 20L STB directly from a 2L STB, surpassing the need for a mid-scale platform and thus reducing seed-train preparation time. The potential of depth filtration for PPRV clarification (upon microcarriers sedimentation) could be confirmed, with yields up to 90%. Process scalability will be validated at the 20L scale in Sartorius BIOSTAT C-Plus (using engineering correlations such as shear stress and Kolmogorov-Eddy size as scale-up criteria) by comparing cells growth, metabolic and PPRV production kinetics to those achieved in 2L STB. In conclusion, the novel/scalable vaccine production process herein proposed has potential to assist the upcoming vaccination program for eradication of PPRV disease

Cuidados paliativos pediátricos domiciliários centrados na família: Contributos para uma Orientação salutogénica.

O presente artigo tem como objetivo abordar os cuidados paliativos pediátricos domiciliários centrados na família numa perspetiva salutogénica. A pesquisa efetuada em plataformas de bases de dados e em revistas científicas revelou um número limitado de estudos centrados no paradigma salutogénico em cuidados paliativos. Os cuidados paliativos pediátricos têm como principal objetivo a prestação de cuidados domiciliários centrados na família, indo ao encontro das necessidades das famílias, com impacte positivo na qualidade de vida da criança. A Associação Europeia de Cuidados Paliativos reconhece a importância de centrar os cuidados nas competências e nos recursos da criança e da família e não exclusivamente nas suas dificuldades, ou seja, da orientação salutogénica nos cuidados paliativos. Esta nova orientação nos cuidados de saúde tem vindo a emergir progressivamente como resultado do desenvolvimento de diversos constructos, nomeadamente do sentido interno de coerência e da resiliência. Conclui-se que esta mudança de paradigma é fundamental para a melhoria da qualidade de vida da criança e da família ao longo do curso da doença