Grapevine response to cold - study of starch metabolism in woody tissues

Dissertação de mestrado Biologia Molecular, Biotecnologia e Bioempreendedorismo em PlantasGrapevine (Vitis vinifera L.) is one of the most important agronomical species in the world, but in the context of the ongoing climate changes, extreme temperatures, high light intensities and water deficit are becoming important threats in many important viticultural areas worldwide. Among these, stress caused by cold has been shown to influence its productivity, metabolism and gene expression, which prompted us to study some molecular mechanisms involved in grapevine response to low temperature. In this regard, cv. Vinhão canes were harvested and incubated at 4 ºC for 7, 14 and 30 days to unravel physiological changes, identify and characterize key genes involved in response to cold. Results showed that starch levels steadily decreased during the incubation period at 4 ºC, and that the expression of the amylolytic gene VvAMY2 increased along time, while the expression of VvBAM5 decreased. Furthermore, HPLC analysis showed an increase in sucrose, fructose and glucose during the incubation at 4 ºC. Additional expression studies of key-genes involved in sugar metabolism showed that the steady-state transcript levels of VvpGT2 and VvSWEET2b transporters as well as VvSusy and VvSPS enzymes increased along time, while the expression of the transporters VvTMT1 and VvSUC27 was downregulated. The VvpGT2 gene was selected for further studies regarding organ specific expression and function. VvpGT2 steady-state transcript levels were high in mature berries and flowers. Following heterologous expression in a mutant yeast (EBY.VW4000) lacking monosaccharide transporters, no VvpGT2 transport activity of fructose, glucose and sucrose could be detected. Raffinose and stachyose levels in the grapevine canes were quantified by HPLC because the metabolism of raffinose has been linked to cold response. Results showed a steadily increase of raffinose levels along incubation at 4 ºC and transcriptional studies also revealed a noticeable increase of the steady-state levels of the galactinol synthase genes VvGolS1 and VvGOLS3 raffinose synthase VvRafS5.A videira (Vitis vinifera L.) é uma das espécies agrícolas mais importantes ao nível global, contudo temperaturas extremas, altas intensidades de luminosas e a secura constituem ameaças importantes em muitas regiões vitivinícolas ao nível mundial, no contexto das alterações climáticas em curso. Em particular, o stresse causado pelo frio afeta a sua produtividade, metabolismo e expressão genética, o que nos motivou a estudar alguns mecanismos moleculares de resposta da planta a temperaturas reduzidas. Varas atempadas de videira da variedade Vinhão foram colhidas e incubadas a 4 ºC durante 7, 14 e 30 dias para se estudar as modificações fisiológicas e identificar e caracterizar genes-chave envolvidos na resposta ao frio. Os níveis de amido quantificados nas varas de videira diminuíram durante o tempo de incubação e, em paralelo, observou-se um aumento da expressão do gene VvAMY2 que codifica uma enzima amilolítica. Contudo, os níveis de transcritos do gene VvBAM5 diminuíram. Estudos de HPLC mostraram que ocorreu um aumento de sacarose, frutose e glucose em resposta ao frio. Estudo da expressão de genes-chave envolvidos no metabolismo dos açúcares mostraram um aumento dos níveis de transcritos dos transportadores VvpGT2 e VvSWEET2b e das enzimas VvSusy e VvSPS, no entanto os níveis de transcritos dos transportadores VvTMT1 e VvSUC27 diminuíram. Quando se estudou o perfil de expressão do gene VvpGT2 em diversos órgãos observou-se que os níveis de transcritos são mais elevados em bagos maduros e flores. Estudos de expressão heteróloga do gene VvpGT2 numa levedura mutante. Estudos de expressão heteróloga do gene VvpGT2 numa levedura mutante (EBY.VW4000) incapaz de transportar açúcares, mostraram que a proteína VvpGT2 é incapaz de transportar glucose, frutose e sacarose. No presente trabalho foram ainda avaliados por HPLC os níveis de rafinose e de estaquiose nas varas de videira uma vez que tem sido observado que o metabolismo dos oligossacarídeos da família da rafinose está envolvido na resposta das plantas ao frio.Com efeito, durante a incubação a 4 ºC observou-se um aumento de rafinose durante o período de incubação a 4 ºC bem como dos níveis de transcritos das enzimas galactinol sintetase, VvGolS1 e VvGolS3 ,e da rafinose sintetase, VvRafS5

Redesign e teste de viga seletora em sistema de transporte de volumes

The parcel and postal market is requesting higher and higher sorting capacities of Vanderlande sorters. One of the companies line of sorters is the Compaxorter which uses a beam to sort a parcel instead of a shoe. Compared to the shoe, a sort beam has some disadvantages regarding machine and human safety. Redesigning the sort beam in order to be flexible would reduce or solve the disadvantages. During the development procedure, two prototypes were created and tested along with a multi-body mathematical model to simulate the motion of one component of the prototype, the bracket. The first prototype was created after several concepts were generated. Once the concept that best fitted the key requirements was chosen, it was improved and turned into the first prototype. After testing prototype alpha, valuable information was created regarding its design and the bracket movement. Afterwards, based on the generated information, either the sort beam and bracket went through a new redesign phase. Therefore, the second prototype was produced and tested. The results were positive for the bending event but the bracket mechanism was still not ideal. Therefore, the multi-body mathematical model was created to predict manually its motion and, then, helping with its design.O mercado de encomendas e correio está a exigir cada vez mais altas capacidades das máquinas sorteadoras da Vanderlande. Uma das máquinas da empresa é o Compaxorter que usa uma viga para sortear uma encomenda em vez de um sapato. Comparando ao sapato, a viga seletora tem certas desvantagens no que toca à segurança da máquina e dos operadores. Redesenhar a viga sorteadora de maneira a torná-la flexível pode reduzir ou até eliminar as desvantagens. Durante o processo de desenvolvimento, dois protótipos foram produzidos e testados juntamento com um modelo matemático multi-corpo que simula o movimento de um componente do protótipo, o bracket. O primeiro protótipo foi produzido após várias ideas serem geradas. O conceito que mais se adequava aos requesitos foi escolhido, melhorado e depois transformado então no primeiro protótipo. Após testar o protótipo alfa, foi gerada informação importante relativamente ao seu design e ao movimento do bracket. Com base nessa informação, a viga e o bracket foram novamente redesenhados. Desta maneira, o segundo protótipo foi produzido e testado. Os resultados foram positivos no que diz respeito à flexibilidade da viga mas o mecanismo do bracket ainda não era ideal. Portanto, o modelo matemático multi-corpo foi criado para prever manualmente o seu movimento, culminando com o seu redesign.Mestrado em Engenharia Mecânic

Utilização de ultrassons para afinação da microestrutura de fundidos de alumínio obtidos por fundição em cera perdida

Dissertação de mestrado integrado em Engenharia MecânicaA realização desta dissertação possui como desígnio o estudo da influência que a aplicação do tratamento físico por ultrassons (US) exerce nos fundidos produzidos através do processo de fundição por cera perdida. Com base neste propósito, são averiguadas as diversas noções inerentes e fundamentais destas temáticas na sua execução relativamente ao processo de manufatura como também à caracterização posterior do fundido produzido. Com este objetivo foi produzido inicialmente um exemplar de um componente com o intuito de avaliar desde logo o impacto na microestrutura deste e consequentemente analisar e determinar as localizações em que a modificação seja evidente. Isto proporciona não só o conhecimento do impacto da utilização do US como também permite balizar essa mesma zona de transformação. Esta fase inicial em conjugação da avaliação deste método com diferentes fundidos em termos dimensionais facultou a conceção de um modelo de cera que alberga as características mais benéficas para as propriedades mecânicas pretendidas que os componentes produzidos obtenham. Nas diversas etapas foram sempre concebidos fundidos com tratamentos do banho metálico diferentes com o intuito de existir um termo de comparação de forma a facilitar a análise da influência que exerce a utilização dos US. A análise das várias fases realizadas possibilitou a perceção de que a potencialidade benéfica deste tratamento é verificada e reflete essa mesma melhoria num superior grau de afinação da microestrutura e também nas propriedades mecânicas com o aumento da extensão e tensão de rotura. Desta forma a realização desta dissertação contribui com a confirmação da potencialidade de aplicação prática no meio industrial desta tecnologia como para o seu contínuo desenvolvimento.The objective of this thesis was to study the effects of the application of ultrasonic vibration (US) on castings obtained by the process of investment casting. Based on this purpose several essential aspects were investigated for implementation of the manufacturing process as well the subsequent characterization of the produced cast. Initially one model of a component was produced in order to know the impact of this treatment on the microstructure and to analyze the occurred change. This provides the chance to delimit the preliminary processing zone. This initial phase in conjunction with the running of some tests in different dimensional models permitted the development of a mold with the best theoretical characteristics to obtain one component with good mechanical properties. In the various stages castings with multiple solidification situations were always produced, with and without acoustic energy to compare the results. The samples produced were analyzed based on the cooling rate, microstructure evaluation and tensile strength. This study confirmed the knowledge of the beneficial effects of using acoustic energy on the treatment of melts. This improvement is visible in mechanical properties of tensile strength and microstructure refinement. This thesis contributed to confirm the potential benefit of US application in this manufacturing process and to add motivation to further research on this subject

Different Physiognomies and the Structure of Euglossini bee (Hymenoptera: Apidae) Communities

Our aim was to evaluate the occurrence of orchid bees in remnants of the Atlantic Forest. We sampled specimens from five regions of Southeast Brazil, covering three different physiognomies of the Atlantic Forest (mixed Araucaria forests with high altitude fields, Atlantic Semi-deciduous Forest and Atlantic Rain Forest). The distances between the sampling sites ranged from 24 to 746 km. Male orchid bees attracted by fragrances were actively sampled monthly during one year using entomological nets and aromatic traps. A total of 1,482 bees were captured, including four genera and at least 31 species. We observed differences in the abundance and richness of species sampled among sites. Climatic variables, mostly relative humidity, explained twice more of the observed differences in the Euglossini communities than simple spatial variation. Our study found differences in the composition of euglossine bee communities as well as in their patterns of abundance and dominance among different vegetation formations. However, the level of pairwise similarity among the Euglossini communities sampled, although highly variable, was not related to climatic factors or geographical distances between the sampling sites. The greatest dissimilarities in the composition of the orchid bee communities were observed between sites around 400 km from each other rather than among sites that were further apart. A possible explanation is that the sampled areas that were 400 km from each other were also highly dissimilar regarding climate, especially altitude

Biology teachers’ conceptions about the origin of life in Brazil, Argentina, And Uruguay: a comparative study

Teachers’ conceptions about the origin of life in three Latin American countries with contrasting levels of secularism were analyzed: Argentina (Catholic constitution), Brazil (formally secular but not in practice), and Uruguay (consolidated secularism). A European survey questionnaire was used and the interpretation of the results drew on Barbour's four categories concerning the relationships of science and religion. A large majority of Argentinian and Uruguayan teachers were clearly evolutionist, even when believing in God (Independence or Dialogue category), with no difference between Argentina and Uruguay. The majority of Brazilian teachers assumed a religious position about the origin of life, being creationist (Conflict or Independence categories) or evolutionary creationist (Dialogue or Integration categories). Differences of Brazilian teachers’ conceptions may result from the higher percentage of evangelicals and lower proportion of agnostics/atheists. Brazilian Catholic teachers were more creationist than their Catholic colleagues in Argentina and Uruguay. Distinct patterns were found, but further research is needed to understand possible classroom impacts.CIEC – Research Centre on Child Studies, IE, UMinho (FCT R&D unit 317), Portugal; National Funds through the FCT (Foundation for Science and Technology) and co-financed by European Regional Development Funds (FEDER) through the Competitiveness and Internationalization Operational Program (POCI) through CIEC (Research Centre on Child Studies, of the University of Minho) with the reference POCI-01-0145-FEDER-007562info:eu-repo/semantics/publishedVersio

Neural gliding versus neural tensioning: effects on heat and cold thresholds, pain thresholds and hand grip strength in asymptomatic individuals

Introduction: Neural mobilization can be performed in a way that facilitates movement through a stretching technique (tensioning) or in a way that maximizes the gliding of peripheral nerves in relation to adjacent structures (gliding). Evidence on how these techniques compare in terms of effects are scarce. The aim of this study is to compare the effects of neural gliding and neural tensioning targeting the median nerve on heat and cold temperature threshold, heat pain threshold, pressure pain thresholds and hand grip strength in asymptomatic participants. Methods: Participants received 4 series of 10 repetitions of either neural gliding (n = 30) or neural tensioning (n = 30) and were assessed for heat and cold temperature threshold, heat pain threshold, pressure pain threshold, and hand grip strength at baseline, immediately after the intervention, and 30 min post-intervention. Results: A significant main interaction between time and intervention was found for the PPT at the forearm (F(2,55) = 5.98; p = 0.004), favouring the tensioning neural mobilization. No significant differences were found for the other variables. Conclusions: Four series of 10 repetitions of neural tensioning targeting the median nerve in asymptomatic subjects seem to be enough to induce hypoalgesia and have no negative effects on A-delta and C mediated sensory function and on hand grip strength production.publishe

Using discrete simulation to support internal logistics process design

The objective of this paper is to present the developments of an ongoing project that aims at implementing an internal materials movement system using autonomous vehicles for supporting logistics processes. In particular the project focus on the movement of final products, from assembly lines to the expedition warehouse and the supply of packaging materials (customer packages), from the raw materials warehouse to the assembly lines. This process is currently carried out by two stackers, in a milk-run environment and, in the context of this project, an innovative solution is proposed to improve the performance of the supply and pickup processes, transforming and optimizing an entirely manual process into an automated one. The main challenge was to investigate the impact on the existing internal logistic system of the new solution and, simultaneously, to explore if new management strategies are needed to satisfy both throughput levels and overall supply chain needs. A simulation model was developed using ARENA Simulation Software to get insights concerning the new challenges posed by the solution to be adopted. Results have revealed that the use of simulation allowed the assessment of the impact of the new internal logistics solution and played a critical role to identify the best dispatching rules to schedule transportation orders in such a complex system. This research shows the flexibility of the simulation tool to address several complex management issues simultaneously that arises in the context of innovative solutions aligned with industry 4.0 challenges.This work has been financially supported by the Portugal Incentive System for Research and Technological Development in scope of the projects in co-promotion nº 002814/2015 (iFACTORY 20152018).info:eu-repo/semantics/publishedVersio

Public preferences for involvement in the governance of health data

Public involvement in the governance of epidemiological and public health studies can foster needs-driven research, enhance participants' recruitment, reduce attrition and improve the quality of and ethics in research and surveillance. However, it can also reinforce health inequalities if it fails to ensure public representation across socioeconomic gradients. This study aimed to assess patients' and carers' preferences for involvement in collective health data governance, and its associated factors, to strengthen the evidence base for policy development