38 research outputs found

    Horizontal transfer of RNAi between honeybees and Varroa destructor

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    Dissertação de mestrado em Biotecnologia Farmacêutica, apresentada à Faculdade de Farmácia da Universidade de CoimbraEste trabalho abrange uma revisão do estado de arte do processo de liofilização e da sua aplicação na área da Nanotecnologia. O foco principal desta revisão é a caracterização dos produtos liofilizados inovadores existentes no mercado e em fase de desenvolvimento clínico, com aplicação em Oncologia. Na área farmacêutica, a exploração das propriedades únicas das nanopartículas tornou-se diferenciadora pela demonstração de efeitos na biodisponibilidade de moléculas ativas e também na proteção contra a degradação enzimática. Mais recentemente, as nanopartículas têm sido usadas para proporcionar um aumento da especificidade e efetividade no direcionamento dos fármacos aos locais alvo, permitindo a diminuição da dose eficaz e da toxicidade geral. Com tudo isto, o desenvolvimento de nanoestruturas para transporte de fármacos tem sido objeto de um interesse crescente, tendo em vista colmatar alguns pontos negativos existentes nos sistemas de dosagem tradicionais. No entanto, a estabilidade das nanopartículas pode constituir um problema, pelo que a sua concepção não é propriamente linear. No caso específico das nanodispersões coloidais, fatores físicos e químicos contribuem para uma baixa estabilidade a longo termo, provocando a destabilização do sistema que limita a sua aplicação clínica. A liofilização surge neste cenário como um processo tecnológico que pode minimizar os problemas de estabilidade das suspensões aquosas de nanopartículas. Trata-se de uma técnica de desidratação muito eficiente quando comparada com as demais, que tem vindo a apresentar melhorias na estabilidade destas suspensões coloidais, e por esta razão merece destaque. Cada produto possui um ciclo de liofilização único em função das suas particularidades, o que faz com que este processo seja uma operação unitária. Para ser percetível a forma como o processo de liofilização é executado cumprindo as boas práticas laboratoriais (BPL/GLP) e de fabrico (BPF/GMP) são explícitos, neste estudo, os requisitos normativos a ter em conta. A liofilização, além de ser útil ao nível da estabilidade, é ainda utilizada para outras finalidades que estão descritas no estudo. De forma a elucidar toda a operação envolvida neste processo é feita uma breve abordagem das etapas envolvidas, dos métodos utilizados, das ix aplicações farmacêuticas e dos principais pros e contras, com o intuito de entender a importância da formulação de nanopartículas liofilizadas. Será ainda dedicado um capítulo ao estudo dos componentes e das condições do processo para conseguir obter resistência às tensões a que as suspensões de nanopartículas são submetidas durante a fase de congelamento na liofilização. Nesta fase, o efeito crioprotetor ou lioprotetor é determinante, dado que na reidratação do produto é mandatório que as propriedades originais das nanopartículas sejam mantidas. A respeito dos avanços recentes com aplicação deste processo, são ainda apresentadas as formulações liofilizadas que estão inseridas no mercado e os ensaios clínicos e pré-clínicos existentes para o tratamento de doenças do foro oncológico, com referência aos produtos protegidos por patente. A relevância deste trabalho traduz-se no facto de o resultado da pesquisa ser orientado em ambiente empresarial e académico, com vista à inserção no processo de valorização da propriedade intelectual de uma spin-off da Universidade de CoimbraThis work aims at conducting a review of the state of art of the lyophilization process and its application in the field of Nanotechnology. The main focus is the characterization of existing innovative freeze-dried products on the market and in clinical evaluation, with applications in Oncology. In the pharmaceutical area, the unique properties of nanoparticles began to demonstrate positive effects on the bioavailability of active molecules and also in protecting against enzymatic degradation. More recently, nanoparticles have been used to provide increased specificity and effectiveness in the targeting of drugs to specific locations, thereby reducing the effective dose and toxicity. With all this, the development of nanostructures for drug delivery has provided the solution for existing weaknesses in traditional dosing systems. However, the stability of the nanoparticles is not exactly a linear topic. In the case of colloidal nanodispersions, physical and chemical factors contribute to a low long term stability, leading to destabilization of the system and thus creating an obstacle to its clinical application. Lyophilization in this scenario appears as a technological process that can minimize stability problems of aqueous suspensions of nanoparticles. It is a very efficient dewatering technique, which has been presenting improvements in the stability of these colloidal suspensions, and therefore noteworthy. Each product has a single lyophilization cycle on the basis of their features, which makes this process a unitary operation. To be noticeable how the freeze-drying process is performed in compliance with the good laboratory practice (BPL/GLP) and manufacturing practice (BPF/GMP), the regulatory requirements to be considered are explicit in this study. This technique, besides being useful for stability issues, it is still used for other purposes which are described in the study. In order to elucidate the entire operation involved in this process, it is made a brief approach of the steps involved, the methods used, the pharmaceutical applications and the main pros and cons of the formulation of lyophilized nanoparticles. The study also comprises a chapter on the components and process conditions to achieve resistance to the stresses to which the nanoparticulate suspensions are subjected during the xi freezing stage of the lyophilization. At this stage, the cryoprotectant or lioprotetor effect is decisive for the rehydration, in order to preserve that the original properties of the nanoparticles. Regarding the recent advances in the application of this process, this study presents the most recent lyophilized formulations on the market and in clinical and preclinical trials for the treatment of oncological diseases, with reference to the proprietary products. The main conclusions of this work will be included in the valuing process of the intellectual property of a spin-off from the University of Coimbra, combining an academic and business perspective on a pharmaceutical process of industrial application

    Individual and Colony Level Foraging Decisions of Bumble Bees and Honey Bees in Relation to Balancing of Nutrient Needs

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    Foraging decisions of social animals occur in the context of social groups, and thus may be subject to considerations of not only an individual's nutritional state and nutrient input, but those of the social group in which they live. In eusocial insects, which live in colonies containing workers that forage for food that is mostly consumed by others, foraging decisions that reflect colony needs may also be considered at both the colony and individual level. If colony energy balance is perturbed, is the counteracting response occurring on the group level (a change in division of labor) or on the individual level (a change in individual foraging choices)? To address this, colony and individual level foraging behaviors were observed in two species of eusocial bees: the highly social honey bee Apis mellifera and the primitively eusocial bumble bee Bombus terrestris. After manipulations of protein (P) and carbohydrate (C) stores in colonies of both species, there were changes in multiple different behavioral responses including colony level (number of foragers, allocation to nectar and pollen foraging, nutrient mass foraged) and individual level (P and C concentration preference and loading during foraging). These results suggest both honey bee and bumble bee colonies balance nutrient needs through a combination of both colony level shifts in foraging allocation, as well as slight modulation of individual nutrient preferences. This study also uncovered colony level differences between the two bee species; honey bees balanced P intake while bumble bees balanced C intake. These patterns may reflect differences in life history traits such as perenniality and hoarding, traits that are developed in more highly social species. Overall, this study highlights the importance of considering both group and individual level behavioral responses in foraging decisions in social animals

    Border collapse and boundary maintenance: militarisation and the micro-geographies of violence in Israel–Palestine

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    This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.Drawing upon subaltern geopolitics and feminist geography, this article explores how militarisation shapes micro-geographies of violence and occupation in Israel–Palestine. While accounts of spectacular and large-scale political violence dominate popular imaginaries and academic analyses in/of the region, a shift to the micro-scale foregrounds the relationship between power, politics and space at the level of everyday life. In the context of Israel–Palestine, micro-geographies have revealed dynamic strategies for ‘getting by’ or ‘dealing with’ the occupation, as practiced by Palestinian populations in the face of spatialised violence. However, this article considers how Jewish Israelis actively shape the spatial micro-politics of power within and along the borders of the Israeli state. Based on 12 months of ethnographic research in Tel Aviv and West Jerusalem during 2010–2011, an analysis of everyday narratives illustrates how relations of violence, occupation and domination rely upon gendered dynamics of border collapse and boundary maintenance. Here, the borders between home front and battlefield break down at the same time as communal boundaries are reproduced, generating conditions of ‘total militarism’ wherein military interests and agendas are both actively and passively diffused. Through gendering the militarised micro-geographies of violence among Jewish Israelis, this article reveals how individuals construct, navigate and regulate the everyday spaces of occupation, detailing more precisely how macro political power endures.This work was supported by the SOAS, University of London; University of London Central Research Fund

    Optic flow informs distance but not profitability for honeybees

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    How do flying insects monitor foraging efficiency? Honeybees (Apis mellifera) use optic flow information as an odometer to estimate distance travelled, but here we tested whether optic flow informs estimation of foraging costs also. Bees were trained to feeders in flight tunnels such that bees experienced the greatest optic flow en route to the feeder closest to the hive. Analyses of dance communication showed that, as expected, bees indicated the close feeder as being further, but they also indicated this feeder as the more profitable, and preferentially visited this feeder when given a choice. We show that honeybee estimates of foraging cost are not reliant on optic flow information. Rather, bees can assess distance and profitability independently and signal these aspects as separate elements of their dances. The optic flow signal is sensitive to the nature of the environment travelled by the bee, and is therefore not a good index of flight energetic costs, but it provides a good indication of distance travelled for purpose of navigation and communication, as long as the dancer and recruit travel similar routes. This study suggests an adaptive dual processing system in honeybees for communicating and navigating distance flown and for evaluating its energetic costs

    The effect of avocado ( Persea americana

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    Omega-6:3 Ratio More Than Absolute Lipid Level in Diet Affects Associative Learning in Honey Bees

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    Floral pollen is a major source of honey bee nutrition that provides them with micro- and macro-nutrients, including proteins, fatty acids, vitamins, and minerals. Different pollens vary in composition, including in the essential fatty acids, alpha-linolenic acid (omega-3) and linoleic acid (omega-6). Monocultures, prevalent in modern agriculture, may expose honey bee colonies to unbalanced omega-6:3 diets. The importance of omega-3 in the diet for adequate learning and cognitive function, with a focus on suitable omega-6:3 ratio, is well documented in mammals. We have recently shown, for the first time in invertebrates, the importance of omega-3 in diets for associative learning ability in honey bees. In the current work, we examine the effect of the absolute amount of omega-3 in diet compared to the omega-6:3 ratio on honey bee associative learning. We fed newly emerged bees for 1 week on different artificial diets, which had lipid concentration of 1, 2, 4, or 8%, with omega-6:3 ratios of 0.3, 1, or 5, respectively. We then tested the bees in a proboscis-extension response olfactory conditioning assay. We found that both omega-6:3 ratio and total lipid concentration affected learning. The most detrimental diet for learning was that with a high omega-6:3 ratio of 5, regardless of the absolute amount of omega-3 in the diet. Bees fed an omega-6:3 ratio of 1, with 4% total lipid concentration achieved the best performance. Our results with honey bees are consistent with those found in mammals. Best cognitive performance is achieved by a diet that is sufficiently rich in essential fatty acids, but as long as the omega-6:3 ratio is not high
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