Nanopartículas lipídicas têm sido desenvolvidas para aplicação tópica de fármacos e ativos cosméticos. Neste trabalho, foi proposta a primeira aplicação de um lipídeo natural não-refinado biodegradável e biocompatível - manteiga de cupuaçu (Theobroma grandiflorum) - para a preparação de nanopartículas lipídicas, as quais foram denominadas teosferas. As teosferas foram preparadas por emulsificação-evaporação do solvente (EES) e por homogeneização à alta pressão (HAP), apresentando tamanho nanométrico e distribuição granulométrica estreita quando preparadas por ambos os métodos. O trabalho teve continuidade com a preparação de teosferas pelo método de EES utilizando manteiga de cupuaçu ou sua mistura com óleo de castanha do Brasil (Bertholletia excelsa) - também derivado da biodiversidade Amazônica - visando a incorporação de antioxidantes. Idebenona (IDB) foi selecionada por sua conhecida ação antioxidante e pela sua utilização em formulações cosméticas antienvelhecimento. IDB foi incorporada nas teosferas com eficiência de encapsulação superior a 99%, sendo que os estudos de liberação in vitro mostraram que a liberação de IDB a partir das teosferas foi mais lenta em comparação à IDB livre. Estes experimentos foram capazes ainda de demonstrar as características elásticas das teosferas. Além disso, foi evidenciada in vitro a atividade antioxidante superior das teosferas contendo IDB em relação ao ativo livre. Visando possibilitar a aplicação tópica de teosferas contendo IDB, em um trabalho subseqüente, suspensões de teosferas preparadas por HAP foram incorporadas em géis hidrofílicos. As formulações apresentaram características pseudoplásticas e demonstraram efeito oclusivo in vitro, o qual foi dependente da composição dos colóides. Finalmente, os estudos de permeação in vitro utilizando pele humana demonstraram que teosferas e nanocápsulas de núcleo lipídico, utilizadas neste estudo de modo comparativo, modificaram a permeção da IDB, permitindo a acumulação do ativo nas camadas superficiais da pele. / Lipid nanoparticles have been developed for administration of active substances to the skin, both for pharmaceutical and cosmetic uses. In the present work, we proposed the first use of a none-refined natural biodegradable and biocompatible lipid – Cupuaçu seed butter (Theobroma grandiflorum) – for the preparation of lipid nanoparticles, which were called theospheres. Theospheres were prepared by emulsification-solvent evaporation (ESE) and by high pressure homogenization technique (HPH), presenting size in nanometrical range and narrow particle size distribution for both methods. Taking these results into account, the next step of this work was the preparation of theospheres by ESE method using Cupuaçu seed butter with or without Brazil nut (Bertholletia excelsa) seed oil - another ingredient derived from an Amazonian fruit - intending the encapsulation of an antioxidant. Idebenone (IDB) has been selected due to its known antioxidant action and because it has been used in antiaging cosmetic formulations. IDB was incorporated in the theospheres presenting encapsulation efficiency higher than 99%. The in vitro release evaluation demonstrated that the release of IDB from theospheres was lower than that of free drug. Besides, the in vitro release study highlighted the elastic characteristics of theospheres. Additionally, IDB-loaded theospheres showed higher antioxidant activity compared to free IDB. Viewing the cutaneous administration, theosphere suspensions prepared by HPH technique were incorporated into hydrogels. The rheograms of the semi-solid formulations exhibited a non-Newtonian behavior presenting pseudoplastic characteristics. In vitro occlusion study highlighted the dependence of the occlusive effect on the lipidic composition of the theospheres. Finally, in vitro human skin permeation studies showed that theospheres and lipid-core nanocapsules, used in this study in a comparative way, changed the permeation of IDB, increasing the accumulative amount of IDB in the upper skin layer.

There is concern that climate change may cause mismatches between timing of flowering and activity of pollinators (phenology). However, concluding that mismatches will occur, and have serious consequences for pollination services, requires assumptions that have not yet been tested. I begin by discussing a set of these assumptions, bringing past research into the context of mismatch. Briefly, the assumptions are that 1) dates of first-flowering or emergence (DFFE) correctly describe phenology (and therefore mismatch); 2) differences in DFFE represent the magnitude of mismatch; 3) advancement of DFFE will be the primary phenological change; 4) shifts will be random and independent for each species; 5) populations of plants and pollinators are “bottom-up” regulated by their mutualistic interactions; 6) all interactions are of similar strength and importance; 7) dispersal, and the spatial context of phenological mismatches can be ignored; and ecological processes including 8) phenotypic plasticity and adaptive evolution of phenology, 9) competition and facilitation, and 10) emergence of novel interactions, will not affect mismatches. I then describe novel experiments, which could help to account for some of these assumptions, clarifying the existence and impacts of mismatches. Next, I present an original field experiment on factors affecting seed set in an alpine meadow in the Coast Range of British Columbia, Canada. I found evidence contradicting the assumption that seed set is primarily limited by pollination. My data highlight the roles of phenology, temperature (degree-days above 15°C, and frost hours), and interactions with pollinators (mutualists) and seed-predators (floral antagonists) in driving patterns of seed set. Seed set of early and late-flowering species responded differently to a 400m elevation gradient, which might be explained by phenology of bumble bees. My data suggest that the consequences of mismatch may be smallest for plants that are fly-pollinated and self-fertile. Non-selfing, bee-pollinated species might be more prone to reproductive limitation through mismatch (affected by snowmelt and cumulative degree-days). Plants that are limited by seed-predators might be negatively affected by warming temperatures with fewer frost hours, and extreme events such as late-season frosts and hail storms can prevent plants from setting seed entirely. Overall, my work emphasizes the importance of complementing theory, data-driven simulations, and meta-analyses with experiments carried out in the field. / Graduate