Bioexploration of wild Alaskan berries: From field screening to functional food

Wild berries are fundamental components of traditional diet and medicine for Native American and Alaska Native tribes and contain a diverse array of phytochemicals, including anthocyanins and proanthocyanidins, with known efficacy against metabolic disorders. Bioexploration represents a new paradigm under which bioactive preparations are screened in coordination with indigenous communities, to prepare for subsequent in-depth chemical and biological analysis. The inclusive, participatory philosophical approach utilized in bioexploration has additional benefits that could be realized in seemingly disparate areas, such as education and economics. Five species of wild Alaskan berries (Vaccinium uliginosum, V. ovalifolium, Empetrum nigrum, Rubus chamaemorus, and R. spectabilis) were tested using “Screens-to-Nature” (STN), a community-participatory approach to screen for potential bioactivity, in partnership with tribal members from three geographically distinct Alaskan villages: Akutan, Seldovia, and Point Hope. Berries were subsequently evaluated via HPLC and LC-MS2, yielding significant species and location-based variation in anthocyanins (0.9-438.6 mg eq /100g fw) and proanthocyanins (73.7-625.2 mg eq /100g fw). A-type proanthocyanidin dimers through tetramers were identified in all species tested. Berries were analyzed for in vitro and in vivo activity related to diabetes and obesity. R. spectabilis samples increased preadipocyte-factor-1 levels by 82% over control, and proanthocyanidin-rich fractions from multiple species reduced lipid accumulation in 3T3-L1 adipocytes. Furthermore, extracts of V. uliginosum and E. nigrum (Point Hope) reduced serum glucose levels in C57bl/6j mice up to 45%. The same precepts of bioexploration, especially the inclusion of indigenous community perspectives and knowledge, have relevance in other areas of study, such as education and economics. Studies have established the apathetic, low-motivational environment characteristic of many introductory science laboratory classes is detrimental to student interest, learning, and continuation in scientific education. A primary means of arresting this decline and stimulating the students’ attention and excitement is via engagement in hands-on experimentation and research. Using field workshops, the STN system is investigated as to its potential as a novel participatory educational tool, using assays centered around bioexploration and bioactive plant compounds that hold the potential to offset human health conditions. This evaluation of the STN system provided ample evidence as to its ability to augment and improve science education. Furthermore, Strengths, Weaknesses, Opportunities, and Threats (SWOT) analysis was employed as a theoretical framework to review the potential benefits and hurdles associated with developing a wild Alaskan berry commodity. Synthesizing various sources of information – including logistics and harvest costs, sources of initial capital, opportunities in the current superfruit industry, and socioeconomic factors – the development of a berry commodity proves to be a complex amalgam of competing factors which would require a delicate balance before proceeding.

Design and fabrication of electro-thermal microcantilevers for ultrafast molecular sorting and delivery

Improvements to the current state of the art in microfabricated cantilevers are investigated in order to realize enhanced functionality and increased versatility for use in ultrafast electrophoretic molecular sorting and delivery. Design rationale and fabrication process flow are described for six types of electro-thermal microcantilevers. Devices have been tailored for the process of separating mixtures of heterogeneous molecules into discrete detectable bands based on electrophoretic mobility, and delivering them to a conductive substrate using electric fields. Four device types include integrated heating elements capable of warming samples to catalyze reactions or cleaning the device for reuse. Similar devices have been shown to be capable of targeting temperatures between ambient conditions and the melting point of silicon, to within 0.1˚C precision or better. All microcantilevers types are equipped with a highly doped conductive silicon tip capable of interacting with a conductive substrate to deliver molecules under the presence of an electric field. Devices are equipped with additional electrodes to aid in sorting molecules on the surface of the probe end. Two designs contain two legs and one additional sorting electrode while four designs contain three legs and have two sorting electrodes. Devices having two sorting electrodes are designed to be capable of sorting three or more molecular species, a distinctive advancement in the state of the art. A detailed process flow of the fabrication process for all six electro-thermal cantilever designs are explained in detail.

Atomic resolution studies of oxide superlattices and ultrathin films

The ability to grow ultrathin films layer-by-layer with well-defined epitaxial relationships has allowed research groups worldwide to grow a range of artificial films and superlattices, first for semiconductors, and now with oxides. In the oxides thin film research community, there have been concerted efforts recently to develop a number of epitaxial oxide systems grown on single crystal oxide substrates that display a wide variety of novel interfacial functionality, such as enhanced ferromagnetic ordering, increased charge carrier density, increased optical absorption, etc, at interfaces. The magnitude of these novel properties is dependent upon the structure of thin films, especially interface sharpness, intermixing, defects, and strain, layering sequence in the case of superlattices and the density of interfaces relative to the film thicknesses. To understand the relationship between the interfacial thin film oxide atomic structure and its properties, atomic scale characterization is required. Transmission electron microscopy (TEM) offers the ability to study interfaces of films at high resolution. Scanning transmission electron microscopy (STEM) allows for real space imaging of materials with directly interpretable atomic number contrast. Electron energy loss spectroscopy (EELS), together with STEM, can probe the local chemical composition as well as local electronic states of transition metals and oxygen. Both techniques have been significantly improved by aberration correctors, which reduce the probe size to 1 Å, or less. Aberration correctors have thus made it possible to resolve individual atomic columns, and possibly probe the electronic structure at atomic scales. Separately, using electron probe forming lenses, structural information such as the crystal structure, strain, lattice mismatches, and superlattice ordering can be measured by nanoarea electron diffraction (NED). The combination of STEM, EELS, and NED techniques allows us to gain a fundamental understanding of the properties of oxide superlattices and ultrathin films and their relationship with the corresponding atomic and electronic structure. In this dissertation, I use the aforementioned electron microscopy techniques to investigate several oxide superlattice and ultrathin film systems. The major findings are summarized below. These results were obtained with stringent specimen preparation methods that I developed for high resolution studies, which are described in Chapter 2. The essential materials background and description of electron microscopy techniques are given in Chapter 1 and 2. In a LaMnO3-SrMnO3 superlattice, we demonstrate the interface of LaMnO3-SrMnO3 is sharper than the SrMnO3-LaMnO3 interface. Extra spectral weights in EELS are confined to the sharp interface, whereas at the rougher interface, the extra states are either not present or are not confined to the interface. Both the structural and electronic asymmetries correspond to asymmetric magnetic ordering at low temperature. In a short period LaMnO3-SrTiO3 superlattice for optical applications, we discovered a modified band structure in SrTiO3 ultrathin films relative to thick films and a SrTiO3 substrate, due to charge leakage from LaMnO3 in SrTiO3. This was measured by chemical shifts of the Ti L and O K edges using atomic scale EELS. The interfacial sharpness of LaAlO3 films grown on SrTiO3 was investigated by the STEM/EELS technique together with electron diffraction. This interface, when prepared under specific conditions, is conductive with high carrier mobility. Several suggestions for the conductive interface have been proposed, including a polar catastrophe model, where a large built-in electric field in LaAlO3 films results in electron charge transfer into the SrTiO3 substrate. Other suggested possibilities include oxygen vacancies at the interface and/or oxygen vacancies in the substrate. The abruptness of the interface as well as extent of intermixing has not been thoroughly investigated at high resolution, even though this can strongly influence the electrical transport properties. We found clear evidence for cation intermixing through the LaAlO3-SrTiO3 interface with high spatial resolution EELS and STEM, which contributes to the conduction at the interface. We also found structural defects, such as misfit dislocations, which leads to increased intermixing over coherent interfaces.