Lolines are specialized metabolites produced by endophytic fungi, such as Neotyphodium and Epichloë species, that are in symbiotic relationships with cool-season grasses. Lolines are vital for the survival of the grasses because their insecticidal and antifeedant properties protect the plant from insect herbivory. Although lolines have various bioactivities, they do not have any concomitant antimammalian activities. Lolines have complex structures that are unique among naturally occurring pyrrolizidine alkaloids. Lolines have four contiguous stereocenters, and they contain an ether bridge connecting C(2) and C(7) of the pyrrolizidine ring. An ether bridge connecting bridgehead C atoms is unusual in natural products and leads to interesting questions about the biosynthesis of lolines in fungal endophytes. Dr. Pan, who was a graduate student in Dr. Schardl Lab at University of Kentucky, isolated a novel metabolite, 1-exo-acetamidopyrrolizidine (AcAP). She observed that AcAP was accumulating in naturally occurring and artificial lolO mutants. I synthesized an authentic sample of (±)-AcAP and compared it spectroscopically with AcAP isolated from a lolO mutant to determine the structure and stereochemistry of the natural product. I was also able to grow crystals of synthetic (±)-AcAP, X-ray analysis of which further supported our structure assignment. There were two possible explanations for the fact that a missing or nonfunctional LolO led to the accumulation of AcAP: that AcAP was the actual substrate of LolO, or that it was a shunt product derived from the real substrate of LolO, 1-exo-aminopyrrolizidine (AP), and that was produced only when LolO was not available to oxidize AP. To distinguish between the two hypotheses, I synthesized 2´,2´,2´,3-[2H4]-AcAP. Dr. Pan used this material to confirm that AcAP was an intermediate in loline alkaloid biosynthesis, not a shunt product. To determine the product of LolO acting on AcAP, Dr. Pan expressed LolO in yeast (Saccharomyces cerevisiae). When Dr. Pan fed AcAP (synthesized by me) to the modified organism, it produced NANL, suggesting that LolO catalyzed two C–H activations of AcAP and the formation of both C–O bonds of the ether bridge in NANL, a highly unusual transformation. Dr. Chang then cloned, expressed, and purified LolO and incubated it with (±)-AcAP, 2-oxoglutarate, and O2. He observed the production of NANL, further confirming the function of LolO. Dr. Chang also observed an intermediate, which we tentatively identified as 2-hydroxy-AcAP. In order to determine whether the initial hydroxylation of AcAP catalyzed by LolO occurred at C(2) or C(7), I prepared (±)-7,7-[2H2]- and (±)-2,2,8-[2H3]-AcAP. When Dr. Pan measured the rate of LolO-catalyzed hydroxylation of these substrates under conditions under which only one C–H activation would occur, she observed a very large kinetic isotope effect when C(2) was deuterated, but not when C(7) was deuterated, establishing that the initial hydroxylation of AcAP occurred at the C(2) position. In order to determine the stereochemical course of C–H bond oxidation by LolO at C(2) and C(7) of AcAP, I synthesized trans- and cis-3-[2H]-Pro and (2S,3R)-3-[2H]- and (2S,3S)-2,3-[2H2]-Asp. Feeding experiments with these substrates carried out by both Dr. Pan (Pro) and me (Asp) showed that at both the C(2) and C(7) positions of AcAP, LolO abstracted the endo H atoms during ether bridge formation. In summary, feeding experiments with deuterated (±)-AcAP derivatives and its amino acid precursors have shown that AcAP is an intermediate in loline biosynthesis. We have shown that LolO catalyzes the four-electron oxidation of AcAP at the endo C(2) position first and then the endo C(7) position to give NANL.

Relationships among the Aesthetic, culture, and music are problematic- Frequently considered as epiphenomenal to culture, music and the arts are typically seen as adjuncts to ceremonial activity- This dissertation examines the nature of the Aesthetic, music and performance in the context of the Lolo, Araigilpua Village, West New Britain Province, Papua New Guinea, in an attempt to develop a definition of the Aesthetic applicable for cross-cultural research and to discover the ways in which the Aesthetic and culture articulate. For the purposes of this dissertation, the Aesthetic is defined as that facet of religion focused on responses to extraordinary powers thought to maintain what are considered to be proper relationships between human members of a community and extraordinary powers. Three forms of aesthetics, social, performance, and musical, are taken as the means and methods of directing interactions between man and extraordinary powers. At present, the Lolo are engaged in a process of secularisation resulting primarily from the introduction of Christianity, Western medicine and money. This dissertation examines the relationship between the Aesthetic and social life, and addresses the impact of changes to the Aesthetic. / Arts, Faculty of / Anthropology, Department of / Graduate

The interest for marine research and exploration has increased rapidly during the past decades and autonomous underwater vehicles (AUV) have been found useful in an increased amount of applications. The demand for versatile platform AUVs, able to perform a wide range of tasks, has become apparent. A vital part of an AUV is its motion control system, and an emerging problem for multipurpose AUVs is that the control performance is affected when the vehicle is configured with different payloads for each mission. Instead of having to manually re-tune the control system between missions, a method for automatic tuning of the control system has been developed in this master’s thesis. A model-based approach was implemented, where the current vehicle dynamics are identified by performing a sequence of excitation maneuvers, generating informative data. The data is used to estimate model parameters in predetermined model structures, and model-based control design is then used to determine an appropriate tuning of the control system. The performance and potential of the suggested approach were evaluated in simulation examples which show that improved control can be obtained by using the developed auto-tuning method. The results are considered to be sufficiently promising to justify implementation and further testing on a real AUV. The automatic tuning process is performed prior to a mission and is meant to compensate for dynamic changes introduced between separate missions. However, the AUV dynamics might also change during a mission which requires an adaptive control system. By using the developed automatic tuning process as foundation, the first steps towards an indirect adaptive control approach have been suggested. Also, the AUV which was studied in the thesis composed another interesting control problem by being overactuated in yaw control, this because yawing could be achieved by using rudders but also by differential drive of the propellers. As an additional and separate part of the thesis, an approach for using both techniques simultaneously have been proposed.