Investigations into the mechanism of orotidine 5'-monophosphate decarboxylase: Sources of substrate destabilization and transition state stabilization

Orotidine 5′-monophosphate decarboxylase (OMPDC) achieves a rarely paralleled rate acceleration, yet the catalytic basis prompting this enhancement have yet to be fully elucidated. To accomplish decarboxylation, OMPDC must overcome the high energy barrier due to the localized anionic charge of the intermediate. Mechanistic studies employing enzyme mutagenesis and product or intermediate analogues were used to investigate possible transition state stabilization by a carbene resonance structure. Viability of the carbene structure depends upon a key hydrogen bond between O4 of the substrate and the amide backbone of a conserved serine or threonine. Substitution of the conserved residue with Pro resulted in a kcat/KM of 1 M-1s-1; deletion of the FUMP O4 resulted in a product analogue that does not undergo H6 exchange or inhibit decarboxylation. Hence, indirect evidence reveals the O4-backbone interaction plays an important role for binding and catalysis. OMPDC likely has honed multiple mechanisms to attain its remarkable catalysis. The successful crystallizations of OMPDC a decade ago sparked hypotheses that structure and sequence conserved residues induced productive strain on the substrate-enzyme complex. Here, we demonstrate a new source of stress: a hydrophobic pocket adjacent to the OMP carboxylate that exhibits kinetic parameters characteristic of substrate destabilization. Substitution of these residues with hydrophilic side-chains, by providing hydrogen-bonding partners, decreased kcat by 10 to 10^4–fold. The same substitutions display very little change in the rate of product H6 exchange, supporting that this hydrophobic pocket affects the substrate-enzyme complex before the transition state. We also provide evidence that hydrophilic residues can insert water molecules into the pocket with detrimental effects to catalysis.

Knowledge management capabilities and organizational performance: an investigation into the effects of knowledge infrastructure and processes on organizational performance

Knowledge is one of the most important assets for surviving in the modern business environment. The effective management of that asset mandates continuous adaptation by organizations, and requires employees to strive to improve the company's work processes. Organizations attempt to coordinate their unique knowledge with traditional means as well as in new and distinct ways, and to transform them into innovative resources better than those of their competitors. As a result, how to manage the knowledge asset has become a critical issue for modern organizations, and knowledge management is considered the most feasible solution. Knowledge management is a multidimensional process that identifies, acquires, develops, distributes, utilizes, and stores knowledge. However, many related studies focus only on fragmented or limited knowledge-management perspectives. In order to make knowledge management more effective, it is important to identify the qualitative and quantitative issues that are the foundation of the challenge of effective knowledge management in organizations. The main purpose of this study was to integrate the fragmented knowledge management perspectives into the holistic framework, which includes knowledge infrastructure capability (technology, structure, and culture) and knowledge process capability (acquisition, conversion, application, and protection), based on Gold's (2001) study. Additionally, because the effect of incentives ̶̶ which is widely acknowledged as a prime motivator in facilitating the knowledge management process ̶̶ was missing in the original framework, this study included the importance of incentives in the knowledge management framework. This study also identified the relationship of organizational performance from the standpoint of the Balanced Scorecard, which includes the customer-related, internal business process, learning & growth, and perceptual financial aspects of organizational performance in the Korean business context. Moreover, this study identified the relationship with the objective financial performance by calculating the Tobin's q ratio. Lastly, this study compared the group differences between larger and smaller organizations, and manufacturing and nonmanufacturing firms in the study of knowledge management. Since this study was conducted in Korea, the original instrument was translated into Korean through the back translation technique. A confirmatory factor analysis (CFA) was used to examine the validity and reliability of the instrument. To identify the relationship between knowledge management capabilities and organizational performance, structural equation modeling (SEM) and multiple regression analysis were conducted. A Student's t test was conducted to examine the mean differences. The results of this study indicated that there is a positive relationship between effective knowledge management and organizational performance. However, no empirical evidence was found to suggest that knowledge management capabilities are linked to the objective financial performance, which remains a topic for future review. Additionally, findings showed that knowledge management is affected by organization's size, but not by type of organization. The results of this study are valuable in establishing a valid and reliable survey instrument, as well as in providing strong evidence that knowledge management capabilities are essential to improving organizational performance currently and making important recommendations for future research.

Formal and philological inquiries into the nature of interrogatives, indefinites, disjunction, and focus in Sinhala and other languages

In this thesis I examine a variety of linguistic elements which involve ``alternative'' semantic values---a class arguably including focus, interrogatives, indefinites, and disjunctions---and the connections between these elements. This study focusses on the analysis of such elements in Sinhala, with comparison to Malayalam, Tlingit, and Japanese. The central part of the study concerns the proper syntactic and semantic analysis of Q[uestion]-particles (including Sinhala "da", Malayalam "-oo", Japanese "ka"), which, in many languages, appear not only in interrogatives, but also in the formation of indefinites, disjunctions, and relative clauses. This set of contexts is syntactically-heterogeneous, and so syntax does not offer an explanation for the appearance of Q-particles in this particular set of environments. I propose that these contexts can be united in terms of semantics, as all involving some element which denotes a set of ``alternatives''. Both wh-words and disjunctions can be analysed as creating Hamblin-type sets of ``alternatives''. Q-particles can be treated as uniformly denoting variables over choice functions which apply to the aforementioned Hamblin-type sets, thus ``restoring'' the derivation to normal Montagovian semantics. The treatment of Q-particles as uniformly denoting variables over choice functions provides an explanation for why these particles appear in just this set of contexts: they all include an element with Hamblin-type semantics. However, we also find variation in the use of Q-particles; including, in some languages, the appearance of multiple morphologically-distinct Q-particles in different syntactic contexts. Such variation can be handled largely by positing that Q-particles may vary in their formal syntactic feature specifications, determining which syntactic contexts they are licensed in. The unified analysis of Q-particles as denoting variables over choice functions also raises various questions about the proper analysis of interrogatives, indefinites, and disjunctions, including issues concerning the nature of the semantics of wh-words and the syntactic structure of disjunction. As well, I observe that indefinites involving Q-particles have a crosslinguistic tendency to be epistemic indefinites, i.e. indefinites which explicitly signal ignorance of details regarding who or what satisfies the existential claim. I provide an account of such indefinites which draws on the analysis of Q-particles as variables over choice functions. These pragmatic ``signals of ignorance'' (which I argue to be presuppositions) also have a further role to play in determining the distribution of Q-particles in disjunctions. The final section of this study investigates the historical development of focus constructions and Q-particles in Sinhala. This diachronic study allows us not only to observe the origin and development of such elements, but also serves to delimit the range of possible synchronic analyses, thus providing us with further insights into the formal syntactic and semantic properties of Q-particles. This study highlights both the importance of considering various components of the grammar (e.g. syntax, semantics, pragmatics, morphology) and the use of philology in developing plausible formal analyses of complex linguistic phenomena such as the crosslinguistic distribution of Q-particles.

Exporting the racial republic: African colonization, national citizenship, and the transformation of U.S. expansion, 1776-1864

The effort to create a colony of African Americans on the west coast of Africa was one of the most celebrated and influential movements in the United States during the first half of the 19th century. While historians have often viewed African colonization through the lens of domestic anti-slavery politics, colonization grew from an imperial impulse which promised to transform the identities of black colonists and indigenous Africans by helping them to build a democratic nation from the foundation of a settler colony. By proposing that persons of African descent could eventually become self-governing subjects, the liberal framework behind colonization offered the possibility of black citizenship rights, but only within racially homogenous nation-states, which some proponents of colonization imagined might lead to a “United States of Africa.” This dissertation examines how the notion of expanding democratic ideals through the export of racial nationhood was crucial to the appeal of colonization. It reveals how colonization surfaced in several crucial debates about race, citizenship, and empire in the antebellum United States by examining discussions about African Americans’ revolutionary claims to political rights, the bounds of US territorial expansion, the removal of native populations in North America, and the racialization of national citizenship, both at home and abroad. By examining African colonization from these perspectives, this dissertation argues that the United States’ efforts to construct a liberal democracy defined by white racial identity were directly connected to the nation’s emerging identity as a defender and exporter of political liberty throughout the world.