Thesis (Ph. D.)--University of Hong Kong, 1988.

Thesis (M. Phil.)--University of Hong Kong, 1978.

The human mitochondrial polymerase (pol γ) is a nuclearly-encoded polymerase that is solely responsible for the faithful replication and repair of the mitochondrial genome. The Y955C mutation in pol γ results in early onset progressive external ophthalmoplegia, premature ovarian failure, and Parkinson’s disease. It is believed that the position of this Y955 residue on the catalytic helix in the polymerase makes it responsible for stabilizing the incoming nucleotide. I have investigated the kinetic effect of the Y955C mutation. Mutation of the tyrosine to a cysteine resulted in a decreased maximum rate of polymerization and increased the dissociation constant for incoming nucleotide. In turn, this decreased catalytic efficiency by 30 to 100-fold. In addition, the polymerase did not incorporate all bases with the same efficiency, it was most efficient when incorporating dGTP opposite a dC, but showed less efficient catalysis when faced with an A:T or T:A base-pair. The polymerase also showed reduced discrimination against misincorporation events. However, when presented with an oxidatively-damaged base, 8-oxo-deoxyguanosine, the polymerase chose to incorporate the base in the correct conformation opposite a dC, discriminating against the mutagenic incorporation of 8-oxo-dGTP opposite a dA. The results presented in this thesis suggest that the severe clinical symptoms of patients with this mutation are at least due in part to the reduced efficiency and discrimination of this polymerase γ mutation. / text

Capillary bridge formation between adjacent surfaces in humid environments is a ubiquitous phenomenon. Capillary forces are important in nature (granular materials, insect locomotion) and in technology (disk drives, adhesion). Although well studied in the equilibrium state, the dynamics of capillary formation merit further investigation. Here, we show that microcantilever crack healing experiments are a viable experimental technique for investigating the influence of capillary nucleation on crack healing between rough surfaces. To demonstrate the effects, a custom micromachine characterization system is built that allows for full environmental control (pressure, humidity, and gas composition) while retaining full micromachine characterization techniques (long working distance interferometry, electrical probe connectivity, actuation scripting capability). The system also includes an effective in situ surface plasma cleaning mechanism. The average spontaneous crack healing velocity, ̅, between plasma-cleaned hydrophilic polycrystalline silicon surfaces of nanoscale roughness is measured. A plot of ̅v versus energy release rate, G, reveals log-linear behavior, while the slope |d[log(v)]/dG| decreases with increasing relative humidity. An interface model that accounts for the nucleation time of water bridges by an activated process is developed to gain insight into the crack healing trends. This methodology enables us to gain insight into capillary bridge dynamics, with a goal of attaining a predictive capability for this important microelectromechanical systems (MEMS) reliability failure mechanism. A variety of alcohol vapors significantly reduce or perhaps eliminate wear in sliding micro-machined contacts. However, these vapors may increase adhesion due to the capillary forces. Equilibrium adhesion energies at various partial pressures are found for n-pentanol (long chain molecule) and ethanol (short chain molecule). For low partial pressures (p/ps=0.3), adhesion energy of n-pentanol is even larger than water.

Not available / text

published_or_final_version / Chemistry / Master / Master of Philosophy

published_or_final_version / Chemistry / Master / Master of Philosophy

A high speed stirring apparatus was constructed for following the kinetics of very rapid solvent extractions. Reactions with half-lives of 15 seconds can be followed with good precision. Experimental data obtained with the device are shown to be superior to kinetic data for the same reaction obtained with batch shakers and cruder stirring devices. The apparatus was used to determine the rate law for the extraction of copper by 2-hydroxyl-5-nonylbenzophenone oxime(I) and 5-dodecyl-2-hydroxylbenzophenone oxime(II). The rate law for the extraction of copper by I catalyzed by 5,8-diethyl-7-hydroxy-6-dodecanone oxime(III) was also determined. Equilibrium data are used to characterize the stoichiometry of the extracted complexes. Compound I, which is an aromatic β-hydroxyoxime, extracts copper as the 2:1 neutral chelate. Compound III, which is an aliphatic α-hydroxyoxime, has a much more complicated extraction chemistry. Experimental evidence indicates the existence of a conventional 2:1 neutral chelate, a neutral (possibly polymeric) complex, and a singly charged 1:1 complex which extracts as an ion pair with a monovalent anion. Distribution constants between chloroform and water were also determined for each ligand. The experimentally observed distribution constants for I and II are much lower than constants derived from theoretical calculations. Compound III has a partitioning constant which is in good agreement with its theoretically calculated value. The rate equations for the extraction of copper by I and II, and for the extraction of copper by I catalyzed by III, are all first order in metal ion, second order in ligand, and inverse first order in hydrogen ion. The catalytic rate law is first order in each ligand. In the case of the reactions which are second order in ligand, the rate determining step is proposed to be the aqueous phase reaction of a 1:1 intermediate complex with the second ligand molecule. Knowledge of the partitioning constants of the ligands, which in turn provides their aqueous phase concentrations, leads to the calculation of second order reaction rate constants of 1.2 x 10('7)M⁻¹s⁻¹ for I and 1.1 x 10('7)M⁻¹ for II. In the case of the catalytic mechanism, the rate law cannot be used to discern which ligand reacts in the rate determining step. If the reactive 1:1 intermediate is assumed to be with I, then a catalytic rate constant of 9.5 x 10('9)M⁻¹s⁻¹ is obtained. A catalytic pathway proceeding through a 1:1 intermediate with III is also possible, but this complex is too poorly defined for calculating a rate constant. A kinetic study of the back extraction of copper into water from an organic solution of its 2:1 complex with I indicates three pathways. The chelate partitions into water and releases its coordinated ligands in a solvolysis reaction. Both hydrogen ion and III catalyze the stripping reaction. In the case of catalysis by III, a mixed ligand complex is proposed to partition. In conclusion, the "homogeneous phase" mechanism is found to be the better mechanistic interpretation of these reaction systems. Unlike the interfacial model, this mechanism can account for all the observed phenomena, is supported by independent measurements, and conforms to the vast body of chemical data already acquired on extraction systems.

No description available.

No description available.