Recombinant giardial arginine deiminase (rADI) was characterized. The enzyme was found to have a specific activity of 12 U (mg protein)-1under at pH 7.4 and 1 mM arginine. The maximum velocity was 14.75 U (mg protein-1) and the KM was 0.167 mM. The specific activity and maximum velocity values are significantly lower than the values reported previously for giardial rADI, while the KM value is quite similar. The optimum pH for the giardial rADI was 6-9, a broad range compared to other arginine deiminases. Recombinant ADI is very specific in its binding specificity, with canavanine (KI 2.4 mM) and ornithine (KI 2.1 mM) being the only substrate analogues giving significant inhibition from the wide variety of analogues tested. None of the analogues could be shown to act as alternative substrates. The contribution of conserved, catalytic and C-terminal residues proposed by previous research towards ADI activity was investigated by site-directed mutagenesis. Mutations of catalytic site residues Asp175, Glu226, His280 and Cys424 decreased the rADI activity to below 2%. Conservative mutations showed significant activity for Asp175 to Glu175 (DE175) and Glu226 to Asp226 (ED226). Site directed mutagenesis on the conserved non-catalytic site Leu46 showed activities below 15%, with the highest activity observed for the mutation to Val46 (LV46), which differs in one CH2 to Leu46 on its side chain. The KM of the mutant LV46 was 3.64 mM while for LA46 (Leu to Ala mutation) was 1.33 mM. Excising 5, 13, 16 amino acids from the C-terminal residues resulted in activity decreasing to 0.8% of the wild type, while excising 54 amino acids caused the rADI to be insoluble. Sequence alignment of Giardia and Dictyostelium suggests a homologous area within the C-terminal extension. Site directed mutagenesis on the Tyr567 residue in this region resulted in a decrease in activity, with the highest activity observed for a Tyr to Phe mutation. Studies using specific cysteine protease inhibitors demonstrated partial protection against proteolysis of ADI against giardial proteases. Degradation of ADI by giardial proteases was more rapid at pH 6 than at pH 7.4.

The optimisation of strength and power through resistance training has been the source of debate amongst health professionals and researchers for many years. As resistance training involves the repeated activation and contraction of skeletal muscles, continuous training will ultimately result in a failure to sustain the training intensity especially when performing multiple sets and / or repetitions. Therefore the prescription of rest periods within the training session becomes an important consideration. Relatively short rest periods (60-90 seconds) have been traditionally used for the maximal strength adaptation involving increases in the cross-sectional area of the muscle, whereas longer rest periods (180-300 seconds) have traditionally been used for the maximal strength adaptation involving enhancement of neural function and maximal power adaptation. However, there is very little scientific evidence to support these current practices. In fact, the effect of different rest periods on maximal strength and power development has received very little research attention. Additionally, research that has been conducted in this area has been typified by a number of methodological inconsistencies, within and between studies, which confound scientific understanding. Although traditionally resistance training has employed continuous training schemes with inter-set rest periods, intra-set rest training methods which distribute rest intervals between groups of repetitions have also been investigated. It has been theorised that the short rest periods within the training set allow partial resynthesis of the intramuscular phosphocreatine stores, potentially allowing an athlete to increase their training volume by training at high intensities for longer durations, or performing additional repetitions (Berg, 2003). This is thought to lead to an increased exposure of the muscle to the kinematic and kinetic stimuli thought important for strength and power adaptation whilst minimising performance-inhibiting metabolic accumulation and substrate depletion. However, research into intra-set rest training schemes is still in its infancy, and many of the theories surrounding intra-set rest training are currently unsubstantiated. It is thought that examinations of the acute kinematic, kinetic and blood lactate profiles of continuous and intra-set rest training schemes may enhance scientific understanding regarding the efficacy of intra-set rest training. The purpose of this study was to investigate and compare the acute kinematic, kinetic and blood lactate responses to continuous and intra-set rest loading schemes. Nine male subjects performed an isoinertial Smith machine bench press task (6RM load) with a continuous loading scheme (CONT), an intra-set rest loading scheme equated by total rest time, volume and load (ISRV) and an intra-set rest loading scheme equated by total rest time and load (ISRR). The order of the loading schemes was assigned in a block randomised order with a minimum of 48 hours recovery between each testing session. Attached to the bar of the Smith machine was a linear position transducer that measured vertical displacement with an accuracy of 0.01cm. Displacement data was sampled at 1000Hz and collected by a laptop computer running custom built data acquisition software. Finger prick blood lactate samples were taken from the non-dominant hand using sterile techniques at the following time points: pre-exercise (Pre), immediately post-exercise (P0), five (P5), fifteen (P15) and thirty minutes (P30) post exercise. Blood glucose samples were taken pre-exercise only. It was observed that manipulating the rest period, by increasing the frequency but decreasing the length of each rest period, did not significantly influence the kinematics and kinetics associated with resistance training, but did have an effect on the post-exercise blood lactate response when the load, rest duration and training volume was equated (ISRV). This finding may be of practical significance if fatigue is important in strength development or conversely if power training needs to be performed with minimal fatigue. It was also observed that increasing the frequency of the rest period enabled the subjects to perform a greater number of repetitions (ISRR), resulting in significantly greater kinematics, kinetics and blood lactate accumulation. It may be speculated, therefore, that ISRR training may offer a superior training stimulus for the development of maximal strength and hypertrophy than CONT training methods, as ISRR loading increased the exposure of the muscle to the kinematic, kinetic and metabolic stimuli thought important for the development of these qualities.

[Truncated abstract] Dyslipidaemia in obesity and the metabolic syndrome is typically characterized by elevated plasma concentrations of apolipoprotein (apo) B and chylomicron remnants, and low apoA-I levels. This may account for the increased risk of cardiovascularrelated diseases. Although the precise mechanisms whereby visceral obesity confers the onset of dyslipidaemia have not been fully established, it may relate chiefly to insulin resistance. Insulin resistance leads to increased hepatic secretion of very low density lipoprotein (VLDL) apoB, as well as impaired catabolism of VLDL, intermediate density lipoprotein (IDL), low-density lipoprotein (LDL) and chylomicron remnants, and high density lipoprotein (HDL) apoA-I. This thesis tests the unifying hypothesis that lipoprotein metabolism, in particular apoB, chylomicron remnants and apoA-I, is abnormal in the metabolic syndrome, and that medical intervention can correct for these abnormalities. The primary objectives were to examine firstly, the kinetics of apoB and apoA-I by stable isotope technology and secondly, chylomicron remnant kinetics by using an indirect assessment of a new breath test. Six observational statements and related hypotheses were constructed and derived from the unifying hypothesis that examine the kinetics of lipoprotein metabolism, adipose tissue mass compartments and liver fat accumulation, as well as the impact of plasma adipocytokines in subjects with visceral adiposity and features of the metabolic syndrome. The first four observational statements related to cross-sectional studies of lipoprotein kinetics, adipose tissue mass distribution and liver fat accumulation as well as plasma adipocytokines in both obese and non-obese men. The latter two observational statements related to the effect of statin therapy and dietary weight loss on the improvement of lipoprotein kinetics in obesity. The findings from these studies collectively support the unifying hypothesis. The kinetics of apoB in VLDL, IDL and LDL, and apoA-I in HDL were assessed by gas-chromatography mass spectrometry following either a primed-constant infusion of 13C-leucine or an intravenous bolus injection of d3-leucine. ... This is the first study to examine the effects of dietary weight loss on LDL and HDL metabolism and the relationships with adipocytokines in men with the metabolic syndrome. The data support the unifying hypothesis that medical intervention with dietary weight loss could correct the kinetic abnormalities in VLDL, LDL and HDL. The aforementioned studies showed that plasma lipid and lipoprotein abnormalities in visceral obesity are chiefly regulated by the combination of hepatic over-secretion of VLDL particles, and catabolic defects in apoB and chylomicron remnants as well as apoA-I-containing lipoprotein particles. These kinetic defects may also relate to low and high plasma adiponectin and RBP-4 levels, respectively. The data arising from the thesis are consistent with the unifying hypothesis and support the role of dietary intervention and pharmacotherapy as a recommended treatment in correcting the abnormalities in lipoprotein metabolism within the metabolic syndrome.

Thesis (M.S.)--University of Nevada, Reno, 2005. / "May, 2005." Includes bibliographical references (leaves 75-78). Online version available on the World Wide Web.

Two liquid-solid reactions, in which the morphology of the solid changes as the reactions proceeds, were examined. One is the NA conversion in oil by decarboxylation on metal oxides and carbonates, and the other is the Kraft pulping in which lignin removal by delignification reaction. In the study of the NA conversion, CaO was chosen as the catalyst for the kinetic study from the tested catalysts based on NA conversion. Two reaction mixtures, carrier oil plus commercial naphthenic acids and heavy vacuum gas oil (HVGO) from Athabasca bitumen, were applied in the kinetic study. The influence of TAN, temperature, and catalyst loading on the NA conversion and decarboxylation were studied systematically. The results showed that the removal rate of TAN and the decarboxylation of NA were both independent of the concentration of NA over the range studied, and significantly dependent on reaction temperature. The data from analyzing the spent catalyst demonstrated that calcium naphthenate was an intermediate of the decarboxylation reaction of NA, and the decomposition of calcium naphthenate was a rate-determining step. In the study on the delignification of the Kraft pulping, a new mechanism was proposed for the heterogeneous delignification reaction during the Kraft pulping process. In particular, the chemical reaction mechanism took into account the heterogeneous nature of Kraft pulping. Lignin reacted in parallel with sodium hydroxide and sodium sulfide. The mechanism consists of three key kinetic steps: (1) adsorption of hydroxide and hydrosulfide ions on lignin; (2) surface reaction on the solid surface to produce degraded lignin products; and (3) desorption of degradation products from the solid surface. The most important step for the delignification process is the surface reaction, rather than the reactions occurring in the liquid phase. A kinetic model has, thus, been developed based on the proposed mechanism. The derived kinetic model showed that the mechanism could be employed to predict the pulping behavior under a variety of conditions with good accuracy. / Chemical Engineering

Graduation date: 1997

We present results of molecular simulations, quantum mechanical calculations, and experimental data aimed towards the rational design of solvent formulations. In particular, we have found that the rate limitation of oxidation of methionine groups is determined by the breaking of O-O bonds in hydrogen peroxide, not by the rate of acidic catalysis as previously thought. We have used this understanding to design molecular level parameters which are correlated to experimental data. Rate data has been determined both for G-CSF and for hPTH(1-34). / Singapore-MIT Alliance (SMA)

Thesis (Ph.D.)--University of Delaware, 2006. / Principal faculty advisor: Jochen Lauterbach, Dept. of Chemical Engineering. Includes bibliographical references.

With increasing rates of obesity, research has begun to focus of co-morbidities of obesity such as osteoarthritis. The majority of existing research has focused on older adults as the group most likely to suffer from osteoarthritis. The purpose of this study was to determine if overweight and obese young adults exhibit biomechanical risk factors for knee osteoarthritis, and to determine if young adults with biomechanical risk factors of osteoarthritis can modify these with instruction. This purpose was divided into two separate studies. Study 1: Thirty adults between 18-35 years old were recruited into three groups according to body mass index: normal, overweight, and obese. Participants walked through the lab while we collected 3-d kinematic and kinetic data. Overweight and obese young adults walked with similar gait compared to normal weight young adults. Study 2: Nine young adults between 18-35 years were recruited who walked with stiff-knee gait. Baseline measures of gait were collected in the form of 3-d kinematics and kinetics as participants walked through the laboratory. They then completed the gait instruction program which consisted of four blocks of training. Each block included ten single steps where the participant was provided feedback, followed by 100 practice steps around the laboratory. Participants were successful in increasing sagittal plane kinematics and kinetics of interest in the study. Conclusion: Identifying individuals who had biomechanical risk factors of osteoarthritis according to body mass index was not possible. According to the results of our study, obese and overweight young adults are not at increased risk of osteoarthritis compared to normal weight young adults. Individuals who may be at increased risk due to stiff-knee gait were able to improve their gait following instruction.

Thesis (Ph. D.)--University of Delaware, 2006. / Principal faculty advisor: Herbert E. Allen, Dept. of Civil & Environmental Engineering. Includes bibliographical references.