<i>Aeromonas hydrophila</i> uses the type II secretion system to transport protein toxins across the outer membrane. The trans-envelope system is comprised of more than ten proteins, including ExeA and ExeB, which form a complex in the inner membrane and are required for assembly of the ExeD secretion channel multimer, called the secretin, into the outer membrane. A putative peptidoglycan binding domain (Pfam protein families database number PF01471) is present in the periplasmic region of ExeA (pExeA), leading to the hypothesis that ExeA generates gaps in peptidoglycan, a barrier for trans-envelope transport and apparatus assembly, to allow ExeD to assemble into the outer membrane.<p> In this study, interactions between ExeA and peptidoglycan were examined both <i>in vivo</i> and <i>in vitro</i>. Wild type ExeA, but not the mutants containing substitution mutations of three highly conserved amino acid residues in the putative peptidoglycan binding domain, was cross-linked to peptidoglycan in vivo with DTSSP. Furthermore, the presence of wild type ExeA was also required for co-crosslinking of ExeB and ExeC to peptidoglycan. <i>In vitro</i> cosedimentation revealed that purified pExeA was able to bind to highly purified peptidoglycan. The protein assembled into large multimers in the presence of peptidoglycan fragments, as shown in cross-linking and co-gel filtration experiments. The requirement of peptidoglycan for multimerization was abrogated when the protein was incubated at temperatures above 25 °C. Two pExeA constructs, which disrupted the putative peptidoglycan binding domain, greatly reduced the cosedimentation, accompanied by decreased multimerization in the presence of peptidoglycan fragments. These results provide evidence that the putative peptidoglycan binding domain of ExeA is involved in physical contact with peptidoglycan. The interactions cause ExeA to multimerize, possibly forming a ring-like structure on the peptidoglycan, to generate a gap large enough to accommodate the secretion apparatus and/or to form an assembly scaffold.<p> The putative peptidoglycan binding domain of ExeA was also analyzed by comparing its amino acid sequence with that of other homologues. The highly conserved amino acid residues were found to cluster at one pocket on the surface in the crystal structure of hydrolase metallo (Zn) DD-peptidase that also contains this domain. We propose that this pocket is the binding site for the peptidoglycan ligand.

<i>Aeromonas hydrophila</i> uses the type II secretion system to transport protein toxins across the outer membrane. The trans-envelope system is comprised of more than ten proteins, including ExeA and ExeB, which form a complex in the inner membrane and are required for assembly of the ExeD secretion channel multimer, called the secretin, into the outer membrane. A putative peptidoglycan binding domain (Pfam protein families database number PF01471) is present in the periplasmic region of ExeA (pExeA), leading to the hypothesis that ExeA generates gaps in peptidoglycan, a barrier for trans-envelope transport and apparatus assembly, to allow ExeD to assemble into the outer membrane.<p> In this study, interactions between ExeA and peptidoglycan were examined both <i>in vivo</i> and <i>in vitro</i>. Wild type ExeA, but not the mutants containing substitution mutations of three highly conserved amino acid residues in the putative peptidoglycan binding domain, was cross-linked to peptidoglycan in vivo with DTSSP. Furthermore, the presence of wild type ExeA was also required for co-crosslinking of ExeB and ExeC to peptidoglycan. <i>In vitro</i> cosedimentation revealed that purified pExeA was able to bind to highly purified peptidoglycan. The protein assembled into large multimers in the presence of peptidoglycan fragments, as shown in cross-linking and co-gel filtration experiments. The requirement of peptidoglycan for multimerization was abrogated when the protein was incubated at temperatures above 25 °C. Two pExeA constructs, which disrupted the putative peptidoglycan binding domain, greatly reduced the cosedimentation, accompanied by decreased multimerization in the presence of peptidoglycan fragments. These results provide evidence that the putative peptidoglycan binding domain of ExeA is involved in physical contact with peptidoglycan. The interactions cause ExeA to multimerize, possibly forming a ring-like structure on the peptidoglycan, to generate a gap large enough to accommodate the secretion apparatus and/or to form an assembly scaffold.<p> The putative peptidoglycan binding domain of ExeA was also analyzed by comparing its amino acid sequence with that of other homologues. The highly conserved amino acid residues were found to cluster at one pocket on the surface in the crystal structure of hydrolase metallo (Zn) DD-peptidase that also contains this domain. We propose that this pocket is the binding site for the peptidoglycan ligand.

Abstract In this work, H2, CH4, and O2 are used as gas sources and C3H903B is used as the doping source, microwave plasma chemical vapor deposition and a two-steps deposition process will be applied to the growth of boron-doped diamond on p-type(111) silicon substrate. In this work, nucleation and growth of diamond film have been studied. A series of experiments are focused on the depenence of experimental pressure, temperature, power, dc bias, flow rates of O2, and doping concentration of C3H903B. The samples are examined by SEM, Raman, XRD, FTIR, and I-V. The results show that if nucleation is assisted by a negative dc bias, it can reach high density. The growth of diamond and the boron-doped diamond film is in multi steps. After 90 minutes of growth, the mechanism of deposition will be changed.

Thermonuclear (Type Ia) supernovae are excellent distance indicators, due to their uniform peak brightness. They are also important contributors to the chemical evolution of galaxies since their explosions supply large amounts of iron peak elements to the interstellar medium. However, there is no consensus on the progenitor systems of these supernovae. As a result, different delay times from the formation of the binary system to the supernova have been proposed. Whether the observed rate of supernova Type Ia in early-type galaxies supports a progenitor channel with one or two degenerate objects has been disputed. While the predominant old population found in early-type galaxies supports longer delay times, the presence of recent star formation might indicate the opposite. In this work, we employ a double-burst model to account for the relative contribution of both populations. We show that for a DTD ∝ t^−1, convolved with star formation histories that are relevant for early-type galaxies, the supernova rate is independent of a host galaxy’s colour. Our results indicate that a DTD with no cutoff is preferred, thus favoring the double-degenerate scenario. / Graduate

published_or_final_version / Psychology / Doctoral / Doctor of Philosophy

Background: Exercise is widely recognized as the cornerstone of management of type II diabetes (T2D). However, it is also known that people with T2D have poor adherence to exercise regimens, which is largely thought to be because of poor exercise tolerance. Recent studies have suggested that this exercise intolerance may be caused by a reduction in exercising muscle blood flow. One physiological mechanism which could potentially contribute is the muscle metaboreflex (MMR). This mechanism is thought to be a pressure-based flow-improving mechanism, but as a result of reduced efficacy of vasodilators and sympatholytic agents, might in fact be restraining the flow-improvement in persons with T2D. Hypothesis: Persons with T2D would not improve exercising muscle blood flow upon MMR activation. This absence of flow-improvement will be due to an augmented vasoconstriction in the exercising muscle. Methods: T2D (n=7) and CTL (n=6) participants performed rhythmic forearm handgrip exercise at an intensity equivalent to 20% MVC for 9 minutes with and without the application of ischemic plantar flexion (IPF). Forearm blood flow (FBF), mean arterial pressure (MAP), cardiac output (CO), heart rate (HR), total peripheral resistance (TPR) and forearm vascular conductance (FVK) were quantified for the last thirty seconds of each of four time points during the protocol. Plasma norepinephrine was measured via deep venous and arterialized venous blood sampling. Results: Steady state exercising FBF was increased in CTL but not in T2D during MMR activation (mean ± SE mL/min: CTLControl 161.16 ± 5.95, CTLMMR 212.72 ± 9.49, T2DControl 156.71 ± 13.08, T2DMMR 144.22 ± 10.55). This occurred despite similar increases in MAP, CO, HR, and TPR (across groups and treatment conditions, NS). FVK increased in CTL during the MMR protocol compared to the Control protocol, but decreased in the T2D group using the same comparison (mean ± SE mL/min/100 mm Hg: CTLControl 144.74 ± 5.63, CTLMMR 176.76 ± 11.99, T2DControl 143.29 ± 13.44, T2DMMR 103.53 ± 8.44). Conclusions: In the exercise model utilized, persons with T2D do not demonstrate the MMR-induced flow improvement seen in CTL. This impaired muscle blood flow in T2D is the result of MMR induced exercising limb vasoconstriction. / Thesis (Master, Kinesiology & Health Studies) -- Queen's University, 2012-01-31 09:30:42.604

Background: Vaccines have been hypothesized in the etiology of autoimmune diseases including type 1 diabetes. There are cases of diabetes reported in the Vaccine Adverse Event Reporting System (VAERS) following the administration of the human papillomavirus (HPV) vaccine, however this potential association has yet to be investigated. The objective of this thesis was to determine whether there is an association between immunization against HPV and the development of type 1 diabetes mellitus in grade 8 girls eligible for Ontario’s vaccination program. Methods: A retrospective, population-based, cohort study of girls residing within an Ontario health unit and eligible for the province’s publicly funded school-based HPV vaccination program between 2007 and 2010 was executed using provincial administrative health databases and the Immunization Recording Information System (IRIS) database. To control for known, unknown and unmeasured time-independent confounders, a self-controlled case series analysis was conducted. The relative incidence and 95% confidence interval were estimated using conditional Poisson regression. Results: The study cohort was comprised of 3465 girls with a mean age of 13.2 years at cohort entry (range 12.7 to 13.6 years). The mean duration of follow up was 2.7 years and ranged from 1.6 to 3.6 years. The proportion of girls who received at least one dose of the qHPV vaccine during the observation period was 58.3% (n=2020). During the study follow-up 15 cases of new onset type 1 diabetes were observed, six of which were classified as etiologically exposed to the qHPV vaccine. Using an indefinite risk window, immunization with the qHPV vaccine was not associated with an increased risk of developing type 1 diabetes (age and season-adjusted RI 0.15; 95% CI 0.02-1.32). Conclusions: The results of this thesis regarding the risk of type 1 diabetes following immunization with the qHPV vaccine are inconclusive as a consequence of the small number of cases identified. However, the random distribution of cases across time and across exposure status suggests that there is no association. Before a definitive conclusion is reached the analysis must be re-conducted on a larger cohort. / Thesis (Master, Community Health & Epidemiology) -- Queen's University, 2012-05-28 10:39:00.73

No description available.

Alzheimer’s disease (AD) is the most reoccurring type of dementia, and remains incurable. Much work has been done to investigate the connections between AD development, type 2 diabetes and insulin receptor signaling abnormalities. Full length amyloid precursor protein (flAPP) is a large transmembrane protein that has significant physiological activities including in utero fetal development. Alpha secretase enzymes cleave flAPP, producing secreted amyloid precursor protein alpha (sAPPα), which has neuroprotective properties, including protection against neuronal apoptosis as well as the induction of neuronal outgrowth. There is no known dedicated receptor for the physiological action of sAPPα. Our data suggest that the physiological actions of sAPPα are a result of the physical interaction between sAPPα and the neuronal insulin receptor. We have shown that sAPPα phosphorylates, and thus activates, the neuronal insulin receptor as well as specific downstream proteins, including insulin receptor substrate (IRS), and protein kinase B (Akt). We have also shown that the observed interaction between sAPPα and neuronal insulin receptors is physical and that sAPPα competes with insulin for the insulin binding site. These findings may have implications for therapies aimed at slowing down the progression of AD through the activation of the insulin receptor pathway, since in neurons, insulin and the insulin receptor pathway are critical to the neuronal health and plasticity.

Proefschrift Universiteit van Amsterdam. / Met lit. opg. - Met samenvatting in het Nederlands.