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- All Subjects: Biochemistry
- Member of: Theses and Dissertations
Description
In somatic cells, the mitotic spindle apparatus is centrosomal and several isoforms of Protein Kinase C (PKC) have been associated with the mitotic spindle, but their role in stabilizing the mitotic spindle is unclear. Other protein kinases such as, Glycogen Synthase Kinase 3â (GSK3â) also have been shown to be associated with the mitotic spindle. In the study in chapter 2, we show the enrichment of active (phosphorylated) PKCæ at the centrosomal region of the spindle apparatus in metaphase stage of 3T3 cells. In order to understand whether the two kinases, PKC and GSK3â are associated with the mitotic spindle, first, the co-localization and close molecular proximity of PKC isoforms with GSK3â was studied in metaphase cells. Second, the involvement of inactive GSK3â in maintaining an intact mitotic spindle was shown. Third, this study showed that addition of a phospho-PKCæ specific inhibitor to cells can disrupt the mitotic spindle microtubules. The mitotic spindle at metaphase in mouse fibroblasts appears to be maintained by PKCæ acting through GSK3â. The MAPK pathway has been implicated in various functions related to cell cycle regulation. MAPKK (MEK) is part of this pathway and the extracellular regulated kinase (ERK) is its known downstream target. GSK3â and PKCæ also have been implicated in cell cycle regulation. In the study in chapter 3, we tested the effects of inhibiting MEK on the activities of ERK, GSK3â, PKCæ, and á-tubulin. Results from this study indicate that inhibition of MEK did not inhibit GSK3â and PKCæ enrichment at the centrosomes. However, the mitotic spindle showed a reduction in the pixel intensity of microtubules and also a reduction in the number of cells in each of the M-phase stages. A peptide activation inhibitor of ERK was also used. Our results indicated a decrease in mitotic spindle microtubules and an absence of cells in most of the M-phase stages. GSK3â and PKCæ enrichment were however not inhibited at the centrosomes. Taken together, the kinases GSK3â and PKCæ may not function as a part of the MAPK pathway to regulate the mitotic spindle.
ContributorsChakravadhanula, Madhavi (Author) / Capco, David G. (Thesis advisor) / Chandler, Douglas (Committee member) / Clark-Curtiss, Josephine (Committee member) / Newfeld, Stuart (Committee member) / Arizona State University (Publisher)
Created2012
Description
The viscous lung mucus of cystic fibrosis (CF) patients is characterized by oxygen gradients, which creates a unique niche for bacterial growth. Pseudomonas aeruginosa and Staphylococcus aureus, two predominant microorganisms chronically infecting the airways of CF patients, typically localize in hypoxic regions of the mucus. While interspecies interactions between P. aeruginosa and S. aureus have been reported, little is known about the role of low oxygen in regulating these interactions. Studying interspecies interactions in CF lung disease is important as evidence suggests that microbial community composition governs disease progression. In this study, P. aeruginosa lab strain PAO1 and two primary clinical isolates from hypoxic tissues were cultured alone, or in combination, with methicillin resistant S. aureus (MRSA) strain N315 under hypoxic or normoxic conditions. Herein, it is shown for the first time that low oxygen conditions relevant to the CF lung affect the competitive behavior between P. aeruginosa and S. aureus. Specifically, S. aureus was able to better survive competition in hypoxic versus normoxic conditions. Competition data from different oxygen concentrations were consistent using PAO1 and clinical isolates even though differences in the level of competition were observed. PAO1 strains carrying mutations in virulence factors known to contribute to S. aureus competition (pyocyanin/phzS, elastase/lasA and lasI quorum sensing/lasI) were used to determine which genes play a role in the differential growth inhibition. The lasA and lasI mutants competed less effectively with S. aureus regardless of the oxygen level present in the culture compared to the isogenic wild type strain. These results are consistent with previous findings that elastase and lasI quorum sensing play a role in competitive behavior of P. aeruginosa and S. aureus. Interestingly, the phzS mutant competed less effectively in hypoxic conditions suggesting that pyocyanin may be important in microaerophilic conditions. This study demonstrates that oxygen plays a role in competition between P. aeruginosa and S. aureus and contributes to understanding CF environmental factors that may regulate microbial community dynamics important for disease progression with potential for development of therapeutic avenues.
ContributorsLedesma Barrera, Maria Alexandra (Author) / Nickerson, Cheryl A. (Thesis advisor) / Reyes del Valle, Jorge (Committee member) / Clark-Curtiss, Josephine (Committee member) / Stout, Valerie (Committee member) / Ott, C M (Committee member) / Arizona State University (Publisher)
Created2014
Description
In competitive Taekwondo, Electronic Body Protectors (EBPs) are used to register hits made by players during sparring. EBPs are comprised of three main components: chest guard, foot sock, and headgear. This equipment interacts with each other through the use of magnets, electric sensors, transmitters, and a receiver. The receiver is connected to a computer programmed with software to process signals from the transmitter and determine whether or not a competitor scored a point. The current design of EBPs, however, have numerous shortcomings, including sensing false positives, failing to register hits, costing too much, and relying on human judgment. This thesis will thoroughly delineate the operation of the current EBPs used and discuss research performed in order to eliminate these weaknesses.
ContributorsSpell, Valerie Anne (Author) / Kozicki, Michael (Thesis director) / Kitchen, Jennifer (Committee member) / Electrical Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05