2009 MU theses - Access restricted to UM
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Item Transcriptional regulation of ADAM-12 expression under normal and osteoarthritic conditions(University of Missouri--Columbia, 2009) Dhar, Srijita; Ray, Bimal K.[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] ADAM 12 overexpression is linked to the pathology of diseases including osteoarthritis, cancer, cardiac hypertrophy, Alzheimers and multiple sclerosis. Degradation of fibrous cap and neo chondrogenesis are key events of OA. Several growth promoting agents including transforming growth factor-[beta] (TGF-[beta]) also play determining roles in the function of the cartilage specific cells. The potential link between growth factors and the pathogenesis of OA still remains unresolved. In the present study, we investigated TGF-[beta] mediated transcriptional induction of ADAM 12 in osteoblasts and synoviocytes, two of the resident cells of the joint. CAT assay revealed both positive and negative functioning elements at the proximal and the first exonic region of human ADAM 12 promoter. By using EMSA we showed increased interaction of NF[kappa]B, SAF-1 and Sp1 with the ADAM 12 promoter in TGF-[beta] stimulated cells. At the exonic region we also identified a highly conserved polydinucleotide repeat sequence that acts as a potent basal transcriptional repressor. These results suggest that the cohort action of NF[kappa]B, Sp1 and SAF-1 lead to the overexpression of ADAM 12 under osteoarthritic conditions.Item Evaluation of stay-in-place PVC-formed concrete walls for blast design(University of Missouri--Columbia, 2009) Saucier, Aaron James, 1983-; Salim, Hani A., 1966-[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] In the event of a blast, concrete might not have the toughness needed to absorb the large amounts of energy imparted, even with the moderate steel reinforcement present in many such walls. If the concrete is used as a composite encased with PVC, it is expected that the large mass of the concrete will absorb much of the energy imparted by the blast, while the PVC will provide the needed ductility and tensile strength. In addition, the PVC skins and webs are expected to provide additional concrete confinement, which will improve the response of the concrete section post ultimate. This additional resistance from the PVC is expected to prevent catastrophic failure of concrete walls during an explosion event. In this thesis, two stay-in-place PVC-formed concrete wall systems are evaluated using laboratory static testing. The results of the loading tree tests for both systems are compared to reinforced concrete wall control samples. PVC-concrete samples exhibited additional strain energy when compared to control samples. Experimental evaluation of the confinement contribution to the compressive strength of concrete was also performed in this study.Item Vascular smooth muscle metabolism and pyruvate dehydrogenase(University of Missouri--Columbia, 2009) Ward, Kathryn R., 1985-; Hardin, Christopher D.[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] In dyslipidemic states, most muscle types preferentially utilize fatty acids rather then glucose largely due to modulation of the pyruvate dehydrogenase complex (PDH) activity by pyruvate dehydrogenase kinase (PDK) (54). We have previously shown that vascular smooth muscle (VSM) does not exhibit a decrease in glucose utilization during exposure to increased levels of either short or long chain fatty acids (2, 3). In order to determine if the metabolic inflexibility of VSM may be due to differences in the PDH regulatory mechanisms compared to other muscles, we examined the effects of the PDK inhibitor dichloroacetate (DCA) and the PDP stimulator D-chiro-inositol (DCI) on glucose utilization and lactate production in hog carotid artery (HCA). We also performed western blot analyses of hog skeletal muscle and hog VSM PDK isoforms. In these experiments glycogen stores were allowed to replete overnight at 37[degree sign]C, VSM was isometrically mounted, and incubated for 6 hours in a physiological saline solution containing 5 mM [1- [superscript13]C] glucose and 1 mM [1,2- [superscript13]C] acetate with or without 2 mM DCA or 25 mU/ml insulin and 200 uM DCI. In the presence of DCA, a small but statistically significant (p=0.01) decrease in glucose utilization was observed in VSM. Glucose utilization was significantly increased (p= 0.04) and lactate production was decreased (p= 0.06) in the presence of insulin and DCI. [superscript13]C-NMR analysis of glutamate in these tissues also showed that glucose oxidation was significantly increased (p=0.03). When experiments were repeated using 0.1 mM [1,2[superscript13]C] acetate, glucose utilization was significantly increased (p=0.0001) and lactate production was decreased (p=0.08) in non-contracted HCA in the presence of insulin and DCI. In contracted HCA, glucose utilization was significantly increased (p=0.001). Western blot analyses revealed that there is 2.8 fold more PDK3 and 3.2 fold less PDK2 in VSM compared to skeletal muscle. The presence of PDK3 and the relative insensitivity of HCA to DCA suggests that the PDK isoforms present in VSM may contribute to the metabolic inflexibility of this tissue. Furthermore, the ability of VSM to increase glucose utilization via PDP stimulation by DCI and insulin suggest that the PDH complex in VSM can be manipulated. Therefore, we conclude that PDH in VSM is chronically inhibited by the presence of PDK-3 and the inhibition of PDH may contribute to the metabolic inflexibility of VSM during dyslipidemia, potentially making VSM susceptible to lipotoxicity and atherosclerosis.Item The art of persuasion : critiquing China's new middle class by multi-perspective storytelling mode(University of Missouri--Columbia, 2009) Zhou, Ming, 1977-; Hawk, William, 1951-[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] My work is a critique of China's New Middle Class's tasteless manners. The New Middle Class in China have attempted to use "good taste" as a means of distinguishing themselves from the masses. However, that 'good taste' has been usually associated with certain status symbols, in particular famous Western brands and popular leisure activities. China's New Middle Class would blindly mimic things from the West in order to present themselves as members of the middle class. Meanwhile, they would pay no attention to the deeper, spiritual aspects of our traditional Chinese culture. They would parade our own traditional culture, wear traditional garments, and stage cultural festivities only for purposes of entertaining and pleasing Western eyes, and no more. And, in doing so, the New Middle Class would devalue the very cultural richness they purport to convey. To enhance this shallow, hasty and boastful mental state of China's New Middle Class, I emphasize the distorted relationship between the activity and its participant: the participant's extraordinary feelings of satisfaction and excitement are caused not by the activity itself but by the external meaning associated with that activity -- the status that this perceived middle class activity provides them. In addition, I enhance color contrasts, especially saturation, to make the elements in my paintings look cheap, unclassified, and mismatched. However, the artistic strategy of my work departs from previous artwork that only satirizes China's New Middle Class through the exaggeration of kitschy features. I attempt to cross the boundaries of different cultures, and avoid using simple dichotomies that would merely provide easy solutions. The judgments and values of the West and East, China's working class, middle class, and intellectuals are starkly different. In my works, I do not reject their impressions. Instead, I provide a forum of debate for those who seek to judge the tastes and manners of China's New Middle Class. My artwork's intention is to bring about self-awareness among China's New Middle Class. I try to tactfully remind them that the cultivation and creation of any culture is long-term project; they should pay attention to a culture's inner spirit instead of mindlessly assuming it and no more.Item A study of the role of the Atg8 protein in autophagic vesicle formation(University of Missouri--Columbia, 2009) Tiwari, Ranjitha; Stromhaug, Per E. (Per Eivind)[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] Autophagy is the term used to refers to an catabolic cellular cleanup process through the lysosomal machinery. During autophagy portions of cytoplasm are non-selectively sequestered in unique double membrane bound vesicles called autophagosomes that are transported to vacuole for degradation. The end products of catabolism are reused by the cell. The formation of autophagosomes is catalyzed by a concerted action of core machinery proteins. Among the core machinery proteins is a ubiquitin like cytosolic protein, Atg8. In yeast, Saccharomyces cerevisiae, Atg8 is induced during starvation. Atg8 is conjugated to a lipid, phosphoethanolamine. (PE). PE conjugated Atg8 tightly binds to the autophagic membrane and is widely used as marker for autophagy. The exact in vivo function of Atg8 is less well understood. In this study, we focused on how Atg8 functions in the formation of autophagosomal membrane and demonstrated that (1) The deconjugation of Atg8 from PE is essential for completion of autophagosome formation (2) Deconjugation of Atg8-PE requires Atg4 protease; (3) presence of Atg18 protein is essential for deconjugation during starvation. Here we present substantial evidence that the delipidation of Atg8 is required for the completion of the formation of autophagosomes. We also show that lipidation and delipidation of Atg8 is a highly dynamic process that is regulated by stress.