Perlecan is an important basement membrane heparan sulfate (HS) proteoglycan that is essential for various cell signaling events involved in tissue development. Heparanase is a lysosomal enzyme involved in the turnover of HS. This project aimed to assist in researching the structure of HS on perlecan and how this structure changes with tissue development. This will be achieved by generating monoclonal antibodies that have an altered affinity for perlecan after heparanase treatment. Recombinant perlecan domain I was characterized by ELISA and western blotting and used as the antigen for two fusions. The first fusion was focused on the production of IgM the common subtype of anti-glycosaminoglycans antibodies. However, no clones were produced, which may have been due to the lack of feeder layers. In order to address this problem, the fibroblast cell line MRC-5 was used as a feeder layer in the second fusion. From this fusion, we obtained 216 positive cultures, which were screened against full length perlecan from endothelial cells. Of these, 26 cultures were tested against heparanase treated perlecan, and then 2 cultures were chosen for subcloning based on the different immunoreactivity between enzyme treated and nontreated perlecan. From the 2 chosen cultures, 13 sub clones were derived and 10 of them were adapted into a serum free culture environment. The 10 monoclonal antibodies displayed strong immunoreactivity with full length perlecan in ELISA and Western Blotting. When they were used as primary antibodies in Immunocytochemistry, they were able to recognize the native perlecan deposited by human chondrocytes. When the cells were incubated with heparanase, antibody 5D7-2E4 and 13E9-3G5 showed an increase in immunoreactivity while antibody 13E9-3B3 gave a decrease. These three antibodies will be the potential tools used in the future to study perlecan turnover in different cells and tissue. The remaining seven antibodies will also be very useful in the research of perlecan as they have been shown to bind to the protein core. In the future, it will be worth subcloning some of the frozen stored stocks of uncloned hybridomas, where there are potential opportunities to select antibodies, which will react with the carbohydrate chains on perlecan.

Bibliography: leaves 119-136.

Late Ordovician Red River strata of southeastern Saskatchewan were deposited in a broad epicontinental sea. In the lower part, the Yeoman and Herald formations comprise two cycles of carbonateevaporite sequences. Although these units possess an overall layer-cake aspect, thickness variations especially in the Herald Formation show that accumulation was affected by syndepositional flexure, differential subsidence and displacement of fault-bounded blocks. The mainly laminated dolomudstones and anhydrites of the Lake Alma and Coronach members of the Herald Formation were deposited under relatively tranquil conditions. These units host different kinds of synsedimentary deformation features, interpreted to have been induced by earthquakes generated because of movements along basement faults thought to have been oriented orthogonally NE−SW and NW−SE. The low-energy environmental setting was conducive to preserving these features, referred to as seismites.<p> The variety of seismites in the Herald Formation is related to the varying rheology of the carbonate or evaporite sediment, as well as shaking intensity. Brittle and quasi-brittle failure is represented by faults, microfaults, shear-vein arrays and pseudo-intraclastic breccias, mostly in dolomudstones which must have been stiff at the time of deformation. Plastic behaviour is recorded by soft-sediment deformation, comprising a family of features that includes loop bedding, folded laminae and convolute bedding. Indeed, these structures in enterolithic anhydrite are more reasonably interpreted as due to deformation than crystal growth, volume expansion and displacement, the more usual explanations. Sediment shrinkage and concomitant fluidization are recorded by dikelets containing injected carbonate mud or granular gypsum, the latter now preserved as anhydrite. Evidence for wholesale liquefaction, however, was not observed. These rheological differences were caused by the primary nature of the sediment plus modifications due to early diagenesis and burial confinement. Shaking intensity is difficult to gauge, but it is presumed that a minimum of VI on the modified Mercalli scale was required to produce these features. Consequently, shaking of lesser magnitude was probably not recorded.<p> The geographic distribution of seismites should reflect the location of basement faults presumed to have been active during deposition, and indeed there is a concentration adjacent to the known location of syndepositonal fault lineaments. In addition, the stratigraphic distribution of seismites records higher frequencies of activity of these same faults. These distributions show that earthquake-induced ground motion was common during deposition of the Lake Alma Member in southeastern Saskatchewan but less so during deposition of the Coronach Member.<p> Seismites serve as proxies for the activity of relatively nearby syndepositional faults making up the tectonic fabric of sedimentary basins. They also point to basement features that, if re-activated, can induce fracture porosity or influence subsurface fluid flow. Syndepositional tectonism undoubtedly had a much more profound influence on many successions than is presently accepted, and its effects are more widespread than currently appreciated.

This paper studies is considered the problem of coherent re ection of an acoustic plane wave from a rough seabed with a randomly inhomogeneous sediment layer overlying a uniform elastic basement. The randomness of the sound eld is attributable to the rough- ness of the seabed and the sound-speed perturbation in the sediment layer, resulting in a joint rough surface and volume scattering problem. An approach based upon perturbation theory, combined with a derived Green's function for a slab bounded above and below by a uid and an elastic half space, respectively, is employed to obtain an analytic solution for the coherent eld in the sediment layer. Furthermore, a boundary perturbation the- ory developed by Kuperman and Schmidt [22] is applied to treat the problem of rough surface scattering. A linear system is then established to facilitate the computation of the coherent re ection eld. The coherent re ection coe cients for various surface roughness, sediment randomness, frequency, sediment thickness, and basement elasticity have been generated numerically and analyzed. It was found that the higher/larger size of surface and/or medium randomness, frequency, thickness, and shear-wave speed, the lower the coherent re ection. Physical interpretations of the various results are provided.

<p>Basement membranes are a dense, sheet-like form of extracellular matrix that underlie epithelia and endothelia, and surround muscle, fat and Schwann cells. Basement membranes separate tissues and protect them from mechanical stresses. Although traditionally thought of as a static support structure, a growing body of evidence suggests that dynamic basement membrane deposition and modification instruct cell behavior and morphogenetic processes. In this thesis, I discuss how changes to basement membrane affect anchor cell (AC) invasion during C. elegans uterine vulval attachment. During AC invasion, the uterine AC breaches two juxtaposed basement membranes to contact the underlying vulval epithelium. Using live-cell imaging, genetics, molecular biology and electron microscopy I identify three modifications to the BM that affect AC invasion. In Chapter 2, I describe a system for linking juxtaposed basement membranes to stably align or connect adjacent tissues. This adhesion system promotes rapid AC invasion and also regulates a more long-term connection between the uterine tissue and the hypodermal seam cell in the adult worm. Chapter 3 elucidates how the BM component SPARC promotes cell invasion. As SPARC overexpression is correlated with cancer metastasis, this aims to understand how SPARC overexpression promote invasion in a pathological situation. In Chapter 4, I discuss preliminary data showing that the AC actively secretes laminin into the basement membrane targeted for invasion. I outline how future studies could elucidate the mechanism by which AC-derived laminin might promote cell invasion. Finally, Chapter 5 discusses conclusions and future directions for these studies.</p> / Dissertation

Perlecan is an important basement membrane heparan sulfate (HS) proteoglycan that is essential for various cell signaling events involved in tissue development. Heparanase is a lysosomal enzyme involved in the turnover of HS. This project aimed to assist in researching the structure of HS on perlecan and how this structure changes with tissue development. This will be achieved by generating monoclonal antibodies that have an altered affinity for perlecan after heparanase treatment. Recombinant perlecan domain I was characterized by ELISA and western blotting and used as the antigen for two fusions. The first fusion was focused on the production of IgM the common subtype of anti-glycosaminoglycans antibodies. However, no clones were produced, which may have been due to the lack of feeder layers. In order to address this problem, the fibroblast cell line MRC-5 was used as a feeder layer in the second fusion. From this fusion, we obtained 216 positive cultures, which were screened against full length perlecan from endothelial cells. Of these, 26 cultures were tested against heparanase treated perlecan, and then 2 cultures were chosen for subcloning based on the different immunoreactivity between enzyme treated and nontreated perlecan. From the 2 chosen cultures, 13 sub clones were derived and 10 of them were adapted into a serum free culture environment. The 10 monoclonal antibodies displayed strong immunoreactivity with full length perlecan in ELISA and Western Blotting. When they were used as primary antibodies in Immunocytochemistry, they were able to recognize the native perlecan deposited by human chondrocytes. When the cells were incubated with heparanase, antibody 5D7-2E4 and 13E9-3G5 showed an increase in immunoreactivity while antibody 13E9-3B3 gave a decrease. These three antibodies will be the potential tools used in the future to study perlecan turnover in different cells and tissue. The remaining seven antibodies will also be very useful in the research of perlecan as they have been shown to bind to the protein core. In the future, it will be worth subcloning some of the frozen stored stocks of uncloned hybridomas, where there are potential opportunities to select antibodies, which will react with the carbohydrate chains on perlecan.

Bibliography: leaves 119-136 / ix, 136 leaves, [9] leaves of plates : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, 1986

Thesis (Ph. D.)--Illinois State University, 1997. / Title from title page screen, viewed June 5, 2006. Dissertation Committee: H. Tak Cheung (chair), Sean Arkins, Herman E. Brockman, Paul A. Garris, Brian J. Wilkinson. Includes bibliographical references (leaves 113-121) and abstract. Also available in print.

Too few documentative descriptions exist from the reactivated Proterozoic Nasa Basement window in Sweden. This re-exposed crustal piece predominantly consisting of 1.8 Ga granite and volcanic basement rock lack structural evidences on maps. The Western Gneiss region gave example of how rocks in extreme cases may experience protolith overprinting through collisional vertical granite basement subduction to eclogite ambient conditions, which would give traces in metamorphic quartz (coesite), muscovite, pyroxene and kyanite, and increased densities. Preliminary indications from literature points towards a locally passive Nasa Basement with limited translation, but because of too little documentation it cannot be assured metamorphic inertia under fluid absence has been unimportant in the basement evolution. To address this highly structural issue, field reconnaissance mapping and sampling was carried out, and aimed to achieve a structural characterisation of historic deformation back to Proterozoic time. Observations were collected from 166 outcrop vicinities, and 17 of 27 carefully examined thin sections aimed to describe microscopic structures, in addition to mineral relation to metamorphic minerals. Collected evidences in this project encompass a Proterozoic macroscopic concentric fold with axis striking NW-SE, as well as, mesoscopic folds striking WNW-ESE. Tectonic movements record syn-collisional top-SE and post- collisional top-SW translation, which truncate weakly shortened and foliated blocks. Basement blocks locally preserve igneous coarse but weakly recrystallized texture fabrics. Overall optically produced paleothermometry indicates low- to medium grade facies conditions of deformation, through widespread bulging recrystallization, occasional subgrain rotation and rare grain boundary migration in quartz. Feldspar response to deformation is brittle and rarely ductile. Hydrous biotite, chlorite and epidote are abundant. Mineralogy together with an observed average density of 2,6 g/cm3 can possibly reject the idea of unreacted local continental subduction to eclogite depths, due to the overall presence of hydrous mineral phases. Top-to-the shear sense proves Caledonian reactivation, but Bourguer anomaly indicates at least locally an important limited translation. NW-SE striking lineaments and strikes of foliation show best accordance with lineaments mapped in Norway and alignment with tectonic contacts in the Baltica interior, and do so show the best examples of candidates for Proterozoic structures.

Master of Science / Department of Geology / Brice LaCroix / Abdelmoneam Raef / South-central Kansas has experienced an increase in seismic activity within the Proterozoic basement over the past 10 years. In 2009, Oklahoma seismic stations recorded 50 earthquakes statewide, a 200% increase from 2008. Oklahoma Geological Survey (OGS) seismograph stations recorded 1,028 in 2010, an increase of over 2000% from 2009. Between 2000-2012, Kansas experienced only 12 earthquakes statewide. Beginning in September 2013, clusters of seismic events in south-central Kansas began to increase. In 2015 alone, Kansas seismograph stations recorded 448 earthquakes, of which 166 resulted in a magnitude 2.0 or greater. Since 2013, United States Geological Survey (USGS) seismograph stations have recorded over 12,000 earthquakes within Kansas and Oklahoma. Pore fluid pressure increases associated with recent high-rate wastewater injection into the dolomitic Arbuckle disposal zone are hypothesized as cause of reactivation of the faulted study region’s Proterozoic basement. Although the magnitude of fluid-pressure change required for reactivation of these faults is likely low given failure equilibrium conditions in the midcontinent, heterogeneities (i.e. permeability, porosity, fluid pressure) in the basement could allow for a range of fluid pressure changes associated with injection. This research aims to quantify the fluid pressure changes responsible for fault reactivation of the Proterozoic basement. To address this issue, we use 97 earthquake focal mechanisms and over 12,000 seismic events, from the USGS catalog, within an area encompassing ~ 4,000 km². Focal mechanism data was utilized to determine the regional stress field present within the study region. Nodal plane data extracted from the focal mechanisms was crucial to identifying lineaments within the underlying basement complex. A 3D seismic dataset covering the Wellington and Anson Bates Fields in north central Sumner County, Kansas was utilized for enhanced structural delineation of an interpreted faultnetwork affecting the Mississippian and Arbuckle Groups, to investigate whether it impacts the underlying granitic basement and its complex network of potentially interconnected fault planes. Smoothed similarity and spectral whitening analyses were applied to the dataset to improve depth of investigation and uncover fault lineaments masked by seismic attenuation due to increasing depth. An interpreted network of fault planes at depths of 3.5 km was uncovered beneath Wellington Field. The lineaments are well aligned with known structural features present within the Proterozoic basement, the Central Kansas Uplift and the Nemaha Ridge-Humboldt fault zone.