Recent advancements in mass spectrometry have facilitated new analytical approaches capable of comprehensively characterizing metabolites in biological samples. Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) combines excellent mass accuracy (ppm<1) and ultra-high resolution, which enables the separation and identification of individual components within complex mixtures, and the determination of elemental composition for each detected mass. FTICR-MS is an ideal method for non-targeted metabolomics as the majority of small molecular compounds (100-1000 Da) in a biological sample can be detected. The objective of this research was to investigate metabolomic alterations associated with key cellular processes deemed fundamental to cancer development and progression. Differentiating U937 cells, fibroblasts synchronously progressing through the cell cycle and a transformed cell line containing a temperature sensitive oncogene were collected and subject to FTICR-MS analysis for non-targeted comprehensive metabolomics. Putative metabolite identifications were confirmed with targeted metabolite analysis using multiple reaction monitoring triple quadrupole mass spectrometry. Analysis of the resulting metabolic profiles revealed robust metabolic alterations associated with fundamental cellular processes. Changes in glycerolipid content were observed in all cellular processes studied. During cell cycle progression, elevated levels of triacylglycerols and vinyl acylglycerols were detected as cells approached mitosis; increased levels of plasmalogens were detected during the induced differentiation of human leukemic cells and activation of the oncogene p130gag-fps in fibroblasts resulted in increased levels of phospholipids, including plasmalogens. When de novo fatty acid synthesis was inhibited in the differentiation cell model, the cells were not able to complete the differentiation process. Removal of the inhibitor resulted in increased lipid content, particularly plasmalogens, and the continuation of differentiation, suggesting a requirement for the de novo synthesis of lipids during this cellular process. This work demonstrates the advantages of non-targeted metabolic profiling for identifying non-intuitive metabolic associations with specific cellular processes. Collectively, the results of this thesis have implicated glycerolipids, in particular phospholipids, in the processes of cell cycle progression, differentiation and tumourigenic transformation. A broadened understanding of the role of global lipid metabolism during fundamental cellular processes may one day lead to new approaches for their modulation, and potentially new therapeutic strategies.

Recent advancements in mass spectrometry have facilitated new analytical approaches capable of comprehensively characterizing metabolites in biological samples. Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) combines excellent mass accuracy (ppm<1) and ultra-high resolution, which enables the separation and identification of individual components within complex mixtures, and the determination of elemental composition for each detected mass. FTICR-MS is an ideal method for non-targeted metabolomics as the majority of small molecular compounds (100-1000 Da) in a biological sample can be detected. The objective of this research was to investigate metabolomic alterations associated with key cellular processes deemed fundamental to cancer development and progression. Differentiating U937 cells, fibroblasts synchronously progressing through the cell cycle and a transformed cell line containing a temperature sensitive oncogene were collected and subject to FTICR-MS analysis for non-targeted comprehensive metabolomics. Putative metabolite identifications were confirmed with targeted metabolite analysis using multiple reaction monitoring triple quadrupole mass spectrometry. Analysis of the resulting metabolic profiles revealed robust metabolic alterations associated with fundamental cellular processes. Changes in glycerolipid content were observed in all cellular processes studied. During cell cycle progression, elevated levels of triacylglycerols and vinyl acylglycerols were detected as cells approached mitosis; increased levels of plasmalogens were detected during the induced differentiation of human leukemic cells and activation of the oncogene p130gag-fps in fibroblasts resulted in increased levels of phospholipids, including plasmalogens. When de novo fatty acid synthesis was inhibited in the differentiation cell model, the cells were not able to complete the differentiation process. Removal of the inhibitor resulted in increased lipid content, particularly plasmalogens, and the continuation of differentiation, suggesting a requirement for the de novo synthesis of lipids during this cellular process. This work demonstrates the advantages of non-targeted metabolic profiling for identifying non-intuitive metabolic associations with specific cellular processes. Collectively, the results of this thesis have implicated glycerolipids, in particular phospholipids, in the processes of cell cycle progression, differentiation and tumourigenic transformation. A broadened understanding of the role of global lipid metabolism during fundamental cellular processes may one day lead to new approaches for their modulation, and potentially new therapeutic strategies.

In den größten Städten Russlands werden aktuell Entwicklungstendenzen sichtbar, welche die Frage nach einem Ende der postsozialistischen Transformation aufwerfen. Ein Beispiel für derartige – posttransformative – Entwicklungstendenzen bildet die Planung und Verwirklichung ikonischer Architekturprojekte. Diese sollen vor allem Innovations- und Wettbewerbsfähigkeit symbolisieren. Vor Ort sind sie aber insbesondere dann umstritten, wenn sie die überkommenen städtebaulichen Traditionen in Frage stellen. Besonders gut zeigt sich das beim Ochta-Zentrum, einem Geschäfts- und Kulturzentrum mit 400 m hoher Dominante, das nach dem Willen des Gazprom-Konzerns am Rande der historischen Innenstadt Sankt Petersburgs errichtet werden sollte. In der Hochphase der Diskussion beschäftigte das Projekt nicht nur die Stadtbevölkerung, sondern auch die Führungsriege der Russischen Föderation und die UNESCO. Die Analyse des Diskurses über das Ochta-Zentrum ermöglicht daher ebenso Erkenntnisse über Tendenzen der Stadtentwicklung Sankt Petersburgs wie über Strategien der russischen Innen- und Außenpolitik. / Recent trends in the development of major Russian cities are raising questions about the end of post-socialist transformation. Post-transformative urban trends have become visible in the context of the planning and realization of iconic architectural projects. Their purpose is to symbolize a capacity for innovation and competitiveness. At the local level, however, they are subject to controversial debates, especially when challenging regional traditions of urban planning. The Okhta-Center, a business and cultural center with a skyscraper reaching a height of 400 m, serves as a good example. The Gazprom Group intended to build the complex at the edge of the historical city center of Saint Petersburg. During the height of the debate the project involved not only the citizens of Saint Petersburg, but also the leadership of the Russian Federation and UNESCO. Analyzing the discourse regarding the Okhta-Center therefore allows insights into current trends of urban development in Saint Petersburg as well as strategies of Russian domestic and foreign policies.

Vi har gjort en studie om hur vi kan förbättra registerkartan med hjälp av mätning med GPS och transformation i området Tobyn i Årjängs kommun. Arbetet innehåller fastighetsbildning förr i tiden och även om hur fastighetsbildning går till i dag. Vidare har vi skrivit om olika mätningsmetoder och fakta om GPS-systemet, exempelvis NRTK, fasmätning, absolutmätning och kort om olika transformationsmetoder. Vi har också skrivit om hur vi gick tillväga när vi först mätte och sedan transformerade. Till detta har vi olika bilagor med jämförelse mellan inmätta och transformerade punkter och kartor som visar hur mycket som skilde mellan transformerade gränser och den befintliga registerkartan. Det finns också en bilaga med flygbild över Tobyn som visar något av hur mätförhållandena är. Vår slutsats är att det gick väldigt smidigt att mäta och transformera och tidsåtgången var inte så stor. Det skilde upp till 75 centimeter mellan transformerad och inmätt punkt på vissa ställen. De största avvikelserna fanns i utkanterna av området Tobyn.

I vårt examensarbete har vi studerat gränspunkter som är skapade genom förrättningar och dokumenterade i koordinatsystem som inte längre är aktuella att använda. Vi har studerat kedjan från inmätning av befintliga gränspunkter, till att de är transformerade och införda i Trollhättans Stads databas. Vi har arbetat med tre olika områden i Trollhättan, vilka är Skoftebyn, centrum och Kalltorps By. De har tillkommit vid olika tidsperioder, har olika typer av bebyggelse samt är redovisade i olika typer av koordinatsystem. Genom vårt arbete har vi sett att olika områden i Trollhättan har olika väl bevarade gränsmarkeringar. I de villaområden vi studerat har flertalet av gränspunkterna återfunnits, medan de i centrum varit obefintliga, sånär som på gränsmarkeringar utvisade med hushörn. För att få ett djupare sammanhang har vi gjort en historisk tillbakablick där vi studerat gränser ur en allmän synvinkel. Vi har även intervjuat mätchefen i Trollhättans Stad, Roger Johansson, för att få information om Trollhättans Stads mäthistorik. Förberedelser inför inmätning av gränspunkter gjordes genom att förrättningsakterna granskades för att få information om gränsernas sträckning och dess utmärkningssätt. Inmätningarna av gränspunkterna utfördes med GPS och fördes därefter in i TopoCad 14. Gränspunkterna från förrättningsakten transformerades från det ursprungliga förrättningssystemet till Sweref 99 och fördes därefter in i databasen. Vårt tillvägagångssätt för koordinattransformationerna och för hur införandet av gränspunkterna i databasen har gått tillväga, finns utförligt dokumenterade. Jämförelsen mellan de radiella avvikelserna och lantmäteriets gränsvärden på 5cm i stadsmiljö och 10 cm på landsbygden visar att mycket noggranna inmätningar kommer att krävas för att införa alla gränspunkter i Trollhättans Stads databas för gränspunkter. / In this thesis, we have studied the boundary points that are created by ordinances and documented in the coordinate system that are no longer relevant to use. We have studied the chain that includes measuring of the current boundary points, transformation of the coordinates into current coordinatesystem and until the boundary points are introduced into the database of Trollhattans Stad. We have worked with three different areas in Trollhättan, which are Skoftebyn, the center area and Kalltorps By. They are generated at different times areas, there are different types of settlements in the different areas, and they are set in different coordinate systems. Through our work we have observed that different areas in Trollhättan have variety preserved boundary markers. In residential areas that we studied, the majority of the markers were found, while in the center area the boundary points were non-existent, except for the boundary markers expelled with "house corners". To get a deeper context, we conducted a retrospective, where we studied the property boundary in an ordinary way. We have also interviewed Roger Johansson, Trollhattans Stad, to obtain information about the measurement history of Trollhattans Stads. Preparation for survey of boundary points was done by ordinance acts that was reviewed to obtain information on property boundaries and which method of marking that was used. Survey of the boundary markers was performed with GPS and then transferred into TopoCad 14. The border points of ordinance file transformed from the original executor system SWEREF 99 and then transferred into the database. The approach for coordinate transformations and how the introduction of boundary points in the database was done, are fully documented. The comparison of the radial deviations and surveying levels of 5cm in urban areas and 10 cm in rural areas shows that highly accurate entries will be required to include all border points in the database of Trollhättan Stad.

Mass tourism is a critical agent of change, with a proven capacity of forging new landscapes and thoroughly modifying the internal structure of a community. Thornbury, a rural settlement located in South-western Ontario, is on the brink of such shift: With the emergence of a new culture of tourism and the introduction of ‘upscale’ resort developments, its physical conditions are rapidly being reconfigured and the character of its social landscape is on the precipice of being irrevocably altered. These luxury vacation facilities occur in a dispersed fashion on the fringe – a condition which has not only initiated a radical modification and decentralization of social and economic activity, but has also resulted in the decline of traditional neighbourhood ties. Likewise, this occurrence has introduced an influx of newcomers who advance an identity and lifestyle clearly distinct from that of Thornbury’s quiet community of rural citizens and retirees. In this context, these newly arrived inhabitants soon become dwellers of “limited liability”– those who reside within a community, but do not identify with it – as Thornbury lacks the infrastructure and amenities to support potential exchanges and direct interest to the qualities of traditional civic life. As such, the thesis will examine the question of: What is the role of architecture in sustaining local identity in an altered social fabric? Or, more specifically, how can social relations in a transformed community be advanced through the use and design of public spaces? The work will attempt to outline the trajectory of urban change and understand the factors that account for present circumstances. The outcome of the research will generate a series of responsive designs that hinge upon the idea of uplifting ordinary landscapes and everyday ruralism. Essentially, the proposed strategy is one in which rurality is integral to the solution: Methods which harness rural assets and refashion them in a progressive manner will be utilized to inspire the formation of a new local culture while establishing an interface between opposing population segments. Overall, this study is intended to serve as a template to be used by communities in Ontario for mitigating the effects of changing rural geographies.

Field pepperweed (<i>Lepidium campestre</i> L.) is a cruciferous winter annual wild edible plant. It has potential medicinal properties as it contains a considerable level of glucoraphanin, which is the precursor for sulforaphane, a phase 2 protein inducer. Phase 2 proteins either directly or indirectly promote the scavenging of strong oxidants, and thus decrease the risk of many complex disorders such as atherosclerosis and Alzheimers disease. However, field pepperweed plants also contain glucosinalbin, an antinutritional compound. For field pepperweed to become a green vegetable crop or source of functional food, it is desirable to reduce or eliminate glucosinalbin. The biosynthesis of glucosinalbin may be down-regulated through biotechnology. To that end, in the present studies, experimental protocols for plant regeneration and Agrobacterium-mediated transformation have been developed for field pepperweed. Establishment of such methods represents a vital first step in the process of engineering field pepperweed for enhanced nutritional value.<p> The effect of explant type and various combinations of growth regulators on regeneration were evaluated in three accessions of field pepperweed (Ames 13179, 13180 and 15718). Among the three genotypes, accession Ames 13179 had the highest regeneration frequency under several conditions. Regeneration from hypocotyl explants was more rapid and prolific than regeneration from either mature leaf or cotyledonary explants. Segments from the acropetal end of the hypocotyls were more regenerable than those from the basipetal end. Evaluation of different hormonal combinations and concentrations identified an optimal growth regulator combination (3 mg L-1 thidiazuron / 0.1 mg L-1 naphthalene acetic acid) for shoot induction.<p> The plant regeneration system established was adopted for field pepperweed transformation using the acropetal segments of hypocotyls as explants. Two plant expression constructs were tested for down-regulating by RNA interference with the expression of a field pepperweed cytochrome P450 gene named LcCYP79B2. This gene may be involved in biosynthesis of glucosinalbin. Conditions for transformation such as pre-culture, co-cultivation time, and antibiotic concentration were evaluated. Transgenic plants were obtained and confirmed by histochemical staining of the reporter â-glucuranidase activity and PCR (polymerase chain reaction) analysis of the NPTII gene. The current study has established efficient plant regeneration and transformation protocols for field pepperweed. They should be useful for future molecular biology studies and biotechnological applications in this species.

<p>The genes encoding the Ras family of small GTPases are mutated to yield constitutively active GTP-bound oncoproteins in one-third of all human cancers. In many other cancers lacking Ras mutations, Ras is activated by other means. One common example of such activation is found in breast cancer, in which epidermal growth factor receptor (EGFR) family receptor tyrosine kinases, including EGFR and HER2 (ErbB-2/Neu), are frequently amplified and overexpressed, which in turn activates Ras. In human cells, activation of the Ral guanine nucleotide exchange factor, or RalGEF, effector pathway is necessary for Ras-mediated tumorigenesis and metastasis. RalGEFs activate the two highly similar Ral GTPases, RalA and RalB. While RalA has been shown to be required for Ras-mediated tumorigenesis, RalB is important for tumor metastasis. Activated Ral GTPases bind to and activate a limited number of effector proteins, including RalBP1, Sec5, and Exo84, to affect numerous diverse activities of the cell. This dissertation research sought to determine which of these well-characterized Ral effector proteins were required for oncogenic mutant Ras-induceded tumorigenesis and metastasis of human cells, as well as to examine the role of RalA in breast cancer cells that can activate Ras through EGFR and HER2 overexpression. </p><p> RNA interference-mediated loss-of-function analysis demonstrated that Sec5 and Exo84 are required for oncogenic Ras-mediated tumorigenesis, and, at least in part, metastasis. Additionally, both gain-of-function and inhibition studies showed that RalA activation is induced by EGFR and HER2 in breast cancer cell lines stimulated with EGF. Furthermore, stable suppression of RalA expression inhibited tumorigenic growth of breast cancer cells, and RalA activation was shown to be higher in a majority of mammary adenocarcinomas versus matched patient normal mammary tissue. These studies provide new insights into the importance of RalA activation in breast cancer, as well as the molecules downstream of RalA and RalB that may be responsible for mediating their effects on tumorigenesis and metastasis.</p> / Dissertation

Extant literature has a controversy over the effects of customer relationship on customer responses to service failure. Buffering perspective suggests that strong relationship can mitigate customers¡¦ unfavorable reactions to service failure; whereas amplifying perspective finds that these negative customer responses are enhanced by strong relationship. Until now, it still lacks a unified framework to explain the mechanisms under each perspective and to reconcile both perspectives. The motivation of this research is to fill up this research gap and to solve the controversy over the effects of customer relationship under service failures. By integrating interdependence theory and reciprocity theory, this research proposes a framework to analyze when and why each of buffering and amplifying effects would occur and how amplifying effects can shift to buffering effects. The major conjectures of the framework proposed by this research are that customer relationship has amplifying effects on customers¡¦ immediate responses while it has buffering effects on their prolonged responses; moreover, customers¡¦ immediate and prolonged responses are guided by different reciprocity motivations and the shift from amplifying effects to buffering effects is triggered by motivation transformation that can be enhanced by increasing a number of customers¡¦ social concerns. This framework is verified through one survey study and two experimental studies. Collectively, the results broadly support that: (a) a customer relationship has amplifying effects on customers¡¦ immediate responses while has buffering effects on their prolonged responses; (b) immediate responses are majorly governed by self-oriented motivations in reciprocation of the firms¡¦ past behavior, whereas to some extent prolonged responses are guided by relationship-oriented motivations in reciprocation of the firms¡¦ anticipated behavior; (c) transformation of reciprocity motivations triggers the shift from amplifying effects into buffering effects; (d) concern about future of relationship with the firm increases the possibility of transformation and thus the occurrence of buffering effects.

The C54 TiSi2 thin films are widely applied in semiconductor devices due to the low electric resistance and high thermal stability. Through the annealing processing in this study, the metastable C49 TiSi2 with an electric resistivity of 219.3 £g£[-cm transforms to the stable C54 TiSi2 phase at a higher annealing temperature, with a resistivity of 30.5 £g£[-cm. Hence the transformation temperature of C49 ¡÷ C54 is of great concern in metallization of gates and local interconnections. In this thesis, it is found that the oxygen content and Ag addition impose significant influence on the transformation temperature of C49 ¡÷ C54. The as-sputtered TiSi2 thin films are confirmed to be amorphous. After annealing at 600oC or 900oC, the silicides would transform to the metastable C49 TiSi2 or C54 TiSi2 phase, respectively. The current transformation temperatures are much higher than 200oC and 600oC for the normal TiSi2 system, due to high oxygen content in the current films (up to 15-20 at% as a result of our old sputtering system). Nevertheless, the co-sputtered TiSi2 thin films with 5 and 20 at% Ag can decrease the formation temperature of C54 TiSi2 phase to 800oC. Compare with the as-sputtered TiSi2 thin films, the desirable electric resistivity of the C54 phase in the 20 at% Ag thin films is also further reduced to 22.9 £g£[-cm. The time-dependent mechanical responses of the amorphous, crystalline C49, and C54 TiSi2 thin films are investigated by room-temperature nanoindentation at the different loading rates ranging from 0.0125 to 5 mN/s. The anelasticity response plays an important role in the current TiSi2 thin films and is found to be sensitive to the loading rate. The displacement of time-dependent anelasticity recorded during the period of hold time increases with increasing loading rate. The anelasticity behavior can be analyzed by the Kelvin model. The as-deposited amorphous phase, with a lower atomic packing density and higher degree of defects and free volumes, exhibits the higher anelasticity deformation and longer relaxation time.