The detection of cellular events is central to understanding biomoleculer processes as well as aid in therapeutic intervention strategies. One of the most fascinating biomoleculer events during the life cycle of a cell is proteolytic cleavage of proteins by enzymes known as proteases. Proteases are ubiquitous and participate in essential functions such as fertilization, embryo development, cell cycle regulation, immune response, tissue remodeling and programmed cell death. As proteases are involved in fundamental cellular processes any dysregulation of protease activity is usually associated with a diseased state. Thus methods for detection of protease activity are desirable as it may facilitate the identification of many pathological conditions which are associated with the aberrant expression and activity of proteases.Towards the goal of a general and modular strategy we have utilized split protein reassembly and coiled coils to develop genetically encoded sensors for detection of proteases. We established our first generation protease design utilizing split firefly luciferase and anti-parallel coiled coils and detected Tobacco Etch Virus (TEV) as a model protease. Two further iterations of the coiled-coil design led to the development of second and third generation of protease sensors which showed substantial improvement in the sensor response and was applied towards detection of therapeutically relevant proteases such as caspase-3, prostate specific antigen (PSA), ß-secretase and calpain-1.We applied our methodolgy to develop protease biosensors for the detection of a family of cysteine protease known as caspases. Caspases are involved in programmed cell death and their misregulation is implicated in cancer as well as neurodegenerative disorders. The panel of caspase biosensors was utilized to investigate caspase cleavage specificity as well as caspase activation in mammalian cytosolic extracts and live mammalian cells. Perhaps more importantly, we discovered cross talk between members of the caspase family which perform different biological functions.Finally, we detail our progress towards mimicking a naturally occurring multicomponent complex formed during programmed cell death, known as the apoptosome which leads to the activation of caspases. We have successfully utilized principles of self assembly and multivalency to assemble multi component complexes which exhibit proteolytic activity similar to the natural apoptosome.

Eukaryotic protein kinases are one of the most important classes of human proteins, and a great deal of research has focused on the development of small molecule inhibitors as biological probes for the determination of their cellular function or as therapeutics for the treatment of disease, such as cancer. The need for new selective inhibitors and a better understanding of the selectivities of existing small molecules is readily apparent. Towards the goal of better understanding protein kinases and the molecules that inhibit them, I have developed a split-protein-based approach for the investigation of these kinase-small molecule interactions. Employing split-firefly luciferase as a reporter domain, we engineered a three-hybrid system capable of determining kinase inhibition through competitive interactions between an active site-directed ligand and a small molecule of interest. This method measures luciferase activity as a function of ligand binding, as opposed to the more traditional assays which quantify kinase activity directly, and alleviates the laborious process of protein purification. The model kinase PKA and the promiscuous ligand staurosporine were used in an initial test case to successfully validate the general design principles of our assay. The modular nature inherent to the assay's design enabled us to adapt it to roughly 300 additional protein kinases and two different ligands. We were able to establish a protocol for rapidly ascertaining the inhibition of a kinase by a library of 80 commercially available kinase inhibitors in a 96-well, high-throughput format. This protocol was then systematically applied to the AGC group of kinases to observe patterns of inhibition across similarly related kinases. We have further shown how these results might be correlated with the sequence identity between kinases to better anticipate inhibitor promiscuity. Finally, we were able to illustrate how a kinase-centric approach could be applied to correlate alterations to the kinase domain with changes in luminescence. This has use for the interrogation of different modes of inhibition as well as in identifying the specific determinants of inhibitor binding. In total, these efforts represent the optimization of a new, general platform for determining kinase inhibitor selectivity across the kinome, and it could potentially be applied universally to the interrogation of protein-ligand interactions.

No description available.

This study investigates the effects of a split injection strategy on combustion performance and exhaust emissions in a high speed direct injection optical diesel engine. The investigation is focused on the effects of injection timing, quantity, and the dwell angle between the injections using commercially available diesel fuel. Three different split injection strategies including 50:50, 30:70, and 70:30 have been investigated. Additionally, the effect of total injected fuel quantity using total fuel quantities of 10 mm3 and 20 mm3 has been investigated. Moreover, the effect of variable and fixed dwell angle in split injections has been examined for five different values between 5o CA and 25o CA in the case of variable and 10o CA for the fixed dwell timing. The last parameter investigated was the injection timing, nine injection timings have been tested for each of the strategies. A Ricardo Hydra single cylinder optical engine running at 1500 rpm was used in this investigation. Conventional methods such as direct in-cylinder pressure measurements and heat release rate analysis have been employed. In addition, optical techniques such as high speed video imaging and two-colour have been applied, aimed at in depth analysis of the effects of the aforementioned parameters on engine performance and emissions. Furthermore, a significant amount of effort was devoted to the development and application of the Laser Induced Excipex Fluorescence (LIEF) technique so that simultaneous fuel liquid and fuel vapour distribution could be visualised. This investigation concludes that split injection strategies have the potential to reduce diesel exhaust emissions while maintaining a good level of fuel economy, provided that injection timings and the dwell angle between injections are appropriately selected. Further investigations are required in order to examine the effect of split injection under different engine operating conditions and speeds. In addition, the effect of alternative fuels must be considered. Moreover, the application of LIEF technique for quantitative fuel vapour concentration measurement should be considered through further optimisation of the LIEF system and careful calibration experiments.

According to the Unaccusative Hypothesis (Perlmutter 1978), intransitive verbs fall in two distinct classes: the <strong>unaccusatives</strong> (whose subjects originate as direct objects) and the <strong>unergatives</strong> (whose subjects originate as subjects). Although there are studies of split intransitivity in Modern Japanese and European languages, very few exist for earlier stages of Japanese. To fill in part of this gap, this thesis presents a comprehensive investigation of split intransitivity in Old Japanese (largely, 8th century Japanese). The descriptive and analytic work of this research is based on the newly developed ‘Oxford Corpus of Old Japanese’ (OCOJ). It consists of original and romanized Old Japanese texts, with a wide range of information including the original orthography, part-of-speech, morphology and syntactic constituency in the form of XML tags following TEI conventions. It is part of a larger collaborative research project: ‘Verb semantics and argument realization in pre-modern Japanese: A comprehensive study of the basic syntax of pre-modern Japanese’, in which my DPhil project is situated. As part of my DPhil project, I took part in the analysis and tagging of the OCOJ, in addition to contributing to translation. My original contribution to knowledge is a comprehensive investigation and in-depth analysis of the lexical-semantic aspects of split intransitivity in relation to its morpho-syntactic expressions in Old Japanese. This includes: exploring to what extent intransitive verbs could be classified as unaccusative and unergative, what factors are involved in the classification, how they interact, what are the possible ways of representation, and the theoretical implications it brings to linguistic theory in general. Syntactically, I looked into manifestations specific to Old Japanese (e.g. perfective auxiliary selection), and also examined to what extent diagnostics – which show split intransitivity in English, Italian and Modern Japanese (e.g. N+V compounding and resultative construction) – could be applied to Old Japanese. Semantically, I investigated various semantic factors and proposed basic and complex models of the interaction between intentionality and affectedness in Old Japanese. I also proposed a ‘complex format for representing simple event structures’ which enhances the understanding of semantic aspects of split intransitivity. As such, the results of my research not only contribute to a detailed understanding of Old Japanese verbs, but also have implications for linguistic theory in general.

Metamaterials offer the freedom to tune the rich electromagnetic coupling between the constituent meta-atoms to tailor their collective electromagnetic response. Therefore, a comprehensive understanding of the nature of electromagnetic interactions between meta-atoms is necessary for novel metamaterial design, which is provided in the first part of this thesis. The subsequent work in the thesis applies the understanding from the first part to design and demonstrate novel one-dimensional metamaterials that overcome the limitations of metamaterials proposed in literature or exhibit electromagnetic responses not previously observed. Split-ring Resonators (SRRs) are a fundamental building block of many electromagnetic metamaterials. In the first part of the work in this thesis, it is shown that bianisotropic SRRs (with magneto-electric cross-polarisation) when in close proximity to each other, exhibit a rich coupling that involves both electric and magnetic interactions. The strength and nature of the coupling between two identical SRRs are studied experimentally and computationally as a function of their separation and relative orientation. The electric and magnetic couplings are characterised and it is found that, when SRRs are close enough to be in each other's near-field, the electric and magnetic couplings may either reinforce each other or act in opposition. At larger separations retardation effects become important. The findings on the electromagnetic interactions between bianisotropic resonators are next applied to developing a one-dimensional ultra-wideband backward-wave metamaterial waveguide. The key concept on which the metamaterial waveguide is built is electro-inductive wave propagation, which has emerged as an attractive solution for designing backward-wave supporting metamaterials. Stacked metasurfaces etched with complementary SRRs (CSRRs) have also been shown to exhibit a broadband negative dispersion. It is demonstrated through experiment and numerical modeling, that the operational bandwidth of a CSRR metamaterial waveguide can be improved by restricting the cross-polarisation effects in the constituent meta-atoms. The metamaterial waveguide constructed using the modified non-bianisotropic CSRRs are found to have a fractional bandwidth of 56.3\% which, based on a thorough search of relevant literature, is the broadest reported value for an electro-inductive metamaterial. A traditional coupled-dipole toy-model is presented as a tool to understand the field interactions in CSRR based metamaterials, and to explain the origin of their negative dispersion response. This metamaterial waveguide should be of assistance in the design of broadband backward-wave metamaterial devices, with enhanced electro-inductive waveguiding effects. In the final part of the thesis, a one-dimensional metamaterial prototype that permits simultaneous forward- and backward-wave propagation is designed. Such a metamaterial waveguide could act as a microwave analogue of nanoparticle chains that support electromagnetic energy transfer with a positive or a negative dispersion due to the excitation of their longitudinal or transverse dipole modes. The symmetry of the designed hybrid meta-atom permits the co-existence of two non-interfering resonances closely separated in frequency. It is experimentally and computationally shown that the metamaterial waveguide supports simultaneous non-interacting forward- and backward-wave propagation in an overlapping frequency band. The proposed metamaterial design should be suitable for realising bidirectional wireless power transfer applications.

The purpose of this study was to assess the acute effects of vibration and stretching on passive and active forward split range of motion in elite adult female synchronized swimmers. Eleven athletes performed a passive forward split test measuring the height of the anterior superior iliac spine on both sides and an active split test on both sides by adopting an inverted split position. Then athletes were assigned randomly by right or left leg to receive vibration on one leg while stretching. The contralateral leg was stretched but did not receive vibration and served as the control. The treatment involved a 40-s exposure to vibration of the forward leg in a split and 40 s of vibration to the rear leg in a split. The athletes were then post-tested using the same protocols. The results indicated that the vibration had a statistically significant influence on passive forward split flexibility, but not active split flexibility. The results of this study confirm earlier work and further demonstrate the efficacy of vibration in enhancing range of motion in a passive split position. Given that it is often difficult to achieve large changes in range of motion with already highly trained elite athletes, this methodology shows considerable promise. Vibration may not be powerful enough to evoke changes in active range of motion in spite of the changes in passive range of motion.

Hintergrund Die Gesundheit und Attraktivität eines Lächelns hängt zum einen von der Beschaffenheit und Form der Zahnhartsubstanz (weiße Ästhetik) ab. Maßgeblich wird sie aber auch durch einen harmonischen Verlauf des Zahnfleisches (rote Ästhetik) geprägt. Dieser Verlauf kann durch Asymmetrien und Zahnfleischrückgang (Rezessionen) gestört werden. Neben dem subjektiven vom Patienten empfundenen, ästhetischen Einbußen können Rezessionen auch ein medizinisches/ funktionelles Risiko für z.B. überempfindliche Zahnhälse, rezidivierende oder dauerhafte mukosale Entzündungen darstellen. Die Prävalenz gingivaler Rezessionen liegt bei Kindern in etwa bei 8% und erhöht sich beim Erwachsenen bis zum Alter von 50 Jahren auf nahezu 100%. Als therapeutische Maßnahmen kann zum einen der entstandene Weichgewebsverlust durch non-chirurgische, konservative Maßnahmen, wie Komposit- Restaurationen(Schmelzlift mittels Schmelz-Ätz-Technik) ausgeglichen werden. 3 4 Ist der Verlust an Weichgewebe zu groß, oder kann der Verlust durch konservative Maßnahmen nicht ästhetisch befriedigend ausgeglichen werden, besteht die Möglichkeit durch einen parodontalchirurgischen Eingriff die verloren gegangene Symmetrie der "rote Ästhetik" wieder herzustellen. Ziel Diese prospektive randomisierte split-mouth Studie vergleicht die Effektivität der Deckung gingivaler Rezessionen mittels koronalem Verschiebelappen mit subepithelialem Bindegewebe (BGT) im Vergleich zur Deckung mit koronalem Verschiebelappen mit Schmelz-Matrix-Proteinen (EMD).Als Zielparameter wurden die Verringerung der Rezessionstiefe und /-breite und der Gewinn an keratinisierter Gingiva erfasst. Alle Parameter wurden zum Zeitpunkt Baseline nach 6, 12, 24, 36, 48 und 60 Monaten dokumentiert. Methode Insgesamt wurden 102 Rezessionen an einwurzeligen Zähnen von 21 Patienten behandelt. Die Auswertung erfolgte patientenbasiert. Ergebnisse Beide Verfahren führten zu signifikanten Verringerungen der Rezessionstiefen von Baseline zur Messung nach 48 Monaten. Zwischen 48 und 60 Monaten konnte eine Zunahme der Rezessionstiefen beobachtet werden. Zu den Untersuchungszeitpunkten von initial 2.8mm bei SCTG und 2.9mm bei EMD kann nach 6 (0.9mm/1.1mm), 12 (0.8mm/0.8mm), 24 (1.0mm/0.9mm), 36 (0.9mm/1.3mm) und 48 Monaten (0.7mm/1.0mm) eine signifikante Reduktion der Rezessionen für beide Behandlungsprocedere nachgewiesen werden. Zum letzten Untersuchungszeitpunkt nach 60 Monaten wurde eine Zunahme der Rezessionstiefe (1.5mm/1.9mm) beobachtet. Die Rezessionsbreite verringerte sich ebenfalls in beiden Gruppen signifikant: EMD von 4.6mm auf 3.2mm (p=0.002), BGT von 4.5mm auf 2.5mm (p<0.001). Alle Vergleiche zwischen beiden Gruppen ergaben keine signifikanten Unterschiede. Die Breite der keratinisierten Gingiva veränderte sich in beiden Gruppen während des Studienverlaufs in beiden Gruppen nicht Fazit Die Ergebnisse zeigen, dass beide Verfahren erfolgreich zur Therapie von Rezessionen der Miller Klasse I und II eingesetzt werden können. / Background The health and attractiveness of a smile depends not only on nature and shape of the tooth-hard substance (white aesthetics), but is significantly influenced by a harmonious course of the gums (red aesthetics). This course can be disrupted by asymmetries and gum decline (recessions). In addition to the subjective aesthetic loss perceived by the patient, recessions can also represent a medical/functional risk for e.g. over-sensitive root, recurrent or permanent mucosal inflammation. The prevalence of gingival recessions in children is about 8% and increases in adults up to the age of 50 years to almost 100%.As therapeutic measures, the resultant soft tissue loss can be compensated by non-surgical, conservative measures such as composite restorations (melting lift using melting etching technology). If loss of soft tissue is too large, or if loss cannot be compensated aesthetically satisfactorily by conservative measures, it is possible to restore the lost symmetry of "Red Aesthetics“. Aim This prospective randomized split-mouth study compares the effectiveness of covering gingival recessions by means of coronally advanced flap with either subepithelial connective tissue graft (BGT) or coronally advanced flap with enamel matrix proteins (EMD). The target parameters were the reduction of recession depth and wide as also the gain of Keratinized Gingiva. All parameters to baseline are documented after 6, 12, 24, 36, 48 and 60 months. Method A total of 102 recessions were treated on single root teeth in 21 patients. The evaluation was based on patients. Conclusion Both procedures led to significant reductions in the depth of recession from baseline to measurement according to 60 months see below. The range of recessions also decreased significantly in both groups: EMD from 4.6 mm to 3.2 mm (P = 0.002), BGT from 4.5 mm to 2.5 mm (p < 0.001). All comparisons between the two groups showed no significant differences. The width of Keratinized Gingiva did not change in both groups during the course of study. Results The results show that both methods can be successfully used for the treatment of recessions of the Miller Class I and II

This thesis investigates the impact of the reform of the split share structure on the Chinese capital market. It adopts an event study methodology to examine the share price performance around the announcements of the reform and its predicted determinants, the type and level of consideration by using a sample of the top 300 companies listed in the combined Shanghai Stock Exchange (SSE) and Shenzhen Stock Exchange (SZSE). I find the three-day cumulative abnormal returns (CAR) to be negative and significant around government announcement of the reform on 29 April 2005, but the three-day CAR around company???s announcement of the reform to be positive and significant. I attribute this change of sentiment by the market to the release of information about the reform process concerning features such as type and level of consideration. I also regress the company???s CAR on the type and level of consideration and find evidence to suggest that type of consideration matters, where investors prefer payment of shares from capital reserves or retained profits, cash, warrants or any combination of these methods as opposed to payment of shares from non-tradable shareholders. However, I observe no relation between level of consideration and CAR. I interpret this reaction to be that investors perceive that the consideration level is fair and reasonable based on the company???s financial and operating conditions. These results imply that the reform of the split share structure exerts a positive impact on a company???s share price and the extent of impact is a function of the type of consideration.

<p>Split-screen innebär att filmduken är uppdelad i flera delar. Uppsatsen behandlar</p><p>filmer där subjektiva upplevelser beskrivs med hjälp av split-screen. Det är ett berättande utifrån en karaktär. En subjektiv upplevelse kan vara en dröm, en hallinucation, ett minne, eller förnimmelse. Split-screen har funnits sen filmens begynnelse och haft sina glansdagar på bland annat 1910-talet och 1970-talet. I och med datorerna är det lättare att skapa split-screeneffekter Vi som publik är också mer vana vid flera datorfönster och bildrutor samtidigt. Split-screen har kommit in i en ny glansdager era. Det finns ett flertal filmer som använder sig av split-screen för att skildra subjektiva upplevelser. Uppsatsens metod är att lyfta fram exempel då det fungerar med split-screen och subjektiva upplevelser.</p>